Clinical Studies

Abstracts are presented below for clinical studies on Grape.

  • Botanical Name: Vitis Vinifera

  • Ayurvedic Name: Draksha

  • Common Name: Grape

Vitis Vinifera

Plant Phytonutrient Profile


1: J Exp Bot. 2007 Feb 24; [Epub ahead of print]

Elicitor and resistance-inducing activities of {beta}-1,4 cellodextrins in
grapevine, comparison with {beta}-1,3 glucans and {alpha}-1,4
oligogalacturonides.

Aziz A, Gauthier A, Bezier A, Poinssot B, Joubert JM, Pugin A, Heyraud A,
Baillieul F.

URVVC-EA 2069, Stress, Defenses et Reproduction des Plantes, Universite de Reims
Champagne-Ardenne, BP 1039, F-51687 Reims cedex 2, France.

Cellodextrins (CD), water-soluble derivatives of cellulose composed of beta-1,4
glucoside residues, have been shown to induce a variety of defence responses in
grapevine (Vitis vinifera L.) cells. The larger oligomers of CD rapidly induced
transient generation of H(2)O(2) and elevation in free cytosolic calcium,
followed by a differential expression of genes encoding key enzymes of the
phenylpropanoid pathway and pathogenesis-related (PR) proteins as well as
stimulation of chitinase and beta-1,3 glucanase activities. Most of these
defence reactions were also induced by linear beta-1,3 glucans (betaGlu) and
alpha-1,4 oligogalacturonides (OGA) of different degree of polymerization (DP),
but the intensity of some reactions induced by CD was different when compared
with betaGlu and OGA effects. Moreover, desensitization assays using H(2)O(2)
production showed that cells treated with CD remained fully responsive to a
second application of OGA, suggesting a different mode of perception of these
oligosaccharides by grape cells. None of CD, betaGlu, or OGA induced HSR gene
expression nor did they induce cell death. In accordance with elicitor activity
in grapevine cells, CD-incubated leaves challenged with Botrytis cinerea also
resulted in a significant reduction of the disease. Data suggest that CD could
operate via other distinct reaction pathways than betaGlu and OGA. They also
highlight the requirement of a specific DP for each oligosaccharide to induce
the defence response.

PMID: 17322548 [PubMed - as supplied by publisher]

2: Bioresour Technol. 2007 Feb 22; [Epub ahead of print]

Volatile components of grape pomaces from different cultivars of Sicilian Vitis
vinifera L.

Ruberto G, Renda A, Amico V, Tringali C.

Istituto del C.N.R. di Chimica Biomolecolare, Via del Santuario 110, I-95028
Valverde, CT, Italy.

The volatile components of grape pomace coming from the processing of some of
the most important varieties of grape (Vitis vinifera L.) cultivated in Sicily,
namely Nero d'Avola, Nerello Mascalese, Frappato and Cabernet Sauvignon, have
been determined by gas-chromatography (GC) and gas-chromatography-mass
spectrometry (GC-MS). According to the winemaking procedure that entails the
removal of stalks before fermentation, two kinds of grape pomace are obtained.
The first consists of skins, pulp residues and seeds, the proper grape pomace,
which is partially used for grappa, a typical Italian spirit, and alcohol
production, the second consists almost exclusively of stalks. On the whole, 38
components have been characterized in the samples of grape pomaces, with
Frappato cv. showing the richest composition; instead, 88 components have been
detected in the stalks of Frappato, Nero d'Avola, Nerello Mascalese and Cabernet
Sauvignon varieties. In order to make a comparison between the grape varieties
easier, the volatile components detected in the two sets of samples (grape
pomaces and stalks) have been grouped in different classes. Significant
differences among varieties have been detected and statistical treatment of data
is also reported. This study is part of a wider project aimed at the possible
exploitation of the main agro-industrial by-products. At the same time it is one
of the first reports on the volatile components of this waste material.

PMID: 17321134 [PubMed - as supplied by publisher]

3: Plant J. 2007 Mar;49(5):772-85.

White grapes arose through the mutation of two similar and adjacent regulatory
genes.

Walker AR, Lee E, Bogs J, McDavid DA, Thomas MR, Robinson SP.

CSIRO Plant Industry, Adelaide Laboratory, PO Box 350, Glen Osmond, SA 5064,
Australia.

Most of the thousands of grapevine cultivars (Vitis vinifera L.) can be divided
into two groups, red and white, based on the presence or absence of anthocyanin
in the berry skin, which has been found from genetic experiments to be
controlled by a single locus. A regulatory gene, VvMYBA1, which could activate
anthocyanin biosynthesis in a transient assay, was recently shown not to be
transcribed in white berries due to the presence of a retrotransposon in the
promoter. We have found that the berry colour locus comprises two very similar
genes, VvMYBA1 and VvMYBA2, located on a single bacterial artificial chromosome.
Either gene can regulate colour in the grape berry. The white berry allele of
VvMYBA2 is inactivated by two non-conservative mutations, one leads to an amino
acid substitution and the other to a frame shift resulting in a smaller protein.
Transient assays showed that either mutation removed the ability of the
regulator to switch on anthocyanin biosynthesis. VvMYBA2 sequence analyses,
together with marker information, confirmed that 55 white cultivars all contain
the white berry allele, but not red berry alleles. These results suggest that
all extant white cultivars of grape vines have a common origin. We conclude that
rare mutational events occurring in two adjacent genes were essential for the
genesis of the white grapes used to produce the white wines and white table
grapes we enjoy today.

PMID: 17316172 [PubMed - in process]

4: Int J Food Microbiol. 2007 Jan 16; [Epub ahead of print]

Formation of vinylphenolic pyranoanthocyanins by selected yeasts fermenting red
grape musts supplemented with hydroxycinnamic acids.

Morata A, Gonzalez C, Suarez-Lepe JA.

Dpto. Tecnologia de Alimentos, Escuela Tecnica Superior de Ingenieros Agronomos,
Universidad Politecnica de Madrid, Ciudad Universitaria S/N, 28040 Madrid,
Spain.

The aim of this work was to study the formation of vinylphenolic pyroanthocyanin
pigments by three strains of Saccharomyces cerevisiae and one strain each of
Saccharomyces uvarum and Saccharomyces montuliensis during the fermentation of
red grape (Vitis vinifera L. cv. Tempranillo) must containing additional
hydroxycinnamic acids (caffeic acid, ferulic acid or p-coumaric acid, 50 mg/l).
These pigments were identified and quantified by high pressure liquid
chromatography-diode array detection, their structures confirmed by liquid
chromatography/electrospray interface-mass spectrometry, and their mechanisms of
formation proposed. Depending on the yeast strain employed, different quantities
of these pigments were formed. These pigments affect the colour of the finished
wine, sometimes greatly intensifying it.

PMID: 17303275 [PubMed - as supplied by publisher]

5: J Agric Food Chem. 2007 Feb 21;55(4):1604-1611.

The Fate of trans-Caftaric Acid Administered into the Rat Stomach.

Vanzo A, Cecotti R, Vrhovsek U, Torres AM, Mattivi F, Passamonti S.

Agricultural Institute of Slovenia, Hacquetova 17, 1001 Ljubljana, Slovenia,
Dipartimento di Biochimica Biofisica e Chimica delle Macromolecole, Universita
di Trieste, via L. Giorgeri 1, 34127 Trieste, Italy, Agrifood Quality
Department, IASMA Research Center, Via E. Mach 1, 38010 San Michele all'Adige,
Italy, and Area Farmacologia, Facultad de Ciencias Bioquimicas y Farmaceuticas,
Universidad Nacional de Rosario, Suipacha 531, 2000 Rosario, Argentina.

trans-Caftaric acid is the most abundant nonflavonoid phenolic compound in
grapes and wines. It occurs in chicory and is one of the bioactive components of
Echinacea purpurea. In order to fill the gap of knowledge about its
bioavailability in mammals, we investigated its absorption, tissue distribution,
and metabolism in rats. Assuming that the stomach is a relevant site of
absorption of dietary polyphenols, a solution of trans-caftaric acid was
maintained in the ligated stomach of anaesthetized rats for 20 min. Intact
trans-caftaric acid was detected in rat plasma at both 10 and 20 min (293 +/- 45
and 334 +/- 49 ng/mL, respectively), along with its O-methylated derivative
trans-fertaric acid, whose concentration rose over time (from 92 +/- 12 to 185
+/- 24 ng/mL). At 20 min, both trans-caftaric acid and trans-fertaric acid were
detected in the kidney (443 +/- 78 and 2506 +/- 514 ng/g, respectively) but not
in the liver. Only trans-fertaric acid was found in the urine (33.3 +/- 12.8
mug/mL). In some rats, trans-caftaric acid was detected in the brain (180 +/- 20
ng/g). Keywords: Trans-caftaric acid; Vitis vinifera; Cichorium intybus;
Echinacea purpurea; gastric absorption; peripheral tissues; metabolism; rat.

PMID: 17300159 [PubMed - as supplied by publisher]

6: Biotechnol Lett. 2007 Feb 13; [Epub ahead of print]

Production of a recombinant carotenoid cleavage dioxygenase from grape and
enzyme assay in water-miscible organic solvents.

Mathieu S, Bigey F, Procureur J, Terrier N, Gunata Z.

UMR IR2B, ENSAM-INRA, Universite Montpellier II, 34060, Montpellier cedex 1,
France, [email protected]

A recombinant carotenoid cleavage dioxygenase from Vitis vinifera L. was
produced by Escherichia coli as a fusion with the glutathione-S-transferase
(GST) protein under different bacterial growth conditions. The enzyme production
was monitored by a GST assay. Addition of Triton X-100 prior to bacterial cell
disruption doubled the release of soluble protein. A simple spectrophotometric
enzyme assay was developed to measure carotenoid cleavage activity using lutein
as substrate. Enzyme activity showed a 26-fold increase with the addition of 10%
(v/v) acetone in the reaction mixture.

PMID: 17295086 [PubMed - as supplied by publisher]

7: New Phytol. 2007;173(4):832-40.

Stomatal deregulation in Plasmopara viticola-infected grapevine leaves.

Allegre M, Daire X, Heloir MC, Trouvelot S, Mercier L, Adrian M, Pugin A.

UMR Plant Microbe Environnement INRA1088-CNRS 5184, Universite de Bourgogne, 17,
rue Sully, BP 86510, 21065 Dijon, France.

* In grapevine, the penetration and sporulation of Plasmopara viticola occur via
stomata, suggesting functional relationships between guard cells and the
pathogen. This assumption was supported by our first observation that grapevine
(Vitis vinifera cv. Marselan) cuttings infected by P. viticola wilted more
rapidly than healthy ones when submitted to water starvation. * Here,
complementary approaches measuring stomatal conductance and infrared
thermographic and microscopic observations were used to investigate stomatal
opening/closure in response to infection. * In infected leaves, stomata remained
open in darkness and during water stress, leading to increased transpiration.
This deregulation was restricted to the colonized area, was not systemic and
occurred before the appearance of symptoms. Cytological observations indicated
that stomatal lock-open was not related to mechanical forces resulting from the
presence of the pathogen in the substomatal cavity. In contrast to healthy
leaves, stomatal closure in excised infected leaves could not be induced by a
water deficit or abscisic acid (ABA) treatment. However, ABA induced stomatal
closure in epidermal peels from infected leaves, indicating that guard cells
remained functional. * These data indicate that the oomycete deregulates guard
cell functioning, causing significant water losses. This effect could be
attributed to a nonsystemic compound, produced by the oomycete or by the
infected plant, which inhibits stomatal closure or induces stomatal opening; or
a reduction of the back-pressure exerted by surrounding epidermal cells. Both
hypotheses are under investigation.

PMID: 17286831 [PubMed - in process]

8: Plant J. 2007 Feb;49(4):579-91. Epub 2007 Jan 18.

Mesocarp localization of a bi-functional resveratrol/hydroxycinnamic acid
glucosyltransferase of Concord grape (Vitis labrusca).

Hall D, De Luca V.

Department of Biological Sciences, Brock University, 500 Glenridge Avenue, St
Catharines, Ontario, Canada, L2S 3A1.

Resveratrol is a stilbene with well-known health-promoting effects in humans
that is produced constitutively or accumulates as a phytoalexin in several plant
species including grape (Vitis sp.). Grape berries accumulate stilbenes in the
exocarp as cis- and trans-isomers of resveratrol, together with their respective
3-O-monoglucosides. An enzyme glucosylating cis- and trans-resveratrol was
purified to apparent homogeneity from Concord (Vitis labrusca) grape berries,
and peptide sequencing associated it to an uncharacterized Vitis vinifera
full-length clone (TC38971, tigr database). A corresponding gene from Vitis
labrusca (VLRSgt) had 98% sequence identity to clone TC38971 and 92% sequence
identity to a Vitis viniferap-hydroxybenzoic acid glucosyltransferase that
produces glucose esters. The recombinant enzyme was active over a broad pH range
(5.5-10), producing glucosides of stilbenes, flavonoids and coumarins at higher
pH and glucose esters of several hydroxybenzoic and hydroxycinnamic acids at low
pH. Vitis labrusca grape berries accumulated both stilbene glucosides and
hydroxycinnamic acid glucose esters, consistent with the bi-functional role of
VLRSgt in stilbene and hydroxycinnamic acid modification. While phylogenetic
analysis of VLRSgt and other functionally characterized glucosyltransferases
places it with other glucose ester-producing enzymes, the present results
indicate broader biochemical activities for this class of enzymes.

PMID: 17270014 [PubMed - in process]

9: Plant Mol Biol. 2007 Feb;63(3):307-23. Epub 2006 Oct 12.

Identification of genes associated with flesh morphogenesis during grapevine
fruit development.

Fernandez L, Torregrosa L, Terrier N, Sreekantan L, Grimplet J, Davies C, Thomas
MR, Romieu C, Ageorges A.

UMR BEPC, Campus Agro-M/INRA, 2 place Viala, 34060, Montpellier, Cedex 01,
France.

Fruit morphogenesis is a process unique to the angiosperms, and yet little is
known about its developmental control. Following fertilization, fruits typically
undergo a dramatic enlargement that is accompanied by differentiation of
numerous distinct cell types. To identify genes putatively involved in the early
development of grapevine fruit, we used the fleshless berry mutant (Vitis
vinifera L. cv Ugni Blanc) that has dramatically reduced fruit size due to a
lack of pericarp development. Using oligo-specific arrays, 53 and 50 genes were
identified as being down- and up-regulated, respectively, in the mutant. In
parallel, Suppression Subtractive Hybridization performed between the mutant and
the wild type (WT) allowed the identification of new transcripts differentially
expressed during the first stages of mutant and WT pericarp development. From
this data, the picture emerged that the mutation promotes the expression of
several genes related to ripening and/or to stress and impairs the expression of
several regulatory genes. Among those, five genes encoding proteins previously
reported to be associated with, or involved in, developmental processes in other
species (a specific tissue protein 2, ATHB13, a BURP domain protein, PISTILLATA,
and YABBY2), were identified and investigated further using real-time PCR and in
situ hybridization. Expression in the pericarp was confirmed, specific spatial
and/or temporal patterns were detected and differences were observed between the
WT and the mutant during fruit development. Expression of these genes appeared
to be affected during young fruit development in the mutant, suggesting that
they may play a role in grape berry morphogenesis.

PMID: 17268889 [PubMed - in process]

10: J Food Prot. 2007 Jan;70(1):157-63.

Effect of chemical treatments on ochratoxigenic fungi and common mycobiota of
grapes (Vitis vinifera).

Belli N, Marin S, Argiles E, Ramos AJ, Sanchis V.

Food Technology Department, CeRTA-UTPV, University of Lleida, Av. Alcalde Rovira
Roure 191, 25198 Lleida, Spain.

The effect of the application of two fungicides (cyprodinil alone and a mixture
of cyprodinil and fludioxonil) on the mycoflora of grapes, especially
ochratoxigenic fungi, was studied. Different doses and application times were
analyzed. Grape mycobiota was isolated and identified, and the classification of
black aspergilli was carried out. We found that 81.7% of the isolates belonged
to Aspergillus niger aggregate and 18.3% to Aspergillus carbonarius. The ability
to produce ochratoxin A (OTA) was studied on Czapek yeast extract agar (CYA)
medium in 238 isolates. Most A. carbonarius (97.2%) produced detectable amounts
of OTA, while only 2.9% of the A. niger aggregate were OTA producers. Most of
the isolates (58%) produced less than 2.5 microg OTA per g of CYA. That,
together with the highest levels of black aspergilli detected near harvest,
proved the reported theory that they are primarily responsible for OTA in
grapes. The fungicides studied had a significant effect on black aspergilli in
three of the four vineyards sampled, as the natural increase of black aspergilli
when approaching harvest was in general lower in all the fields treated with
fungicides as compared to the control treatment. A mixture of cyprodinil (37.5%)
and fludioxonil (25%) applied at veraison and 21 days before harvest was the
most effective treatment to prevent black aspergilli in grapes, together with a
single application of this mixture at veraison followed by an application of
cyprodinil (50%) 21 days before harvest. No OTA was detected in musts (n=112)
produced from either the control treatment or the treated grapes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17265875 [PubMed - indexed for MEDLINE]

11: J Agric Food Chem. 2007 Feb 7;55(3):992-1002.

Flavonol profiles of Vitis vinifera red grapes and their single-cultivar wines.

Castillo-Munoz N, Gomez-Alonso S, Garcia-Romero E, Hermosin-Gutierrez I.

Area de Tecnologia de Alimentos, Escuela Universitaria de Ingenieria Tecnica
Agricola, Ronda de Calatrava 7, 13071 Ciudad Real, Spain.

The main flavonols found in seven widespread Vitis vinifera red grape cultivars
include the 3-glucosides and 3-glucuronides of myricetin and quercetin and the
3-glucosides of kaempferol and isorhamnetin. In addition, the methoxylated
trisubstituted flavonols, laricitrin and syringetin, were predominantly found as
3-glucosides. As minority flavonols, the results suggest the detection of the
3-galactosides of kaempferol and laricitrin, the 3-glucuronide of kaempferol,
and the 3-(6' '-acetyl)glucosides of quercetin and syringetin. The flavonol
profiles based on the eight above-mentioned flavonols allowed the cultivar
differentiation of the grape samples. With regard to flavonol biosynthesis in
the berry skin, quercetin 3-glucuronide predominated at veraison, followed by
quercetin 3-glucoside, and only trace amounts of trisubstituted flavonols were
detected. The proportion of quercetin 3-glucoside remained almost constant
during berry ripening, whereas the proportion of quercetin 3-glucuronide
decreased and the other flavonols, especially myricetin 3-glucoside, increased
their importance. In wines, flavonol 3-glycosides coexisted with their
corresponding free aglycones released by hydrolysis. The presence of laricitrin,
syringetin, and laricitrin 3-glucoside in red wines is reported here for the
first time. The extent of hydrolysis was widely variable among wines made from
the same grape cultivar, and the results suggest the influence of the type of
aglycone and glycoside on the rate of hydrolysis. Due to hydrolysis, the
differentiation of single-cultivar wines gave acceptable results only when
aglycone-type flavonol profiles were used.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17263504 [PubMed - in process]

12: J Agric Food Chem. 2007 Feb 7;55(3):779-86.

Effect of soil type on wines produced from Vitis vinifera L. cv. Grenache in
commercial vineyards.

de Andres-de Prado R, Yuste-Rojas M, Sort X, Andres-Lacueva C, Torres M,
Lamuela-Raventos RM.

Bodegas Miguel Torres, Miquel Torres i Carbo 6, 08720 Vilafranca del Penedes,
Barcelona, Spain.

In recent years, the wine industry has become increasingly interested in the
influence of the terroir characteristics on the features of a wine and, in
particular, the mechanisms by which a soil influences wine quality. Among
published papers on this topic, most merely describe the effect of the soil; few
explain it. In this study were conducted a sensory evaluation and phenolic
composition and stilbene concentration tests in order to analyze the effects of
soil on wine. Significant differences were found in the results of the tests
conducted on two vineyards during two consecutive harvests in 2004 and 2005. The
results, in line with previous reports, show that the more fertile of the two
vineyards, which was also the one with the greatest water-holding capacity,
produced wines that presented significantly lower color intensity and shade, as
well as lower total phenolic composition and a smaller quantity of
hydroxycinnamic compounds. In 2004, these wines presented significantly higher
trans-resveratrol content, due to a fungal attack that was favored by the
vineyard's soil characteristics. Extreme drought conditions in 2005 had a marked
impact on the characteristics of the wines, increasing wavelength measurements
significantly and reducing stilbene concentrations. Finally, sensory evaluations
revealed significant differences between the wines produced on the two vineyards
in both years for five of the seven attributes evaluated. No significant
differences were found from one year to the next between the wines produced from
the same vineyard, indicating that the attributes of these wines were maintained
despite markedly different vintage conditions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17263474 [PubMed - in process]

13: J Agric Food Chem. 2007 Feb 21;55(4):1053-1061. Epub 2007 Jan 30.

Assessment of Anthocyanins in Grape (Vitis vinifera L.) Berries Using a
Noninvasive Chlorophyll Fluorescence Method.

Agati G, Meyer S, Matteini P, Cerovic ZG.

Istituto di Fisica Applicata 'Nello Carrara', CNR, Via Madonna del Piano 10
(Edificio C), I-50019 Sesto Fiorentino, Italy, and Equipe de Biospectroscopie
Vegetale, Laboratoire d'Ecologie Systematique et Evolution, CNRS UMR 8079, Bat.
362, Universite Paris-Sud, 91405 Orsay Cedex, France.

Anthocyanins (Anths) in grape (Vitis vinifera L.) berries harvested at veraison
from Pinot Noir and Pinot Meunier cultivars were assessed nondestructively by
measuring chlorophyll fluorescence (ChlF) excitation spectra. With increasing
Anth content, less excitation light was transmitted to the deeper Chl layers,
and thus the ChlF signal decreased proportionally. By applying Beer-Lambert's
law, the logarithm of the ratio between the fluorescence excitation spectra (log
FER) from a green and a red berry gave the in vivo absorption spectrum of Anths,
which peaked at about 540 nm. Absolute quantitative nondestructive determination
of Anths for each berry was obtained by the log FER calculated for two
excitation wavelengths, 540 and 635 nm (absorbed and not-absorbed by Anths,
respectively) of ChlF at 685 nm. Over a range of skin colors going from green to
purple, the relationship between the log [ChlF(635)/ChlF(540)] and the Anth
concentration of berry extracts was fairly well fitted (r 2 = 0.92) using a
power function. Reflectance spectra on the same berry samples were also
measured, and Anth reflectance indices, which were originally developed for
apples and table grapes, were derived. The log FER Anth index was superior to
the reflectance-ratio-based index, but was as good as the color index for red
grapes (CIRG) calculated from the whole visible reflectance spectrum. The
proposed log FER method, applied by means of suitable portable devices, may
represent a new, rapid, and noninvasive tool for the assessment of grape
phenolic maturity in vineyards. Keywords: Anthocyanins; chlorophyll
fluorescence; color index; grape berry; fluorescence excitation spectra;
nondestructive determination; reflectance; ripening; Vitis vinifera.

PMID: 17261018 [PubMed - as supplied by publisher]

14: Theor Appl Genet. 2007 Feb;114(4):723-30. Epub 2007 Jan 13.

Wine grape (Vitis vinifera L.) color associates with allelic variation in the
domestication gene VvmybA1.

This P, Lacombe T, Cadle-Davidson M, Owens CL.

Institut National de la Recherche Agronomique, Centre de Montpellier, UMR,
Diversite et Genomes des Plantes Cultivees, Montpellier, France.

During the process of crop domestication and early selection, numerous changes
occur in the genetic and physiological make-up of crop plants. In grapevine
(Vitis vinifera) numerous changes have occurred as a result of human selection,
including the emergence of hermaphroditism and greatly increased variation in
berry color. This report examines the effect of human selection on variable skin
color by examining the variation present in the gene VvmybA1, a transcriptional
regulator of anthocyanin biosynthesis. In over 200 accessions of V. vinifera,
the insertion of the retroelement Gret1 in the promoter region of VvmybA1 was in
strong association with the white-fruited phenotype. This retroelement was
inserted at the same location for each individual in which it was present.
Additional polymorphisms in the VvmybA1 gene were also strongly associated with
red or pink fruited accessions, including variation that was generated by the
excision of Gret1 from the promoter of VvmybA1. Differences in nucleotide
diversity were observed between the white and pigmented alleles of VvmybA1,
suggesting that the white allele arose only once or a limited number of times.
Rarely, association of Gret1 with the white fruited phenotype was not observed,
suggesting that the white phenotype can also be obtained through mutation in
additional genes. These results provide evidence that variation in one
transcriptional regulator has generated an allelic series strongly associated
with fruit color variation in cultivated grapevine. These findings provide
information about the evolution of grapes since domestication and have direct
implications for the regulation of fruit and wine quality of this important crop
plant.

PMID: 17221259 [PubMed - in process]

15: Plant Physiol. 2007 Jan 5; [Epub ahead of print]

The Grapevine Transcription Factor VvMYBPA1 regulates Proanthocyanidin Synthesis
during Fruit Development.

Bogs J, Jaffe FW, Takos AM, Walker AR, Robinson SP.

CSIRO Plant Industry, PO Box 350, Glen Osmond, 5064, Australia; CRC for
Viticulture, PO Box 154, Glen Osmond, 5064, Australia; Heidelberger Institut fur
Pflanzenwissenschaften, Im Neuenheimer Feld 360, 69120 Heidelberg, Germany.

Proanthocyanidins (PAs or condensed tannins) can protect plants against
herbivores, contribute to the taste of many fruits and act as dietary
antioxidants beneficial for human health. We have previously shown that in
grapevine (Vitis vinifera L.) PA synthesis involves both leucoanthocyanidin
reductase (LAR) and anthocyanidin reductase (ANR). Here we report the
characterization of a grapevine MYB transcription factor VvMYBPA1, which
controls expression of PA pathway genes including both LAR and ANR. Expression
of VvMYBPA1 in grape berries correlated with PA accumulation during early berry
development and in seeds. In a transient assay, VvMYBPA1 activated the promoters
of LAR and ANR, as well as the promoters of several of the general flavonoid
pathway genes. VvMYBPA1 did not activate the promoter of VvUFGT which encodes
the anthocyanin specific enzyme UDP glucose:flavonoid-3-O-glucosyltransferase,
suggesting VvMYBPA1 is specific to regulation of PA biosynthesis in grapes. The
Arabidopsis MYB transcription factor TT2 (TRANSPARENT TESTA2) regulates PA
synthesis in the seed coat of Arabidopsis. By complementing the PA-deficient
seed phenotype of the Arabidopsis tt2 mutant with VvMYBPA1, we confirmed the
function of VvMYBPA1 as a transcriptional regulator of PA synthesis. In contrast
to ectopic expression of TT2 in Arabidopsis, constitutive expression of VvMYBPA1
resulted in accumulation of PAs in cotyledons, vegetative meristems, leaf hairs
and roots in some of the transgenic seedlings. This is the first report of a MYB
factor that controls genes of the PA pathway in fruit, including both LAR and
ANR, and this single MYB factor can induce ectopic PA accumulation in
Arabidopsis.

PMID: 17208963 [PubMed - as supplied by publisher]

16: Ann Bot (Lond). 2007 Jan 4; [Epub ahead of print]

Quantitative Analysis of the Phenotypic Variability of Shoot Architecture in Two
Grapevine (Vitis vinifera) Cultivars.

Louarn G, Guedon Y, Lecoeur J, Lebon E.

INRA, Montpellier SupAgro, UMR759 LEPSE, 2 place Viala, F-34060 Montpellier,
France.

Background and Aims Plant architecture and its interaction with agronomic
practices and environmental constraints are determinants of the structure of the
canopy, which is involved in carbon acquisition and fruit quality development. A
framework for the quantitative analysis of grapevine (Vitis vinifera) shoot
architecture, based on a set of topological and geometrical parameters, was
developed for the identification of differences between cultivars and the
origins of phenotypic variability. Methods Two commercial cultivars ('Grenache
N', 'Syrah') with different shoot architectures were grown in pots, in
well-irrigated conditions. Shoot topology was analysed, using a hidden
semi-Markov chain and variable-order Markov chains to identify deviations from
the normal pattern of succession of phytomer types (P0-P1-P2), together with
kinematic analysis of shoot axis development. Shoot geometry was characterized
by final internode and individual leaf area measurements. Key Results Shoot
architecture differed significantly between cultivars. Secondary leaf area and
axis length were greater for 'Syrah'. Secondary leaf area distribution along the
main axis also differed between cultivars, with secondary leaves preferentially
located towards the basal part of the shoot in 'Syrah'. The main factors leading
to differences in leaf area between the cultivars were: (a) slight differences
in main shoot structure, with the supplementary P0 phytomer on the lower part of
the shoot in 'Grenache N', which bears a short branch; and (b) an higher rate
and duration of development of branches bearing by P1-P2 phytomers related to P0
ones at the bottom of the shoot in 'Syrah'. Differences in axis length were
accounted for principally by differences in individual internode morphology,
with 'Syrah' having significantly longer internodes. This trait, together with a
smaller shoot diameter, may account for the characteristic 'droopy' habit of
'Syrah' shoots. Conclusions This study highlights the architectural parameters
involved in the phenotypic variability of shoot architecture in two grapevine
cultivars. Differences in primary shoot structure and in branch development
potential accounted for the main differences in leaf area distribution between
the two cultivars. By contrast, shoot shape seemed to be controlled by
differences in axis length due principally to differences in internode length.

PMID: 17204533 [PubMed - as supplied by publisher]

17: J Chromatogr A. 2006 Dec 29; [Epub ahead of print]

Headspace-solid phase microextraction-gas chromatography as a tool to define an
index that establishes the retention capacity of the wine polymeric fraction
towards ethyl esters.

Rocha SM, Coutinho P, Delgadillo I, Coimbra MA.

Departamento de Quimica, Universidade de Aveiro, 3810-193 Aveiro, Portugal.

A headspace-solid phase microextraction followed by gas chromatographic analysis
(HS-SPME-GC) was developed to be applied in the study of the interactions
between the wine polymeric fraction and the ethyl esters: ethyl hexanoate, ethyl
octanoate, and ethyl decanoate. Wine models (WM) were prepared with 10% (v/v)
aqueous ethanol at pH 3.5 with distinct wine polymeric concentrations prepared
from white wine of Vitis vinifera L. var. Fernao-Pires: 1.0gL(-1) (PWM(1)), with
a polymeric concentration approaching the real one in wine; 10.0gL(-1)
(PWM(10)); and 30.0gL(-1) (PWM(30)), saturated with polymeric fraction. A
reference wine model (RWM) was prepared without polymeric fraction. Each
volatile compound (4.0mgL(-1)) was added separately to the RWM and to the WM
with the three levels of polymeric material (PWM). From the retention index (RI)
calculated for each compound using the formula: [RI=1-(C(RWM)-C(PWM))/C(RWM)],
where C(RWM) is the concentration of the compound in the RWM and C(PWM) is the
concentration of the compound in the given PWM, the retention capacity of each
wine polymeric fraction towards the three esters was established. The higher
retention indexes were observed for ethyl decanoate, the more hydrophobic
compound, and for the PWM with higher concentration. Furthermore, this study
also suggested that the retained compounds are dosed to the headspace, which may
promote the perception of their aroma for a longer period of time.

PMID: 17196969 [PubMed - as supplied by publisher]

18: Plant Physiol. 2007 Feb;143(2):1024-36. Epub 2006 Dec 22.

Xylella fastidiosa Infection and Ethylene Exposure Result in Xylem and Water
Movement Disruption in Grapevine Shoots.

Perez-Donoso AG, Greve LC, Walton JH, Shackel KA, Labavitch JM.

Department of Plant Sciences , University of California, Davis, California
95616.

It is conventionally thought that multiplication of the xylem-limited bacterium
Xylella fastidiosa (Xf) within xylem vessels is the sole factor responsible for
the blockage of water movement in grapevines (Vitis vinifera) affected by
Pierce's disease. However, results from our studies have provided substantial
support for the idea that vessel obstructions, and likely other aspects of the
Pierce's disease syndrome, result from the grapevine's active responses to the
presence of Xf, rather than to the direct action of the bacterium. The use of
magnetic resonance imaging (MRI) to observe the distribution of water within the
xylem has allowed us to follow nondestructively the development of vascular
system obstructions subsequent to inoculation of grapevines with Xf. Because we
have hypothesized a role for ethylene produced in vines following infection, the
impact of vine ethylene exposure on obstruction development was also followed
using MRI. In both infected and ethylene-exposed plants, MRI shows that an
important proportion of the xylem vessels become progressively air embolized
after the treatments. The loss of xylem water-transporting function, assessed by
MRI, has been also correlated with a decrease in stem-specific hydraulic
conductivity (K(S)) and the presence of tyloses in the lumens of obstructed
water conduits. We have observed that the ethylene production of leaves from
infected grapevines is greater than that from healthy vines and, therefore,
propose that ethylene may be involved in a series of cellular events that
coordinates the vine's response to the pathogen.

PMID: 17189331 [PubMed - in process]

19: Chemosphere. 2007 Apr;67(6):1243-51. Epub 2006 Dec 20.

Effect of the herbicide flumioxazin on photosynthetic performance of grapevine
(Vitis vinifera L.).

Bigot A, Fontaine F, Clement C, Vaillant-Gaveau N.

Laboratoire de Stress, Defenses et Reproduction des Plantes, URVVC EA 2069,
Universite de Reims Champagne-Ardenne, UFR Sciences Exactes et Naturelles,
Batiment 18, Moulin de la Housse - BP 1039, F-51687 Reims Cedex 2, France.

Among the herbicides used in vineyards, the pre-emergence soil-applied
flumioxazin (fmx) is a recently used molecule that inhibits chlorophyll
biosynthesis in weed species. The aim of this work is to further characterize
the effects of fmx on the non-target grapevine (Vitis vinifera L. cv.
Chardonnay) using cutting as a model. Several photosynthesis parameters were
estimated during 25 days after treatment with various fmx concentrations (from
0.5mM to 50mM). Measuring chlorophyll fluorescence it appeared that fmx or a
by-product penetrated the plant throughout roots and spread throughout vessels.
Besides the initial target, protox, fmx affected other functions related to
photosynthesis. Fmx induced a simultaneous drop of both P(n), g(s) and T. Fmx
caused stomatal closure, which partially explains the decrease of the net
photosynthesis. The decline in F(v)/F(m) indicates that the photochemistry of
PSII and its ability to reduce the primary acceptor Q(A) are also affected by
fmx in grapevine. Fmx leads to a decrease in the coefficients of both
photochemical and non-photochemical quenching. Simultaneous stomatal closure and
decrease in the quantum yield of CO(2) assimilation indicate a change in energy
metabolism following fmx stress. After ten days of fmx treatment, analyses of
the response of net carbon assimilation in leaves to different intercellular
CO(2) concentrations have shown a decrease of the maximum carboxylation velocity
of RuBP. Stomatal closure, PSII photochemistry, change in energy metabolism and
RuBP activity were affected by fmx treatment.

PMID: 17184818 [PubMed - in process]

20: Bull Environ Contam Toxicol. 2006 Nov;77(5):665-70.

Lead, mercury, and nickel in grapevine, Vitis vinifera L., in polluted and
nonpolluted regions.

Todic S, Beslic Z, Lakic N, Tesic D.

University of Belgrade, Nemanjina 6, 11080 Zemun, Serbia and Montenegro.

PMID: 17176992 [PubMed - indexed for MEDLINE]

21: Plant Biotechnol J. 2005 Jan;3(1):57-69.

Antioxidant metabolite profiles in tomato fruit constitutively expressing the
grapevine stilbene synthase gene.

Giovinazzo G, d'Amico L, Paradiso A, Bollini R, Sparvoli F, Degara L.

Istituto di Scienze delle Produzioni Alimentari-CNR, via Prov. Lecce-Monteroni,
73100 Lecce, Italy.

Tomato (Lycopersicon esculentum Mill.) tissues were transformed with a grape
(Vitis vinifera L.) stilbene synthase cDNA, transcriptionally regulated by the
cauliflower mosaic virus (CaMV) 35S promoter. Transgenic plants accumulated new
compounds, not present in either wild-type or vector-transformed plants. These
were identified, by high-pressure liquid chromatography, as trans-resveratrol
and trans-resveratrol-glucopyranoside. The amounts of trans-resveratrol and its
piceid form were evaluated in the transgenic fruit. It was found that the
content of the metabolite varied during fruit maturation to up to 53 microg/g
fresh weight of total trans-resveratrol at the red stage of ripening. This
metabolite accumulation was possibly dependent on a combination of sufficiently
high levels of stilbene synthase and the availability of substrates. With the
aim of verifing the metabolic impairment, the amounts of chlorogenic acid and
naringenin in both transgenic and wild-type ripening fruit were compared and no
dramatic variation in the synthesis profile of the two metabolites was noted. To
our knowledge, no data are available on the assessment of the effects of the
expression of the StSy gene on other antioxidant compounds present in tomato
fruit. To establish whether the presence of a novel antioxidant molecule
affected the redox regulation in transgenic tomato fruit cells, the effect of
resveratrol accumulation on the naturally present antioxidant pool was analysed.
We showed that, in transgenic fruit which accumulate trans-resveratrol, there is
an increase in the levels of ascorbate and glutathione, the soluble antioxidants
of primary metabolism, as well as in the total antioxidant activity. Conversely,
the content of tocopherol and lycopene, which are membrane-located antioxidants,
is not affected. Consistent with the increased antioxidant properties, the lipid
peroxidation was lower in transformed than in wild-type fruit.

PMID: 17168899 [PubMed - in process]

22: Sheng Wu Gong Cheng Xue Bao. 2006 Nov;22(6):984-9.

[Impact of subculture cycles and inoculum sizes on suspension cultures of Vitis
vinifera]

[Article in Chinese]

Qu JG, Zhang W, Hu QL, Jin MF.

Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian
116023, China.

The commercial application of plant cell cultures is often hindered by the
instability of secondary metabolite biosynthesis, where the metabolite yield
fluctuates and decline dramatically over subcultures. This study proposed that
such instability is due to the fluctuations of culture variables. To validate
this hypothesis, the effects of the fluctuations of two culture variables
(subculture cycle and inoculum size) on the biomass, anthocyanin biosynthesig,
intracellular carbon, nitrogen and phosphate during continuous 10 subculture
cycles were investigated. The subculture cycle was fluctuated for 12h in a 7 day
cycle (6.5, 7 and 7.5 d), and the inoculum size was fluctuated by 20% on basis
of 2.00 g (1.60, 2.00 and 2.40 g). It was found that all the measured culture
parameters fluctuated over the 10 subculture cycles. The fluctuation in terms of
inoculum sizes had a greater effect on the stability of anthocyanin biosynthesis
in suspension cultures of V. vinifera. Among all the subculture conditions
investigated, 7d-subculture cycle and 1.60 g-inoculum size was the best one to
hold the relatively stable anthocyanin production. The anthocyanin yield
presented a negative correlation with intracellular sucrose content or
intracellular total phosphate content.

Publication Types:
English Abstract

PMID: 17168324 [PubMed - in process]

23: Plant Mol Biol. 2007 Mar;63(5):637-50. Epub 2006 Dec 10.

The FT/TFL1 gene family in grapevine.

Carmona MJ, Calonje M, Martinez-Zapater JM.

Departamento de Biotecnologia, Escuela Tecnica Superior de Ingenieros Agronomos,
Universidad Politecnica de Madrid, Ciudad Universitaria, 28040, Madrid, Spain,
[email protected]

The FT/TFL1 gene family encodes proteins with similarity to
phosphatidylethanolamine binding proteins which function as flowering promoters
and repressors. We show here that the FT/TFL1 gene family in Vitis vinifera is
composed of at least five genes. Sequence comparisons with homologous genes
identified in other dicot species group them in three major clades, the FT, MFT
and TFL1 subfamilies, the latter including three of the Vitis sequences. Gene
expression patterns are in agreement with a role of VvFT and VvMFT as flowering
promoters; while VvTFL1A, VvTFL1B and VvTFL1C could be associated with
vegetative development and maintenance of meristem indetermination.
Overexpression of VvFT in transgenic Arabidopsis plants generates early
flowering phenotypes similar to those produced by FT supporting a role for this
gene in flowering promotion. Overexpression of VvTFL1A does not affect flowering
time but the determination of flower meristems, strongly altering inflorescence
structure, which is consistent with the biological roles assigned to similar
genes in other species.

PMID: 17160562 [PubMed - in process]

24: J Agric Food Chem. 2006 Dec 13;54(25):9465-71.

Composition and cellular localization of tannins in Cabernet Sauvignon skins
during growth.

Gagne S, Saucier C, Geny L.

Laboratoire de Biochimie et de Physiologie de la Vigne and Laboratoire de Chimie
Appliquee, UMR 1219 Oenologie-Ampelologie, Faculte d'Oenologie-Universite Victor
Segalen, Bordeaux 2/INRA/ENITA, Institut des Sciences de la Vigne et du Vin,
Talence, France.

For two successive years, cell walls were isolated from the internal part of
skin cells of Vitis vinifera L. cv. Cabernet Sauvignon grape berries grown in a
vineyard. Procyanidin localization and composition were determined over the
course of development. Tannins were mainly localized in the inner cell fraction,
due to their biosynthesis and storage. Cell wall tannins always exhibited a
higher mean degree of polymerization as compared to the internal cell fraction,
which had a constant mDP. The mDP of cell wall tannins also tended to increase
at the end of maturation. Our results suggest tannin polymerization near the
cell wall but an aggregation in the vacuole during growth. The tannin
composition was typical of skins, and small differences were noted between the
two cell parts. Surprisingly, epigallocatechin-3-gallate was also detected,
although in a very small amount. Epicatechin was present in significant
proportions in both fractions, especially as an extension subunit, while
epigallocatechin was likewise abundant, also as a terminal subunit. Last,
procyanidin composition and organization seemed to be characteristic of the
Cabernet Sauvignon variety.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17147434 [PubMed - in process]

25: Funct Integr Genomics. 2007 Apr;7(2):111-34. Epub 2006 Nov 29.

Water and salinity stress in grapevines: early and late changes in transcript
and metabolite profiles.

Cramer GR, Ergul A, Grimplet J, Tillett RL, Tattersall EA, Bohlman MC, Vincent
D, Sonderegger J, Evans J, Osborne C, Quilici D, Schlauch KA, Schooley DA,
Cushman JC.

Department of Biochemistry and Molecular Biology, MS200, University of Nevada,
Reno, NV, 89557-0014, USA, [email protected]

Grapes are grown in semiarid environments, where drought and salinity are common
problems. Microarray transcript profiling, quantitative reverse
transcription-PCR, and metabolite profiling were used to define genes and
metabolic pathways in Vitis vinifera cv. Cabernet Sauvignon with shared and
divergent responses to a gradually applied and long-term (16 days) water-deficit
stress and equivalent salinity stress. In this first-of-a-kind study, distinct
differences between water deficit and salinity were revealed. Water deficit
caused more rapid and greater inhibition of shoot growth than did salinity at
equivalent stem water potentials. One of the earliest responses to water deficit
was an increase in the transcript abundance of RuBisCo activase (day 4), but
this increase occurred much later in salt-stressed plants (day 12). As water
deficit progressed, a greater number of affected transcripts were involved in
metabolism, transport, and the biogenesis of cellular components than did
salinity. Salinity affected a higher percentage of transcripts involved in
transcription, protein synthesis, and protein fate than did water deficit.
Metabolite profiling revealed that there were higher concentrations of glucose,
malate, and proline in water-deficit-treated plants as compared to salinized
plants. The metabolite differences were linked to differences in transcript
abundance of many genes involved in energy metabolism and nitrogen assimilation,
particularly photosynthesis, gluconeogenesis, and photorespiration.
Water-deficit-treated plants appear to have a higher demand than salinized
plants to adjust osmotically, detoxify free radicals (reactive oxygen species),
and cope with photoinhibition.

PMID: 17136344 [PubMed - in process]

26: Antiviral Res. 2007 Mar;73(3):206-11. Epub 2006 Nov 3.

Antiviral activity of tiazofurin and mycophenolic acid against Grapevine
Leafroll-associated Virus 3 in Vitis vinifera explants.

Panattoni A, D'Anna F, Triolo E.

Department of Tree Science Entomology and Plant Pathology "G. Scaramuzzi",
University of Pisa, Via del Borghetto 80, 56124 Pisa, Italy.

The ability to control plant viral diseases with chemicals has great potential
value for agriculture, but few chemicals are available to date due to the
difficulty in obtaining effective drugs. IMP dehydrogenase is an enzyme which
catalyzes the conversion of inosine 5'-monophosphate to xanthosine
5'-monophosphate in the de novo purine nucleotide synthetic pathway, and is
considered a sensitive target for antiviral drugs. Two IMPDH inhibitors,
tiazofurin (TR) and mycophenolic acid (MPA), were tested for their inhibitory
effect on Grapevine leafroll-associated virus 3 (GLRaV-3) in in vitro grapevine
explants. TR administration produced plantlets characterized by negative ELISA
readings. No PCR products were obtained from these samples. This was confirmed
by the absence of viral particles. MPA was essentially ineffective against
GLRaV-3 replication in Sangiovese explants. This is the first report of GLRaV-3
eradication in grapevine explants following TR administration.

PMID: 17125850 [PubMed - in process]

27: J Agric Food Chem. 2006 Nov 29;54(24):9206-15.

Anatomical, histological, and histochemical changes in grape seeds from Vitis
vinifera L. cv Cabernet franc during fruit development.

Cadot Y, Minana-Castello MT, Chevalier M.

Grapevine and Wine Experimental Unit, National Institute for Agricultural
Research, Angers Research Centre, 42 rue George Morel, B.P. 60057, 49071
Beaucouze cedex, France. [email protected]

Cabernet franc berries were sampled at five stages from berry set to harvest
from an experimental vineyard in mid Loire Valley. Seeds were collected from
representative berries in term of stage of development. The evolution of seed
was followed both macro- and microscopically. For microscopy analysis, seeds
were cut, put in a fixation solution, and cut into thin sections with a
microtome. Five staining solutions were used for each seed sample: toluidine
blue O, phloroglucinol, periodic acid-Schiff's reagent and naphtol blue black,
vanillin, and p-dimethylaminocinnamaldehyde. Toluidine blue O staining revealed
the evolution of tissue structures during grape seed development. We studied the
changes in chemical compounds (lignin, polysaccharides, proteins, and tannins)
with the other reagents. Seed lignification was achieved at veraison.
Proanthocyanidins were localized in epidermis, inner cells of the soft seed
coat, and inner cell layer of the inner integument. Finally, the localization of
flavan-3-ols was linked with changes in cell walls of the outer integument.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17117811 [PubMed - indexed for MEDLINE]

28: J Agric Food Chem. 2006 Nov 29;54(24):9119-25.

Methyl jasmonate induces defense responses in grapevine and triggers protection
against Erysiphe necator.

Belhadj A, Saigne C, Telef N, Cluzet S, Bouscaut J, Corio-Costet MF, Merillon
JM.

Laboratoire de Mycologie et Biotechnologie Vegetale, EA 3675, Universite Victor
Segalen Bordeaux 2, UFR Sciences Pharmaceutiques, 146 rue Leo Saignat, 33076
Bordeaux Cedex, France.

Grapevine is subject to a number of diseases that affect yield and wine quality.
To limit the excessive use of phytochemicals in the vineyard, alternative
strategies have to be developed. Plant treatment with signaling molecules like
elicitors stimulates their natural defense mechanisms. To improve grapevine
tolerance against fungal pathogens, Vitis vinifera plants were treated with a
natural exogenous elicitor, methyl jasmonate (MeJA). MeJA-treated leaves
(Cabernet Sauvignon foliar cuttings) reacted by increasing transcript levels
coding pathogenesis-related proteins (acidic class IV chitinase, serine protease
inhibitor, polygalacturonase-inhibiting protein, and beta-1,3-glucanase) and
coding enzymes involved in phytoalexin biosynthesis (one phenylalanine ammonia
lyase and one stilbene synthase). This was correlated with the accumulation of
stilbenes (antimicrobial compounds). The eliciting activity of MeJA was
confirmed by enhanced tolerance of grapevine foliar cuttings and vineyard
against powdery mildew (75% and 73%, respectively). On the basis of these
original results, MeJA can therefore act as an efficient elicitor in an
alternative strategy of grapevine protection.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17117799 [PubMed - indexed for MEDLINE]

29: Plant Physiol Biochem. 2006 Nov-Dec;44(11-12):724-31. Epub 2006 Oct 27.

Accumulation of tocopherols and tocotrienols during seed development of grape
(Vitis vinifera L. cv. Albert Lavallee).

Horvath G, Wessjohann L, Bigirimana J, Monica H, Jansen M, Guisez Y, Caubergs R,
Horemans N.

Research Group of Plant Physiology, Department of Biology, University of
Antwerp, Groenenborgerlaan 171, 2020 Antwerp, Belgium. [email protected]

Tocopherols and tocotrienols are present in mature seeds. Yet, little is known
about the physiological role and the metabolism of these compounds during seed
development. Here we present data on tocopherol and tocotrienol accumulation
during seed development in Vitis vinifera L. cv. Albert Lavallee (Royal). This
species was chosen for its ability to synthesize both tocopherols and
tocotrienols. It is shown here for the first time that during seed development
there are significant differences in localization and accumulation kinetics of
tocopherols and tocotrienols. Tocopherols are found homogeneously dispersed
throughout all tissues of the seed, in concentrations ranging from 20 to 100
microg tocopherol per g dry weight. Tocopherol levels decrease gradually during
seed development. In contrast, tocotrienols are only found in the endosperm of
the seeds, accumulating in a sigmoid fashion during the maturation period of
seed development. Tocotrienol levels were found to be (54+/-7.4) microg/g dry
seed in 90-day-old seeds of V. vinifera L. Furthermore, tocotrienol biosynthesis
is demonstrated in these seeds during tocotrienol accumulation and in an
endosperm fraction isolated at 75 days after flowering.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17110122 [PubMed - indexed for MEDLINE]

30: BMC Plant Biol. 2006 Nov 14;6:27.

An optimized grapevine RNA isolation procedure and statistical determination of
reference genes for real-time RT-PCR during berry development.

Reid KE, Olsson N, Schlosser J, Peng F, Lund ST.

Faculty of Land and Food Systems, University of British Columbia, Vancouver,
Canada. [email protected]

BACKGROUND: Accuracy in quantitative real-time RT-PCR is dependent on high
quality RNA, consistent cDNA synthesis, and validated stable reference genes for
data normalization. Reference genes used for normalization impact the results
generated from expression studies and, hence, should be evaluated prior to use
across samples and treatments. Few statistically validated reference genes have
been reported in grapevine. Moreover, success in isolating high quality RNA from
grapevine tissues is typically limiting due to low pH, and high polyphenolic and
polysaccharide contents. RESULTS: We describe optimization of an RNA isolation
procedure that compensates for the low pH found in grape berries and improves
the ability of the RNA to precipitate. This procedure was tested on pericarp and
seed developmental series, as well as steady-state leaf, root, and flower
tissues. Additionally, the expression stability of actin, AP47
(clathrin-associated protein), cyclophilin, EF1-alpha (elongation factor
1-alpha), GAPDH (glyceraldehyde 3-phosphate dehydrogenase), MDH (malate
dehydrogenase), PP2A (protein phosphatase), SAND, TIP41, alpha-tubulin,
beta-tubulin, UBC (ubiquitin conjugating enzyme), UBQ-L40 (ubiquitin L40) and
UBQ10 (polyubiquitin) were evaluated on Vitis vinifera cv. Cabernet Sauvignon
pericarp using three different statistical approaches. Although several of the
genes proved to be relatively stable, no single gene outperformed all other
genes in each of the three evaluation methods tested. Furthermore, the effect of
using one reference gene versus normalizing to the geometric mean of several
genes is presented for the expression of an aquaporin and a sucrose transporter
over a developmental series. CONCLUSION: In order to quantify relative
transcript abundances accurately using real-time RT-PCR, we recommend that
combinations of several genes be used for normalization in grape berry
development studies. Our data support GAPDH, actin, EF1-alpha and SAND as the
most relevant reference genes for this purpose.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17105665 [PubMed - indexed for MEDLINE]

31: Transgenic Res. 2006 Nov 14; [Epub ahead of print]

Silencing of chaperonin 21, that was differentially expressed in inflorescence
of seedless and seeded grapes, promoted seed abortion in tobacco and tomato
fruits.

Hanania U, Velcheva M, Or E, Flaishman M, Sahar N, Perl A.

Department of Fruit Tree Sciences, Institute of Plant Sciences, Agricultural
Research Organization, P.O. Box 6, 50250, Bet-Dagan, Israel, [email protected]

Vitis vinifera L. cv. 'Thompson Seedless' presents a type of stenospermocarpy in
grape where fertilization occurs but seeds abort and fail to develop. To unravel
the molecular basis for stenospermocarpy in grapes, subtractive hybridization
was carried out in order to isolate differentially regulated genes that
participate in the seedlessness machinery. Two 'Thompson' lines, a seeded and a
seedless, were screened during different flower developmental stages. One of the
genes, that was differentially expressed between the seeded and seedless lines,
was the chloroplast chaperonin 21 (ch-Cpn21). ch-Cpn21 is a 21-kDa co-chaperonin
polypeptide formed by two GroES-like domains fused together in tandem. Silencing
of ch-Cpn21 in Nicotiana benthamiana plants resulted in leaf stunting,
chlorosis, as well as ovary necrogenesis leading to seed abortion. Moreover,
organ-specific silencing of ch-Cpn21 only in Lycopersicum esculentum fruits
resulted in the development of seedless tomatoes. These results suggest that
ch-Cpn21 may play a role in seed abortion in stenospermocarpic grapes.

PMID: 17103240 [PubMed - as supplied by publisher]

32: J Agric Food Chem. 2006 Nov 15;54(23):8775-81.

Extraction of polyphenols from vine shoots of Vitis vinifera by superheated
ethanol-water mixtures.

Luque-Rodriguez JM, Perez-Juan P, Luque de Castro MD.

Department of Analytical Chemistry, University of Cordoba, Campus of Rabanales,
Annex C-3, 14071 Cordoba, Spain.

A study of the nonvolatile fraction of extracts from vine shoots obtained by
superheated ethanol-water mixtures is presented. The influence of the
temperature, extraction time, and percentage of ethanol on extraction was
investigated by a multivariate experimental design to maximize the yield of
total phenolic compounds, measured by using the Folin-Ciocalteu method. The best
values found for these variables were 80% (v/v) ethanol, 240 degrees C, and 60
min. Under these conditions, the effect of pH was also investigated, and a
strong improvement of yield was observed by decreasing the pH. The extracts were
subject to liquid-liquid extraction with n-hexane. The remaining polar phase was
dried in a rotary evaporator and then reconstituted in 10 mL of water. The
insoluble residue was dissolved in 10 mL of methanol. Both fractions (aqueous
and methanolic) were analyzed by HPLC, and the differences in composition
according to the extraction conditions were studied. Compounds usually present
in commercial wood extracts were identified (mainly benzoic and hydroxycinnamic
acids and aldehydes); the most abundant were quantified, and the stability of
the identified phenolic families under different extraction conditions was also
investigated. Finally, the superiority of the superheated liquid extraction over
conventional solid-liquid extraction was demonstrated.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17090121 [PubMed - indexed for MEDLINE]

33: Plant Cell Environ. 2006 May;29(5):993-1001.

Direct in situ measurement of cell turgor in grape (Vitis vinifera L.) berries
during development and in response to plant water deficits.

Thomas TR, Matthews MA, Shackel KA.

Department of Viticulture and Enology, University of California, Davis, CA
95616-8683, USA.

Vitis vinifera L. berries are non-climacteric fruits that exhibit a
double-sigmoid growth pattern, and at the point known as 'veraison', which is
just before the beginning of the second period of rapid fruit growth, these
berries undergo several abrupt physiological changes. Cell pressure probe was
used to examine the in situ turgor (P) of cells in the mesocarp during berry
development and in response to plant water deficits. Initial tests comparing
attached and detached berries demonstrated that cell P was stable for up to 48 h
after detachment from the vine, provided that water loss from the berry was
prevented. Cell P at pre-dawn was on the order of 0.25 MPa pre-veraison (PreV)
and was reduced by an order of magnitude to 0.02 MPa post veraison (PostV). Cell
P declined slightly but significantly with depth from the berry surface PreV,
but not PostV. When water was withheld from potted vines, cell P declined about
0.2 Mpa, as pre-dawn vine water potential declined about 0.6 MPa over 12 d,
whereas cell P was completely insensitive to a 1.10 MPa decrease in pre-dawn
vine water potential after veraison. Rewatering of stressed plants also resulted
in a 24 h recovery of cell P before, but not after veraison. The substantial
decline in cell P around veraison is consistent with the decline in berry
firmness that is known to occur at this time, and the PostV insensitivity of P
to changes in vine water status is consistent with current hypotheses that the
PostV berry is hydraulically isolated from the vine. The fact that a measurable
P of about 0.02 MPa and typical cell hydraulic/osmotic behaviour were exhibited
in PostV berries, however, indicates that cell membranes remain intact after
veraison, contrary to many current hypotheses that veraison is associated with a
general loss of membrane function and cellular compartmentation in the grape
berry. We hypothesize that cell P is low in the PostV berry, and possibly other
fleshy fruits, because of the presence of regulated quantities of apoplastic
solutes.

PMID: 17087481 [PubMed - indexed for MEDLINE]

34: New Phytol. 2006;172(3):469-78.

Mercury hinders recovery of shoot hydraulic conductivity during grapevine
rehydration: evidence from a whole-plant approach.

Lovisolo C, Schubert A.

Dipartimento Colture Arboree, Universita di Torino, Via Leonardo da Vinci 44,
I-10095 Grugliasco, Italy. [email protected]

This experiment aimed to test whether recovery of shoot hydraulic conductivity
after drought depends on cellular metabolism in addition to xylem hydraulics. We
rehydrated droughted grapevines (Vitis vinifera) after treating intact plants
through the root with 0.5 mm mercuric chloride (a metabolic inhibitor) at the
end of the stress period, before rehydration. The contribution of
mercury-inhibited water transport in both shoot and root, and the extent of
shoot vessel embolization, were assessed. Drought stress decreased plant water
potential and induced embolization of the shoot vessels. The rehydration in
Hg-untreated plants re-established both shoot water potential and specific shoot
hydraulic conductivity (Kss) at levels comparable with watered controls, and
induced recovery of most of the embolisms formed in the shoot during the
drought. In contrast, in plants treated with HgCl2, recovery of Kss and root
hydraulic conductance were impaired. In rehydrated, Hg-treated plants, the
effects of Hg on Kss were reversed when either the shoot or the root was treated
with 60 mM beta-mercaptoethanol as a mercuric scavenger. This work suggests that
plant cellular metabolism, sensitive to mercuric chloride, affects the recovery
of shoot hydraulic conductivity during grapevine rehydration by interfering with
embolism removal, and that it involves either the root or the shoot level.

PMID: 17083677 [PubMed - indexed for MEDLINE]

35: Plant Cell Environ. 2006 Jul;29(7):1410-21.

Three grape CBF/DREB1 genes respond to low temperature, drought and abscisic
acid.

Xiao H, Siddiqua M, Braybrook S, Nassuth A.

Department of Molecular and Cellular Biology, University of Guelph, Guelph,
Ontario N1G 2W1, Canada.

The C-repeat (CRT)-binding factor/dehydration-responsive element (DRE) binding
protein 1 (CBF/ DREB1) transcription factors control an important pathway for
increased freezing and drought tolerance in plants. Three CBF/DREB1-like genes,
CBF 1-3, were isolated from both freezing-tolerant wild grape (Vitis riparia)
and freezing-sensitive cultivated grape (Vitis vinifera). The deduced proteins
in V. riparia are 63-70% identical to each other and 96-98% identical to the
corresponding proteins in V. vinifera. All Vitis CBF proteins are 42-51%
identical to AtCBF1 and contain CBF-specific amino acid motifs, supporting their
identification as CBF proteins. Grape CBF sequences are unique in that they
contain 20-29 additional amino acids and three serine stretches.
Agro-infiltration experiments revealed that VrCBF1b localizes to the nucleus.
VrCBF1a, VrCBF1b and VvCBF1 activated a green fluorescent protein (GFP) or
glucuronidase (GUS) reporter gene behind CRT-containing promoters. Expression of
the endogenous CBF genes was low at ambient temperature and enhanced upon low
temperature (4 degrees C) treatment, first for CBF1, followed by CBF2, and about
2 d later by CBF3. No obvious significant difference was observed between V.
riparia and V. vinifera genes. The expression levels of all three CBF genes were
higher in young tissues than in older tissues. CBF1, 2 and 3 transcripts also
accumulated in response to drought and exogenous abscisic acid (ABA) treatment,
indicating that grape contains unique CBF genes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17080962 [PubMed - indexed for MEDLINE]

36: Transgenic Res. 2006 Dec;15(6):687-702. Epub 2006 Oct 27.

The grapevine polygalacturonase-inhibiting protein (VvPGIP1) reduces Botrytis
cinerea susceptibility in transgenic tobacco and differentially inhibits fungal
polygalacturonases.

Joubert DA, Slaughter AR, Kemp G, Becker JV, Krooshof GH, Bergmann C, Benen J,
Pretorius IS, Vivier MA.

Department of Viticulture & Oenology, Institute for Wine Biotechnology,
Stellenbosch University, Stellenbosch, 7600, South Africa, [email protected]

Polygalacturonase-inhibiting proteins (PGIPs) selectively inhibit
polygalacturonases (PGs) secreted by invading plant pathogenic fungi. PGIPs
display differential inhibition towards PGs from different fungi, also towards
different isoforms of PGs originating from a specific pathogen. Recently, a
PGIP-encoding gene from Vitis vinifera (Vvpgip1) was isolated and characterised.
PGIP purified from grapevine was shown to inhibit crude polygalacturonase
extracts from Botrytis cinerea, but this inhibitory activity has not yet been
linked conclusively to the activity of the Vvpgip1 gene product. Here we use a
transgenic over-expression approach to show that the PGIP encoded by the Vvpgip1
gene is active against PGs of B. cinerea and that over-expression of this gene
in transgenic tobacco confers a reduced susceptibility to infection by this
pathogen. A calculated reduction in disease susceptibility of 47-69% was
observed for a homogeneous group of transgenic lines that was statistically
clearly separated from untransformed control plants following infection with
Botrytis over a 15-day-period. VvPGIP1 was subsequently purified from transgenic
tobacco and used to study the specific inhibition profile of individual PGs from
Botrytis and Aspergillus. The heterologously expressed and purified VvPGIP1
selectively inhibited PGs from both A. niger and B. cinerea, including BcPG1, a
PG from B. cinerea that has previously been shown to be essential for virulence
and symptom development. Altogether our data confirm the antifungal nature of
the VvPGIP1, and the in vitro inhibition data suggest at least in part, that the
VvPGIP1 contributed to the observed reduction in disease symptoms by inhibiting
the macerating action of certain Botrytis PGs in planta. The ability to
correlate inhibition profiles to individual PGs provides a more comprehensive
analysis of PGIPs as antifungal genes with biotechnological potential, and adds
to our understanding of the importance of PGIP:PG interactions during disease
and symptom development in plants.

PMID: 17072564 [PubMed - in process]

37: J Agric Food Chem. 2006 Nov 1;54(22):8510-20.

Effect of shading on accumulation of flavonoid compounds in (Vitis vinifera L.)
pinot noir fruit and extraction in a model system.

Cortell JM, Kennedy JA.

Department of Food Science and Technology, Oregon State University, Corvallis,
Oregon, 97331, USA.

Accumulation and compositional changes of flavonols, proanthocyanidins, and
anthocyanins were measured in Vitis vinifera L. cv. Pinot noir in shaded and
exposed treatments. In addition, extraction of these compounds into a model wine
solution was measured. The study was conducted in a commercial vineyard within a
uniform zone of relatively low vigor vines. Light exclusion boxes were installed
on pairs of clusters on the same shoot (shaded treatment), and a second set of
clusters on an adjacent shoot were labeled as the exposed treatment. Fruit
samples were harvested at the onset of ripening (veraison) and at commercial
harvest. Cluster shading resulted in a substantial decrease in mg/berry
accumulation of flavonols and skin proanthocyanidins and minimal differences in
anthocyanins. In analyzing seed proanthocyanidins by phloroglucinolysis, shaded
and exposed treatments were similar at veraison; however, by harvest, the shaded
treatment had higher extension and terminal subunits (nmol/seed) as compared to
the exposed treatment. For skin proanthocyanidins, shaded fruit was lower for
all subunits (nmol/berry) at both veraison and harvest. Shading caused an
increase in the proportion of (-)-epicatechin and a decrease in
(-)-epigallocatechin at harvest in skin extension subunits. Seed
proanthocyanidins in shaded fruit contained a lower proportion of (+)-catechin
and a higher proportion of (-)-epicatechin-3-O-gallate in extension subunits and
a lower proportion of (+)-catechin and (-)-epicatechin-3-O-gallate and a higher
proportion of (-)-epicatechin in terminal subunits. For anthocyanins, the shaded
treatment had a proportional reduction in delphinidin, cyanidin, petunidin, and
malvidin and a large increase in peonidin glucosides. The model extractions from
the two treatments paralleled differences in the fruit with a lower
concentration of flavonols, anthocyanins, and proanthocyanidins in the shaded
treatment. The skin proanthocyanidin percent extraction was found to be
approximately 17% higher in the exposed model extraction than the shaded
treatment.

PMID: 17061828 [PubMed - indexed for MEDLINE]

38: Plant Cell. 2006 Oct;18(10):2767-81. Epub 2006 Oct 13.

Abiotic stress generates ROS that signal expression of anionic glutamate
dehydrogenases to form glutamate for proline synthesis in tobacco and grapevine.

Skopelitis DS, Paranychianakis NV, Paschalidis KA, Pliakonis ED, Delis ID,
Yakoumakis DI, Kouvarakis A, Papadakis AK, Stephanou EG, Roubelakis-Angelakis
KA.

Department of Biology, University of Crete, 71409 Heraklion, Greece.

Glutamate dehydrogenase (GDH) may be a stress-responsive enzyme, as GDH exhibits
considerable thermal stability, and de novo synthesis of the alpha-GDH subunit
is induced by exogenous ammonium and senescence. NaCl treatment induces reactive
oxygen species (ROS), intracellular ammonia, expression of tobacco (Nicotiana
tabacum cv Xanthi) gdh-NAD;A1 encoding the alpha-subunit of GDH, increase in
immunoreactive alpha-polypeptide, assembly of the anionic isoenzymes, and in
vitro GDH aminating activity in tissues from hypergeous plant organs. In vivo
aminating GDH activity was confirmed by gas chromatorgraphy-mass spectrometry
monitoring of (15)N-Glu, (15)N-Gln, and (15)N-Pro in the presence of methionine
sulfoximine and amino oxyacetic acid, inhibitors of Gln synthetase and
transaminases, respectively. Along with upregulation of alpha-GDH by NaCl,
isocitrate dehydrogenase genes, which provide 2-oxoglutarate, are also induced.
Treatment with menadione also elicits a severalfold increase in ROS and
immunoreactive alpha-polypeptide and GDH activity. This suggests that ROS
participate in the signaling pathway for GDH expression and protease activation,
which contribute to intracellular hyperammonia. Ammonium ions also mimic the
effects of salinity in induction of gdh-NAD;A1 expression. These results,
confirmed in tobacco and grape (Vitis vinifera cv Sultanina) tissues, support
the hypothesis that the salinity-generated ROS signal induces alpha-GDH subunit
expression, and the anionic iso-GDHs assimilate ammonia, acting as antistress
enzymes in ammonia detoxification and production of Glu for Pro synthesis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17041150 [PubMed - indexed for MEDLINE]

39: Sheng Wu Gong Cheng Xue Bao. 2006 Sep;22(5):805-10.

[Effect of homogeneity on cell growth and anthocyanin biosynthesis in suspension
cultures of Vitis vinifera]

[Article in Chinese]

Qu JG, Zhang W, Jin MF, Yu XJ.

Dalian Institute of Chemical Physics, Chinese Academy of Sciences, China.

The instability of secondary metabolite production is a ubiquitous problem in
plant cell culture. To understand the instability, the investigation of
anthocyanin accumulation in suspension cultures of Vitis vinifera, as a model
system, has been initiated in our laboratory. Suspension culture of a relatively
homogeneous cell line E of V. vinifera, was established by long-term cell line
selection by anthocyanin content differentiation. The aggregate size of E was
smaller than that of other cell lines obtained by routine screening method. The
variation coefficients of anthocyanin content in suspension cultures of E were
8.7% in long-term subcultures and 5% in repeated flasks, respectively. The
effects of elicitor, precursor feeding and light irridiation on biomass and
anthocyanin accumulation in suspension cultures of E had been investigated and
the results showed that all the variation coefficients were lower than 12% and
this indicated the importance of homogeneity on stable production in plant cell
culture. With the combination treatment of 30micromol/L phenylalanine and
218micromol/L methyl jasmonate in the dark in suspension cultures of E, the
anthocyanin content and production in suspension culture of E was 5.89-fold and
4.30-fold of the controls, respectively, and all the variation coefficients of
biomass and anthocyanin accumulation were lower than those of the controls in 5
successive subcultures.

Publication Types:
English Abstract

PMID: 17037206 [PubMed - in process]

40: Methods Mol Biol. 2006;344:273-85.

Grapevine (Vitis vinifera L.).

Bouquet A, Torregrosa L, Iocco P, Thomas MR.

UMR Biology of Development of Cultivated Perennial Plants, Agro-M/INRA, 2, place
Pierre Viala, 34060 Montpellier Cedex1, France.

Grapevine (Vitis) is considered to be one of the major fruit crops in the world
based on hectares cultivated and economic value. Grapes are used not only for
wine but also for fresh fruit, dried fruit, and juice production. Wine is by far
the major product of grapes and the focus of this chapter is on wine grape
cultivars. Grapevine cultivars of Vitis vinifera L. have a reputation for
producing premium quality wines. These premium quality wines are produced from a
small number of cultivars that enjoy a high level of consumer acceptance and are
firmly entrenched in the market place because of varietal name branding and the
association of certain wine styles and regions with specific cultivars. In light
of this situation, grapevine improvement by a transgenic approach is attractive
when compared to a classical breeding approach. The transfer of individual
traits as single genes with a minimum disruption to the original genome would
leave the traditional characteristics of the cultivar intact. However, a
reliable transformation system is required for a successful transgenic approach
to grapevine improvement. There are three criteria for achieving an efficient
Agrobacterium-mediated transformation system: (1) the production of highly
regenerative transformable tissue, (2) optimal co-cultivation conditions for
both grapevine tissue and Agrobacterium, and (3) an efficient selection regime
for transgenic plant regeneration. In this chapter, we describe a grapevine
transformation system which meets the above mentioned criteria.

PMID: 17033070 [PubMed - indexed for MEDLINE]

41: J Exp Bot. 2006 Oct 10; [Epub ahead of print]

Exploring thermal imaging variables for the detection of stress responses in
grapevine under different irrigation regimes.

Grant OM, Tronina L, Jones HG, Chaves MM.

Laboratorio de Ecofisiologia Molecular, Instituto de Tecnologia Quimica e
Biologica, Apartado 127, 2781-901 Oeiras, Portugal.

Temperatures of leaves or canopies can be used as indicators of stomatal closure
in response to soil water deficit. In 2 years of field experiments with
grapevines (Vitis vinifera L., cvs Castelao and Aragones), it was found that
thermal imaging can distinguish between irrigated and non-irrigated canopies,
and even between deficit irrigation treatments. Average canopy temperature was
inversely correlated with stomatal conductance measured with a porometer.
Variation of the distribution of temperatures within canopies was not found to
be a reliable indicator of stress. A large degree of variation between images
was found in reference 'wet' and 'dry' leaves used in the first year for the
calculation of an index proportional to stomatal conductance. In the second
year, fully irrigated (FI) (100% Et(c)) and non-irrigated (NI) canopies were
used as alternatives to wet and dry leaves. A crop water stress index utilizing
these FI and NI 'references', where stressed canopies have the highest values
and non-stressed canopies have the lowest values, was found to be a suitable
measure for detecting stress. It is suggested that the average temperatures of
areas of canopies containing several leaves may be more useful for
distinguishing between irrigation treatments than the temperatures of individual
leaves. Average temperatures over several leaves per canopy may be expected to
reduce the impact of variation in leaf angles. The results are discussed in
relation to the application of thermal imaging to irrigation scheduling and
monitoring crop performance.

PMID: 17032729 [PubMed - as supplied by publisher]

42: Mol Ecol. 2006 Oct;15(12):3707-14.

Multiple origins of cultivated grapevine (Vitis vinifera L. ssp. sativa) based
on chloroplast DNA polymorphisms.

Arroyo-Garcia R, Ruiz-Garcia L, Bolling L, Ocete R, Lopez MA, Arnold C, Ergul A,
Soylemezoglu G, Uzun HI, Cabello F, Ibanez J, Aradhya MK, Atanassov A, Atanassov
I, Balint S, Cenis JL, Costantini L, Goris-Lavets S, Grando MS, Klein BY,
McGovern PE, Merdinoglu D, Pejic I, Pelsy F, Primikirios N, Risovannaya V,
Roubelakis-Angelakis KA, Snoussi H, Sotiri P, Tamhankar S, This P, Troshin L,
Malpica JM, Lefort F, Martinez-Zapater JM.

Departamento de Genetica Molecular de Plantas, Centro Nacional de Biotecnologia,
CSIC, C/Darwin 3, 28049 Madrid, Spain.

The domestication of the Eurasian grape (Vitis vinifera ssp. sativa) from its
wild ancestor (Vitis vinifera ssp. sylvestris) has long been claimed to have
occurred in Transcaucasia where its greatest genetic diversity is found and
where very early archaeological evidence, including grape pips and artefacts of
a 'wine culture', have been excavated. Whether from Transcaucasia or the nearby
Taurus or Zagros Mountains, it is hypothesized that this wine culture spread
southwards and eventually westwards around the Mediterranean basin, together
with the transplantation of cultivated grape cuttings. However, the existence of
morphological differentiation between cultivars from eastern and western ends of
the modern distribution of the Eurasian grape suggests the existence of
different genetic contribution from local sylvestris populations or multilocal
selection and domestication of sylvestris genotypes. To tackle this issue, we
analysed chlorotype variation and distribution in 1201 samples of sylvestris and
sativa genotypes from the whole area of the species' distribution and studied
their genetic relationships. The results suggest the existence of at least two
important origins for the cultivated germplasm, one in the Near East and another
in the western Mediterranean region, the latter of which gave rise to many of
the current Western European cultivars. Indeed, over 70% of the Iberian
Peninsula cultivars display chlorotypes that are only compatible with their
having derived from western sylvestris populations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17032268 [PubMed - indexed for MEDLINE]

43: Mol Plant Microbe Interact. 2006 Oct;19(10):1103-12.

Molecular basis of ergosterol-induced protection of grape against botrytis
cinerea: induction of type I LTP promoter activity, WRKY, and stilbene synthase
gene expression.

Laquitaine L, Gomes E, Francois J, Marchive C, Pascal S, Hamdi S, Atanassova R,
Delrot S, Coutos-Thevenot P.

Laboratoire de Physiologie et Biochimie Vegetales, UMR CNRS 6161, Universite de
Poitiers, Batiment Botanique, 40 Avenue du Recteur Pineau, 86022 Poitiers Cedex,
France.

Type I lipid transfer proteins (LTPs) are basic, 9-kDa cystein-rich proteins
believed to be involved in plant defense mechanisms. A 2,100-bp fragment
containing the coding region of Vitis vinifera lipid transfer protein 1 (VvLTP1)
and 1,420-bp of its promoter region was isolated by screening a grape genomic
library. In silico analysis revealed several putative, defense-related,
cis-regulatory elements such as W- and MYB-boxes, involved in the binding of
WRKY and MYB transcription factors, respectively. The 5'-truncated versions of
the VvLTP1 promoter were generated, cloned in front of the beta-glucuronidase
(GUS) reporter gene, and introduced in tobacco plants and grapevine cell
suspensions using Agrobacterium spp. Single MYB- and the W-boxes identified on
the 0.250-kbp fragment were sufficient to induce GUS activity in transgenic
tobacco plants after transient expression of MYB and WRKY. Ergosterol, a
nonspecific fungal elicitor, induced GUS activity in transgenic grapevine cell
suspensions transformed with the 1,420- and 750-bp promoter containing a
palindromic arrangement of two W-boxes but not the 650- or 250-bp fragment,
where only one W-box was present. Moreover, ergosterol triggered WRKY, VvLTP1,
and stilbene synthase gene expression in grape plantlets and enhanced protection
against Botrytis cinerea. The molecular basis of ergosterol-induced protection
is discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17022174 [PubMed - indexed for MEDLINE]

44: Plant Physiol. 2006 Nov;142(3):1216-32. Epub 2006 Sep 29.

Light-induced expression of a MYB gene regulates anthocyanin biosynthesis in red
apples.

Takos AM, Jaffe FW, Jacob SR, Bogs J, Robinson SP, Walker AR.

Commonwealth Scientific and Industrial Research Organization, Plant Industry,
Adelaide Laboratory, Waite Campus, Urrbrae, SA 5064, Australia.

Anthocyanins are secondary metabolites found in higher plants that contribute to
the colors of flowers and fruits. In apples (Malus domestica Borkh.), several
steps of the anthocyanin pathway are coordinately regulated, suggesting control
by common transcription factors. A gene encoding an R2R3 MYB transcription
factor was isolated from apple (cv Cripps' Pink) and designated MdMYB1. Analysis
of the deduced amino acid sequence suggests that this gene encodes an ortholog
of anthocyanin regulators in other plants. The expression of MdMYB1 in both
Arabidopsis (Arabidopsis thaliana) plants and cultured grape cells induced the
ectopic synthesis of anthocyanin. In the grape (Vitis vinifera) cells MdMYB1
stimulated transcription from the promoters of two apple genes encoding
anthocyanin biosynthetic enzymes. In ripening apple fruit the transcription of
MdMYB1 was correlated with anthocyanin synthesis in red skin sectors of fruit.
When dark-grown fruit were exposed to sunlight, MdMYB1 transcript levels
increased over several days, correlating with anthocyanin synthesis in the skin.
MdMYB1 gene transcripts were more abundant in red skin apple cultivars compared
to non-red skin cultivars. Several polymorphisms were identified in the promoter
of MdMYB1. A derived cleaved amplified polymorphic sequence marker designed to
one of these polymorphisms segregated with the inheritance of skin color in
progeny from a cross of an unnamed red skin selection (a sibling of Cripps'
Pink) and the non-red skin cultivar Golden Delicious. We conclude that MdMYB1
coordinately regulates genes in the anthocyanin pathway and the expression level
of this regulator is the genetic basis for apple skin color.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 17012405 [PubMed - indexed for MEDLINE]

45: J Agric Food Chem. 2006 Oct 4;54(20):7692-702.

Metabolite profiling of grape: Flavonols and anthocyanins.

Mattivi F, Guzzon R, Vrhovsek U, Stefanini M, Velasco R.

IASMA Research Centre, Via E. Mach 1, 38010 San Michele all'Adige, Italy.
[email protected]

Flavonols are products of the flavonoid biosynthetic pathway, which also give
rise to anthocyanins and condensed tannins in grapes. We investigated their
presence in the berry skins of 91 grape varieties (Vitis vinifera L.), in order
to produce a classification based on the flavonol profile. The presence of
laricitrin 3-O-galactoside and syringetin 3-O-galactoside in red grapes is
reported here for the first time. In red grapes, the main flavonol was quercetin
(mean = 43.99%), followed by myricetin (36.81%), kaempferol (6.43%), laricitrin
(5.65%), isorhamnetin (3.89%), and syringetin (3.22%). In white grapes, the main
flavonol was quercetin (mean = 81.35%), followed by kaempferol (16.91%) and
isorhamnetin (1.74%). The delphinidin-like flavonols myricetin, laricitrin, and
syringetin were missing in all white varieties, indicating that the enzyme
flavonoid 3',5'-hydroxylase is not expressed in white grape varieties. The
pattern of expression of flavonols and anthocyanins in red grapes was compared,
in order to gain information on the substrate specificity of enzymes involved in
flavonoid biosynthesis.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 17002441 [PubMed - indexed for MEDLINE]

46: J Agric Food Chem. 2006 Oct 4;54(20):7488-94.

Survey of grapevine Vitis vinifera stem polyphenols by liquid
chromatography-diode array detection-tandem mass spectrometry.

Pussa T, Floren J, Kuldkepp P, Raal A.

Department of Food Hygiene and Control, Estonian University of Life Sciences,
Kreutzwaldi 58A, 51014 Tartu, Estonia.

Grapes and red wine prepared from Vitis vinifera L. contain a variety of
polyphenols. Some information is available about the polyphenols of the seeds
and leaves of grapevine, but considerably less is known about the polyphenols of
woody stems. In this paper, we describe the results of a study of polyphenolic
compounds in grapevine stems. We demonstrate how a combination of reversed phase
high-performance liquid chromatography with ultraviolet-diode array detection
and electrospray ionization-tandem mass spectrometry ion-trap detection enables
characterization of a phytochemical mixture of considerable complexity. As the
polyphenol source, the stems of three frost-hardy grapevine varieties [Hasaine
(Hasansky) sladki, Zilga, and Yubilei Novgoroda] were used. The main group of
methanol-extractable polyphenols of stems consists of trans-resveratrol and its
derivatives including oligomers and glucosides. As minor components of the
extract, stilbenoid piceatannol as well as a number of nonstilbenoid
polyphenols, mostly flavan-3-ols and phenolic acids glucosides, were determined.
The total polyphenol content of the grapevine stems depends on the variety,
whereby the stems of cultivar Yubilei Novgoroda with white grapes contain
significantly less of both groups of polyphenols.

PMID: 17002412 [PubMed - indexed for MEDLINE]

47: Food Addit Contam. 2006 Oct;23(10):994-9.

Rate of degradation of lambda-cyhalothrin and methomyl in grapes (Vitis vinifera
L.).

Banerjee K, Upadhyay AK, Adsule PG, Patil SH, Oulkar DP, Jadhav DR.

National Research Centre for Grapes, P. O. Manjri Farm, P. B. No. 3, Solapur
Road, Pune, 412 307, Maharashtra, India. [email protected]

Rates of degradation of lambda-cyhalothrin and methomyl residues in grape are
reported. The dissipation behavior of both insecticides followed first-order
rate kinetics with similar patterns at standard and double-dose applications.
Residues of lambda-cyhalothrin were lost with pre-harvest intervals (PHI) of
12.0-12.5 and 15.0-15.5 days, corresponding to the applications at 25 and 50 g
a.i. ha-1, respectively. In the case of methomyl, residues were lost with PHI of
55.0 and 61.0 days, following applications at 1 and 2 kg a.i. ha-1,
respectively. The PHI, recommended on the basis of the experimental results, was
shown to be effective in minimizing residue load of these insecticides below
their maximum residue limits (MRLs) in vineyard samples.

PMID: 16982521 [PubMed - in process]

48: J Plant Physiol. 2006 Sep 16; [Epub ahead of print]

Cloning, functional identification and structural modelling of Vitis vinifera
S-adenosylmethionine decarboxylase.

Tassoni A, Franceschetti M, Tasco G, Casadio R, Bagni N.

Department of Biology e.s. and Interdepartmental Centre for Biotechnology,
University of Bologna, Via Irnerio 42, Bologna 40126, Italy.

In this paper we report the cloning and full sequencing of S-adenosylmethionine
decarboxylase (SAMDC, EC 4.1.1.50) cDNA from Vitis vinifera L. (VV) leaves, an
enzyme belonging to the polyamine biosynthetic pathway, which appears to play an
important role in the regulation of plant growth and development. The presence
of two overlapping ORFs (tiny ORF and small ORF) upstream of the main ORF is
reported in the Vitis cDNA. When the Vitis SAMDC cDNA was expressed in yeast
without the two upstream ORFs, the resulting activity was about 50 times higher
than the activity obtained with the full cDNA. These results demonstrated the
strong regulatory activity of the tiny and small ORFs. RT-PCR expression
analysis showed evidence of a similar mRNA level in all the tissues tested, with
the exception of the petioles. The VV SAMDC was also modelled using its
homologues from Solanum tuberosum and Homo sapiens as template. The present work
confirmed, for the first time in a woody plant of worldwide economic interest
such as grapevine, the presence of a regulatory mechanism of SAMDC, enzyme that
has a well-established importance in the modulation of plant growth and
development.

PMID: 16982115 [PubMed - as supplied by publisher]

49: Appl Environ Microbiol. 2006 Nov;72(11):7246-52. Epub 2006 Sep 15.

Enhancement of chilling resistance of inoculated grapevine plantlets with a
plant growth-promoting rhizobacterium, Burkholderia phytofirmans strain PsJN.

Ait Barka E, Nowak J, Clement C.

Laboratoire de Stress, Defenses et Reproduction des Plantes, Unite de Recherche
Vignes et Vins de Champagne, UPRES EA 2069, UFR Sciences, Universite de Reims
Champagne-Ardenne, 51687 Reims Cedex 2, France. [email protected]

In vitro inoculation of Vitis vinifera L. cv. Chardonnay explants with a plant
growth-promoting rhizobacterium, Burkholderia phytofirmans strain PsJN,
increased grapevine growth and physiological activity at a low temperature.
There was a relationship between endophytic bacterial colonization of the
grapevine plantlets and their growth at both ambient (26 degrees C) and low (4
degrees C) temperatures and their sensitivities to chilling. The major benefits
of bacterization were observed on root growth (11.8- and 10.7-fold increases at
26 degrees C and 4 degrees C, respectively) and plantlet biomass (6- and
2.2-fold increases at 26 degrees C and 4 degrees C, respectively). The
inoculation with PsJN also significantly improved plantlet cold tolerance
compared to that of the nonbacterized control. In nonchilled plantlets,
bacterization enhanced CO(2) fixation and O(2) evolution 1.3 and 2.2 times,
respectively. The nonbacterized controls were more sensitive to exposure to low
temperatures than were the bacterized plantlets, as indicated by several
measured parameters. Moreover, relative to the noninoculated controls,
bacterized plantlets had significantly increased levels of starch, proline, and
phenolics. These increases correlated with the enhancement of cold tolerance of
the grapevine plantlets. In summary, B. phytofirmans strain PsJN inoculation
stimulates grapevine growth and improves its ability to withstand cold stress.

PMID: 16980419 [PubMed - indexed for MEDLINE]

50: J Exp Bot. 2006 Oct 4; [Epub ahead of print]

Use of thermal and visible imagery for estimating crop water status of irrigated
grapevine.

Moller M, Alchanatis V, Cohen Y, Meron M, Tsipris J, Naor A, Ostrovsky V,
Sprintsin M, Cohen S.

Institute of Soil, Water and Environmental Sciences, Agricultural Research
Organization (ARO), The Volcani Center, PO Box 6, 50250 Bet Dagan, Israel.

Achieving high quality wine grapes depends on the ability to maintain mild to
moderate levels of water stress in the crop during the growing season. This
study investigates the use of thermal imaging for monitoring water stress.
Experiments were conducted on a wine-grape (Vitis vinifera cv. Merlot) vineyard
in northern Israel. Irrigation treatments included mild, moderate, and severe
stress. Thermal and visible (RGB) images of the crop were taken on four days at
midday with a FLIR thermal imaging system and a digital camera, respectively,
both mounted on a truck-crane 15 m above the canopy. Aluminium crosses were used
to match visible and thermal images in post-processing and an artificial wet
surface was used to estimate the reference wet temperature (T(wet)). Monitored
crop parameters included stem water potential (Psi(stem)), leaf conductance
(g(L)), and leaf area index (LAI). Meteorological parameters were measured at 2
m height. CWSI was highly correlated with g(L) and moderately correlated with
Psi(stem). The CWSI-g(L) relationship was very stable throughout the season, but
for that of CWSI-Psi(stem) both intercept and slope varied considerably. The
latter presumably reflects the non-direct nature of the physiological
relationship between CWSI and Psi(stem). The highest R(2) for the CWSI to g(L)
relationship, 0.91 (n=12), was obtained when CWSI was computed using
temperatures from the centre of the canopy, T(wet) from the artificial wet
surface, and reference dry temperature from air temperature plus 5 degrees C.
Using T(wet) calculated from the inverted Penman-Monteith equation and estimated
from an artificially wetted part of the canopy also yielded crop water-stress
estimates highly correlated with g(L) (R(2)=0.89 and 0.82, respectively), while
a crop water-stress index using 'theoretical' reference temperatures computed
from climate data showed significant deviations in the late season. Parameter
variability and robustness of the different CWSI estimates are discussed. Future
research should aim at developing thermal imaging into an irrigation scheduling
tool applicable to different crops.

PMID: 16968884 [PubMed - as supplied by publisher]

51: J Appl Microbiol. 2006 Oct;101(4):764-74.

Isolation of an ethanol-tolerant endospore-forming Gram-negative Brevibacillus
sp. as a covert contaminant in grape tissue cultures.

Thomas P.

Division of Biotechnology, Indian Institute of Horticultural Research,
Bangalore, India. [email protected]

AIMS: To characterize the alcohol-surviving bacterial isolate ARBG1 from in
vitro grapes (Vitis vinifera). METHODS AND RESULTS: Two bacterial strains that
survived in covert form in grape cultures were isolated from the spent alcohol
used for disinfecting the tools of which one (ARBG2) was characterized earlier.
The present study describes characterization of the second isolate, ARBG1.
Nutrient agar (NA)-derived colonies of ARBG1 displayed consistently
Gram-negative staining rods (2-4x0.5-0.6 micro) substantiated by KOH mucoid
thread test. Older cultures (3-7 days) showed emergence of Gram-negative
staining, oblong, phase-refractile cells with ellipsoidal spores. The growth and
sporulation were modified by growth medium and incubation temperature with the
optimum around 37 degrees C. Identification attempts involving microscopic,
biochemical, Biolog or fatty acid profiling approaches brought in mixed and
inconclusive results. PCR amplification of 16S rDNA was not successful with the
standard primers 27F and 1492R but with 27F and a modified primer ARBG1-RP1. The
identity of the isolate was established as Brevibacillus sp. based on partial
16S rDNA sequence data from eight single colonies with Gram-positive
Brevibacillus choshinensis as the closest match (99.5%). Spotting tests on NA
employing spore suspension in aqueous ethanol (0%, 25%, 50%, 60%, 70%, 80% or
90%, v/v) indicated unhindered bacterial-survival in alcohol for 1 month, and
that at 2 or 4 months revealed 90% ethanol as more sporicidal than lower levels,
corroborated by plating results. Grape microcuttings inoculated with ARBG1
showed substantial general colonization of shoots, roots and medium but low
endophytic colonization. CONCLUSIONS: The rare type of spore-producing
consistently Gram-negative bacterial isolate ARBG1 was identified as
Brevibacillus sp. based on 16S rDNA sequence similarity. The alcohol-defying
organism was nonpathogenic and survived in covert form in grape cultures.
Aqueous 90% ethanol appeared more sporicidal than lower levels. SIGNIFICANCE AND
IMPACT OF THE STUDY: Characterization of an unusual endospore-forming
Gram-negative bacterium, observation that some bacteria may fall outside the
purview of standard 16S rDNA primers, elucidation of the threats of covert
bacteria in plant tissue cultures and alcohol-mediated lateral transmission of
spore formers, and the revelation that 70-80% ethanol may not be the most
effective bactericidal concentration for all bacteria.

PMID: 16968288 [PubMed - in process]

52: J Agric Food Chem. 2006 Sep 20;54(19):7251-5.

Stereoisomeric distribution of 3-mercaptohexan-1-ol and 3-mercaptohexyl acetate
in dry and sweet white wines made from Vitis vinifera (Var. Sauvignon Blanc and
Semillon).

Tominaga T, Niclass Y, Frerot E, Dubourdieu D.

Universite Bordeaux 2, UMR1219, Faculte d'Oenologie, ISVV, 351 Cours de la
Liberation, 33405 Talence Cedex, France.
[email protected]

The enantiomeric distribution of 3-mercaptohexan-1-ol (3MH) and 3-mercaptohexyl
acetate (3MHA) in Vitis vinifera wines was determined by combining two
techniques: specific purification of volatile thiols from the wines using
p-hydroxymercuribenzoate and separation of the chiral molecules by gas-phase
chromatography on a cyclodextrin capillary column. The R and S enantiomer ratios
of these two thiols in dry white Sauvignon blanc and Semillon wines are
approximately 30:70 for A3MH and 50:50 for 3MH. However, in sweet white wines
made from grapes affected by "noble rot" due to the development of Botrytis
cinerea on ripe grapes, the proportion of the R and S forms of 3MH is in the
vicinity of 30:70. During alcoholic fermentation, a change in the ratio of the
two enantiomers of 3MH in dry white wines was observed. At the beginning of
fermentation (around density 1.08), the S form represented over 60%; then, at
lower density, as fermentation proceeded, the enatiomeric ratio approached
50:50. The ratio of the two 3MHA enantiomers remained constant throughout
fermentation. On the contrary, the distribution of the two 3MH enantiomers
changed very little during fermentation of the botrytized sweet wines. The
perception thresholds for the R and S forms of 3MH in hydroalcoholic model
solution are similar (50 and 60 ng/L). These two enantiomers have quite
different aromas: The R form is fruitier, with a zesty aroma reminiscent of
grapefruit, while the S form smells more of passion fruit. The perception
thresholds of the R and S enantiomers of 3MHA are slightly different (9 and 2.5
ng/L). The less odoriferous R form is reminiscent of passion fruit, while the S
form has a more herbaceous odor of boxwood.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16968090 [PubMed - indexed for MEDLINE]

53: J Agric Food Chem. 2006 Sep 6;54(18):6692-704.

Aging effect on the pigment composition and color of Vitis vinifera L. Cv.
Tannat wines. Contribution of the main pigment families to wine color.

Boido E, Alcalde-Eon C, Carrau F, Dellacassa E, Rivas-Gonzalo JC.

Seccion de Enologia, Facultad de Quimica, Universidad de la Republica, Gral.
Flores 2124, 11800 Montevideo, Uruguay.

Red wines made from Vitis vinifera L. cv. Tannat grapes are known to possess
high contents of tannins and intense color, features that are responsible for
the originality of these wines. This work aimed to study the evolution of the
pigment composition and CIELAB color parameters as Tannat wines become older, as
well as to establish the contribution to wine color of the main pigment
families. Tannat wines produced in Uruguay from grapes of the same vineyard in
six consecutive vintages (1998-2003) and Tannat grapes of the 2003 harvest were
analyzed by means of HPLC-DAD-MS and UV-vis spectrometric techniques. The
correlations between the different pigment families and the CIELAB parameters
revealed the importance of the variations of the percentage, found in
anthocyanins and flavanol-anthocyanin acetaldehyde-mediated condensation
products (decrease) and pyranoanthocyanins and direct condensation products
(increase), in the modification of the color from purple-red hues to more
orange-red ones. The color suffered qualitative rather than quantitative
changes, that is, the hue (h*ab) increased, whereas the chroma (C*ab) and
lightness (L) did not show a defined trend with time.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16939328 [PubMed - indexed for MEDLINE]

54: Protoplasma. 2006 Aug;228(1-3):131-5. Epub 2006 Aug 31.

Ultrastructure and germination of Vitis vinifera cv. Loureiro pollen.

Abreu I, Costa I, Oliveira M, Cunha M, de Castro R.

Instituto de Biologia Molecular e Celular, Universidade do Porto, Porto,
Portugal. [email protected]

The cultivar Loureiro of Vitis vinifera is one of the most economically
important, recommended in almost the totality of the Regiao Demarcada dos Vinhos
Verdes. In vineyards, the grape productivity of this cultivar is normal while in
others it is extremely low. The aim of this work was to study the morphology and
germination of Vitis vinifera cv. Loureiro pollen with high and low
productivity. The pollen grain was examined under light, transmission and
scanning electron microscopy. Typically V. vinifera pollen present three furrows
but in the cultivar Loureiro we found tricolporated and acolporated (without
furrows or pores) pollen grains. Both pollen types present generative and
vegetative cells with the usual aspect and a dense cytoplasm rich in organelles.
In the acolporated pollen a continuous exine layer and an irregular intine layer
were observed. Differences were found in the starch accumulation, since only in
tricolporated pollen abundant plastids filled with numerous starch granules were
observed. To determine the causes of the low productivity of this cultivar we
tested pollen viability by the fluorochromatic reaction and pollen germinability
by in vitro assays. We observed that the acolporated pollen grain is viable, but
no germination was recorded.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16937066 [PubMed - indexed for MEDLINE]

55: Genome. 2006 Jul;49(7):721-8.

Somatic chimerism, genetic inheritance, and mapping of the fleshless berry (flb)
mutation in grapevine (Vitis vinifera L.).

Fernandez L, Doligez A, Lopez G, Thomas MR, Bouquet A, Torregrosa L.

UMR BEPC, campus-Agro-M/INRA, Montpellier, CEDEX 01, France.

The fleshless berry (flb) mutation of grapevine (Vitis vinifera L. 'Ugni Blanc')
impairs the differentiation and division of inner mesocarp cells responsible for
flesh in grapevine berries. In order to study the inheritance of the mutation
and to map the flb locus, 5 segregating populations were created. Progeny plants
were classified as mutant or wild type by scoring for the presence of an ovary
phenotype associated with the Flb- phenotype at anthesis. Phenotypic segregation
revealed the involvement of a single dominant allele that was heterozygous in
the original mutant. Through bulk segregant analysis, microsatellite (simple
sequence repeat (SSR)) markers linked to the mutation were identified, and the
flb locus was assigned to linkage group 18. The locus position was then refined
by analyzing individual progeny and the segregation of SSR markers in the target
region with the closest marker 5.6 cM distant from the flb locus. All progeny
with the Flb- ovary phenotype differed from the original fleshless berry mutant
in that no berries formed after anthesis. Our data suggest that the original
mutant plant was a chimera with the mutated allele present in only 1 cell layer
(L2 layer) of the ovary and berry.

PMID: 16936780 [PubMed - indexed for MEDLINE]

56: J Exp Bot. 2006;57(12):3209-16. Epub 2006 Aug 25.

Transporters expressed during grape berry (Vitis vinifera L.) development are
associated with an increase in berry size and berry potassium accumulation.

Davies C, Shin R, Liu W, Thomas MR, Schachtman DP.

CSIRO Plant Industry and Cooperative Research Centre for Viticulture, PO Box
350, Glen Osmond, SA 5064, Australia. [email protected]

Potassium accumulation is essential for grapevine (Vitis vinifera L.) growth and
development, but excessive levels in berries at harvest may reduce wine quality
particularly for red wines. In addition to decreasing the free acid levels,
potassium also combines with tartaric acid to form largely insoluble potassium
bitartrate. This precipitates during winemaking and storage, resulting in an
increase in wine pH that is associated with negative impacts on wine colour,
flavour, and microbiological stability. For these reasons, a better
understanding of potassium transport and accumulation within the vine and
berries is important for producing fruit with improved winemaking
characteristics. Here two genes encoding KUP/KT/HAK-type potassium transporters
that are expressed in grape berries are described. Their function as potassium
transporters was demonstrated by complementation of an Escherichia coli mutant.
The two transporters are expressed most highly in the berry skin during the
first phase of berry development (pre-veraison), with similar patterns in two
grapevine varieties. The timing and location of expression of these transporters
are consistent with an involvement in potassium accumulation in grape berries.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16936223 [PubMed - indexed for MEDLINE]

57: Mutat Res. 2006 Oct 30;609(2):165-75. Epub 2006 Aug 28.

Activity of grape extracts from Greek varieties of Vitis vinifera against
mutagenicity induced by bleomycin and hydrogen peroxide in Salmonella
typhimurium strain TA102.

Stagos D, Kazantzoglou G, Theofanidou D, Kakalopoulou G, Magiatis P, Mitaku S,
Kouretas D.

Department of Biochemistry and Biotechnology, University of Thessaly, Ploutonos
26 & Aiolou, GR-41221 Larissa, Greece.

Several in vivo and in vitro studies have shown that grape extracts could
prevent certain steps in carcinogenesis and a few mechanisms have been proposed
for this activity. In this study, the potential antimutagenic activity of
methanolic and aqueous extracts from two Greek grape varieties of Vitis vinifera
against DNA damage induced by reactive oxygen species (ROS) was assessed as a
potential novel chemopreventive mechanism, using Salmonella typhimurium strain
TA102. The two grape varieties were Assyrtiko (white grapes) and Mandilaria (red
grapes), while the oxidant mutagens used were bleomycin (BLM) and hydrogen
peroxide (H(2)O(2)). Since it has been considered that polyphenols present in
grapes are their most potent biologically active compounds, we also tested the
effects of polyphenol-rich fractions as well as some of the more common grape
polyphenols on the activity of the two test mutagens. These polyphenols were
quercetin, (+)-catechin, (-)-epicatechin, trans-resveratrol, gallic acid and
protocatechuic acid. Almost all extracts showed inhibitory activity against both
mutagens. On the other hand, polyphenol-rich fractions as well as individual
polyphenols at concentrations found in the extracts either did not diminish or
did enhance the activity of the mutagens. These results suggest that the
protection of DNA from mutations induced by ROS may be one of the mechanisms
accounting for the chemopreventive activity of grape extracts. However, it seems
that this protective activity may not be attributed to polyphenols but rather to
a synergism of many compounds in the grapes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16935024 [PubMed - indexed for MEDLINE]

58: Plant Mol Biol. 2006 Nov;62(4-5):623-35. Epub 2006 Aug 24.

Two new grape cultivars, bud sports of Cabernet Sauvignon bearing pale-coloured
berries, are the result of deletion of two regulatory genes of the berry colour
locus.

Walker AR, Lee E, Robinson SP.

CSIRO Plant Industry, Adelaide Laboratory, PO Box 350, Glen Osmond, SA 5064,
Australia. [email protected]

Bud sports are infrequent changes in phenotype affecting shoots of woody
perennials but the molecular basis of these mutations has rarely been
identified. In this report, we show that the bronze-coloured berries of the
Malian cultivar, a documented bud sport of the wine grape Cabernet Sauvignon
(Vitis vinifera L.), lack anthocyanins in the subepidermal cells compared to the
red/black berried Cabernet Sauvignon in which both the epidermis and several
subepidermal cell layers contain anthocyanin. The Malian phenotype is correlated
with an alteration in the genome indicated by a reduction of hybridisation
signal using a MYBA probe. In Shalistin, a white-berried bud sport of Malian,
the red allele at the berry colour locus appears to have been deleted
completely. These data suggest that Malian could be a L1/L2 periclinal chimera,
which gave rise to Shalistin by an invasion of epidermal cells (L1) by the
mutated subepidermal cells (L2). The red grape Pinot Noir has given rise to a
number of pale coloured sports, although the provenance of the extant sports is
not known. We show that a clone of Pinot Blanc (white-berried) does not have a
deletion of the red allele of the same dimensions as that in Shalistin, though a
small deletion is a likely explanation for the altered phenotype. However, the
mechanism of deletion of the red allele of the berry colour locus is a possible
means by which other red to white clonal mutations of grapevines have occurred.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16932847 [PubMed - indexed for MEDLINE]

59: J Plant Res. 2006 Sep;119(5):469-78. Epub 2006 Aug 19.

Sequence analysis and transcriptional profiling of two vacuolar H+
-pyrophosphatase isoforms in Vitis vinifera.

Venter M, Groenewald JH, Botha FC.

Institute for Plant Biotechnology, Stellenbosch University, Private Bag X1,
Matieland, South Africa. [email protected]

Gene expression of grapevine vacuolar H(+)-pyrophosphatase (V-PPase EC 3.6.1.1.)
during fruit ripening has previously been reported. Here we report on putative
multiple V-PPase isoforms in grapevine. In this study a full-length cDNA
sequence with an open reading frame of 2,295 nucleotides encoding a V-PPase gene
(vpp2: acc. nr. AJ557256) was cloned. Sequence analyses of the deduced amino
acid residues and RT-PCR experiments indicated that Vitis vinifera L. has at
least two distinct isoforms of the V-PPase gene. Bioinformatic analyses of 13
V-PPase protein sequences revealed two highly conserved motifs associated with
pyrophosphate (PPi) binding and response to stress, respectively. Both V-PPase
isoforms were expressed at higher levels in the late post-veraison stage of
grape berry ripening. Results also showed that the expression of grapevine
V-PPase was induced by cold stress.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16924561 [PubMed - indexed for MEDLINE]

60: J Agric Food Chem. 2006 Aug 23;54(17):6322-31.

Hydrolysis and transformation of grape glycosidically bound volatile compounds
during fermentation with three Saccharomyces yeast strains.

Ugliano M, Bartowsky EJ, McCarthy J, Moio L, Henschke PA.

Dipartimento di Scienze degli Alimenti, Universita di Foggia, via Napoli 25,
Foggia, Italy.

The ability of three Saccharomyces wine yeasts (S. cerevisiae AWRI 838, S.
cerevisiae AWRI 1537, and S. bayanus AWRI 1375) to liberate volatile compounds
from sugar-bound aroma precursors was investigated using synthetic and grape
glycosides under different experimental conditions. In model systems involving
the incubation of yeast cells with either synthetic or grape-derived glycosides
under conditions more favorable for glycosidase activities and less favorable
for acid-catalyzed hydrolysis (pH 5.0 and 30 degrees C), all yeast strains
studied proved to be capable of hydrolyzing glycosides, with S. bayanus AWRI
1375 displaying greater hydrolytic activity than S. cerevisiae AWRI 838 and AWRI
1537. During the fermentation of a chemically defined grape juice-like medium
containing glycosidic precursors extracted from Vitis vinifera cv. White
Frontignac (synonym Muscat a Petit Grains Blanc), all yeasts promoted a
significant hydrolysis of different precursors, which varied according to the
chemical structures of both the sugar and the aglycon moieties, as determined by
GC-MS analysis of trifluoroacetylated derivatives. Hydrolysis of the White
Frontignac derived glycosidic precursors during fermentation resulted in the
release of monoterepene alcohols, terpene oxides, terpene diols, and
3-oxo-alpha-ionol, demonstrating the significant potential of these yeast
strains to contribute to wine varietal volatile composition during alcoholic
fermentation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16910726 [PubMed - indexed for MEDLINE]

61: J Chromatogr A. 2006 Sep 29;1129(1):14-20. Epub 2006 Aug 8.

Application of sample disruption methods in the extraction of anthocyanins from
solid or semi-solid vegetable samples.

Manhita AC, Teixeira DM, da Costa CT.

Departamento de Quimica da Universidade de Evora, CLAV, Rua Romao Ramalho 59,
7000-617 Evora, Portugal.

Cassis, a commercial powder obtained from blackcurrant juice (Ribes nigrum L.),
was used to optimize the anthocyanin extraction by matrix solid-phase dispersion
(MSPD) and by sea sand disruption method (SSDM). The optimum eluent conditions
were 5.0 mL of MeOH/H2O (1:1, v/v) at pH 2. The extracts were analyzed by HPLC
with diode-array detection (DAD) and the extraction efficiency was evaluated by
statistical comparison (analysis of variance, ANOVA-single factor) of the
compounds peak areas. The extraction efficiency for the MSPD procedure was
dependent on the chemical nature of the C18 material but the optimized procedure
yielded results similar to those obtained by the inexpensive sea sand disruption
method (SSDM). This procedure also compared favorably to a previously published
liquid-liquid extraction method. The optimized sea sand method was then used in
the anthocyanin extraction from red grapes (Vitis vinifera L.) and strawberries
(Fragaria ananassa D.) yielding chromatographic profiles similar to those
reported in the literature for these products.

PMID: 16893546 [PubMed - indexed for MEDLINE]

62: Cell Microbiol. 2006 Aug;8(8):1310-21.

Botrytis cinerea virulence is drastically reduced after disruption of chitin
synthase class III gene (Bcchs3a).

Soulie MC, Perino C, Piffeteau A, Choquer M, Malfatti P, Cimerman A, Kunz C,
Boccara M, Vidal-Cros A.

Interactions plantes-pathogenes, UMR 217 INRA/INA-PG/UPMC, 16 rue Claude
Bernard, 75005 Paris, France. [email protected]

Botrytis cinerea is an important phytopathogenic fungus requiring new methods of
control. Chitin biosynthesis, which involves seven classes of chitin synthases,
could be an attractive target. A fragment encoding one of the class III enzymes
was used to disrupt the corresponding Bcchs3a gene in the B. cinerea genome. The
resulting mutant exhibited a 39% reduction in its chitin content and an 89%
reduction in its in vitro chitin synthase activity, compared with the wild-type
strain. Bcchs3a mutant was not affected in its growth in liquid medium, neither
in its production of sclerotia, micro- and macroconidia. In contrast, the mutant
Bcchs3a was severely impaired in its growth on solid medium. Counterbalancing
this defect in radial growth, Bcchs3a mutant presented a large increase in
hyphal ramification, resulting in an enhanced aerial growth. Observations by
different techniques of microscopy revealed a thick extracellular matrix around
the hyphal tips. Moreover, Bcchs3a mutant had a largely reduced virulence on
Vitis vinifera and Arabidopsis thaliana leaves.

PMID: 16882034 [PubMed - indexed for MEDLINE]

63: Trends Genet. 2006 Sep;22(9):511-9. Epub 2006 Jul 26.

Historical origins and genetic diversity of wine grapes.

This P, Lacombe T, Thomas MR.

INRA, UMR Diversite et Genomes des Plantes Cultivees, 2 place P. Viala, 34060
Montpellier, France.

The genomic resources that are available to the grapevine research community
have increased enormously during the past five years, in parallel with a renewed
interest in grapevine (Vitis vinifera L.) germplasm resources and analysis of
genetic diversity in grapes. Genetic variation, either natural or induced, is
invaluable for crop improvement and understanding gene function, and the same is
true for the grapevine. The history and vineyard cultural practices have largely
determined the genetic diversity that exists today in grapevines. In this
article, we provide a synopsis of what is known about the origin and genetics of
grapes and how molecular genetics is helping us understand more about this
plant: its evolution, historical development, genetic diversity and potential
for genetic improvement.

Publication Types:
Historical Article
Research Support, Non-U.S. Gov't
Review

PMID: 16872714 [PubMed - indexed for MEDLINE]

64: Anal Bioanal Chem. 2006 Sep;386(2):306-12. Epub 2006 Jul 26.

Quantification of abscisic acid in grapevine leaf (Vitis vinifera) by
isotope-dilution liquid chromatography-mass spectrometry.

Vilaro F, Canela-Xandri A, Canela R.

Centre UdL-IRTA, Rovira Roure 191, 25198 Lleida, Spain.

A specific, sensitive, precise, and accurate method for the determination of
abscisic acid (ABA) in grapevine leaf tissues is described. The method employs
high-performance liquid chromatography and electrospray ionization-mass
spectrometry (LC-ESI-MS) in selected ion monitoring mode (SIM) to analyze ABA
using a stable isotope-labeled ABA as an internal standard. Absolute recoveries
ranged from 72% to 79% using methanol/water pH 5.5 (50:50 v/v) as an extraction
solvent. The best efficiency was obtained when the chromatographic separation
was carried out by using a porous graphitic carbon (PGC) column. The statistical
evaluation of the method was satisfactory in the work range. A relative standard
deviation (RDS) of < 5.5% and < 6.0% was obtained for intra-batch and
inter-batch comparisons, respectively. As for accuracy, the relative error (%Er)
was between -2.7 and 4.3%, and the relative recovery ranged from 95% to 107%.

PMID: 16868725 [PubMed - in process]

65: J Exp Bot. 2006;57(11):2577-87. Epub 2006 Jul 25.

Ripening grape berries remain hydraulically connected to the shoot.

Keller M, Smith JP, Bondada BR.

Irrigated Agriculture Research and Extension Center, Washington State
University, 24106 N. Bunn Road, Prosser, 99350, USA. [email protected]

Berry diameter was monitored during dry-down and rewatering cycles and
pressurization of the root system of Vitis vinifera (cv. Merlot) and Vitis
labruscana (cv. Concord) to test changes in xylem functionality during grape
ripening. Prior to veraison (onset of ripening), berries maintained their size
under declining soil moisture until the plants had used 80% of the transpirable
soil water, began to shrink thereafter, and recovered rapidly after rewatering.
By contrast, berry diameter declined slowly but steadily during post-veraison
water stress and did not recover after rewatering; irrigation merely prevented
further shrinking. Preconditioning vines with a period of water stress after
flowering did not influence the berries' reaction to subsequent changes in
transpirable soil water. Pressurizing the root system led to concomitant changes
in berry diameter only prior to veraison, although some post-veraison Concord,
but not Merlot, berries cracked under root pressurization. The xylem-mobile dye
basic fuchsin, infused via the shoot base, moved throughout the berry
vasculature before veraison, but became gradually confined to the brush area
during ripening. When the dye was infused through the stylar end of attached
berries, it readily moved back to the plant both before and after veraison. Our
work demonstrated that berry-xylem conduits retain their capacity for water and
solute transport during ripening. It is proposed here that apoplastic phloem
unloading coupled with solute accumulation in the berry apoplast may be
responsible for the decline in xylem water influx into ripening grape berries.
Instead, the xylem may serve to recycle excess phloem water back to the shoot.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16868045 [PubMed - indexed for MEDLINE]

66: Plant Physiol. 2006 Sep;142(1):220-32. Epub 2006 Jul 21.

A shift of Phloem unloading from symplasmic to apoplasmic pathway is involved in
developmental onset of ripening in grape berry.

Zhang XY, Wang XL, Wang XF, Xia GH, Pan QH, Fan RC, Wu FQ, Yu XC, Zhang DP.

China State Key Laboratory of Plant Physiology and Biochemistry, China
Agricultural University, 100094 Beijing, China.

It remains unclear whether the phloem unloading pathway alters to adapt to
developmental transition in fleshy fruits that accumulate high level of soluble
sugars. Using a combination of electron microscopy, transport of the
phloem-mobile symplasmic tracer carboxyfluorescein, movement of the companion
cell-expressed and the green fluorescent protein-tagged viral movement protein,
and assays of the sucrose cleavage enzymes, the pathway of phloem unloading was
studied in the berries of a hybrid grape (Vitis vinifera x Vitis labrusca).
Structural investigations showed that the sieve element-companion cell complex
is apparently symplasmically connected through plasmodesmata with surrounding
parenchyma cells throughout fruit development, though a small portion of
plasmodesmata are apparently blocked in the ripening stage. Both
carboxyfluorescein and the green fluorescent protein-tagged viral movement
protein were released from the functional phloem strands during the early and
middle stages of fruit development, whereas the two symplasmic tracers were
confined to the phloem strands during the late stage. This reveals a shift of
phloem unloading from symplasmic to apoplasmic pathway during fruit development.
The turning point of the phloem unloading pathways was further shown to be at or
just before onset of ripening, an important developmental checkpoint of grape
berry. In addition, the levels of both the expression and activities of cell
wall acid invertase increased around the onset of ripening and reached a high
level in the late stage, providing further evidence for an operation of the
apoplasmic unloading pathway after onset of ripening. These data demonstrate
clearly the occurrence of an adaptive shift of phloem unloading pathway to
developmental transition from growing phase to ripening in grape berry.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16861573 [PubMed - indexed for MEDLINE]

67: J Agric Food Chem. 2006 Jul 26;54(15):5422-9.

Influence of different phenolic copigments on the color of malvidin 3-glucoside.

Gomez-Miguez M, Gonzalez-Manzano S, Escribano-Bailon MT, Heredia FJ,
Santos-Buelga C.

Laboratory of Food Color and Quality, Department of Food Science and Nutrition,
Faculty of Pharmacy, University of Seville, Seville, Spain.

The effectiveness of seven phenolic compounds (catechin, epicatechin,
procyanidin B2, caffeic acid, p-coumaric acid, myricitrin, and quercitrin) as
copigments of malvidin 3-glucoside, the major anthocyanin in red wines from
Vitis vinifera, using a copigment/pigment molar ratio of 1:1 was assayed in
model wine solutions under the same conditions (pH=3.6, 12% ethanol). The
stability of the copigment-pigment complexes formed was studied during a storage
period of 60 days at 25 degrees C. Tristimulus colorimetry was applied for color
characterization of the copigmentation process, and HPLC-DAD-MS was used to
monitor changes in the composition of the samples. Copigmentation has been found
to occur in all cases despite the low copigment/pigment molar ratio used,
although the effect was different depending on the compound. Flavan-3-ols
appeared as the less effective copigments, procyanidin B2 being even worse than
monomeric flavanols, whereas flavonols behaved as the best ones. These latter
copigments also induced the most statistically significant bathochromic shift in
lambdamax. In the colorimetric analysis, it was observed that the lightness L*
of the copigmented solutions increased with time, chroma C*ab decreased, and the
hue hab increased. The copigments that produced a greater increase in the hue
angle were the monomeric flavan-3-ols, which therefore showed to be the least
protective cofactors against browning of the anthocyanin solution during the
storage. With the time of storage, the formation of new pigments was observed in
the solutions containing flavanols (xanthylium structures) and hydroxycinnamic
acids (pyranoanthocyanins), which explains some of the color modifications
produced in these solutions. Another relevant observation was that the stability
of the anthocyanin was not much improved by most of the assayed copigments,
since quite similar degradation rates were observed in the presence and absence
of those cofactors.

PMID: 16848527 [PubMed - indexed for MEDLINE]

68: Coll Antropol. 2006 Jun;30(2):429-36.

Comparative analysis of plant finds from Early Roman graves in Ilok (Cuccium)
and Scitarjevo (Andautonia), Croatia--a contribution to understanding burial
rites in southern Pannonia.

Sostaric R, Dizdar M, Kusan D, Hrsak V, Marekovic S.

Department of Botany, Faculty of Science, University of Zagreb, Croatia.
[email protected]

A comparative archaeobotanical analysis of the plant remains from the Early
Roman incineration graves in Ilok and Scitarjevo shows the existence of a
complex burial ritual, but at the same time enables a better understanding of
the agriculture and trade of the 1st/early 2nd century AD in southern Pannonia.
Most of the cereals found (Hordeum vulgare, Panicum miliaceum, Triticum
monococcum, T. dicoccon, T. aestivum i T. cf. spelta), the legumes (Lens
culinaris, Vicia ervilia) and the fruit contributions (Cucumis melo/sativus,
Malus/Pyrus sp., the Prunus avium group, P. domestica, Vitis vinifera) were
probably grown in the vicinity of the investigated localities, but they might at
the same time have been trade goods. Trade was undoubtedly well developed at
that period, as shown by the remains of the fig (Ficus carica) and olive (Olea
europaea), typically Mediterranean crops, in the finds. All the species of
cereals, except millet (Panicum miliaceum) in Scitarjevo, and of bitter vetch
(Vicia ervilia) found in the Ilok grave were carbonised and were probably placed
on the funeral pyre with the departed. The lentil (Lens culinaris) and the other
fruit remains were non-carbonised and mineralised, which means that they were
placed in the grave in fresh, dried or cooked form as food for the deceased
(belief in an immortal soul), as remains of the funerary feast, or as a
sacrifice to the goods.

Publication Types:
Comparative Study
Historical Article
Research Support, Non-U.S. Gov't

PMID: 16848163 [PubMed - indexed for MEDLINE]

69: Biocell. 2006 Apr;30(1):1-7.

Argentinean cultivars of Vitis vinifera grow better than European ones when
cultured in vitro under salinity.

Cavagnaro JB, Ponce MT, Guzman J, Cirrincione MA.

Laboratorio de Fisiologia Vegetal. Facultad Ciencias Agrarias. Univ. Nacional de
Cuyo. Mendoza, Argentina. [email protected]

Argentinean Vitis vinifera cultivars although originated from Europe, have clear
ampelographic and genotypic differences as compared with the European cultivars
currently used in wine making. In vitro evaluation of salt tolerance has been
used in many species. Our hypothesis was that Argentinean cultivars are more
tolerant to salinity than European ones. Three European cultivars, Malbec,
Cabernet Sauvignon and Chardonnay and four Argentincan cultivars, Cereza,
Criolla Chica, Pedro Gimcnez and Torrontes Riojano were tested by in vitro
culture. Treatments included: 1) Control, 2) 60 mEq/L of a mixture of three
parts of NaCl and one part of CaCl2 and 3) 90 mEq/L of the salt mixture. Results
from two experiments (I and II) are reported. No differences were found in plant
survival, expressed as % of the respective control, among cultivars. Leaf area,
leaf, stem and total dry matter (DM) in Experiment I and leaf area, leaf number
and leaf, stem, root and total DM in Experiment II, were higher in Argentinean
cultivars than in European ones. We conclude that Argentinean cultivars show
better performance in growing under salinity, especially in the highest salt
concentration. Differences among cultivars, inside each group, were found for
most of the measured variables.

Publication Types:
Comparative Study

PMID: 16845822 [PubMed - indexed for MEDLINE]

70: J Exp Bot. 2006;57(11):2687-95. Epub 2006 Jul 12.

Relationships between leaf conductance to CO2 diffusion and photosynthesis in
micropropagated grapevine plants, before and after ex vitro acclimatization.

Fila G, Badeck FW, Meyer S, Cerovic Z, Ghashghaie J.

Consiglio per la Ricerca e Sperimentazione in Agricoltura-Istituto Sperimentale
per le Colture Industriali, via di Corticella 133, I-40128 Bologna, Italy.
[email protected]

In vitro-cultured plants typically show a low photosynthetic activity, which is
considered detrimental to subsequent ex vitro acclimatization. Studies conducted
so far have approached this problem by analysing the biochemical and
photochemical aspects of photosynthesis, while very little attention has been
paid to the role of leaf conductance to CO(2) diffusion, which often represents
an important constraint to CO(2) assimilation in naturally grown plants.
Mesophyll conductance, in particular, has never been determined in in vitro
plants, and no information exists as to whether it represents a limitation to
carbon assimilation during in vitro growth and subsequent ex vitro
acclimatization. In this study, by means of simultaneous gas exchange and
chlorophyll fluorescence measurements, the stomatal and mesophyll conductance to
CO(2) diffusion were assessed in in vitro-cultured plants of the grapevine
rootstock '41B' (Vitis vinifera 'Chasselas'xVitis berlandieri), prior to and
after ex vitro acclimatization. Their impact on electron transport rate
partitioning and on limitation of potential net assimilation rate was analysed.
In vitro plants had a high stomatal conductance, 155 versus 50 mmol m(-2) s(-1)
in acclimatized plants, which ensured a higher CO(2) concentration in the
chloroplasts, and a 7% higher electron flow to the carbon reduction pathway. The
high stomatal conductance was counterbalanced by a low mesophyll conductance, 43
versus 285 mmol m(-2) s(-1), which accounted for a 14.5% estimated relative
limitation to photosynthesis against 2.1% estimated in acclimatized plants. It
was concluded that mesophyll conductance represents an important limitation for
in vitro plant photosynthesis, and that in acclimatization studies the correct
comparison of photosynthetic activity between in vitro and acclimatized plants
must take into account the contribution of both stomatal and mesophyll
conductance.

Publication Types:
Comparative Study

PMID: 16837534 [PubMed - indexed for MEDLINE]

71: J Ethnopharmacol. 2006 Nov 24;108(2):280-6. Epub 2006 May 20.

In-vivo assessment of antidiabetic and antioxidant activities of grapevine
leaves (Vitis vinifera) in diabetic rats.

Orhan N, Aslan M, Orhan DD, Ergun F, Yesilada E.

Department of Pharmacognosy, Faculty of Pharmacy, Gazi University, Hipodrom
6330, Ankara, Turkey. [email protected]

The acute and the subacute (15 days) hypoglycaemic and antihyperglycaemic effect
of the two different doses (250, 500 mg/kg) of the aqueous extract from the
leaves of Vitis vinifera L. were evaluated in this study. The aqueous extract
was further fractionated through successive solvent extractions and the acute
effect of different doses of its subfractions, 25 mg/kg for ethylacetate
fraction, 80 mg/kg for n-butanol fraction and 375 mg/kg for remaining aqueous
fraction were investigated using normal, glucose-hyperglycaemic and
streptozotocin-induced diabetic rats. Blood glucose levels were measured
according to the glucose oxidase method. Tolbutamide was used as a reference
drug at a dose of 100 mg/kg. The antioxidant activity of the test samples was
studied in the liver, kidney and heart tissues of diabetic rats by measuring
malondialdehyde (MDA) and glutathion (GSH) levels. All results were compared to
the diabetic control groups. The results showed that EtOAc Fr. was rich in
polyphenolics and possessed a significant antihyperglycaemic and antioxidant
activity equipotent with the reference hypoglycaemic agent (tolbutamide), when
evaluated in diabetic rats.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16824713 [PubMed - in process]

72: Arzneimittelforschung. 2006;56(5):317-21.

Effect of oligomer procyanidins on reperfusion arrhythmias and lactate
dehydrogenase release in the isolated rat heart.

Al-Makdessi S, Sweidan H, Jacob R.

Institute of Physiology II, University of Tubingen, Germany.
[email protected]

The antiarrhythmic effect of an oral 3-week-pretreatment with oligomer
procyanidins derived from Vitis vinifera was investigated on the isolated
perfused heart after global no-flow ischemia (procyanidin-treated group: n = 9,
control group: n = 13). Hearts were perfused with a modified Krebs-Henseleit
solution in which the K+ content was reduced to 3.0 mmol/l in order to lower the
fibrillation threshold. Monophasic action potentials in addition to ECG were
recorded. The durations of ischemia and reperfusion were 20 and 30 min,
respectively. Arrhythmias including ventricular fibrillation (VF), ventricular
tachycardia (VT), flutter (Fl) and bradycardia were evaluated. During the
reperfusion, irreversible VF occurred in most of control hearts. The incidence
of VF (percentage of the hearts in which VF occurred) was lowered by oligomer
procyanidins from 84.6 to 55.6 %, and the duration of the episodes of VF
(expressed as percentage relative to the total duration) was significantly
shortened from 76.1 +/- 27.9 % to 36.6 +/- 40.6 % (p = 0.036). Simultaneously,
the percentage of duration of normal sinus rhythm (NSR) increased from 19.5 +/-
30.3 % to 46.2 +/- 35.9 % (n.s.). VF occuring in the procyanidin-treated hearts
could be reversed in two hearts within few minutes to a stage of "reversible
arrhythmias" consisting of short episodes (1 to 60 s) of either Fl or VT or
bradycardia or NSR alternating with each other. LDH (lactate dehydrogenase)
release in the first drops appearing from the reperfused heart was significantly
reduced in the procyanidin-treated rats (66.7 +/- 36.2 mU/min, n = 8) in
comparison to controls (159.7 +/- 79.0 mU/min, n = 10; p = 0.010). These results
demonstrate an antiarrhythmic and cytoprotective effect of oral pretreatment
with oligomer procyanidins under the given experimental conditions.

Publication Types:
In Vitro

PMID: 16821641 [PubMed - indexed for MEDLINE]

73: J Agric Food Chem. 2006 Jul 12;54(14):5077-86.

Influence of vineyard location and vine water status on fruit maturation of
nonirrigated cv. Agiorgitiko (Vitis vinifera L.). Effects on wine phenolic and
aroma components.

Koundouras S, Marinos V, Gkoulioti A, Kotseridis Y, van Leeuwen C.

School of Agriculture, Aristotle University, 54124 Thessaloniki, Greece.
[email protected]

The influence of site on grape and wine composition was investigated for Vitis
vinifera L. cv. Agiorgitiko in the Nemea appellation area in southern Greece.
Three nonirrigated plots were studied during the 1997 and 1998 vintages, which
were typically very hot and without summer rainfall. Vines were subjected to
different water regimens as a result of the variation of soil water-holding
capacity and evaporative demand. Vine water status was determined by means of
predawn leaf water potential. Differences in vine water status between sites
were highly correlated with the earliness of shoot growth cessation and
veraison. Grape composition was monitored during fruit ripening. Water deficit
accelerated sugar accumulation and malic acid breakdown in the juice. Early
water deficit during the growth period was demonstrated to have beneficial
effects on the concentration of anthocyanins and total phenolics in berry skins.
A similar pattern was observed for the phenolic content of wines elaborated
after vinification of grapes harvested on each plot, in both seasons. Limited
water availability seemed to increase glycoconjugates of the main aromatic
components of grapes as a quantitative increase in levels of bound volatile
compounds of the experimental wines was observed under water deficit in both
years. Wines produced from grapes of stressed vineyards were also preferred in
tasting trials.

PMID: 16819919 [PubMed - indexed for MEDLINE]

74: Plant Physiol Biochem. 2006 May-Jun;44(5-6):323-34. Epub 2006 Jun 13.

Biosynthesis of flavan 3-ols by leucoanthocyanidin 4-reductases and
anthocyanidin reductases in leaves of grape (Vitis vinifera L.), apple (Malus x
domestica Borkh.) and other crops.

Pfeiffer J, Kuhnel C, Brandt J, Duy D, Punyasiri PA, Forkmann G, Fischer TC.

Chair of Floriculture Crops and Horticultural Plant Breeding, Technical
University Munich, Am Hochanger 4, 85350 Freising, Germany.

Catechin and epicatechin biosyntheses were studied of grape (Vitis vinifera L.),
apple (Malus x domestica Borkh.) and other crop leaves, since these monomers and
the derived proanthocyanidins are important disease resistance factors. Grape
and apple leucoanthocyanidin 4-reductase (LAR; EC 1.17.1.3) enzymes were
characterized on basis of plant and recombinant enzymes. In case of grape, two
LAR cDNAs were cloned by assembling available EST sequences. Grape and apple
leaf anthocyanidin reductase (ANR; EC 1.3.1.77) cDNAs were also obtained and the
respective plant and recombinant enzymes were characterized. Despite general low
substrate specificity, within the respective flavonoid biosyntheses of grape and
apple leaves, both enzyme types deliver differently hydroxylated catechins and
epicatechins, due to substrate availability in vivo. Furthermore, for LAR
enzymes conversion of 3-deoxyleucocyanidin was shown. Beside relevance for plant
protection, this restricts the number of possible reaction mechanisms of LAR.
ANR enzyme activity was demonstrated for a number of other crop plants and its
correlation with (-)-epicatechin and obvious competition with
UDP-glycosyl:flavonoid-3-O-glycosyltransferases was considered.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16806954 [PubMed - indexed for MEDLINE]

75: Plant Cell Rep. 2006 Nov;25(11):1166-73. Epub 2006 Jun 27.

Transgenic peas (Pisum sativum) expressing polygalacturonase inhibiting protein
from raspberry (Rubus idaeus) and stilbene synthase from grape (Vitis vinifera).

Richter A, Jacobsen HJ, de Kathen A, de Lorenzo G, Briviba K, Hain R, Ramsay G,
Kiesecker H.

Department of Molecular Genetics, University of Hannover, Herrenhauserstr 2,
30419, Hannover, Germany.

The pea (Pisum sativum L.) varieties Baroness (United Kingdome) and Baccara
(France) were transformed via Agrobacterium tumefaciens-mediated gene transfer
with pGPTV binary vectors containing the bar gene in combination with two
different antifungal genes coding for polygalacturonase-inhibiting protein
(PGIP) from raspberry (Rubus idaeus L.) driven by a double 35S promoter, or the
stilbene synthase (Vst1) from grape (Vitis vinifera L.) driven by its own
elicitor-inducible promoter. Transgenic lines were established and transgenes
combined via conventional crossing. Resveratrol, produced by Vst1 transgenic
plants, was detected using HPLC and the PGIP expression was determined in
functional inhibition assays against fungal polygalacturonases. Stable
inheritance of the antifungal genes in the transgenic plants was demonstrated.

PMID: 16802117 [PubMed - indexed for MEDLINE]

76: Nutr Cancer. 2006;54(1):84-93.

Anthocyanin-rich extracts inhibit multiple biomarkers of colon cancer in rats.

Lala G, Malik M, Zhao C, He J, Kwon Y, Giusti MM, Magnuson BA.

Department of Nutrition and Food Science, University of Maryland, College Park
20742, USA.

The aim of the present study was to investigate the chemoprotective activity of
anthocyanin-rich extracts (AREs) from bilberry (Vaccinium myrtillus L.),
chokeberry (Aronia meloncarpa E.), and grape (Vitis vinifera) by assessing
multiple biomarkers of colon cancer in male rats treated with a colon
carcinogen, azoxymethane. Fischer 344 male rats were fed the AIN-93 diet
(control) or AIN-93 diet supplemented with AREs for 14 wk. Biomarkers that were
evaluated included the number and multiplicity of colonic aberrant crypt foci
(ACF), colonic cell proliferation, urinary levels of oxidative DNA damage, and
expression of cyclooxygenase (COX) genes. To assess the bioavailability, levels
of anthocyanins in serum, urine, and feces were evaluated. Total ACF were
reduced (P<0.05) in bilberry, chokeberry, and grape diet groups compared with
the control group. The number of large ACF was also reduced (P<0.05) in bilberry
and chokeberry ARE-fed rats. Colonic cellular proliferation was decreased in
rats fed bilberry ARE and chokeberry ARE diets. Rats fed bilberry and grape ARE
diets had lower COX-2 mRNA expression of gene. High levels of fecal anthocyanins
and increased fecal mass and fecal moisture occurred in ARE-fed rats. There was
also a significant reduction (P<0.05) in fecal bile acids in ARE-fed rats. The
levels of urinary 8-hydroxyguanosine were similar among rats fed different
diets. These results support our previous in vitro studies suggesting a
protective role of AREs in colon carcinogenesis and indicate multiple mechanisms
of action.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16800776 [PubMed - indexed for MEDLINE]

77: Theor Appl Genet. 2006 Aug;113(3):369-82. Epub 2006 Jun 24.

An integrated SSR map of grapevine based on five mapping populations.

Doligez A, Adam-Blondon AF, Cipriani G, Di Gaspero G, Laucou V, Merdinoglu D,
Meredith CP, Riaz S, Roux C, This P.

INRA, UMR DGPC 1097, equipe Genetique Vigne, batiment 6, 2 place Viala, 34060,
Montpellier Cedex 1, France. [email protected]

A grapevine (mainly Vitis vinifera L., 2n = 38) composite genetic map was
constructed with CarthaGene using segregation data from five full-sib
populations of 46, 95, 114, 139 and 153 individuals, to determine the relative
position of a large set of molecular markers. This consensus map comprised 515
loci (502 SSRs and 13 other type PCR-based markers), amplified using 439 primer
pairs (426 SSRs and 13 others) with 50.1% common markers shared by at least two
crosses. Out of all loci, 257, 85, 74, 69 and 30 were mapped in 1, 2, 3, 4 and 5
individual mapping populations, respectively. Marker order was generally well
conserved between maps of individual populations, with only a few significant
differences in the recombination rate of marker pairs between two or more
populations. The total length of the integrated map was 1,647 cM Kosambi
covering 19 linkage groups, with a mean distance between neighbour loci of 3.3
cM. A framework-integrated map was also built, with marker order supported by a
LOD of 2.0. It included 257 loci spanning 1,485 cM Kosambi with a mean
inter-locus distance of 6.2 cM over 19 linkage groups. These integrated maps are
the most comprehensive SSR-based maps available so far in grapevine and will
serve either for choosing markers evenly scattered over the whole genome or for
selecting markers that cover particular regions of interest. The framework map
is also a useful starting point for the integration of the V. vinifera physical
and genetic maps.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16799809 [PubMed - indexed for MEDLINE]

78: Ann Bot (Lond). 2006 Sep;98(3):483-94. Epub 2006 Jun 21.

Xylem structure and connectivity in grapevine (Vitis vinifera) shoots provides a
passive mechanism for the spread of bacteria in grape plants.

Chatelet DS, Matthews MA, Rost TL.

Section of Plant Biology, University of California, Davis, CA 95616, USA.
[email protected]s.edu

BACKGROUND AND AIMS: Bacterial leaf scorch occurring in a number of economically
important plants is caused by the xylem-limited bacterium Xylella fastidiosa
(Xf). In grapevine, Xf systemic infection causes Pierce's disease and is lethal.
Traditional dogma is that Xf movement between vessels requires the digestion of
inter-vessel pit membranes. However, Yersinia enterocolitica (Ye) (a bacterium
found in animals) and fluorescent beads moved rapidly within grapevine xylem
from stem into leaf lamina, suggesting open conduits consisting of long,
branched xylem vessels for passive movement. This study builds on and expands
previous observations on the nature of these conduits and how they affect Xf
movement. METHODS: Air, latex paint and green fluorescence protein (GFP)-Xf were
loaded into leaves and followed to confirm and identify these conduits. Leaf
xylem anatomy was studied to determine the basis for the free and sometimes
restricted movement of Ye, beads, air, paint and GFP-Xf into the lamina. KEY
RESULTS: Reverse loading experiments demonstrated that long, branched xylem
vessels occurred exclusively in primary xylem. They were observed in the stem
for three internodes before diverging into mature leaves. However, this
stem-leaf connection was an age-dependent character and was absent for the first
10-12 leaves basal to the apical meristem. Free movement in leaf blade xylem was
cell-type specific with vessels facilitating movement in the body of the blade
and tracheids near the leaf margin. Air, latex paint and GFP-Xf all moved about
50-60% of the leaf length. GFP-Xf was never observed close to the leaf margin.
CONCLUSIONS: The open vessels of the primary xylem offered unimpeded long
distance pathways bridging stem to leaves, possibly facilitating the spread of
bacterial pathogens in planta. GFP-Xf never reached the leaf margins where
scorching appeared, suggesting a signal targeting specific cells or a toxic
build-up at hydathodes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16790469 [PubMed - indexed for MEDLINE]

79: J Agric Food Chem. 2006 Jun 28;54(13):4839-48.

Simulated digestion of Vitis vinifera seed powder: polyphenolic content and
antioxidant properties.

Janisch KM, Olschlager C, Treutter D, Elstner EF.

Lehrstuhl fur Phytopathologie, Labor fur Angewandte Biochemie, WZW TU Munchen,
Germany.

There is increasing evidence that reactive oxygen species arising from several
enzymatic reactions are mediators of inflammatory events. Plant preparations
have the potential for scavenging such reactive oxygen species. Flavans and
procyanidins are bioavailable and stable during the process of cooking. This
study used conditions that mimicked digestion of Vitis vinifera seed powder in
the stomach (acidic preparation) and small intestine (neutral preparation). The
flavonoids of these two preparations were released during simulated digestion
and were determined with HPLC analysis. Biochemical model reactions relevant for
the formation of reactive oxygen species in vivo at inflammatory sites were used
to determine the antioxidant properties of the two preparations. The inhibition
of the indicator reaction for the formation of reactive oxygen species
represents a potential mechanism of the physiological activity of the
corresponding preparation. The results of this work show clearly that the
polyphenols released during the simulated digestion of the two preparations have
good scavenging potential against superoxide radicals, hydroxyl radicals, and
singlet oxygen. They protect low-density lipoprotein against copper-induced
oxidation due to the copper-chelating properties and their chain-breaking
abilities in lipid peroxidation.

Publication Types:
In Vitro

PMID: 16787037 [PubMed - indexed for MEDLINE]

80: Funct Integr Genomics. 2007 Apr;7(2):95-110. Epub 2006 Jun 15.

Gene expression associated with compatible viral diseases in grapevine
cultivars.

Espinoza C, Vega A, Medina C, Schlauch K, Cramer G, Arce-Johnson P.

Departamento de Genetica Molecular y Microbiologia, Facultad de Ciencias
Biologicas, Pontificia Universidad Catolica de Chile, Casilla 114-D, Alameda
340, Santiago, Chile, [email protected]

Viral diseases affect grapevine cultures without inducing any resistance
response. Thus, these plants develop systemic diseases and are chronically
infected. Molecular events associated with viral compatible infections
responsible for disease establishment and symptoms development are poorly
understood. In this study, we surveyed viral infection in grapevines at a
transcriptional level. Gene expression in the Vitis vinifera red wine cultivars
Carmenere and Cabernet-Sauvignon naturally infected with GLRaV-3 were evaluated
using a genome-wide expression profiling with the Vitis vinifera GeneChip((R))
from Affymetrix. We describe numerous genes that are induced or repressed in
viral infected grapevines leaves. Changes in gene expression involved a wide
spectrum of biological functions, including processes of translation and protein
targeting, metabolism, transport, and cell defense. Considering cellular
localization, the membrane and endomembrane systems appeared with the highest
number of induced genes, while chloroplastic genes were mostly repressed. As
most induced genes associated with the membranous system are involved in
transport, the possible effect of virus in this process is discussed. Responses
of both cultivars are analyzed and the results are compared with published data
from other species. This is the first study of global gene profiling in
grapevine in response to viral infections using DNA microarray.

PMID: 16775684 [PubMed - in process]

81: Genome. 2006 May;49(5):467-75.

AFLP analysis of genetic variation within the two economically important
Anatolian grapevine (Vitis vinifera L.) varietal groups.

Ergul A, Kazan K, Aras S, Cevik V, Celik H, Soylemezoglu G.

Ankara University, Institute of Biotechnology, Besevler-Ankara, Turkey,

The Anatolian region of modern-day Turkey is believed to have played an
important role in the history of grapevine (Vitis vinifera L.) domestication and
spread. Despite this, the rich grape germplasm of this region is virtually
uncharacterized genetically. In this study, the amplified fragment length
polymorphisms (AFLP)-based genetic relations of the grapevine accessions
belonging to the 2 economically important Anatolian table grape varietal groups
known as V. vinifera 'Misket' (Muscat) and V. vinifera 'Parmak' were studied.
Thirteen AFLP primer combinations used in the analyses revealed a total of 1495
(35.5% polymorphic) and 1567 (34.6% polymorphic) DNA fragments for the 'Misket'
and 'Parmak' varietal groups, respectively. The unweighted pair-group method
with arthimetic averaging (UPGMA) cluster analysis and principal coordinate
analysis (PCA) conducted on polymorphic AFLP markers showed that both varietal
groups contain a number of synonymous (similar genotypes known by different
names) as well as homony mous (genetically different genotypes known by the same
name) accessions. Our results also showed that 6 of the Anatolian 'Misket'
genotypes were genetically very similar to V. vinifera 'Muscat of Alexandria',
implying that these genotypes might have played some role in the formation of
this universally known grape cultivar. Finally, the close genetic similarities
found here between 'Muscat of Alexandria' and V. vinifera 'Muscat of Hamburg'
support the recent suggestion that 'Muscat of Hamburg' probably originated from
'Muscat of Alexandria' through spontaneous hybridizations. Overall, the results
of this study have implications for not only preservation and use of the
Anatolian grape germplasm, but also better understanding of the historical role
that this region has played during the domestication of grapes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16767171 [PubMed - indexed for MEDLINE]

82: Plant Physiol. 2006 Aug;141(4):1563-77. Epub 2006 Jun 9.

Pathways of glucose regulation of monosaccharide transport in grape cells.

Conde C, Agasse A, Glissant D, Tavares R, Geros H, Delrot S.

Departamento de Biologia, Universidade do Minho, Campus de Gualtar, 4710-057
Braga, Portugal.

Grape (Vitis vinifera) heterotrophic suspension-cultured cells were used as a
model system to study glucose (Glc) transport and its regulation. Cells
transported D-[14C]Glc according to simple Michaelis-Menten kinetics
superimposed on first-order kinetics. The saturating component is a
high-affinity, broad-specificity H+ -dependent transport system (Km = 0.05 mm).
Glc concentration in the medium tightly regulated the transcription of VvHT1
(Vitis vinifera hexose transporter 1), a monosaccharide transporter previously
characterized in grape berry, as well as VvHT1 protein amount and monosaccharide
transport activity. All the remaining putative monosaccharide transporters
identified so far in grape were poorly expressed and responded weakly to Glc.
VvHT1 transcription was strongly repressed by Glc and 2-deoxy-D-Glc, but not by
3-O-methyl-D-Glc or Glc plus mannoheptulose, indicating the involvement of a
hexokinase-dependent repression. 3-O-Methyl-D-Glc, which cannot be
phosphorylated, and Glc plus mannoheptulose induced a decrease of transport
activity caused by the reduction of VvHT1 protein in the plasma membrane without
affecting VvHT1 transcript levels. This demonstrates hexokinase-independent
posttranscriptional regulation. High Glc down-regulated VvHT1 transcription and
Glc uptake, whereas low Glc increased those parameters. Present data provide an
example showing control of plant sugar transporters by their own substrate both
at transcriptional and posttranscriptional levels. VvHT1 protein has an
important role in the massive import of monosaccharides into mesocarp cells of
young grape berries because it was localized in plasma membranes of the early
developing fruit. Protein amount decreased abruptly throughout fruit development
as sugar content increases, consistent with the regulating role of Glc on VvHT1
expression found in suspension-cultured cells.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16766675 [PubMed - indexed for MEDLINE]

83: J Exp Bot. 2006;57(9):2025-35. Epub 2006 May 23.

The contribution of extensin network formation to rapid, hydrogen
peroxide-mediated increases in grapevine callus wall resistance to fungal lytic
enzymes.

Ribeiro JM, Pereira CS, Soares NC, Vieira AM, Feijo JA, Jackson PA.

Instituto de Tecnologia Quimica e Biologica, Oeiras, Portugal.

Grapevine (Vitis vinifera cv. Touriga) callus cell walls contain a high level of
the monomeric extensin, GvP1. Hydrogen peroxide stimulus of these cultures
causes the rapid loss of monomeric GvP1, concomitant with marked increases in
insoluble GvP1 amino acids and wall resistance to digestion by fungal lytic
enzymes. JIM11 immunolocalization studies indicated that monomeric and network
GvP1 were evenly distributed in the callus cell wall. These primary cell walls
were used to investigate the specific contribution of extensin and other
ionically bound cell-wall proteins to hydrogen peroxide-mediated increases in
resistance to fungal lytic enzymes. This was performed by removing
ionically-bound proteins and assaying for hydrogen peroxide-enhanced resistance
after the addition of selected protein fractions. The results indicate that
hydrogen peroxide-induced increases in resistance to digestion by fungal lytic
enzymes require a co-operative action between network extensin formation and the
electrostatic interaction of additional wall proteins with the extracellular
matrix.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16720614 [PubMed - indexed for MEDLINE]

84: J Agric Food Chem. 2006 May 31;54(11):3882-6.

Use of Aloe vera gel coating preserves the functional properties of table
grapes.

Serrano M, Valverde JM, Guillen F, Castillo S, Martinez-Romero D, Valero D.

Department of Food Technology, and Department of Applied Biology, EPSO,
University Miguel Hernandez, Ctra. Beniel km. 3.2, 03312 Orihuela, Alicante,
Spain.

Table grapes (Vitis vinifera L. cv. Crimson Seedless) were coated with Aloe vera
gel according to our developed patent (SP Patent P200302937) and then stored for
35 days at 1 degrees C, and the subsequent shelf life (SL) was monitored at 20
degrees C. Uncoated clusters showed a rapid loss of functional compounds, such
as total phenolics and ascorbic acid. These changes were accompanied by
reduction of the total antioxidant activity (TAA) and increases in total
anthocyanins, showing an accelerated ripening process. On the contrary, table
grapes coated with Aloe vera gel significantly delayed the above changes, such
as the retention of ascorbic acid during cold storage or SL. Consequently, Aloe
vera gel coating, a simple and noncontaminating treatment, maintained the
functional properties during postharvest storage of table grapes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16719510 [PubMed - indexed for MEDLINE]

85: J Plant Physiol. 2006 May 19; [Epub ahead of print]

Study on symptomless leaf infection in grapevine plants grown in vitro with
grapewine leafroll associated virus 3.

Christov I, Stefanov D, Velinov T, Goltsev V, Georgieva K, Abracheva P, Genova
Y, Christov N.

Institute of Viticulture and Enology, 5800 Pleven, Bulgaria.

The photosynthetic changes evaluated by oxygen evolution, chlorophyll
fluorescence, photoacoustics, and delayed fluorescence (DF) were studied in
leaves of grown in vitro for 8 weeks grapevine plants (Vitis vinifera) infected
by grapevine leafroll-associated virus 3 (GLRaV-3). The infected leaves were
characterized during the viral infection without visible disease symptoms. The
symptomless infection led to a decrease in plant biomass. The non-photochemical
fluorescence quenching, qN, declined, whereas the photochemical quenching, qP,
and the Chl a/b ratio were not significantly affected. Photoacoustic and oxygen
evolution measurements showed that the energy storage and oxygen evolution rate
decreased in the infected leaves. Enhanced alternative electron sinks during the
symptomless viral infection were also estimated. The changes in fluorescence and
DF temperature curves demonstrated an enhanced stability of the thylakoid
membranes in the infected leaves. This effect was clearly expressed at high
actinic light intensities. The viral infected in vitro grown grapevine plants
were used in the present study as a simplified model system that allow to avoid
the involvement of different environmental factors that could interfere with the
GLRaV infection and the virus-grapevine interactions. Thus, the 'pure' impact of
the viral infection on photosynthesis could be investigated.

PMID: 16716452 [PubMed - as supplied by publisher]

86: Ann Bot (Lond). 2006 Jul;98(1):257-65. Epub 2006 May 16.

The photoprotective role of epidermal anthocyanins and surface pubescence in
young leaves of grapevine (Vitis vinifera).

Liakopoulos G, Nikolopoulos D, Klouvatou A, Vekkos KA, Manetas Y, Karabourniotis
G.

Laboratory of Plant Physiology, Department of Agricultural Biotechnology,
Agricultural University of Athens, Iera Odos 75, 118 55, Botanikos, Athens,
Greece. [email protected]

BACKGROUND AND AIMS: Depending on cultivar, surfaces of young leaves of Vitis
vinifera may be glabrous-green ('Soultanina') or transiently have anthocyanins
('Siriki') or pubescence ('Athiri'). A test is made of the hypothesis that
anthocyanins and pubescence act as light screens affording a photoprotective
advantage to the corresponding leaves, and an assessment is made of the
magnitude of their effect. METHODS: Measurements were made on young leaves of
the three cultivars in spring under field conditions. Photosynthetic
gas-exchange and in vivo chlorophyll fluorescence were measured. Photosynthetic
and photoprotective pigments were analysed by HPLC. KEY RESULTS: Compared with
glabrous-green leaves, both anthocyanic and pubescent leaves had greater
dark-adapted PSII photochemical efficiency and net photosynthesis. In leaves
possessing either anthocyanins or pubescence, the ratio of xanthophyll cycle
components to total chlorophyll, and mid-day de-epoxidation state of the
xanthophyll cycle were considerably smaller, than in glabrous-green leaves.
These differences were more evident in pubescent leaves, probably indicating
that trichomes were more effective in decreasing light stress than anthocyanins
in the epidermis. CONCLUSIONS: Light screens, especially in the form of
pubescence, decrease the risk of photoinhibition whilst allowing leaves to
maintain a smaller content of xanthophyll cycle components and depend less on
xanthophyll cycle energy dissipation. This combination of photoprotective
features, i.e. decreased photon flux to the photosynthetic apparatus and lower
xanthophyll cycle utilization rates may be particularly advantageous under
stressful conditions.

PMID: 16704996 [PubMed - indexed for MEDLINE]

87: Ann Bot (Lond). 2006 Jul;98(1):175-85. Epub 2006 May 5.

Branch development controls leaf area dynamics in grapevine (Vitis vinifera)
growing in drying soil.

Lebon E, Pellegrino A, Louarn G, Lecoeur J.

Institut National de la Recherche Agronomique, Ecole Nationale Superieure
Agronomique, Laboratoire d'Ecophysiologie des Plantes sous Stress
Environnementaux, UMR 759, INRA-ENSA.M, 2 place Viala, 34060 Montpellier cedex
01, France. [email protected]

BACKGROUND AND AIMS: Soil water deficit is a major abiotic stress with severe
consequences for the development, productivity and quality of crops. However, it
is considered a positive factor in grapevine management (Vitis vinifera), as it
has been shown to increase grape quality. The effects of soil water deficit on
organogenesis, morphogenesis and gas exchange in the shoot were investigated.
METHODS: Shoot organogenesis was analysed by distinguishing between the various
steps in the development of the main axis and branches. Several experiments were
carried out in pots, placed in a greenhouse or outside, in southern France. Soil
water deficits of various intensities were imposed during vegetative development
of the shoots of two cultivars ('Syrah' and 'Grenache N'). KEY RESULTS: All
developmental processes were inhibited by soil water deficit, in an
intensity-dependent manner, and sensitivity to water stress was
process-dependent. Quantitative relationships with soil water were established
for all processes. No difference was observed between the two cultivars for any
criterion. The number of leaves on branches was particularly sensitive to soil
water deficit, which rapidly and strongly reduced the rate of leaf appearance on
developing branches. This response was not related to carbon availability,
photosynthetic activity or the soluble sugar content of young expanding leaves.
The potential number of branches was not a limiting factor for shoot
development. CONCLUSIONS: The particularly high sensitivity to soil water
deficit of leaf appearance on branches indicates that this process is a major
determinant of the adaptation of plant leaf area to soil water deficit. The
origin of this particular developmental response to soil water deficit is
unclear, but it seems to be related to constitutive characteristics of branches
rather than to competition for assimilates between axes differing in sink
strength.

PMID: 16679414 [PubMed - indexed for MEDLINE]

88: J Agric Food Chem. 2006 May 3;54(9):3334-40.

Metabolic changes of Malvasia grapes for wine production during postharvest
drying.

Costantini V, Bellincontro A, De Santis D, Botondi R, Mencarelli F.

Department of Food Science and Technology, University of Viterbo, via S. Camillo
de Lellis, snc 01100 Viterbo, Italy.

Malvasia (Vitis vinifera L.) grapes were harvested at 17.8% of soluble solids
content (SSC) and placed inside an innovative dehydration room where
temperature, relative humidity, and air flow were maintained, respectively, at
15 degrees C, 40%, and 1-1.5 m s(-1). Weight loss of bunches reached
approximately 33% in 29 days. SSC increased inversely proportionally with the
weight decrease, reaching at the end of experiment 23%. Abscisic acid (ABA)
increased rapidly from around 29 to 80 microg g(-1) of dry weight at 11.7% of
bunch weight loss and then declined gradually. Lipoxygenase (LOX) showed the
same behavior as ABA, whereas alcohol dehydrogenase (ADH), read in the way of
ethanol oxidation, increased continuously when the weight loss reached
approximately 19.5%. In parallel with the activity of LOX, C6 compound [hexanal,
hex-1-enol, (E)-hex-2-enal] concentrations reached a peak at 11.7% of weight
loss, whereas ethanol and acetaldehyde increased with the increase of ADH and
successively decrease and ethyl acetate increased. Proline increased initially
as ABA and successively with the increase of ADH, 5.3-fold increase versus
4.2-fold increase of proteins. Postharvest dehydration of Malvasia grapes shows
a biphasic pattern: a first metabolic stress response up to 11.7% of bunch
weight loss and a second stress response beyond 19.5% of weight loss. The
metabolic mechanism of these postharvest water stress responses is discussed.

PMID: 16637693 [PubMed - indexed for MEDLINE]

89: J Appl Microbiol. 2006 May;100(5):946-54.

Isolation and characterization of rhizosphere bacteria with potential for
biological control of weeds in vineyards.

Flores-Vargas RD, O'Hara GW.

Centre for Rhizobium Studies, School of Biological Sciences and Biotechnology,
Division of Science and Engineering, Murdoch University, WA, Australia.
[email protected]

AIMS: Deleterious rhizosphere inhabiting bacteria (DRB) have potential to
suppress plant growth. This project focuses on the isolation of DRB with
potential for development as commercial products for weed control. METHODS AND
RESULTS: Bacteria were isolated from the rhizosphere, rhizoplane, and
endorhizosphere of seedlings and mature plants of wild radish (Raphanus
raphanistrum), annual ryegrass (Lolium rigidum) and capeweed (Arctotheca
calendula) growing in vineyards in the Swan Valley, Western Australia. A
majority (81.5%) of the 442 strains was obtained from either rhizospheres or
rhizoplanes. Rapid screening techniques were developed to evaluate in the
laboratory and glasshouse the effects of bacteria on plants. Strains were
screened in the glasshouse for deleterious effects on annual ryegrass, wild
radish, grapevine rootlings (Vitis vinifera) and the legume cover crop
subterranean clover (Trifolium subterraneum). Three strains were identified
using the Biolog system and 16S rRNA gene sequencing as two strains of
Pseudomonas fluorescens (WSM3455 and WSM3456) and one strain of Alcaligenes
xylosoxidans (WSM3457). One of the P. fluorescens (WSM3455) strain produced
hydrogen cyanide, an inhibitor of plant roots and a broad-spectrum antimicrobial
compound. CONCLUSIONS: Three strains specifically inhibited wild radish but had
no significant deleterious effects on either grapevine rootlings or subterranean
clover. SIGNIFICANCE AND IMPACT OF THE STUDY: This study suggested manipulation
of the weed seedling rhizosphere using identified DRB as a potential biocontrol
agent for wild radish.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16629995 [PubMed - indexed for MEDLINE]

90: Int J Food Microbiol. 2006 May 25;109(1-2):97-102. Epub 2006 Apr 19.

Yeasts isolated from three varieties of grapes cultivated in different locations
of the Dolenjska vine-growing region, Slovenia.

Raspor P, Milek DM, Polanc J, Mozina SS, Cadez N.

University of Ljubljana, Biotechnical Faculty, Food Science and Technology
Department, Jamnikarjeva 101, 1000 Ljubljana, Slovenia.
[email protected]

The number and diversity of yeasts on grape berry surfaces are influenced by
several factors, such as grape variety, degree of grape maturity at harvest,
climatological conditions, geographic location, physical damage of grapes, the
intensity of pest management etc. Cvicek is a typical Slovene wine, which has
obtained a special protection under the Slovene Wine Law for its geographical
origin. This blended red wine is produced from different grape varieties (Vitis
vinifera L.), mostly from red grapes of Zametovka and Modra frankinja and from
white grapes of Kraljevina. The aim of this study was to evaluate the impact of
geographical locations in the Dolenjska vine-growing region and to obtain
precise information about the influence of different grape varieties on the
composition of yeast community on grape berries. The restriction fragment length
polymorphism of PCR-amplified fragments from the rDNA gene cluster (PCR RFLP of
rDNA) has been used for the differentiation of yeast species. The standard
identification procedure has been performed on representative strains that
shared identical RFLP profiles. The number of yeasts and yeast species isolated
varied according to different grape varieties, Zametovka, Modra frankinja and
Kraljevina (V. vinifera L.) and according to different sampling location. On the
surface of grape berries 13 different yeast species have been identified.
Saccharomyces cerevisiae has not been found.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16626833 [PubMed - indexed for MEDLINE]

91: New Phytol. 2006;170(3):537-50.

Characterization of a new, nonpathogenic mutant of Botrytis cinerea with
impaired plant colonization capacity.

Kunz C, Vandelle E, Rolland S, Poinssot B, Bruel C, Cimerman A, Zotti C, Moreau
E, Vedel R, Pugin A, Boccara M.

Laboratoire Interactions Plantes-Pathogenes (UMR 217 INRA/INA-PG/Universite
Pierre et Marie Curie), INA-PG, 16 rue Claude Bernard, 75231 Paris Cedex 05,
France. [email protected]

Botrytis cinerea is a necrotrophic pathogen that attacks more than 200 plant
species. Here, the nonpathogenic mutant A336, obtained via insertional
mutagenesis, was characterized. Mutant A336 was nonpathogenic on leaves and
fruits, on intact and wounded tissue, while still able to penetrate the host
plant. It grew normally in vitro on rich media but its conidiation pattern was
altered. The mutant did not produce oxalic acid and exhibited a modified
regulation of the production of some secreted proteins (acid protease 1 and
endopolygalacturonase 1). Culture filtrates of the mutant triggered an important
oxidative burst in grapevine (Vitis vinifera) suspension cells, and the
mutant-plant interaction resulted in the formation of hypersensitive
response-like necrosis. Genetic segregation analyses revealed that the
pathogenicity phenotype was linked to a single locus, but showed that the
mutated gene was not tagged by the plasmid pAN7-1. Mutant A336 is the first
oxalate-deficient mutant to be described in B. cinerea and it differs from all
the nonpathogenic B. cinerea mutants described to date.

PMID: 16626475 [PubMed - indexed for MEDLINE]

92: J Chromatogr A. 2006 May 12;1114(2):188-97. Epub 2006 Apr 18.

Rapid tool for distinction of wines based on the global volatile signature.

Rocha SM, Coutinho P, Barros A, Delgadillo I, Coimbra MA.

Departamento de Quimica, Universidade de Aveiro, 3810-193 Aveiro, Portugal.
[email protected]

This work describes a novel methodology for the rapid distinction of wines by
headspace solid-phase microextraction coupled to gas chromatography-mass
spectrometry, followed by principal component analysis of the data
(HS-SPME-GC-MS-PCA). Headspace SPME is used to extract and concentrate the
volatile and semi-volatile fractions. A DB-FFAP fused silica GC capillary column
of 30 m at 220 degrees C was used acting as a transfer line of the components
sorbed by the Carbowax-divinylbenzene coating fibre to the mass spectrometer,
which acts as a sensor (30<m/z<300). In this methodology, which does not require
any pre-treatment of the sample, the global volatile signature of the wine
headspace (chromatographic profile and m/z pattern of fragmentation in each
scan) is evaluated without complete chromatographic separation of its
components. In order to retrieve from the data as much chemical information as
possible and to extract m/z fragments (markers) for the characterisation and
distinction of the wines varieties, a PCA was applied to the data resultant from
the unresolved volatile fraction. In the present study, two different
monovarietal white wines (Vitis vinifera L. var. Fernao-Pires and Arinto) were
tested. Associated to the fast character of the proposed methodology and
robustness taking into account the extraction time, it is also important to
focus the higher sensibility and the lower effect of the sample moisture of the
MS sensor response when compared to the conventional e-noses.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16620852 [PubMed - indexed for MEDLINE]

93: Plant Cell Rep. 2006 Sep;25(9):978-85. Epub 2006 Apr 14.

Nuclear DNA content of Vitis vinifera cultivars and ploidy level analyses of
somatic embryo-derived plants obtained from anther culture.

Leal F, Loureiro J, Rodriguez E, Pais MS, Santos C, Pinto-Carnide O.

Center of Genetics and Biotechnology, University of Tras-os-Montes and Alto
Douro, Vila Real, Portugal.

Flow cytometry was employed to determine the ploidy level of Vitis vinifera L.
somatic embryo-derived plants obtained from anther culture. Only one among the
41 analysed plants (2.4%) presented somaclonal variation (tetraploidy); the
other plants were diploid. No significant differences (P<or=0.05) were detected
between diploid and parental field plants. No haploid or aneuploid plants were
observed. The nuclear DNA content of nine V. vinifera cultivars was also
estimated using flow cytometry. A non-significant variation was found among the
cultivars, with DNA content ranging from 1.17 pg/2C (cv. 'Tinta Barroca' and
'Viosinho') to 1.26 pg/2C (cv. 'Cabernet Sauvignon'). These results and previous
studies on other Vitis species suggest that Vitis genome is stable with regard
to nuclear DNA content.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16614867 [PubMed - indexed for MEDLINE]

94: Sheng Wu Gong Cheng Xue Bao. 2006 Mar;22(2):299-305.

[Significant improved anthocyanins biosynthesis in suspension cultures of Vitis
vinifera by process intensification]

[Article in Chinese]

Qu JG, Yu XJ, Zhang W, Jin MF.

Dalian Institute of Chemical Physics, Chinese Academy of Sciences, Dalian
116023, China.

The low-production is a ubiquitous problem and has prevented the
commercialization of secondary metabolite production in plant cell culture. In
order to examine the effective approaches to improvement of secondary metabolite
production in plant cell culture, the investigation of anthocyanins accumulation
in suspension cultures of Vitis vinifera, as a model system, had been initiated
in our laboratory. In this present research, various elicitors and the precursor
of phenylalanine were used in combination to enhance the anthocyanins production
in suspension cultures of Vitis vinifera. And an integrated process with the
combination of elicitation, precursor feeding and light irradiation was reported
for rational bioprocess design. Among the combination treatment of phenylalanine
feeding and several elicitors (methyl-beta-cyclodextrin, dextran T-40, methyl
jasmonate, extracts of Aspergillus niger and Fusarium orthoceras), the
combination with methyl jasmonate gave the highest anthocyanins production in
suspension cultures of Vitis vinifera. When compared to the controls, the
anthocyanins content (CV/g, FCW) and production (CV/L) increased by 2.7-fold and
3.4-fold, respectively. The optimum time for the addition of phenylalanine and
methyl jasmonate was 4 days after inoculation. Two cell lines with different
anthocyanins-producing capacity responded differently to the optimum combination
treatment of 30 micromol/L phenylalanine feeding, 218 micromol/L methyl
jasmonate elicitation and 3000 to approximately 4000 1x light illumination. The
high-and low-anthocyanins-producing cell lines of VV05 and VV06 produced the
maximum of 2975 and 4090 CV/L of anthocyanins that were 2.5- and 5.2-fold of the
controls, respectively.

Publication Types:
English Abstract
Research Support, Non-U.S. Gov't

PMID: 16607960 [PubMed - in process]

95: Transgenic Res. 2006 Apr;15(2):181-95.

Consequences of transferring three sorghum genes for secondary metabolite
(cyanogenic glucoside) biosynthesis to grapevine hairy roots.

Franks TK, Powell KS, Choimes S, Marsh E, Iocco P, Sinclair BJ, Ford CM, van
Heeswijck R.

Cooperative Research Centre for Viticulture, SA, Australia.
[email protected]

A multigenic trait (biosynthesis of the secondary metabolite, dhurrin cyanogenic
glucoside) was engineered de novo in grapevine (Vitis vinifera L.). This follows
a recent report of transfer of the same trait to Arabidopsis (Arabidopsis
thaliana) using three genetic sequences from sorghum (Sorghum bicolor): two
cytochrome P450-encoding cDNAs (CYP79A1 and CYP71E1) and a
UDPG-glucosyltransferase-encoding cDNA (sbHMNGT). Here we describe the two-step
process involving whole plant transformation followed by hairy root
transformation, which was used to transfer the same three sorghum sequences to
grapevine. Transgenic grapevine hairy root lines that accumulated transcript
from none, one (sbHMNGT), two (CYP79A1 and CYP71E1) or all three transgenes were
recovered and characterisation of these lines provided information about the
requirements for dhurrin biosynthesis in grapevine. Only lines that accumulated
transcripts from all three transgenes had significantly elevated cyanide
potential (up to the equivalent of about 100 mg HCN kg(-1) fresh weight), and
levels were highly variable. One dhurrin-positive line was tested and found to
release cyanide upon maceration and can therefore be considered 'cyanogenic'. In
in vitro dual co-culture of this cyanogenic hairy root line or an acyanogenic
line with the specialist root-sucking, gall-forming, aphid-like insect,
grapevine phylloxera (Daktulosphaira vitifoliae, Fitch), there was no evidence
for protection of the cyanogenic plant tissue from infestation by the insect.
Consistently high levels of dhurrin accumulation may be required for this to
occur. The possibility that endogenous grapevine gene expression is modulated in
response to engineered dhurrin biosynthesis was investigated using microarray
analysis of 1225 grapevine ESTs, but differences in patterns of gene expression
associated with dhurrin-positive and dhurrin-negative phenotypes were not
identified.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16604459 [PubMed - in process]

96: BMC Evol Biol. 2006 Apr 9;6:32.

Phylogenetic analyses of Vitis (Vitaceae) based on complete chloroplast genome
sequences: effects of taxon sampling and phylogenetic methods on resolving
relationships among rosids.

Jansen RK, Kaittanis C, Saski C, Lee SB, Tomkins J, Alverson AJ, Daniell H.

Section of Integrative Biology and Institute of Cellular and Molecular Biology,
Patterson Laboratories 141, University of Texas, Austin, TX 78712, USA.
[email protected]

BACKGROUND: The Vitaceae (grape) is an economically important family of
angiosperms whose phylogenetic placement is currently unresolved. Recent
phylogenetic analyses based on one to several genes have suggested several
alternative placements of this family, including sister to Caryophyllales,
asterids, Saxifragales, Dilleniaceae or to rest of rosids, though support for
these different results has been weak. There has been a recent interest in using
complete chloroplast genome sequences for resolving phylogenetic relationships
among angiosperms. These studies have clarified relationships among several
major lineages but they have also emphasized the importance of taxon sampling
and the effects of different phylogenetic methods for obtaining accurate
phylogenies. We sequenced the complete chloroplast genome of Vitis vinifera and
used these data to assess relationships among 27 angiosperms, including nine
taxa of rosids. RESULTS: The Vitis vinifera chloroplast genome is 160,928 bp in
length, including a pair of inverted repeats of 26,358 bp that are separated by
small and large single copy regions of 19,065 bp and 89,147 bp, respectively.
The gene content and order of Vitis is identical to many other unrearranged
angiosperm chloroplast genomes, including tobacco. Phylogenetic analyses using
maximum parsimony and maximum likelihood were performed on DNA sequences of 61
protein-coding genes for two datasets with 28 or 29 taxa, including eight or
nine taxa from four of the seven currently recognized major clades of rosids.
Parsimony and likelihood phylogenies of both data sets provide strong support
for the placement of Vitaceae as sister to the remaining rosids. However, the
position of the Myrtales and support for the monophyly of the eurosid I clade
differs between the two data sets and the two methods of analysis. In parsimony
analyses, the inclusion of Gossypium is necessary to obtain trees that support
the monophyly of the eurosid I clade. However, maximum likelihood analyses place
Cucumis as sister to the Myrtales and therefore do not support the monophyly of
the eurosid I clade. CONCLUSION: Phylogenies based on DNA sequences from
complete chloroplast genome sequences provide strong support for the position of
the Vitaceae as the earliest diverging lineage of rosids. Our phylogenetic
analyses support recent assertions that inadequate taxon sampling and incorrect
model specification for concatenated multi-gene data sets can mislead
phylogenetic inferences when using whole chloroplast genomes for phylogeny
reconstruction.

Publication Types:
Research Support, N.I.H., Extramural
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16603088 [PubMed - indexed for MEDLINE]

97: Mycologia. 2005 Sep-Oct;97(5):1111-21.

Botryosphaeria viticola sp. nov. on grapevines: a new species with a Dothiorella
anamorph.

Luque J, Martos S, Phillips AJ.

Departament de Proteccio Vegetal, Institut de Recerca i Tecnologia
Agroalimentaries, Centre de Cabrils, Barcelona, Spain.

Botryosphaeria viticola sp. nov., isolated from pruned canes of Vitis vinifera
in NE Spain, is described and illustrated. Phylogenetic analysis based on ITS
and EF1-alpha sequences and morphological characters of both anamorph and
teleomorph confirmed this taxon to be included within the group of
Botryosphaeria species with Dothiorella anamorphs. It is related most closely to
B. sarmentorum and B. iberica from which it differs in morphological characters
of the teleomorph and DNA sequences.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16596961 [PubMed - indexed for MEDLINE]

98: Nutr Cancer. 2005;53(2):244-54.

Quality assessment of commercial dietary antioxidant products from Vitis
vinifera L. grape seeds.

Monagas M, Hernandez-Ledesma B, Garrido I, Martin-Alvarez PJ, Gomez-Cordoves C,
Bartolome B.

Instituto de Fermentaciones Industriales (CSIC), Madrid, Spain.

Phenolic preparations from Vitis vinifera L. grape seeds are products commonly
used in the formulation of dietary antioxidant supplements. In this article, we
used a methodology (the oxygen radical absorbance capacity, ORAC) to evaluate
the in vitro antioxidant capacity of commercial dietary grape seed products and
studied the relationship of the antioxidant capacity with the phenolic
composition of these products. The ORAC value of the different brands of
commercial products studied (n = 16) varied from 2.71 to 26.4 micromol Trolox
equivalents/mg (approximately equal to 10-fold difference). For four of these
products, the batch-to-batch ORAC variation, expressed as a coefficient of
variation of the mean, was 10.5% (n = 6), 13.1% (n = 3), 19.4% (n = 4), and 7.8%
(n = 4). Analysis of monomeric and oligomeric flavan-3-ols by liquid
chromatography-diode array detection (LC-DAD)/electrospray-mass spectrometry and
procyanidins by thiolysis-LC-DAD also revealed large differences among the
commercial grape seed products. Moreover, the ORAC value could be fitted to a
regression model using variables from contents of individual phenolic compounds
and procyanidins. The product-to-product and batch-to-batch variation in ORAC
values and flavan-3-ol composition found among the commercial products studied
demonstrated that they are poorly standardized, resulting in inconsistent
composition and biological activity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16573385 [PubMed - indexed for MEDLINE]

99: Biosci Biotechnol Biochem. 2006 Mar;70(3):632-8.

Organ-specific transcription of putative flavonol synthase genes of grapevine
and effects of plant hormones and shading on flavonol biosynthesis in grape
berry skins.

Fujita A, Goto-Yamamoto N, Aramaki I, Hashizume K.

National Research Institute of Brewing, Hiroshima, Japan. [email protected]

In order to investigate the control mechanism of flavonol biosynthesis of
grapevine, we obtained five genomic sequences (FLS1 to FLS5) of putative
flavonol synthase genes from Vitis vinifera cv. Cabernet Sauvignon. The mRNA of
five FLSs accumulated in flower buds and flowers, while the mRNA of FLS2, FLS4,
and FLS5 accumulated in small berry skins and then decreased toward veraison. At
the ripening stage, the mRNA of only FLS4 and FLS5 accumulated again. This
change in mRNA accumulation did not contradict the flavonol accumulation in the
berry skins. Shading of the berries completely inhibited the increase in
flavonol content and mRNA accumulation of FLS4, but did not affect the mRNA
accumulation of FLS5. The effects of light and plant hormones on flavonol
accumulation were different from those on anthocyanin accumulation. Thus
flavonol biosynthesis appears to be under a different control system from that
of anthocyanin biosynthesis.

PMID: 16556978 [PubMed - indexed for MEDLINE]

100: Plant Cell Rep. 2006 Sep;25(9):968-77. Epub 2006 Mar 22.

Stress- and development-induced expression of spliced and unspliced transcripts
from two highly similar dehydrin 1 genes in V. riparia and V. vinifera.

Xiao H, Nassuth A.

Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON,
Canada.

Dehydrins are proteins that accumulate in vegetative tissues subjected to
various dehydrating stress conditions such as cold, drought, and salinity and in
seeds at later stages of embryogenesis. Here, we report on two highly identical
dehydrin genes, DHN1a and DHN1b, in wild and cultivated grapes, Vitis riparia
and Vitis vinifera, and their expression in different tissues and under
different environmental conditions. The two genes and their transcripts can
easily be distinguished by RT-PCR because DHN1b has an 18 bp deletion compared
to DHN1a. V. riparia expressed only DHN1a; V. vinifera expressed both DHN1a and
DHN1b. Spliced transcripts, DHN1-S, encoding a putative YSK(2)-type dehydrin
were present in low amounts in control leaves, but in high amounts in buds and
seeds. Unspliced transcripts, DHN1-U, accumulated to high levels in buds and
seeds. Cold, drought, and ABA treatment increased accumulation of both DHN1-S
and DHN1-U in leaves, whereas short-day treatment increased only DHN1-S. The
possible relation of these results with the difference in freezing stress
tolerance between V. riparia and V. vinifera is discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16552595 [PubMed - indexed for MEDLINE]

101: Plant Physiol Biochem. 2006 Jan;44(1):58-67. Epub 2006 Feb 21.

Expression analysis of defence-related genes in grapevine leaves after
inoculation with a host and a non-host pathogen.

Kortekamp A.

Institute for Phytomedicine, University of Hohenheim, 70593 Stuttgart, Germany.
[email protected]

The expression of PR protein encoding genes and genes involved in the
phenylpropanoid metabolism was analysed on grapevine leaves of susceptible and
resistant cvs. in response to inoculation with the host-pathogen Plasmopara
viticola and the non-host pathogen Pseudoperonospora cubensis, the downy mildew
pathogen of cucumber. These experiments were conducted to elucidate whether or
not grapevine plants susceptible to downy mildew exhibit an identical defence
response after inoculation with the non-host pathogen. Expression analysis of
defence-related genes revealed marked differences between the susceptible
cultivar "Riesling" (Vitis vinifera) and the resistant cultivar "Gloire de
Montpellier" (Vitis riparia). Whereas some genes seem to be expressed
constitutively in "Gloire" or induced after an inoculation with both pathogens,
expression of defence-related genes in Riesling was influenced mainly after
inoculation with the non-host pathogen: PR-2, PR-3, PR-4, a PGIP gene, and
especially genes encoding enzymes involved in anthocyanin biosynthesis (DFR,
F3H, LDOX) were affected. Therefore, the occurrence of the respective products
(flavans and other phenolics) in inoculated leaves was investigated with
appropriate histological staining techniques. These stainings revealed a
production of catechins and related phenolic compounds within the first 48 hai
(hours after inoculation) with Ps. cubensis but not with P. viticola in
Riesling, whereas in Gloire no further production was seen, which may be due to
the high content of polyphenolics as observed in control leaves. In addition to
the staining procedures, sporulation intensity was monitored on leaf discs.
Pretreatments of leaf discs with Ps. cubensis led to a reduced browning reaction
(as a result of a hypersensitive reaction) in Gloire and significantly reduced
the intensity of sporulation in Riesling after a subsequent inoculation with P.
viticola.

Publication Types:
Comparative Study

PMID: 16531058 [PubMed - indexed for MEDLINE]

102: Plant Cell Rep. 2006 Aug;25(8):807-14. Epub 2006 Mar 10.

Biolistic transformation of grapevine using minimal gene cassette technology.

Vidal JR, Kikkert JR, Donzelli BD, Wallace PG, Reisch BI.

Department of Horticultural Sciences, New York State Agricultural Experiment
Station, Cornell University, Geneva, NY, 14456, USA.

The use of minimal gene cassettes (MCs), which are linear DNA fragments
(promoter+open reading frame+terminator) lacking the vector backbone sequence,
was compared to the traditional use of whole circular plasmids (CPs) for
transformation of grapevine. Embryogenic cell suspensions of 'Chardonnay' (Vitis
vinifera L.) were transformed via particle co-bombardment using two nonlinked
genes in either MCs or CPs. One construct contained the npt-II selectable marker
and the second construct contained the MSI99 antimicrobial peptide gene. A total
of five lines each from MC and CP treatments that showed positive signals by PCR
for both the npt-II and MSI99 genes were selected. Southern blot analyses
revealed up to five integration events in the DNA treatments. Transcription
levels determined by semi-quantitative RT-PCR varied among transgenic lines. No
significant differences were found in transgene transcription between lines from
MC and CP transformation. The correlation between npt-II and MSI99 transcription
levels was positive (P<0.05), however, no correlation between the transcription
level and the number of integration events was observed. Transgenic lines
presented a similar phenotype in leaf morphology and plant vigor compared to
non-transgenic lines. Moreover, transgenic lines from both MC and CP DNA
treatments produced fruit as did the non-transgenic lines in the third year of
growth in the greenhouse. Our data confirm the effectiveness of the minimal
cassette technology for genetic transformation of grapevine cultivars.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16528564 [PubMed - indexed for MEDLINE]

103: Transgenic Res. 2006 Feb;15(1):69-82.

Evaluation of transgenic 'Chardonnay' (Vitis vinifera) containing magainin genes
for resistance to crown gall and powdery mildew.

Vidal JR, Kikkert JR, Malnoy MA, Wallace PG, Barnard J, Reisch BI.

Department of Horticultural Sciences, New York State Agricultural Experiment
Station, Cornell University, Geneva, NY 14456, USA.

Magainins, short peptides with broad-spectrum antimicrobial activity in vitro,
were assayed for their ability to confer resistance to pathogens in transgenic
grapevines. Embryogenic cell suspensions of 'Chardonnay' (Vitis vinifera L.)
were co-transformed by microprojectile bombardment with a plasmid carrying the
npt-II gene and a second plasmid harboring either a natural magainin-2 (mag2) or
a synthetic derivative (MSI99) gene. Magainin genes and the marker gene were
driven by Arabidopsis ubiquitin-3 and ubiquitin-11 promoters, respectively. A
total of 10 mag2 and 9 MSI99 regenerated lines were studied by Southern blot
hybridization, which showed 1-6 transgene integration events into the plant
genome. Semi-quantitative reverse transcription-polymerase chain reaction
(RT-PCR) revealed a variable range in transcription levels among mag2 and MSI99
lines. A positive correlation between number of integration events and
transcription level was observed (p<0.05). Plants were acclimated and challenged
in the greenhouse with either Agrobacterium vitis strains (bacterial crown gall
pathogen) at 10(8) cfu/ml or Uncinula necator (fungal powdery mildew pathogen)
at 10(5) conidia/ml for evaluation of disease resistance. A total of 6 mag2 and
5 MSI99 lines expressing the antimicrobial genes exhibited significant
reductions of crown gall symptoms as compared to non-transformed controls.
However, only two mag2 lines showed measurable symptom reductions in response to
U. necator, but not strong resistance. Our results suggest that the expression
of magainin-type genes in grapevines may be more effective against bacteria than
fungi. Additional strategies to enhance transgene expression and the spectrum of
resistance to grape diseases are suggested.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16475011 [PubMed - in process]

104: Appl Environ Microbiol. 2006 Feb;72(2):1467-75.

A novel Bacteroidetes symbiont is localized in Scaphoideus titanus, the insect
vector of Flavescence doree in Vitis vinifera.

Marzorati M, Alma A, Sacchi L, Pajoro M, Palermo S, Brusetti L, Raddadi N,
Balloi A, Tedeschi R, Clementi E, Corona S, Quaglino F, Bianco PA, Beninati T,
Bandi C, Daffonchio D.

Dipartimento di Scienze e Tecnologie Alimentari e Microbiologiche, Universita
degli Studi di Milano, 20133 Milan, Italy.

Flavescence doree (FD) is a grapevine disease that afflicts several wine
production areas in Europe, from Portugal to Serbia. FD is caused by a
bacterium, "Candidatus Phytoplasma vitis," which is spread throughout the
vineyards by a leafhopper, Scaphoideus titanus (Cicadellidae). After collection
of S. titanus specimens from FD-contaminated vineyards in three different areas
in the Piedmont region of Italy, we performed a survey to characterize the
bacterial microflora associated with this insect. Using length heterogeneity PCR
with universal primers for bacteria we identified a major peak associated with
almost all of the individuals examined (both males and females).
Characterization by denaturing gradient gel electrophoresis confirmed the
presence of a major band that, after sequencing, showed a 97 to 99% identity
with Bacteroidetes symbionts of the "Candidatus Cardinium hertigii" group. In
addition, electron microscopy of tissues of S. titanus fed for 3 months on
phytoplasma-infected grapevine plants showed bacterial cells with the typical
morphology of "Ca. Cardinium hertigii." This endosymbiont, tentatively
designated ST1-C, was found in the cytoplasm of previtellogenic and vitellogenic
ovarian cells, in the follicle cells, and in the fat body and salivary glands.
In addition, cell morphologies resembling those of "Ca. Phytoplasma vitis" were
detected in the midgut, and specific PCR assays indicated the presence of the
phytoplasma in the gut, fat body and salivary glands. These results indicate
that ST1-C and "Ca. Phytoplasma vitis" have a complex life cycle in the body of
S. titanus and are colocalized in different organs and tissues.

PMID: 16461701 [PubMed - indexed for MEDLINE]

105: Curr Med Chem. 2006;13(1):87-98.

Comparative studies of the antioxidant effects of cis- and trans-resveratrol.

Orallo F.

Departamento de Farmacologia, Facultad de Farmacia, Universidad de Santiago de
Compostela, Santiago de Compostela (La Coruna) Spain. [email protected]

Resveratrol (3,4',5-trihydroxystilbene, RESV) is a natural phenolic compound
that exists as cis and trans isomers [c-RESV or (Z)-RESV and t-RESV or (E)-RESV,
respectively]. t-RESV is a natural component of Vitis vinifera L. (Vitaceae),
abundant in the skin of grapes (but not in the flesh) and in the leaf epidermis,
and present in wines, especially red wines. In in vitro, ex vivo and in vivo
experiments t-RESV exhibits a number of biological activities, including
anti-inflammatory and anticarcinogenic properties. RESV also exists in wines as
a cis isomer, which (unlike t-RESV) is not currently available commercially; as
a result, little is known about this isomer's pharmacological activity. In this
review, I will focus on the few comparative studies of the antioxidant effects
of the two RESV isomers in different experimental models.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't
Review

PMID: 16457641 [PubMed - indexed for MEDLINE]

106: BMC Genomics. 2006 Jan 24;7:12.

Colour variation in red grapevines (Vitis vinifera L.): genomic organisation,
expression of flavonoid 3'-hydroxylase, flavonoid 3',5'-hydroxylase genes and
related metabolite profiling of red cyanidin-/blue delphinidin-based
anthocyanins in berry skin.

Castellarin SD, Di Gaspero G, Marconi R, Nonis A, Peterlunger E, Paillard S,
Adam-Blondon AF, Testolin R.

Dipartimento di Scienze Agrarie e Ambientali, University of Udine, via delle
Scienze 208, 33100 Udine, Italy. [email protected]

BACKGROUND: Structural genes of the phenyl-propanoid pathway which encode
flavonoid 3'- and 3',5'-hydroxylases (F3'H and F3'5'H) have long been invoked to
explain the biosynthesis of cyanidin- and delphinidin-based anthocyanin pigments
in the so-called red cultivars of grapevine. The relative proportion of the two
types of anthocyanins is largely under genetic control and determines the colour
variation among red/purple/blue berry grape varieties and their corresponding
wines. RESULTS: Gene fragments of VvF3'H and VvF3'5'H, that were isolated from
Vitis vinifera 'Cabernet Sauvignon' using degenerate primers designed on plant
homologous genes, translated into 313 and 239 amino acid protein fragments,
respectively, with up to 76% and 82% identity to plant CYP75 cytochrome P450
monooxygenases. Putative function was assigned on the basis of sequence
homology, expression profiling and its correlation with metabolite accumulation
at ten different ripening stages. At the onset of colour transition,
transcriptional induction of VvF3'H and VvF3'5'H was temporally coordinated with
the beginning of anthocyanin biosynthesis, the expression being 2-fold and
50-fold higher, respectively, in red berries versus green berries. The peak of
VvF3'5'H expression was observed two weeks later concomitantly with the increase
of the ratio of delphinidin-/cyanidin-derivatives. The analysis of structural
genomics revealed that two copies of VvF3'H are physically linked on linkage
group no. 17 and several copies of VvF3'5'H are tightly clustered and embedded
into a segmental duplication on linkage group no. 6, unveiling a high complexity
when compared to other plant flavonoid hydroxylase genes known so far, mostly in
ornamentals. CONCLUSION: We have shown that genes encoding flavonoid 3'- and
3',5'-hydroxylases are expressed in any tissues of the grape plant that
accumulate flavonoids and, particularly, in skin of ripening red berries that
synthesise mostly anthocyanins. The correlation between transcript profiles and
the kinetics of accumulation of red/cyanidin- and blue/delphinidin-based
anthocyanins indicated that VvF3'H and VvF3'5'H expression is consistent with
the chromatic evolution of ripening bunches. Local physical maps constructed
around the VvF3'H and VvF3'5'H loci should help facilitate the identification of
the regulatory elements of each isoform and the future manipulation of grapevine
and wine colour through agronomical, environmental and biotechnological tools.

PMID: 16433923 [PubMed - indexed for MEDLINE]

107: J Agric Food Chem. 2006 Jan 25;54(2):319-27.

Commercial dietary ingredients from Vitis vinifera L. leaves and grape skins:
antioxidant and chemical characterization.

Monagas M, Hernandez-Ledesma B, Gomez-Cordoves C, Bartolome B.

Instituto de Fermentaciones Industriales (CSIC), Juan de la Cierva 3, 28006
Madrid, Spain.

This paper reports an attempt to functionally and chemically characterize
commercial ingredients from Vitis vinifera L. grape skins, grape pomace, and
leaves, which are used in the formulation of dietary antioxidant supplements.
The antioxidant capacity of these ingredients was assessed for the first time by
the oxygen radical absorbance capacity (ORAC) methodology. Ingredients from
grape skins and pomace (n = 17) showed ORAC values from 1.38 to 21.4 mumol
Trolox equivalents/mg whereas ingredients from leaves (n = 4) showed ORAC values
from 1.52 to 2.55 mumol Trolox equivalents/mg. The high-performance liquid
chromatography-diode array detection/electrospray ionization-mass sprectrometry
analysis of anthocyanins and flavonols revealed the authenticity of the
ingredients as derived from V. vinifera L. and confirmed large differences in
their phenolic content and distribution. A progressive decline in both
antioxidant capacity and total anthocyanin content of a grape skin ingredient
(43 and 40% decrease, respectively) was observed over a 60 day storage period
(45 degrees C and 75% relative humidity), demonstrating its poor stability under
these conditions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16417286 [PubMed - indexed for MEDLINE]

108: J Agric Food Chem. 2006 Jan 25;54(2):299-305.

Noninvasive evaluation of the degree of ripeness in grape berries (vitis
vinifera L. Cv. Bacchus and silvaner) by chlorophyll fluorescence.

Kolb CA, Wirth E, Kaiser WM, Meister A, Riederer M, Pfundel EE.

Julius-von-Sachs-Institut fur Biowissenschaften, Universitat Wurzburg,
Julius-von-Sachs-Platz, D-97082 Wurzburg, Germany.

The use of chlorophyll fluorescence measurements to noninvasively evaluate
degrees of ripeness was investigated in berries at various stages of ripening
from two white grapevine cultivars (Vitis vinifera L. Cv. Bacchus and Silvaner).
Berries were characterized by diameter, weight, and density and by
concentrations of fructose, glucose, sucrose, and total sugars, as well as
fructose/glucose ratios, and also by chlorophyll fluorescence at F(0) and F(M)
levels and the fluorescence ratio F(V)/F(M). Pearson product moment correlation
analysis on data from both cultivars revealed clear negative associations
between F(0) and concentrations of fructose, glucose, and total sugars, and
fructose/glucose ratios (correlation coefficient < -0.89). Curvilinear
trend-lines were established for plots of F(0) versus concentrations of
fructose, glucose, and total sugars, but a linear relationship between F(0) and
fructose/glucose ratios was found: the corresponding coefficients of
determination were always >0.82. Therefore, chlorophyll fluorescence
measurements are well-suited to determine noninvasively sugar accumulation in
grape berries during ripening.

PMID: 16417283 [PubMed - indexed for MEDLINE]

109: Microb Ecol. 2006 Jan;51(1):109-16. Epub 2006 Jan 11.

Interactions between yeasts and grapevines: filamentous growth,
endopolygalacturonase and phytopathogenicity of colonizing yeasts.

Gognies S, Barka EA, Gainvors-Claisse A, Belarbi A.

Laboratoire de Microbiologie Generale et Moleculaire, Universite de Reims, UFR
Sciences, URVVC, UPRES EA 2069, B.P. 1039, 51687 Reims Cedex 2, France.

It has been clearly established that phytopathogenic fungi, bacteria, and
viruses exert biotic stresses on plants. Much less is known, however, about the
interactions between enological species of yeast and their host plants. In a
previous study, we described how Saccharomyces cerevisiae, the most common
enological yeast, can act as a grapevine (Vitis vinifera L.) pathogen, causing
growth retardation or plant death. In the present in vitro study on 11 strains
of yeast belonging to different genera, which often occur on the surfaces of
vineyard grapes and V. vinifera, a link was found to exist between strain
phytopathogenecity and pseudohyphal growth habits and/or endopolygalacturonase
activity. The results obtained here are consistent with earlier findings showing
that the phytopathogenicity of yeast strains depends on the filamentous growth
process, and show that endopolygalacturonase alone is not responsible for the
invasion of plants tissues. The mechanisms observed here may be of significant
ecological importance and may help to explain the long periods of yeast survival
found to occur in vineyards.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16408245 [PubMed - indexed for MEDLINE]

110: Plant Cell Rep. 2006 Jun;25(6):546-53. Epub 2006 Jan 12.

Molecular characterization of grapevine plants transformed with GFLV resistance
genes: II.

Maghuly F, Leopold S, da Camara Machado A, Borroto Fernandez E, Ali Khan M,
Gambino G, Gribaudo I, Schartl A, Laimer M.

Plant Biotechnology Unit, Institute of Applied Microbiology BOKU, Nussdorfer
Lande 11, A-1190 Vienna, Austria.

A collection of 127 putatively transgenic individuals of Vitis vinifera cv.
Russalka was characterized by PCR and Southern hybridization. Six different
constructs containing the neomycin phosphotransferase (nptII) marker gene and
sequences of the Grapevine Fanleaf Virus Coat Protein (GFLV CP) gene including
non-translatable and truncated forms were transferred via Agrobacterium-mediated
transformation. Detection of transgenic sequences by PCR was positive in all
lines. Southern blot analysis revealed that the number of inserted T-DNA copies
ranged from 1 to 6. More than 46% of the tested transgenic lines contain one
copy of the inserted T-DNA, qualifying them as interesting candidates for
further breeding programs. Southern data of one line indicate the presence of an
incomplete copy of the T-DNA, thus confirming previous PCR results. Since many
putative transgenic lines shared identical hybridization patterns, they were
clustered into 39 lines and considered as having originated from independent
transformation events. The detection of the tetracycline (TET) resistance genes
in 15% of the lines shows that an integration of plasmid backbone sequences
beyond the T-DNA borders occurred. Enzyme-linked immunosorbent assay (ELISA)
performed on leaf tissue did not show any accumulation of the GFLV CP in the 39
transgenic lines analyzed. Reverse transcription polymerase chain reaction
(RT-PCR) and Northern blot were carried out; RT-PCR analyses showed that the
GFLV CP mRNA was expressed at variable levels.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16408176 [PubMed - indexed for MEDLINE]

111: Plant Physiol. 2006 Feb;140(2):558-79. Epub 2006 Jan 11.

Abscisic acid stimulates a calcium-dependent protein kinase in grape berry.

Yu XC, Li MJ, Gao GF, Feng HZ, Geng XQ, Peng CC, Zhu SY, Wang XJ, Shen YY, Zhang
DP.

China State Key Laboratory of Plant Physiology and Biochemistry, China
Agricultural University, 100094 Beijing, China.

It has been demonstrated that calcium plays a central role in mediating abscisic
acid (ABA) signaling, but many of the Ca2+-binding sensory proteins as the
components of the ABA-signaling pathway remain to be elucidated. Here we
identified, characterized, and purified a 58-kD ABA-stimulated calcium-dependent
protein kinase from the mesocarp of grape berries (Vitis vinifera x Vitis
labrusca), designated ACPK1 (for ABA-stimulated calcium-dependent protein
kinase1). ABA stimulates ACPK1 in a dose-dependent manner, and the ACPK1
expression and enzyme activities alter accordantly with the endogenous ABA
concentrations during fruit development. The ABA-induced ACPK1 stimulation
appears to be transient with a rapid effect in 15 min but also with a slow and
steady state of induction after 60 min. ABA acts on ACPK1 indirectly and
dependently on in vivo state of the tissues. Two inactive ABA isomers,
(-)-2-cis, 4-trans-ABA and 2-trans, 4-trans-(+/-)-ABA, are ineffective for
inducing ACPK1 stimulation, revealing that the ABA-induced effect is stereo
specific to physiological active (+)-2-cis, 4-trans-ABA. The other phytohormones
such as auxin indoleacetic acid, gibberellic acid, synthetic cytokinin
N-benzyl-6-aminopurine, and brassinolide are also ineffective in this ACPK1
stimulation. Based on sequencing of the two-dimensional electrophoresis-purified
ACPK1, we cloned the ACPK1 gene. The ACPK1 is expressed specifically in grape
berry covering a fleshy portion and seeds, and in a developmental
stage-dependent manner. We further showed that ACPK1 is localized in both plasma
membranes and chloroplasts/plastids and positively regulates plasma membrane
H+-ATPase in vitro, suggesting that ACPK1 may be involved in the ABA-signaling
pathway.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16407437 [PubMed - indexed for MEDLINE]

112: Theor Appl Genet. 2006 Feb;112(4):708-16. Epub 2006 Jan 10.

Linkage disequilibrium in cultivated grapevine, Vitis vinifera L.

Barnaud A, Lacombe T, Doligez A.

INRA, UMR DGPC Equipe Genetique Vigne, 2 place Viala, 34060, Montpellier Cedex
1, France.

We present here the first study of linkage disequilibrium (LD) in cultivated
grapevine, Vitis vinifera L. subsp. vinifera (sativa), an outcrossing highly
heterozygous perennial species. Our goal was to characterize the amount and
pattern of LD at the scale of a few centiMorgans (cM) between 38 microsatellite
loci located on five linkage groups, in order to assess its origin and potential
applications. We used a core collection of 141 cultivars representing the
diversity of the cultivated compartment. LD was evaluated with both independence
tests and multilocus r2, both on raw genotypic and reconstructed haplotypic
data. Significant genotypic LD was found only within linkage groups, extending
up to 16.8 cM. It appeared not to be influenced by the weak structure of the
sample and seemed to be mainly of haplotypic origin. Significant haplotypic LD
was found over 30 cM. Both genotypic and haplotypic r2 values declined to around
0.1 within 5-10 cM, suggesting a rather narrow genetic base of the cultivated
compartment and limited recombination since domestication events. These first
results open up a few application opportunities for association mapping of QTLs
and marker assisted selection.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16402190 [PubMed - indexed for MEDLINE]

113: J Plant Physiol. 2006 Feb;163(2):115-27. Epub 2005 Sep 9.

Changes and subcellular localizations of the enzymes involved in phenylpropanoid
metabolism during grape berry development.

Chen JY, Wen PF, Kong WF, Pan QH, Wan SB, Huang WD.

Center for Viticulture and Enology, 301#, College of Food Science and
Nutritional Engineering, China Agricultural University, Beijing 100083, PR
China.

The phenylpropanoid pathway yields a variety of phenolics that are closely
associated with fruit qualities in addition to structural and defense-related
functions. However, very little has been reported concerning its metabolism in
fruit. This experiment was designed to assess changes of eleven phenolic acids
in grape berry (Vitis vinifera L. cv. Cabernet Sauvignon) and explore both the
activities and amounts of three key enzymes--phenylalanine ammonia-lyase (PAL),
cinnamate-4-hydroxylase (C4H) and 4-coumarate:coenzyme A ligase
(4CL)--catalyzing the biosynthesis of these compounds during berry development.
Finally, the subcellular localizations of the enzymes within berry tissues were
also investigated using immuno-gold electron microscopic technique. The results
indicated that the contents of gallic, protocatechuic, gentisic and caffeic acid
all changed drastically during berry development, while other compounds
containing p-hydroxybenzoic, vanillic, syringic, chlorogenic, p-coumaric,
ferulic and sinapic acid varied only slightly. Activities of PAL, C4H and 4CL
showed similar pattern changes with two accumulated peaks throughout berry
development. In addition, their activities all showed a highly positive
correlation with the total contents of phenolic acids, whereas the
immunoblotting analysis showed that changes in enzyme activities were
independent of the enzyme amounts. Results from the subcellular-localization
study revealed that PAL was mainly present in the cell walls, secondarily
thickened walls, and the parenchyma cells of the berry mesocarp cells, C4H was
found primarily in the chloroplast (plastid) and nucleus and 4CL predominantly
in the secondarily thickened walls and the parenchyma cells of mesocarp vascular
tissue.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16399002 [PubMed - indexed for MEDLINE]

114: J Chromatogr A. 2005 Oct 14;1091(1-2):72-82.

Screening for anthocyanins using high-performance liquid chromatography coupled
to electrospray ionization tandem mass spectrometry with precursor-ion analysis,
product-ion analysis, common-neutral-loss analysis, and selected reaction
monitoring.

Tian Q, Giusti MM, Stoner GD, Schwartz SJ.

Department of Food Science and Technology, The Ohio State University, 2015 Fyffe
Road, Columbus, Ohio 43210, USA.

A systematic method for anthocyanin identification using tandems mass
spectrometry (MS/MS) coupled to high-performance liquid chromatography (HPLC)
with photo-diode array detection (PDA) was developed. Scan for the precursor
ions of commonly found anthocyanidins (cyanidin, delphinidin, malvidin,
pelargonidin, petunidin, and peonidin) using LC/MS/MS on a triple quadrupole
instrument allows for the specific determination of each category of
anthocyanins. Further characterization of each anthocyanin was performed using
MS/MS product-ion analysis, common-neutral-loss analysis, and selected reaction
monitoring (SRM). The method was demonstrated for analysis of anthocyanins in
black raspberries, red raspberries, highbush blueberries, and grapes (Vitis
vinifera). Previous reported anthocyanins in black raspberries and red
raspberries are confirmed and characterized. Common-neutral-loss analysis allows
for the distinction of anthocyanin glucosides or galactoside and arabinosides in
highbush blueberries. Separation and identification of anthocyanin glucosides
and galactosides were achieved by LC/MS/MS using SRM. Anthocyanin isomers such
as cyanidin sophoroside and 3,5-diglucoside were differentiated by their
fragmentation pattern during product-ion analysis. Fifteen anthocyanins (all
possible combinations of five anthocyanidins and three sugars) were
characterized in highbush blueberries. Pelargonidin 3-glucoside and pelargonidin
3,5-diglucoside were detected and characterized for the first time in grapes.
The present approach allows mass spectrometry to be used as a highly selective
detector for rapid identification and characterization of anthocyanins and can
be used as a sensitive procedure for screening anthocyanins in fruits and
vegetables.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16395794 [PubMed - indexed for MEDLINE]

115: J Agric Food Chem. 2006 Jan 11;54(1):65-71.

Modified cyclodextrins are chemically defined glucan inducers of defense
responses in grapevine cell cultures.

Bru R, Selles S, Casado-Vela J, Belchi-Navarro S, Pedreno MA.

Departamento de Agroquimica y Bioquimica, Facultad de Ciencias, Universidad de
Alicante, Apartado 99, E-03080 Alicante, Spain. [email protected]

In grapevine (Vitis vinifera L.), defense responses after microbial infection or
treatment with elicitors involve accumulation of phytoalexins, oxidative burst,
and the synthesis of pathogenesis-related proteins. Oligosaccharide fractions
from fungal or algal cell walls efficiently induce the defense responses, but a
detailed analysis of the elicitor-plant cell surface interaction at the
molecular level is precluded by the lack of chemically pure oligosaccharide
elicitors. A grapevine liquid cell culture system was used to examine the
properties of cyclodextrins (CDs) as inducers of defense responses. This work
shows that the chemically pure heptakis(2,6-di-O-methyl)-betaCD caused a
dramatic extracellular accumulation of the phytoalexin resveratrol and changes
in peroxidase activity and isoenzymatic pattern. Other modified CDs tested on
several grapevine cell lines resulted in different eliciting capacities of CDs
and different sensibilities of the cell lines. The spent medium of elicited
cultures was shown to disturb Botrytis cinerea growth in a plate assay.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16390179 [PubMed - indexed for MEDLINE]

116: J Agric Food Chem. 2006 Jan 11;54(1):29-33.

A novel method for quantification of 2-methyl-3-furanthiol and
2-furanmethanethiol in wines made from Vitis vinifera grape varieties.

Tominaga T, Dubourdieu D.

Faculte d'oenologie, Universite Victor Segalen Bordeaux 2, 351, Cours de la
liberation, 33405 Talence, France. [email protected]

A rapid, easy method has been developed for isolating and quantifying
2-methyl-3-furanthiol (2M3F) in wines. Until now, it was not possible to
quantify this highly odoriferous compound, with a smell reminiscent of cooked
meat, in wine. The original aspect of this method is the specific release of
volatile thiols using a cysteamine solution applied in reverse flow to sample
percolation on the basis of a p-hydroxymercuribenzoate (pHMB)-volatile thiol
conjugate formed by the direct addition of pHMB to 50 mL of wine. Purification
of volatile thiols in wines is much faster and easier than our previous method.
This method may also be used to assay 2-furanmethanethiol in wine. This thiol's
strong aroma of roasted coffee has been shown to contribute to the "roast
coffee" aroma of certain wines. Assaying 2M3F by this method showed that it was
present in the wines analyzed (red and white Bordeaux, Loire Valley Sauvignon
blanc, white Burgundy, and Champagne) at concentrations up to 100 ng/L, i.e.,
significantly above the olfactory perception threshold for this compound in
model dilute alcohol solution.

PMID: 16390173 [PubMed - indexed for MEDLINE]

117: Int J Food Microbiol. 2006 Apr 1;107(3):281-6. Epub 2006 Jan 4.

Antilisterial activity of grape juice and grape extracts derived from Vitis
vinifera variety Ribier.

Rhodes PL, Mitchell JW, Wilson MW, Melton LD.

Food Science Postgraduate Programme, Department of Chemistry, The University of
Auckland, Private Bag 92019, Auckland, New Zealand.

Grape juice and skin and seed extracts of Vitis vinifera var. Ribier black table
grapes were found to be highly inhibitory towards Listeria monocytogenes. This
grape juice was also active against all other Listeria species tested but not
against Bacillus cereus, Salmonella Menston, Escherichia coli, Staphylococcus
aureus or Yersinia enterocolitica. Fractionation of the extracts showed that the
antilisterial activity was strongest in the polymeric phenolic fractions. Two
different types of active compounds were identified: the red-pigmented polymeric
phenolics from juice and skin showed pH-dependent antilisterial activity, while
the unpigmented polymeric phenolics from the seed showed antilisterial activity
which was independent of pH.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16386816 [PubMed - indexed for MEDLINE]

118: Plant Physiol. 2006 Feb;140(2):499-511. Epub 2005 Dec 29.

Characterization of a grapevine R2R3-MYB transcription factor that regulates the
phenylpropanoid pathway.

Deluc L, Barrieu F, Marchive C, Lauvergeat V, Decendit A, Richard T, Carde JP,
Merillon JM, Hamdi S.

Unite Mixte de Recherche 619, Physiologie et Biotechnologie Vegetales,
Universite Bordeaux 1, Universite Bordeaux 2, Institut National de la Recherche
Agonomique, Centre de Recherche de Bordeaux, 33883 Villenave d'Ornon, France.

The ripening of grape (Vitis vinifera) berry is characterized by dramatic
changes in gene expression, enzymatic activities, and metabolism that lead to
the production of compounds essential for berry quality. The phenylpropanoid
metabolic pathway is one of the components involved in these changes. In this
study, we describe the cloning and functional characterization of VvMYB5a, a
cDNA isolated from a grape L. cv Cabernet Sauvignon berry library. VvMYB5a
encodes a protein belonging to a small subfamily of R2R3-MYB transcription
factors. Expression studies in grapevine indicate that the VvMYB5a gene is
mainly expressed during the early steps of berry development in skin, flesh, and
seeds. Overexpression of VvMYB5a in tobacco (Nicotiana tabacum) affects the
expression of structural genes controlling the synthesis of phenylpropanoid and
impacts on the metabolism of anthocyanins, flavonols, tannins, and lignins.
Overexpressing VvMYB5a induces a strong accumulation of several phenolic
compounds, including keracyanin (cyanidin-3-rhamnoglucoside) and
quercetin-3-rhamnoglucoside, which are the main anthocyanin and flavonol
compounds in tobacco. In addition, VvMYB5a overexpression increases the
biosynthesis of condensed tannins and alters lignin metabolism. These findings
suggest that VvMYB5a may be involved in the control of different branches of the
phenylpropanoid pathway in grapevine.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16384897 [PubMed - indexed for MEDLINE]

119: Plant Physiol. 2006 Feb;140(2):537-47. Epub 2005 Dec 29.

The grapevine fleshless berry mutation. A unique genotype to investigate
differences between fleshy and nonfleshy fruit.

Fernandez L, Romieu C, Moing A, Bouquet A, Maucourt M, Thomas MR, Torregrosa L.

Unite Mixte de Recherche Biologie des Especes Perennes Cultivees,
Agro-Montpellier/Institut National de la Recherche Agronomique, 34060
Montpellier cedex 01, France.

In flowering plants, fruit morphogenesis is a distinct process following
fertilization resulting in the formation of a specialized organ associated with
seeds. Despite large variations in types and shapes among species, fleshy fruits
share common characteristics to promote seed dispersal by animals such as organ
growth and metabolite accumulation to attract animal feeding. The molecular
biology of fruit ripening has received considerable attention, but little is
known about the determinism of early fruit morphogenesis and why some fruits are
fleshy while others lack flesh. We have identified in grapevine (Vitis vinifera)
a mutation we have named fleshless berry (flb) that reduces by 20 times the
weight of the pericarp at ripening without any effect on fertility or seed size
and number. The flb mutation strongly impaired division and differentiation of
the most vacuolated cells in the inner mesocarp. The timing of ripening was not
altered by the mutation although the accumulation of malic acid in the green
stage was noticeably reduced while sucrose content (instead of hexoses)
increased during ripening. The mutation segregates as a single dominant locus.
These results indicate that the Flb- mutant is suitable material to advance our
understanding of the genetic and developmental processes involved in the
differentiation of an ovary into a fruit.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16384896 [PubMed - indexed for MEDLINE]

120: Plant Physiol. 2006 Jan;140(1):279-91. Epub 2005 Dec 23.

Identification of the flavonoid hydroxylases from grapevine and their regulation
during fruit development.

Bogs J, Ebadi A, McDavid D, Robinson SP.

Commonwealth Scientific and Industrial Research Organization, Plant Industry,
Horticulture Unit, Glen Osmond, South Australia 5064, Australia.

Flavonoids are important secondary metabolites in many fruits, and their
hydroxylation pattern determines their color, stability, and antioxidant
capacity. Hydroxylation of the B-ring of flavonoids is catalyzed by flavonoid
3'-hydroxylase (F3'H) and flavonoid 3',5'-hydroxylase (F3'5'H), and may also
require cytochrome b5. We report the identification of genes encoding F3'H,
F3'5'H, and a putative cytochrome b5 from grapevine (Vitis vinifera L. cv
Shiraz) and their transcriptional regulation in fruit. Functionality of the
genes VvF3'H and VvF3'5'H1 was demonstrated by ectopic expression in petunia
(Petunia hybrida), which altered flower color and flavonoid composition as
expected. VvF3'H was expressed in grapes before flowering, when 3'-hydroxylated
flavonols are made, and all three genes were expressed after flowering, when
proanthocyanidins (PAs) are synthesized. In berry skin, expression of all three
genes was low at the onset of ripening (veraison) but increased after veraison
concomitant with the accumulation of 3'- and 3',5'-hydroxylated anthocyanins.
VvF3'H and VvCytoB5 were expressed in seeds but not VvF3'5'H1, consistent with
the accumulation of 3'-hydroxylated PAs in this tissue. VvCytoB5 expression was
correlated with expression of both VvF3'H and VvF3'5'H1 in the different grape
tissues. In contrast to red grapes, where VvF3'H, VvF3'5'H1, and VvCytoB5 were
highly expressed during ripening, the expression of VvF3'5'H1 and VvCytoB5 in
white grapes during ripening was extremely low, suggesting a difference in
transcriptional regulation. Our results show that temporal and tissue-specific
expression of VvF3'H, VvF3'5'H1, and VvCytoB5 in grapes is coordinated with the
accumulation of the respective hydroxylated flavonols and PAs, as well as
anthocyanins. Understanding the regulation of flavonoid hydroxylases could be
used to modify flavonoid composition of fruits.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16377741 [PubMed - indexed for MEDLINE]

121: J Agric Food Chem. 2005 Dec 28;53(26):10034-41.

Carotenoid, chlorophyll, and chlorophyll-derived compounds in grapes and port
wines.

Mendes-Pinto MM, Silva Ferreira AC, Caris-Veyrat C, Guedes de Pinho P.

Escola Superior de Biotecnologia, Universidade Catolica Portuguesa, R. Dr.
Antonio Bernardino de Almeida, 4200-072 Porto, Portugal.

Carotenoids and chlorophyll-derived compounds in grapes and Port wines were
investigated by HPLC-DAD and HPLC-DAD-MS (ESP+) analysis. A total of 13
carotenoid and chlorophyll-derived compounds are formally reported in grapes, 3
are identified for the first time, pheophytins a and b and (13Z)-beta-carotene,
and 3 others remain unknown. In Port wines 19 compounds with carotenoid or
chlorophyll-like structures are present, 8 still unidentified. The young wines
showed higher total carotenoid content and chlorophyll-like compounds compared
to aged Ports, with lutein and beta-carotene as major carotenoids. Among samples
analyzed of monovarietal Vitis vinifera L. cultivar wines produced with the five
most important Douro varieties, Tinta Roriz contained the highest levels of
carotenoids and Touriga Franca the lowest. The forced-aging study indicated that
lutein was more sensitive to temperature than beta-carotene. Additionally, aged
wines showed higher ratios of beta-carotene/lutein concentrations compared to
new Ports. Rates of degradation of chlorophyll derivative compounds were higher
than those for carotene and lutein.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16366691 [PubMed - indexed for MEDLINE]

122: J Chromatogr A. 2006 Apr 21;1112(1-2):165-70. Epub 2005 Dec 20.

Densitometric thin-layer chromatographic determination of aescin in a herbal
medicinal product containing Aesculus and Vitis dry extracts.

Apers S, Naessens T, Pieters L, Vlietinck A.

Department of Pharmaceutical Sciences, University of Antwerp, Universiteitsplein
1, B-2610 Wilrijk, Antwerp, Belgium. [email protected]

A thin-layer chromatographic (TLC) method is developed to analyze the total
saponin content, also referred to as the aescin content, in a herbal medicinal
product (HMP) containing two dry extracts in capsules. The capsules contain 250
mg of Aesculus hippocastanum dry extract, 120 mg of Vitis vinifera dry extract
and 50mg of excipients. After a purification step using C(18) solid phase
extraction (SPE) cartridges, the samples are analyzed on a silica-gel HPTLC
plate with the upper layer of a mixture of acetic acid/water/butanol (10/40/50
v/v/v) as the mobile phase. Spots are visualized by spraying with anisaldehyde
reagent and heating the plate for 5-10 min (100-105 degrees C) and measured at a
wavelength of 535 nm. This method, applicable for the quality control and
stability investigation of both the Aesculus dry extract and HMP capsules
thereof containing Vitis dry extract in combination with the Aesculus dry
extract, is validated according to the International Conference on Harmonization
(ICH) guidelines. The proposed assay method is specific for aescin in the
presence of Vitis dry extract and formulation excipients. Analysis of stressed
samples in forced degradation tests proves the method to be applicable for
stability evaluation. The standard aescin curve is linear (r > 0.99) over a
concentration range of 0.16-0.80 microg/spot. Recovery from the HMP capsules is
statistically equal to 100%. The precision of the method with respect to time
and concentration is acceptable, with relative standard deviation (RSD) values
of 1.28 and 1.49%, respectively.

Publication Types:
Research Support, Non-U.S. Gov't
Validation Studies

PMID: 16364347 [PubMed - indexed for MEDLINE]

123: Plant Physiol. 2006 Jan;140(1):150-8. Epub 2005 Dec 16.

Grapes on steroids. Brassinosteroids are involved in grape berry ripening.

Symons GM, Davies C, Shavrukov Y, Dry IB, Reid JB, Thomas MR.

School of Plant Science, University of Tasmania, Hobart, Tasmania 7005,
Australia.

Fruit ripening is a unique plant developmental process with direct implications
for our food supply, nutrition, and health. In contrast to climacteric fruit,
where ethylene is pivotal, the hormonal control of ripening in nonclimacteric
fruit, such as grape (Vitis vinifera), is poorly understood. Brassinosteroids
(BRs) are steroidal hormones, essential for normal plant growth and development
but not previously implicated in the ripening of nonclimacteric fruit. Here we
show that increases in endogenous BR levels, but not indole-3-acetic acid (IAA)
or GA levels, are associated with ripening in grapes. Putative grape homologs of
genes encoding BR biosynthesis enzymes (BRASSINOSTEROID-6-OXIDASE and DWARF1)
and the BR receptor (BRASSINOSTEROID INSENSITIVE 1) were isolated, and the
function of the grape BRASSINOSTEROID-6-OXIDASE gene was confirmed by transgenic
complementation of the tomato (Lycopersicon esculentum) extreme dwarf (dx/dx)
mutant. Expression analysis of these genes during berry development revealed
transcript accumulation patterns that were consistent with a dramatic increase
in endogenous BR levels observed at the onset of fruit ripening. Furthermore, we
show that application of BRs to grape berries significantly promoted ripening,
while brassinazole, an inhibitor of BR biosynthesis, significantly delayed fruit
ripening. These results provide evidence that changes in endogenous BR levels
influence this key developmental process. This may provide a significant insight
into the mechanism controlling ripening in grapes, which has direct implications
for the logistics of grape production and down-stream processing.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16361521 [PubMed - indexed for MEDLINE]

124: Appl Environ Microbiol. 2005 Dec;71(12):8292-300.

Bleeding sap and old wood are the two main sources of contamination of merging
organs of vine plants by Xylophilus ampelinus, the causal agent of bacterial
necrosis.

Grall S, Roulland C, Guillaumes J, Manceau C.

UMR PaVe, Centre INRA d'Angers, 42 rue Georges Morel, BP 60071, F-49071
Beaucouze Cedex, France.

The spatial distribution of vine plants contaminated by Xylophilus ampelinus,
the agent responsible for bacterial necrosis, was studied over a 5-year period
within two vineyards in the Cognac area. Both vineyards were planted with Vitis
vinifera cv. Ugni blanc but were different in age and agronomic location. The
emission of X. ampelinus in contaminated bleeding sap was observed during vine
sprouting. Contaminated bleeding sap is an important source of inoculum for
external contamination due to the high susceptibility of young merging shoots to
the pathogen. X. ampelinus emission by bleeding sap was not affected by the age
of the plants or the location of the vineyards. However, its emission was
irregular with time, and it varied between two fruit canes from individual
plants and between plants as well as between years. Moreover, the two vineyards
appeared to be entirely contaminated. Consequently, the behavior of the pathogen
is not predictable. The distribution of the pathogen inside vine plant organs
was analyzed through the four growing seasons. The old wood was contaminated
throughout the year and constituted a stock inoculum for endophytic
contamination of crude sap during the winter and the spring. Despite the fact
that most of the young green shoots were contaminated in May, X.ampelinus was
not found in green shoots in June and September, refuting the hypothesis of an
epiphytic life of the pathogen under natural conditions. Although all plants
were entirely contaminated in both vineyards, symptoms were rare and were
observed on different plants each year.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16332815 [PubMed - indexed for MEDLINE]

125: J Sep Sci. 2005 Nov;28(16):2050-6.

Supercritical fluid extraction and HPLC determination of relevant polyphenolic
compounds in grape skin.

Chafer A, Pascual-Marti MC, Salvador A, Berna A.

Departamento de Ingenieria Quimica, Universitat de Valencia, Burjassot,
Valencia, Spain.

The polyphenols determined are: (+)-catechin, (-)-epicatechin, rutin, quercetin
and trans-resveratrol. Suitable conditions of supercritical fluid extraction
were established using ethanol as a modifier of the polarity solvent
(supercritical carbon dioxide). Final extraction conditions were: 20% v/v
ethanol, 60degreesC, 250 bars and flow rate 2 mL/min. Static step time and
dynamic step time were established using a selected grape skin sample. The
extract was collected in water; the more polar polyphenols ((+)-catechin and
(-)-epicatechin) remain in solution but rutin, quercetin and trans-resveratrol
precipitate in this medium, thereby the solution of the extracted polyphenols
was filtered. (+)-Catechin and (-)-epicatechin were determined in the liquid
fraction, while the solid fraction, containing rutin, quercetin and
trans-resveratrol, was solved with ethanol/H20 (40:60). HPLC determination was
carried out at C18 stationary phase, with ethanol/water/acetic acid as mobile
phases and UV-visible diode array detection. Due to the significant differences
between the polarity of the polyphenols, two different mobile phases were used.
An ethanol/water/acetic acid (5:93:2) mobile phase was used to determine
(+)-catechin (280 nm) and (-)-epicatechin (280 nm). On the other hand, rutin
(254 nm), quercetin (254 nm) and trans-resveratrol (306 nm) were resolved using
ethanol/water/acetic acid (40:58:2) as mobile phase. Instrumental parameters
were optimised and analytical parameters obtained. The analytical method was
validated and applied to five different varieties of Vitis vinifera from the
geographical area of Valencia.

PMID: 16318198 [PubMed - in process]

126: Phytother Res. 2005 Nov;19(11):924-7.

Studies on antidiarrhoeal activity of an extract of wine from Jacquez grapes in
mice.

Vitali F, Bonina FP, Saija A, Tomaino A, Fonte G, Pennisi C, Tita B.

Department Farmaco-Biologico, University of Messina, Villaggio SS Annunziata,
Italy.

The present study was designed to verify the antidiarrhoeal effects of a
lyophilized extract of wine from Jacquez grapes (Ord. Rhamnales; Fam. Vitaceae;
Sp. Vitis aestivalis M.-cinerea E. x Vitis vinifera L.), studying its influence
on castor oil-induced diarrhoea and enteropooling, and on gastrointestinal
transit (measured by a charcoal marker) in mice. The pre-treatment of the
animals with the JWE (Jacquez wine extract) produced a significant inhibition
against castor oil induced-diarrhoea and intestinal fluid accumulation;
furthermore the extract significantly decreased the propulsive movement of the
charcoal meal. These findings suggest a potential beneficial use of the JWE in
the treatment of diarrhoeal diseases.

PMID: 16317645 [PubMed - indexed for MEDLINE]

127: Biochimie. 2006 May;88(5):431-5. Epub 2005 Oct 27.

Editing of the grapevine mitochondrial cytochrome b mRNA and molecular modeling
of the protein.

Islas-Osuna MA, Silva-Moreno B, Caceres-Carrizosa N, Garcia-Robles JM,
Sotelo-Mundo RR, Yepiz-Plascencia GM.

Direccion de Tecnologia de Alimentos de Origen Vegetal, Centro de Investigacion
en Alimentacion y Desarrollo, A.C. Apdo. Postal 1735, Hermosillo, Sonora 83000,
Mexico. [email protected]

Cytochrome b (COB), the central catalytic subunit of ubiquinol cytochrome c
reductase, is a component of the transmembrane electron transfer chain that
generates proton motive force. Some plant COB mRNAs are processed by RNA
editing, which changes the gene coding sequence. This report presents the
sequences of the grapevine (Vitis vinifera L.) mitochondrial gene for
apocytochrome b (cob), the edited mRNA and the deduced protein. Grapevine COB is
393 amino acids long and is 98% identical to homologs in rapeseed, Arabidopsis
thaliana and Oenothera sp. Twenty-one C-U editing sites were identified in the
grapevine cob mRNA, resulting in 20 amino acid changes. These changes increase
the overall hydrophobicity of the protein and result in a more conserved
protein. Molecular modeling of grapevine COB shows that residues changed by RNA
editing fit the secondary structure characteristic of an integral membrane
protein. This is the first complete mitochondrial gene reported for grapevine.
Novel RNA editing sites were identified in grapevine cob, which have not been
previously reported for other plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16310923 [PubMed - indexed for MEDLINE]

128: Plant J. 2005 Nov;44(4):606-19.

The biosynthesis and regulation of biosynthesis of Concord grape fruit esters,
including 'foxy' methylanthranilate.

Wang J, Luca VD.

Department of Biological Sciences, 500 Glenridge Avenue, Brock University, St
Catharines, ON, Canada L2S 3A1.

The biosynthesis of methyl anthranilate, the volatile compound responsible for
the distinctive 'foxy' aroma and flavor of the Washington Concord grape (Vitis
labrusca), involves an alcohol acyltransferase that catalyzes the formation of
methyl anthranilate from anthraniloyl-coenzyme A (CoA) and methanol. Although
methanol is a poor substrate in comparison with the co-substrate, high levels of
this acyltransferase (0.5% of the total protein) combined with relatively high
levels of this alcohol make this reaction possible in grapes. This 449 amino
acid protein belongs to the BAHD family of acyltransferases, having 58% identity
with the benzoyl CoA:benzyl alcohol benzoyl transferase from Clarkia. Both
native and recombinant enzymes can use a broad range of acyl-CoAs and alcohols
as substrates. The ability of Concord grape alcohol acyltransferase to accept a
range of different CoA esters and alcohols suggests this to be a versatile
ester-forming enzyme, similar to those of other fruits that than can produce a
range of fruit esters based on the supply of appropriate substrates. Expression
is coordinately regulated, with transcript, protein and enzyme activities
coinciding with the accumulation of methyl anthranilate that occurs after the
initiation of berry ripening. The majority of acyltransferase protein in grape
tissues is localized to the outer fruit mesocarp, a result consistent with the
fact that methyl anthranilate is released to the external environment throughout
the ripening process. Wine grapes (Vitis vinifera) that accumulate neither
anthranilate nor methyl anthranilate do not express this enzyme activity nor do
they accumulate this protein.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16262710 [PubMed - indexed for MEDLINE]

129: J Nutr Sci Vitaminol (Tokyo). 2005 Aug;51(4):283-6.

Attenuation of renal ischemia-reperfusion injury by proanthocyanidin-rich
extract from grape seeds.

Nakagawa T, Yokozawa T, Satoh A, Kim HY.

Institute of Natural Medicine, Toyama Medical and Pharmaceutical University,
Toyama, Japan.

The effects of proanthocyanidin-rich extract in rats subjected to renal
ischemia-reperfusion were examined. Proanthocyanidin-rich extract, which is
prepared from grape seeds (Vitis vinifera L.), was given orally at doses of 5
and 10 mg/kg body weight/d for 20 consecutive days prior to
ischemia-reperfusion. Administration of proanthocyanidin-rich extract attenuated
renal dysfunction, as indicated by serum urea nitrogen and creatinine levels.
Additionally, in the ischemic-reperfused kidneys, increased levels of
thiobarbituric acid (TBA)-reactive substance and alterations of antioxidant
enzyme activities such as superoxide dismutase, catalase and glutathione
peroxidase (GSH-Px) were observed. Proanthocyanidin-rich extract-treated groups
showed significantly reduced renal TBA-reactive substance levels and enhanced
catalase and GSH-Px activities. These results suggest that proanthocyanidin-rich
extract has protective effects against ischemia-reperfusion-induced renal damage
associated with oxidative stress.

PMID: 16262003 [PubMed - indexed for MEDLINE]

130: Photosynth Res. 2003;75(1):29-39.

Action of UV and visible radiation on chlorophyll fluorescence from dark-adapted
grape leaves (Vitis vinifera L.).

Pfundel EE.

Lehrstuhl fur Botanik II, Universitat Wurzburg, Julius-von-Sachs-Platz 3,
D-97082, Wurzburg, Germany, [email protected]

Grapevine plants (Vitis vinifera L. cv. Silvaner) were cultivated under shaded
conditions in the absence of UV radiation in a greenhouse, and subsequently
placed outdoors under filters transmitting natural radiation, or screening out
the UV-B (280 to 315 nm), or screening out the UV-A (315 to 400 nm) and the UV-B
spectral range. All conditions decreased maximum chlorophyll fluorescence (F(M))
and increased minimum chlorophyll fluorescence (F(0)) from dark-adapted leaves;
however, with increasing UV, F(M) quenching was stimulated but increases in F(0)
were reduced. The F(V)/F(M) ratio (where F(V)=F(M)-F(0)) was clearly reduced by
visible radiation (VIS): UV-B caused a moderate extra-reduction in F(V)/F(M).
Exposure of leaves (V. vinifera L. cv. Bacchus) to UV or VIS lamps quenched the
F(M) to similar extents; further, UV-B doses comparable to the field, quenched
F(0). A model was developed to describe how natural radiation intensities affect
PS II and thereby change leaf fluorescence. Fitting theory to experiment was
successful when the same F(M) yield for UV- and VIS-inactivated PS II was
assumed, and for lower F(0) yields of UV- than for VIS-inactivated PS II. It is
deduced, that natural UV can produce inactivated PS II exhibiting relatively
high F(V)/F(M). The presence of UV-inactivated PS II is difficult to detect by
measuring F(V)/F(M) in leaves. Hence, relative concentrations of intact PS II
during outdoor exposure were derived from F(M). These concentrations, but not
F(V)/F(M), correlated reasonably well with CO(2) gas exchange measurements.
Consequently, PS II inhibition by natural UV could be a main factor for UV
inhibition of photosynthesis.

PMID: 16245091 [PubMed - as supplied by publisher]

131: Plant Cell Rep. 2005 Dec;24(11):655-62. Epub 2005 Oct 21.

Molecular characterization of grapevine plants transformed with GFLV resistance
genes: I.

Gambino G, Gribaudo I, Leopold S, Schartl A, Laimer M.

Istituto Virologia Vegetale C.N.R.-Unita staccata Viticoltura-Grugliasco, Via
Leonardo da Vinci 44, I-10095 Grugliasco (Turin), Italy.

The Grapevine FanLeaf Virus-Coat Protein (GFLV CP) gene was inserted through
Agrobacterium-mediated transformation in Vitis vinifera "Nebbiolo", "Lumassina"
and "Blaufrankisch". Two plasmids were used: pGA-CP+ (full-length GFLV CP gene
with an introduced start codon) and pGA-AS (same gene in antisense orientation).
Forty-three transgenic lines were regenerated. As several lines in Southern
blots share same hybridization patterns, eight independent line groups resulted
for "Nebbiolo", one for "Lumassina", and two for "Blaufrankisch". Inserted T-DNA
copies ranged from one to three; one line probably contains an incomplete copy
of T-DNA. Except for one "Nebbiolo" line, no evidence for methylation of the
transgene at cytosine residues was found by Southern analyses. Specific mRNA was
present at variable expression levels; some lines accumulated the coat protein
while in others the protein was not detectable by ELISA.

PMID: 16240119 [PubMed - indexed for MEDLINE]

132: J Biosci Bioeng. 2003;95(6):645-7.

Characterization of a class III chitinase from Vitis vinifera cv. Koshu.

Ano A, Takayanagi T, Uchibori T, Okuda T, Yokotsuka K.

The Institute of Enology and Viticulture, Yamanashi University, 1-13-1 Kitashin,
Kofu 400-0005, Japan.

A chitinase gene (Chi3K) was cloned from the genomic DNA of Vitis vinifera cv.
Koshu. The structural gene comprised 891 by without introns and encoded 297
amino acids. The Chi3K product showed high similarity to the class III chitinase
of V. vinifera cv. Pinot noir. Chi3K was expressed using a bacterial expression
vector for purification and enzymatic characterization of its gene product. The
recombinant chitinase exhibited hydrolytic activity toward glycol chitin and its
optimum pH was 4.0. It also inhibited the growth of Botrytis cinerea, which
causes grey mold disease in grapes.

PMID: 16233474 [PubMed]

133: J Biosci Bioeng. 2002;94(2):135-9.

Enhanced anthocyanin production from grape callus in an air-lift type bioreactor
using a viscous additive-supplemented medium.

Honda H, Hiraoka K, Nagamori E, Omote M, Kato Y, Hiraoka S, Kobayashi T.

Department of Biotechnology, School of Engineering, Nagoya University,
Chikusa-ku, Nagoya 464-8603, Japan. [email protected]

An N-medium containing carboxymethyl cellulose (CMC) was applied to an air-lift
type bioreactor culture of grape (Vitis vinifera cv. Bailey alicant A.) callus,
and anthocyanin production was investigated. Grape callus grew well at an air
flow rate of 80 ml/min and anthocyanin production was significantly increased in
the N-medium, reaching 17 mg/l after 7 d of culture. The anthocyanin content of
the N-medium was about two times higher than that of the conventional medium
without CMC. The effect of air flow rate was also investigated within the range
from 40 to 160 ml/min. A twofold increase in anthocyanin content was obtained at
all the air flow rates tested in the N-medium. The distribution of grape callus
size obtained after 7 d of the bioreactor culture was investigated. The average
callus size was 490 mum which was 1.6 times larger than that obtained in the
conventional medium. It was found that large calli with a relatively high
anthocyanin pigment content were formed in the bioreactor culture using the
N-medium. The fluid dynamics in the bioreactor was also investigated at three
points (top, middle and bottom) in the bioreactor by laser doppler velocimetry.
The average axial velocity of the circulated medium was 0.4 times lower than
that of the conventional medium while their average radial velocities were
almost the same (zero). The standard deviation of radial velocity fluctuation in
the N-medium was also 0.4 times less than that in the conventional medium. These
results suggest that turbulent flow occurred in the bioreactor culture using the
conventional medium and the degree of turbulent flow decreased significantly
when 0.8% CMC was added to the medium to prepare the N-medium. A change of the
flow pattern is considered to be the cause of the decrease in hydrodynamic
stress, resulting in enhanced pigment production due to the enlargement of the
callus.

PMID: 16233283 [PubMed]

134: Plant Physiol. 2005 Oct;139(2):574-97.

Characterizing the grape transcriptome. Analysis of expressed sequence tags from
multiple Vitis species and development of a compendium of gene expression during
berry development.

da Silva FG, Iandolino A, Al-Kayal F, Bohlmann MC, Cushman MA, Lim H, Ergul A,
Figueroa R, Kabuloglu EK, Osborne C, Rowe J, Tattersall E, Leslie A, Xu J, Baek
J, Cramer GR, Cushman JC, Cook DR.

Department of Plant Pathology, University of California, Davis, 95616, USA.

We report the analysis and annotation of 146,075 expressed sequence tags from
Vitis species. The majority of these sequences were derived from different
cultivars of Vitis vinifera, comprising an estimated 25,746 unique contig and
singleton sequences that survey transcription in various tissues and
developmental stages and during biotic and abiotic stress. Putatively homologous
proteins were identified for over 17,752 of the transcripts, with 1,962
transcripts further subdivided into one or more Gene Ontology categories. A
simple structured vocabulary, with modules for plant genotype, plant
development, and stress, was developed to describe the relationship between
individual expressed sequence tags and cDNA libraries; the resulting vocabulary
provides query terms to facilitate data mining within the context of a
relational database. As a measure of the extent to which characterized metabolic
pathways were encompassed by the data set, we searched for homologs of the
enzymes leading from glycolysis, through the oxidative/nonoxidative pentose
phosphate pathway, and into the general phenylpropanoid pathway. Homologs were
identified for 65 of these 77 enzymes, with 86% of enzymatic steps represented
by paralogous genes. Differentially expressed transcripts were identified by
means of a stringent believability index cutoff of > or =98.4%. Correlation
analysis and two-dimensional hierarchical clustering grouped these transcripts
according to similarity of expression. In the broadest analysis, 665
differentially expressed transcripts were identified across 29 cDNA libraries,
representing a range of developmental and stress conditions. The groupings
revealed expected associations between plant developmental stages and tissue
types, with the notable exception of abiotic stress treatments. A more focused
analysis of flower and berry development identified 87 differentially expressed
transcripts and provides the basis for a compendium that relates gene expression
and annotation to previously characterized aspects of berry development and
physiology. Comparison with published results for select genes, as well as
correlation analysis between independent data sets, suggests that the inferred
in silico patterns of expression are likely to be an accurate representation of
transcript abundance for the conditions surveyed. Thus, the combined data set
reveals the in silico expression patterns for hundreds of genes in V. vinifera,
the majority of which have not been previously studied within this species.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 16219919 [PubMed - indexed for MEDLINE]

135: J Exp Bot. 2005 Nov;56(421):2783-95. Epub 2005 Oct 10.

Proteomic analysis of grapevine (Vitis vinifera L.) tissues subjected to
herbicide stress.

Castro AJ, Carapito C, Zorn N, Magne C, Leize E, Van Dorsselaer A, Clement C.

Laboratoire de Stress, Defenses et Reproduction des Plantes, URVVC UPRES EA
2069, Universite de Reims Champagne Ardenne, UFR Sciences, BP 1039 Moulin de la
Housse, F-51687 Reims cedex 2, France.

Two-dimensional gel electrophoresis coupled to mass spectrometry analysis was
used to examine for the first time the effect of a herbicide (flumioxazin) on a
crop species (Vitis vinifera L.) at the proteome level. Examination of 2-D maps
derived from chemically stressed tissues revealed the presence of 33 spots
displaying a differential expression pattern. The presence of stress responsive
proteins in the different plant organs analysed suggests that flumioxazin could
act systemically. Among the responsive proteins, some photosynthesis-related
proteins, including several fragments of the enzyme Rubisco, were identified.
This effect suggests that photosynthesis could be impaired by the herbicide. The
induction of several enzymatic antioxidant systems was also observed, probably
as a result of an oxidative stress. Moreover, the photorespiration pathway was
stimulated, as suggested by the induction of some key enzymes involved in this
process. Changes in carbon metabolism-associated proteins presumably reflect
altered patterns of carbon flux in response to impaired photosynthesis and an
increased need for osmotic adjustment in affected tissues. Finally, plant
defences were stimulated as revealed by the induction of a set of proteins
belonging to the pathogenesis-related 10 class, suggesting that they could play
an essential role in cell defence mechanisms against flumioxazin.

PMID: 16216849 [PubMed - indexed for MEDLINE]

136: J Exp Bot. 2005 Nov;56(421):2949-57. Epub 2005 Oct 5.

Functional xylem in the post-veraison grape berry.

Bondada BR, Matthews MA, Shackel KA.

University of California, Department of Viticulture and Enology, One Shields
Avenue, Davis, CA 95616, USA.

A number of studies have shown a transition from a primarily xylem to a
primarily phloem flow of water as fleshy fruits develop, and the current
hypothesis to explain this transition, particularly in grape (Vitis vinifera L.)
berries, is that the vascular tissue (tracheids) become non-functional as a
result of post-veraison berry growth. In most studies, pedicels have been dipped
in a vial containing an apoplastic dye, which was taken up into the entire
peripheral and axial xylem vasculature of pre-veraison, but not post-veraison
berries. The pressure plate/pressure membrane apparatus that is commonly used to
study soil moisture characteristics was adapted and the pre- to post-veraison
change in xylem functionality in grape berries was re-evaluated by establishing
a hydrostatic (tension) gradient between the pedicel and a cut surface at the
stylar end of the berry. Under the influence of this applied hydrostatic
gradient, movement of the apoplastic tracer dye, basic fuchsin, was found in the
pedicel and throughout the axial and peripheral xylem of the berry mesocarp. A
similar movement of dye could be obtained by simply adjoining the stylar cut
surface to a dry, hydrophilic wicking material. Since both pre- and
post-veraison berries hydrate when the pedicel is dipped in water, it is
hypothesized that the absence of dye movement into the vasculature of
post-veraison berries indicates not a loss of xylem function, but rather the
loss of an appropriate driving force (hydrostatic gradient) in the berry
apoplast. Based on this hypothesis, and the substantial decrease in xylem flows
that occur in intact grape berries at veraison, it is suggested that there may
be significant changes in the pattern of solute partitioning between the fruit
symplast and apoplast at veraison. It is further suggested that diurnal patterns
in symplast/apoplast solute partitioning in grapes and other fleshy fruit, may
explain the observed minimal xylem contribution to the water budgets of these
fruits.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16207748 [PubMed - indexed for MEDLINE]

137: Planta Med. 2005 Sep;71(9):888-90.

New stilbenoid glucosides isolated from Vitis vinifera cell suspension cultures
(cv. Cabernet Sauvignon).

Larronde F, Richard T, Delaunay JC, Decendit A, Monti JP, Krisa S, Merillon JM.

Groupe d'Etude des Substances Vegetales a Activite Biologique, EA 3675,
Universite de Bordeaux 2, Bordeaux, France.

Three new monomeric stilbenoid glucosides, (Z)- and (E)-resveratrol 3,5-
O-beta-diglucosides (1 and 2, respectively) and (Z)-resveratrol 3,5,4'-
O-beta-triglucoside (3), were isolated from an extract of Vitis vinifera cell
cultures (Cabernet Sauvignon) together with the known (E)- and (Z)-piceids and
(E)- and (Z)-resveratrol 3,4'- O-beta-diglucosides that have already been
identified in a Gamay cell culture extract. The structure determinations were
based on spectroscopic data analysis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16206048 [PubMed - indexed for MEDLINE]

138: BMC Plant Biol. 2005 Sep 29;5:20.

Genetic chimerism of Vitis vinifera cv. Chardonnay 96 is maintained through
organogenesis but not somatic embryogenesis.

Bertsch C, Kieffer F, Maillot P, Farine S, Butterlin G, Merdinoglu D, Walter B.

Laboratoire Vigne Biotechnologies et Environnement, Universite de Haute-Alsace,
33 Rue de Herrlisheim, 68000 Colmar, France. [email protected]

BACKGROUND: Grapevine can be a periclinal chimera plant which is composed at
least of two distinct cell layers (L1, L2). When the cell layers of this plant
are separated by passage through somatic embryogenesis, regenerated plants could
show distinct DNA profiles and a novel phenotype which proved different from
that of the parent plant. RESULTS: Genetically Chardonnay clone 96 is a
periclinal chimera plant in which is L1 and L2 cell layers are distinct. Plants
obtained via organogenesis through meristematic bulks are shown to be composed
of both cell layers. However, plants regenerated through somatic embryogenesis
starting from anthers or nodal explants are composed only of L1 cells. These
somaclones do not show phenotypic differences to the parental clone up to three
years after regeneration. Interestingly, the only somaclone showing an atypical
phenotype (asymmetric leave) shows a genotypic modification. CONCLUSION: These
results suggest that the phenotype of Chardonnay 96 does not result from an
interaction between the two distinct cell layers L1 and L2. If phenotype
conformity is further confirmed, somatic embryogenesis will result in
true-to-type somaclones of Chardonnay 96 and would be well suitable for gene
transfer.

PMID: 16194273 [PubMed - indexed for MEDLINE]

139: J Plant Physiol. 2005 Sep;162(9):985-1002.

Expression of transgenic stilbene synthases in wheat causes the accumulation of
unknown stilbene derivatives with antifungal activity.

Serazetdinova L, Oldach KH, Lorz H.

Institute of General Botany and Botanical Garden, AMP II, University of Hamburg,
Ohnhorststrasse 18, D-22609 Hamburg, Germany.
[email protected]

The expression of foreign phytoalexins in a new host is thought to increase
fungal resistance, since host-specific pathogens have not experienced selection
for detoxifying or metabolising the novel antifungal compounds. Two resveratrol
synthase genes vst1 and vst2 from grapevine (Vitis vinifera L.) and the
pinosylvin synthase gene pss from pine (Pinus sylvestris L.) were stably
transformed into bread wheat. The expression of the target genes is regulated by
stress-inducible grapevine promoters. The vst1 and vst2 promoters were
functional in wheat and retained their expression profiles described for
grapevine. ALL vst and pss transgenic lines accumulated stilbene derivatives
upon induction by UV light. The detected stilbenes showed a remarkable
similarity to resveratrol and pinosylvin, however were found to be more
hydrophilic than resveratrol and pinosylvin. Upon inoculation with the
biotrophic pathogen Puccinia recondita f.sp. tritici several vst expressing
wheat lines showed a significant reduction of disease symptoms (19 +/- 9% to 27
+/- 8%) compared to wild-type plants. The reduction of disease symptoms was even
more obvious after inoculation with the facultative biotrophic pathogen Septoria
nodorum Berk. and ranged from 42 +/- 13% to 71 +/- 4%. None of the four tested
pss expressing lines showed a reduction in disease incidence.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16173460 [PubMed - indexed for MEDLINE]

140: Plant Physiol. 2005 Oct;139(2):652-63. Epub 2005 Sep 16.

Proanthocyanidin synthesis and expression of genes encoding leucoanthocyanidin
reductase and anthocyanidin reductase in developing grape berries and grapevine
leaves.

Bogs J, Downey MO, Harvey JS, Ashton AR, Tanner GJ, Robinson SP.

Commonwealth Scientific and Industrial Research Organization, Plant Industry,
Horticulture Unit, Glen Osmond, South Australia.

Proanthocyanidins (PAs), also called condensed tannins, can protect plants
against herbivores and are important quality components of many fruits. Two
enzymes, leucoanthocyanidin reductase (LAR) and anthocyanidin reductase (ANR),
can produce the flavan-3-ol monomers required for formation of PA polymers. We
isolated and functionally characterized genes encoding both enzymes from
grapevine (Vitis vinifera L. cv Shiraz). ANR was encoded by a single gene, but
we found two highly related genes encoding LAR. We measured PA content and
expression of genes encoding ANR, LAR, and leucoanthocyanidin dioxygenase in
grape berries during development and in grapevine leaves, which accumulated PA
throughout leaf expansion. Grape flowers had high levels of PA, and accumulation
continued in skin and seeds from fruit set until the onset of ripening. VvANR
was expressed throughout early flower and berry development, with expression
increasing after fertilization. It was expressed in berry skin and seeds until
the onset of ripening, and in expanding leaves. The genes encoding LAR were
expressed in developing fruit, particularly in seeds, but had low expression in
leaves. The two LAR genes had different patterns of expression in skin and
seeds. During grape ripening, PA levels decreased in both skin and seeds, and
expression of genes encoding ANR and LAR were no longer detected. The results
indicate that PA accumulation occurs early in grape development and is completed
when ripening starts. Both ANR and LAR contribute to PA synthesis in fruit, and
the tissue and temporal-specific regulation of the genes encoding ANR and LAR
determines PA accumulation and composition during grape berry development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16169968 [PubMed - indexed for MEDLINE]

141: Theor Appl Genet. 2005 Nov;111(8):1553-62. Epub 2005 Nov 10.

Resveratrol glucoside (Piceid) synthesis in seeds of transgenic oilseed rape
(Brassica napus L.).

Husken A, Baumert A, Milkowski C, Becker HC, Strack D, Mollers C.

Institute of Agronomy and Plant Breeding, Georg-August-University,
Von-Siebold-Str. 8, 37075 Gottingen, Germany.

Resveratrol is a phytoalexin produced in various plants like wine, peanut or
pine in response to fungal infection or UV irradiation, but it is absent in
members of the Brassicaceae. Moreover, resveratrol and its glucoside (piceid)
are considered to have beneficial effects on human health, known to reduce heart
disease, arteriosclerosis and cancer mortality. Therefore, the introduction of
the gene encoding stilbene synthase for resveratrol production in rapeseed is a
tempting approach to improve the quality of rapeseed products. The stilbene
synthase gene isolated from grapevine (Vitis vinifera L.) was cloned under
control of the seed-specific napin promotor and introduced into rapeseed
(Brassica napus L.) by Agrobacterium-mediated co-transformation together with a
ds-RNA-interference construct deduced from the sequence of the key enzyme for
sinapate ester biosynthesis, UDP-glucose:sinapate glucosyltransferase (BnSGT1),
assuming that the suppression of the sinapate ester biosynthesis may increase
the resveratrol production in seeds through the increased availability of the
precursor 4-coumarate. Resveratrol glucoside (piceid) was produced at levels up
to 361 microg/g in the seeds of the primary transformants. This value exceeded
by far piceid amounts reported from B. napus expressing VST1 in the wild type
sinapine background. There was no significant difference in other important
agronomic traits, like oil, protein, fatty acid and glucosinolate content in
comparison to the control plants. In the third seed generation, up to 616
microg/g piceid was found in the seeds of a homozygous T3-plant with a single
transgene copy integrated. The sinapate ester content in this homozygous
T3-plant was reduced from 7.43 to 2.40 mg/g. These results demonstrate how the
creation of a novel metabolic sink could divert the synthesis towards the
production of piceid rather than sinapate ester, thereby increasing the value of
oilseed products.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 16160820 [PubMed - indexed for MEDLINE]

142: Photosynth Res. 2004;82(2):141-50.

Tissue specific protochlorophyll(ide) forms in dark-forced shoots of grapevine
(Vitis viniferaL.).

Boddi B, Boka K, Sundqvist C.

Department of Plant Anatomy, Eotvos University, Pazmany P. s. 1/C, Hungary.

Cuttings of grapevine (Vitis vinifera L. cv. Chardonnay) were dark-forced at
least three weeks. Pigment contents, 77 K fluorescence emission, excitation
spectra of the leaves, petioles, stems, transmission electron micrographs of the
etioplasts from leaves, the chlorenchyma tissues of the stems were analysed. The
dark-grown leaves, stems contained 8 to 10, 3 to 5 mug/g fresh weight
protochlorophyllide, its esters, respectively. HPLC analysis showed that the
molar ratio of the unesterified, esterified pigments was 7:3 in the shoot
developed in darkness. The dark-forced leaves contained carotenoids identified
as: neoxanthin, violaxanthin, antheraxanthin, lutein, beta-carotene. Detailed
analyses of the fluorescence spectra proved that all tissues of the dark-forced
shoots had protochlorophyllide or protochlorophyll forms with emission maxima at
628, 636, 644, 655, 669 nm. The 628, 636 nm emitting forms were present in all
parts of the dark-forced shoot, but dominated in the stems, which may indicate
an organ specificity of the etioplast development. Variations in the
distribution of the pigment forms were even found in the different tissues of
the stem. The subepidermal layers were more abundant in the 655 nm form than the
parenchyma cells of the inner part of the cortex, the pith. In the latter cells,
the plastid differentiation stopped in intermediary stages between proplastids,
etioplasts. The plastids in the subepidermal layers had developed prolamellar
body structures, which were similar to those of etiolated leaves. The results
highlight the importance of organ-, tissue specificity of plastid
differentiation for chlorophyll biosynthesis, greening of different plant
organs.

PMID: 16151870 [PubMed - as supplied by publisher]

143: Planta. 2005 Nov;222(5):832-47. Epub 2005 Sep 6.

Isogene specific oligo arrays reveal multifaceted changes in gene expression
during grape berry (Vitis vinifera L.) development.

Terrier N, Glissant D, Grimplet J, Barrieu F, Abbal P, Couture C, Ageorges A,
Atanassova R, Leon C, Renaudin JP, Dedaldechamp F, Romieu C, Delrot S, Hamdi S.

Unite Mixte de Recherche S.P.O., Biologie Integrative de la Vigne et du Raisin,
I.N.R.A., 2 Place Viala, 34060, Montpellier Cedex 1, France.

The transition from a green, hard, and acidic pericarp to a sweet, soft,
coloured, and sugar-rich ripe fruit occurs in many unrelated fruit species. High
throughput identification of differentially expressed genes in grape berry has
been achieved by the use of 50-mers oligoarrays bearing a set of 3,200 Unigenes
from Vitis vinifera to compare berry transcriptome at nine developmental stages.
Analysis of transcript profiles revealed that most activations were triggered
simultaneously with softening, occurring within only 24 h for an individual
berry, just before any change in colouration or water, sugar, and acid content
can be detected. Although most dramatically induced genes belong to unknown
functional categories, numerous changes occur in the expression of isogenes
involved in primary and secondary metabolism during ripening. Focusing on
isogenes potentially significant in development regulation (hormonal control of
transcription factor) revealed a possible role for several hormones (cytokinin,
gibberellin, or jasmonic acid). Transcription factor analysis revealed the
induction of RAP2 and WRKY genes at veraison, suggesting increasing biotic and
abiotic stress conditions during ripening. This observation was strengthened by
an increased expression of multiple transcripts involved in sugar metabolism and
also described as induced in other plant organs during stress conditions. This
approach permitted the identification of new isogenes as possible control
points: a glutathione S-transferase exhibits the same expression profile as
anthocyanin accumulation and a new putative sugar transporter is induced in
parallel with sugar import.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16151847 [PubMed - indexed for MEDLINE]

144: Ann Chim. 2005 Jun;95(6):415-9.

Determination of furaneol (4-hydroxy-2,5-dimethyl-3(2H)-furanone) in some wines
from Italian native grapes by Gas-Chromatography-SIM/MASS spectrometry.

Genovese A, Piombino P, Lisanti MT, Moio L.

Dipartimento di Scienze degli Alimenti, Universita degli Studi di Foggia,
Facolta di Agraria Via Napoli 25, 71100 Foggia, Italy. [email protected]

Gas Chromatography-Mass Spectrometry (GC-MS) analysis by Selective Ion
Monitoring (SIM) was applied to quantify 4-Hydroxy-2,5-dimethyl-3(2H)-furanone
(HDMF) in both red and white wines obtained from some Italian cultivar of Vitis
vinifera. Wines were extracted by liquid-liquid extraction performed with
1,1,2-trichlorotrifluoroethane (Freon 113). The ion m/z 128 was used for
quantification while the ion m/z 129 as qualifier. Precision, linearity and
accuracy of the method resulted satisfactory. Results showed a significant
variation in the concentration of furaneol in wine with grape variety.
Generally, HDMF concentrations in white wines were lower than in red wines.
Among white wines, Chardonnay resulted characterized by the highest
concentration of HDMF. Among red wines the highest concentrations of HDMF were
detected in Primitivo and Refosco varieties.

PMID: 16136836 [PubMed - indexed for MEDLINE]

145: Mol Plant Microbe Interact. 2005 Aug;18(8):819-29.

beta-Aminobutyric acid-induced resistance against downy mildew in grapevine acts
through the potentiation of callose formation and jasmonic acid signaling.

Hamiduzzaman MM, Jakab G, Barnavon L, Neuhaus JM, Mauch-Mani B.

University of Neuchatel, Institute of Botany, Department of Biochemistry, Rue
Emile-Argand 11, Case Postale 2, CH-2007 Neuchatel, Switzerland.

beta-Aminobutyric acid (BABA) was used to induce resistance in grapevine (Vitis
vinifera) against downy mildew (Plasmopara viticola). This led to a strong
reduction of mycelial growth and sporulation in the susceptible cv. Chasselas.
Comparing different inducers, the best protection was achieved with BABA
followed by jasmonic acid (JA), whereas benzo (1,2,3)-thiadiazole-7-carbothionic
acid-S-methyl ester (a salicylic acid [SA] analog) and abscisic acid (ABA)
treatment did not increase the resistance significantly. Marker genes for the SA
and JA pathways showed potentiated expression patterns in BABA-treated plants
following infection. The callose synthesis inhibitor 2-deoxy-D-glucose partially
suppressed BABA- and JA-induced resistance against P viticola in Chasselas.
Application of the phenylalanine ammonia lyase inhibitor
2-aminoindan-2-phosphonic acid and the lipoxygenase (LOX) inhibitor 5, 8, 11,
14-eicosatetraynoic acid (ETYA) also led to a reduction of BABA-induced
resistance (BABA-IR), suggesting that callose deposition as well as defense
mechanisms depending on phenylpropanoids and the JA pathways all contribute to
BABA-IR. The similar phenotype of BABA- and JA-induced resistance, the
potentiated expression pattern of JA-regulated genes (LOX-9 and PR-4) following
BABA treatment, and the suppression of BABA-IR with ETYA suggest an involvement
of the JA pathway in BABA-IR of grapevine leading to a primed deposition of
callose and lignin around the infection sites.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16134894 [PubMed - indexed for MEDLINE]

146: J Exp Bot. 2005 Oct;56(420):2721-31. Epub 2005 Aug 30.

A carotenoid cleavage dioxygenase from Vitis vinifera L.: functional
characterization and expression during grape berry development in relation to
C13-norisoprenoid accumulation.

Mathieu S, Terrier N, Procureur J, Bigey F, Gunata Z.

UMR IR2B, ENSAM-INRA-Universite Montpellier II, F-34060 Montpellier cedex 1,
France.

A potential Carotenoid Cleavage Dioxygenase (CCD) gene was identified among a
Vitis vinifera L. EST collection and a full-length cDNA (VvCCD1) was isolated.
Recombinant expression of VvCCD1 confirmed that the gene encoded a functional
CCD. Experimental evidence was obtained that VvCCD1 cleaves zeaxanthin
symmetrically yielding 3-hydroxy-beta-ionone, a C(13)-norisoprenoidic compound,
and a C(14)-dialdehyde. Expression of the gene was studied by real-time PCR at
different developmental stages of grape berries from Muscat of Alexandria and
Shiraz cultivars. A significant induction of the gene expression approaching
veraison was observed in both cultivars. In parallel, the C(13)-norisoprenoid
level increased from veraison to maturity in both cultivars.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16131507 [PubMed - indexed for MEDLINE]

147: J Agric Food Chem. 2005 Sep 7;53(18):7043-51.

Transformation of Eutypa dieback and esca disease pathogen toxins by
antagonistic fungal strains reveals a second detoxification pathway not present
in Vitis vinifera.

Christen D, Tharin M, Perrin-Cherioux S, Abou-Mansour E, Tabacchi R, Defago G.

Phytopathology Group, Institute of Plant Sciences, Swiss Federal Institute of
Technology, 8092 Zurich, Switzerland.

Eutypine, 4-hydroxybenzaldehyde, and 3-phenyllactic acid are some of the
phytotoxins produced by the pathogens causing Eutypa dieback and esca disease,
two trunk diseases of grapevine (Vitis vinifera). Known biocontrol agents such
as Fusarium lateritium and Trichoderma sp. were screened for their ability to
consume these toxins. Transformation time courses were performed, and an
high-performance liquid chromatography-based method was developed to analyze
toxin metabolism and to identify and quantify the converted products. The
results show that the aldehyde function of eutypine was reduced to eutypinol, as
by V. vinifera cv. Merlot, the cultivar tolerant to Eutypa dieback. We revealed
a supplementary detoxification pathway, not known in Merlot, where the aldehyde
function was oxidized to eutypinic acid. Moreover, some strains tested could
further metabolize the transformation products. Every strain tested could
transform 4-hydroxybenzaldehyde to the corresponding alcohol and acid, and these
intermediates disappeared totally at the end of the time courses. When
biological assays on cells of V. vinifera cv. Chasselas were carried out, the
transformation products exhibited a lower toxicity than the toxins. The
possibility of selecting new biocontrol agents against trunk diseases of
grapevine based on microbial detoxification is discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16131109 [PubMed - indexed for MEDLINE]

148: J Agric Food Chem. 2005 Sep 7;53(18):7029-34.

Fast access of some grape pigments to the brain.

Passamonti S, Vrhovsek U, Vanzo A, Mattivi F.

Dipartimento di Biochimica Biofisica e Chimica delle Macromolecole, Universita
di Trieste, via L. Giorgeri 1, 34127 Trieste, Italy.

Anthocyanins represent the main flavonoid pigments in red grape and wine, in red
berries, and in many other fruits and vegetables and are widespread in the human
diet. After ingestion, these complex, hydrophilic compounds quickly appear as
intact molecules in the plasma. This study investigated their presence in the
brain of anesthetized rats that received 8 mg/kg of body weight of a pure
anthocyanin mixture extracted from Vitis vinifera grapes. The mixture was
maintained in the stomach for 10 min. After this time, intact anthocyanins were
detected by HPLC-DAD-MS not only in the plasma (176.4 +/- 50.5 ng/mL, mean +/-
SEM) but also in the brain (192.2 +/- 57.5 ng/g). These results demonstrate for
the first time that grape pigments can reach the mammalian brain within minutes
from their introduction into the stomach.

PMID: 16131107 [PubMed - indexed for MEDLINE]

149: J Chem Ecol. 2005 May;31(5):1051-63.

Vitex agnus-castus is a preferred host plant for Hyalesthes obsoletus.

Sharon R, Soroker V, Wesley SD, Zahavi T, Harari A, Weintraub PG.

Northern Research and Development, Kiryat Sh 'mona, Israel.

Hyalesthes obsoletus Signoret (Homoptera: Cixiidae) is a polyphagous planthopper
that transmits stolbur phytoplasma (a causative agent of "yellows" disease) to
various weeds, members of the Solanaceae, and wine grapes (Vitis vinifera L.) in
Europe and the Middle East. Planthoppers were collected by hand vacuuming eight
native plant species. Vitex agnus-castus L., a shrub in the Verbenaceae, hosted
the largest number of H. obsoletus, although Olea europaea L. also served as a
host for adults. Using a Y-olfactometer, we compared the planthoppers relative
preference for V. agnus-castus, Convolvulus arvensis, and V. vinifera. V.
agnus-castus was more attractive to both male and female H. obsoletus than the
other plants. H. obsoletus antennal response was stronger to volatiles collected
from V. agnuscastus than from Cabernet Sauvignon variety of V. vinifera. To
determine if V. agnus-castus would serve as a reservoir for the pathogen, H.
obsoletus were collected from leaf and stem samples of native V. agnus-castus,
and were tested by polymerase chain reaction (PCR) for the presence of
phytoplasma DNA. While 14% and 25% (2003 and 2004, respectively) of the insects
tested positive for phytoplasma DNA, none of the plant samples tested positive.
To determine if V. agnus-castus could serve as a host plant for the development
of the planthopper, we placed emergence cages beneath field shrubs and enclosed
wild-caught H. obsoletus in a cage with a potted young shrub. We found adult H.
obsoletus in the emergence cases and planthopper nymphs in the soil of the
potted plant. We concluded that V. agnus-castus is attractive to H. obsoletus,
which seems to be refractory to phytoplasma infections and warrants further
testing as a trap plant near vineyards.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16124232 [PubMed - indexed for MEDLINE]

150: Pathol Biol (Paris). 2005 Sep;53(7):411-5. Epub 2005 Jan 22.

Effect of procyanidolic oligomers of Vitis vinifera on the biosynthesis and
excretion of corneal glycosaminoglycans.

Robert AM, Robert L, Renard G.

Laboratoire de Recherche Ophtalmologique, Hopital Hotel Dieu, Universite Paris
5, France. [email protected]

Glycosaminoglycans (GAG) as hyaluronan and proteoglycans (PG) as dermatan
sulfate (DSPG) and keratan sulfate (KSPG) play important role in the structure
and physiological functions of cornea. Several corneal pathologies are
associated with qualitative and quantitative alterations of GAG and/or PG
biosynthesis. For these reasons we investigated the effect of procyanidolic
oligomers (PCO) on the biosynthesis and excretion of GAG-s in bovine corneal
explant cultures. This was accomplished by the determination of their chemical
components, hexoses, hexosamines and uronic acids as well as the incorporation
of (35)S-sulfate in the presence and without PCO in the culture medium. In
presence of 1 mg/ml PCO total hexoses, hexosamines and uronic acids increased at
5 and 24 h of incubation, hexoses less than the two other components.
Stimulation of biosynthesis concerned essentially the uronic acid containing
GAG-s, DS and hyaluronan. (35)S-sulfate incorporation decreased in presence of
PCO, showing a decreased sulfation of GAG-s. The most striking effect was
however the strongly increased excretion of neosynthesized GAG-s in the culture
medium. This effect could be beneficial in decreasing corneal hydration in
inflammatory conditions, together with the previously demonstrated protection of
corneal macromolecules against proteolytic degradation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16085118 [PubMed - indexed for MEDLINE]

151: Pharmacol Res. 2005 Oct;52(4):353-66.

Understanding local Mediterranean diets: a multidisciplinary pharmacological and
ethnobotanical approach.

The Local Food-Nutraceuticals Consortium.

Epidemiological data indicate a beneficial effect of Mediterranean diets on
human health, especially on the prevalence of cardiovascular disease. These
observations are supported by recent intervention studies. However, very little
is known about the current role of local Mediterranean food products, which are
consumed on a less regular basis and their contribution to a healthy diet. The
European consortium "Local Food-Nutraceuticals" collected 127 locally consumed
wild or semi-wild plants in three Mediterranean countries, i.e. Greece, Italy,
and Spain, in order to assess their ethnobotanical features as well as their
biological activities. The project also includes a second line of research, the
study of local conceptions about these food resources. All pharmacological
assays were conducted with ethanolic extracts prepared from the dried plant
material. The biological activities of the extracts were assessed with the
following 12 different assays covering a broad range of mechanisms considered
crucial in the pathology of chronic, aging-related diseases. Four antioxidant
tests: DPPH scavenging, prevention of oxyhaemoglobin bleaching, prevention of
lipid peroxidation (malondialdehyde formation), and protection from DNA damage
(Comet assay); three enzyme inhibition tests: inhibition of xanthine oxidase,
inhibition of myeloperoxidase-catalysed guaiacol oxidation as well as the
inhibition of acetylcholine esterase; one test investigating the inhibition of
cytokine-induced cell activation (including the extracts' potential
cytotoxicity); one assay measuring the anti-proliferation potential; one test
assessing the anti-diabetic activity (PPARgamma) as well as one assay
investigating the extracts' effect on mood disorder-related biochemical
parameters (hSERT). Furthermore, the polyphenol content of all extracts was
determined using the Folin-Ciocalteaus method. The assays revealed diverse
biological effects for the tested extracts ranging from no activity to almost
complete inhibition/activation. Moreover, the experimental matrix led to the
identification of a sub-set of extracts, i.e. Berberis vulgaris, Reichardia
picroides, Scandix australis, Satureja montana, Thymus piperella, Lythrum
salicaria and Vitis vinifera, showing high activity in a broad range of assays.
In summary, the in vitro observed modulations and effects exerted by extracts
derived from local food plants suggest that these plants may contribute to the
observed better aging of rural Mediterranean populations.

Publication Types:
Comparative Study
In Vitro
Research Support, Non-U.S. Gov't

PMID: 16051496 [PubMed - indexed for MEDLINE]

152: Theor Appl Genet. 2005 Sep;111(5):871-8. Epub 2005 Oct 18.

Genomic analysis of Grapevine Retrotransposon 1 (Gret 1) in Vitis vinifera.

Pereira HS, Barao A, Delgado M, Morais-Cecilio L, Viegas W.

Seccao de Genetica, Instituto Superior de Agronomia, Centro de Botanica Aplicada
a Agricultura, Tapada da Ajuda, 1349-017 Lisboa, Portugal.
[email protected]

The complete sequence of the first retrotransposon isolated in Vitis vinifera,
Gret 1, was used to design primers that permitted its analysis in the genome of
grapevine cultivars. This retroelement was found to be dispersed throughout the
genome with sites of repeated insertions. Fluorescent in situ hybridization
indicated multiple Gret 1 loci distributed throughout euchromatic portions of
chromosomes. REMAP and IRAP proved to be useful as molecular markers in
grapevine. Both of these techniques showed polymorphisms between cultivars but
not between clones of the same cultivar, indicating differences in Gret 1
distribution between cultivars. The combined cytological and molecular results
suggest that Gret 1 may have a role in gene regulation and in explaining the
enormous phenotypic variability that exists between cultivars.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16025304 [PubMed - indexed for MEDLINE]

153: Plant Physiol Biochem. 2005 Jul;43(7):693-9.

Low-night temperature (LNT) induced changes of photosynthesis in grapevine
(Vitis vinifera L.) plants.

Bertamini M, Muthuchelian K, Rubinigg M, Zorer R, Nedunchezhian N.

Istituto Agrario di San Michele all' Adige, 38010, San Michele all' Adige,
Italy.

Changes of leaf pigments, ribulose-1,5-bisphosphate carboxylase (RuBPC) and
photosynthetic efficiency were examined in grapevine (Vitis vinifera L.) plants
grown under ambient irradiation (maximum daily PAR = 1500 micromol m(-2) s(-1))
for 7 days to low night temperature (LNT) of 5 degrees C (daily from 18:00 to
06:00). The contents of chlorophyll (Chl) and carotenoids (Car) per fresh mass
were lower in LNT leaves than in control leaves. The contents of alpha + beta
carotene and lutein-5,6-epoxide remained unaffected, but the de-epoxidation
state involving the components of xanthophyll cycle increased. RuBPC activity
and soluble proteins were also significantly reduced in LNT leaves. In isolated
thylakoids, a marked inhibition of whole chain (PS I + PS II) and PS II activity
were observed in LNT leaves. Smaller inhibition of PS I activity was observed in
LNT leaves. The artificial exogenous electron donors, MnCl2, DPC and NH2OH did
not restored the loss of PS II activity in LNT leaves. The same results were
obtained when F(v)/F(m) was evaluated by Chl fluorescence measurements. The
marked loss of PS II activity in LNT leaves was due to the marked loss of D1
protein which was determined by immunological studies.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 16023356 [PubMed - indexed for MEDLINE]

154: J Agric Food Chem. 2005 Jul 13;53(14):5798-808.

Influence of vine vigor on grape (Vitis vinifera L. Cv. Pinot Noir) and wine
proanthocyanidins.

Cortell JM, Halbleib M, Gallagher AV, Righetti TL, Kennedy JA.

Department of Food Science and Technology, Oregon State University, Corvallis,
Oregon 97331, USA.

The relationships between variations in grapevine (Vitis vinifera L. cv. Pinot
noir) growth and resulting fruit and wine phenolic composition were
investigated. The study was conducted in a commercial vineyard consisting of the
same clone, rootstock, age, and vineyard management practices. The experimental
design involved monitoring soil, vine growth, yield components, and fruit
composition (soluble solids, flavan-3-ol monomers, proanthocyanidins, and
pigmented polymers) on a georeferenced grid pattern to assess patterns in growth
and development. Vine vigor parameters (trunk cross-sectional area, average
shoot length, and leaf chlorophyll) were used to delineate zones within both
blocks to produce research wines to investigate the vine-fruit-wine continuum.
There was no significant influence of vine vigor on the amount of
proanthocyanidin per seed and only minimal differences in seed proanthocyanidin
composition. However, significant increases were found in skin proanthocyanidin
(mg/berry), proportion of (-)-epigallocatechin, average molecular mass of
proanthocyanidins, and pigmented polymer content in fruit from zones with a
reduction in vine vigor. In the wines produced from low-vigor zones, there was a
large increase in the proportion of skin tannin extracted into the wine, whereas
little change occurred in seed proanthocyanidin extraction. The level of
pigmented polymers and proanthocyanidin molecular mass were higher in wines made
from low-vigor fruit compared to wines made from high-vigor fruit, whereas the
flavan-3-ol monomer concentration was lower.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 15998151 [PubMed - indexed for MEDLINE]

155: Theor Appl Genet. 2005 Aug;111(4):658-64. Epub 2005 Jul 2.

QTL analysis for fruit yield components in table grapes (Vitis vinifera).

Fanizza G, Lamaj F, Costantini L, Chaabane R, Grando MS.

DIBCA, Genetics and Plant Breeding Unit, University of Bari, Via Amendola 165/A,
70126 Bari, Italy. [email protected]

A segregation population of 184 genotypes derived from a pseudo-testcross of
table grapes (Vitis vinifera), together with 203 AFLP and 110 SSR markers was
used to detect quantitative trait loci (QTLs) for fruit yield components.
Diffferent QTLs, a low percentage of phenotypic variance explained by the QTLs
detected and QTL instability over years were detected for each fruit yield
component. These results confirm the complex genetic architecture of the yield
components in grapevine due to the perennial nature of this species, which has
to adapt to yearly variations in climate. Phenotypic correlation analyses
between fruit yield components were also performed. The negative correlation
between berry weight and the number of berries per cluster seems to have an
indirect negative effect on cluster weight, as revealed by the path coefficient
analysis; however, this negative correlation was not supported at the molecular
level because no coincident QTLs were observed between these traits.
Nonetheless, the possibility to select seedless genotypes with large berries
without affecting cluster weight needs to be substantiated in future experiments
because factors such as sample size and heritability might influence QTL
identification in table grapes.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 15995866 [PubMed - indexed for MEDLINE]

156: Chemosphere. 2006 Jan;62(4):650-7. Epub 2005 Jun 28.

Transient effect of the herbicide flazasulfuron on carbohydrate physiology in
Vitis vinifera L.

Magne C, Saladin G, Clement C.

LEBHAM, UPRES EA 3877, Institut Universitaire Europeen de la Mer, Universite de
Bretagne Occidentale, Technopole Brest Iroise, 29280 Plouzane, France.

In the Champagne vineyard, most of the areas treated in early 2000 with the
newly approved herbicide flazasulfuron had vines with altered growth and yellow
leaves throughout the growing season. In order to clarify the physiological
perturbations caused on the non-target grapevine and their potential
consequences, C nutrition of grape plants grown in vineyards treated or not with
flazasulfuron in 2000 was characterized during the following season. Vines from
treated areas exhibited yellow leaves and an alteration of photosynthetic
activity, characterized by declines in leaf gas exchanges (by 85%) and
photosynthetic pigment concentrations (by 88%), and a marked disorganization of
the leaf plastids. The herbicide also caused a decrease in leaf starch and
soluble carbohydrate levels (-74% and -90%, respectively). Surprisingly, some
vines re-greened after bloom, then exhibiting similar carbohydrate physiology to
those grown in a non-treated area. Thus, recovery of CO(2) fixation rates,
plastid ultra-structure, pigment concentrations and carbohydrate levels was
found in re-greening leaves. Unlike the informations available in the
literature, our results showed that flazasulfuron may be phytotoxic for
grapevine. However, this toxicity was overcome the following year, indicating
that vines have the potential to recover from this herbicide stress after one
season.

PMID: 15987652 [PubMed - indexed for MEDLINE]

157: J Virol Methods. 2005 Aug;127(2):126-32. Epub 2005 Apr 19.

RT-PCR-RFLP for genetic diversity analysis of Tunisian Grapevine fanleaf virus
isolates in their natural host plants.

Fattouch S, Acheche H, M'hirsi S, Mellouli L, Bejar S, Marrakchi M, Marzouki N.

Laboratory of Biological Engineering, Institut National des Sciences Appliquees
et de Technologie, Centre Urbain Nord, BP No. 676, Tunis 1080, Tunisia.
[email protected]

Genetic diversity was characterized in 20 isolates of Grapevine fanleaf virus
(GFLV) recovered from naturally infected grapevine plants (Vitis vinifera) in
the North of Tunisia. Viral RNAs were isolated by oligoprobe capture, and a 605
bp fragment containing a part of the viral coat protein gene was amplified by
RT-PCR. Sequence variation among isolates was characterized by restriction
fragment length polymorphism (RFLP) analysis and confirmed by sequencing. The
GFLV infections are found as a complex mixture of closely related genomes. In
further studies, RFLP analyses of virus isolates using AluI showed that GFLV
populations in Tunisian vineyards consist of two restrictotypes corresponding to
distinct sub-populations Sp1 and Sp2. The relative field distribution of these
sub-populations showed that Sp2 was more abundant. Individual genomes were
recovered by cloning the RT-PCR products. The sequences were found to vary from
each other by as much as 11%. Cloning from mixed infections showed that Sp2 are
also predominant.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15967236 [PubMed - indexed for MEDLINE]

158: Virus Genes. 2005 Aug;31(1):31-41.

Genome Sequences and Structures of Two Biologically Distinct Strains of
Grapevine leafroll-associated virus 2 and Sequence Analysis.

Meng B, Li C, Goszczynski DE, Gonsalves D.

Department of Molecular and Cellular Biology, College of Biological Science,
University of Guelph, N1G 2W1, Ontario, Canada.

Grapevine leafroll-associated virus 2 (GLRaV-2), a member of the genus
Closterovirus within Closteroviridae, is implicated in several important
diseases of grapevines including "leafroll", "graft-incompatibility", and "quick
decline" worldwide. Several GLRaV-2 isolates have been detected from different
grapevine genotypes. However, the genomes of these isolates were not sequenced
or only partially sequenced. Consequently, the relationship of these viral
isolates at the molecular level has not been determined. Here, we group the
various GLRaV-2 isolates into four strains based on their coat protein gene
sequences. We show that isolates "PN" (originated from Vitis vinifera cv. "Pinot
noir"), "Sem" (from V. vinifera cv. "Semillon") and "94/970" (from V. vinifera
cv. "Muscat of Alexandria") belong to the same strain, "93/955" (from hybrid
"LN-33") and "H4" (from V. rupestris "St. George") each represents a distinct
strain, while Grapevine rootstock stem lesion-associated virus.

PMID: 15965606 [PubMed - in process]

159: Theor Appl Genet. 2005 Jul;111(2):370-7. Epub 2005 May 18.

Genetic and physical mapping of the grapevine powdery mildew resistance gene,
Run1, using a bacterial artificial chromosome library.

Barker CL, Donald T, Pauquet J, Ratnaparkhe MB, Bouquet A, Adam-Blondon AF,
Thomas MR, Dry I.

CSIRO Plant Industry, Glen Osmond, SA, Australia. [email protected]

Resistance to grapevine powdery mildew is controlled by Run1, a single dominant
gene present in the wild grapevine species, Muscadinia rotundifolia, but absent
from the cultivated species, Vitis vinifera. Run1 has been introgressed into V.
vinifera using a pseudo-backcross strategy, and genetic markers have previously
been identified that are linked to the resistance locus. Here we describe the
construction of comprehensive genetic and physical maps spanning the resistance
locus that will enable future positional cloning of the resistance gene.
Physical mapping was performed using a bacterial artificial chromosome (BAC)
library constructed using genomic DNA extracted from a resistant V. vinifera
individual carrying Run1 within an introgression. BAC contig assembly has
enabled 20 new genetic markers to be identified that are closely linked to Run1,
and the position of the resistance locus has been refined, locating the gene
between the simple sequence repeat (SSR) marker, VMC4f3.1, and the BAC end
sequence-derived marker, CB292.294. This region contains two multigene families
of resistance gene analogues (RGA). A comparison of physical and genetic mapping
data indicates that recombination is severely repressed in the vicinity of Run1,
possibly due to divergent sequence contained within the introgressed fragment
from M. rotundifolia that carries the Run1 gene.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 15902396 [PubMed - indexed for MEDLINE]

160: J Plant Physiol. 2005 Apr;162(4):365-74.

Monitoring the stability of Rubisco in micropropagated grapevine (Vitis vinifera
L.) by two-dimensional electrophoresis.

Carvalho LC, Esquivel MG, Martins I, Ricardo CP, Amancio S.

DBEB/CBAA, Instituto Superior de Agronomia, UTL, Tapada da Ajuda, 1349-017
Lisboa, Portugal.

Plants cultured in vitro suffer from several physiological and biochemical
impairments due to the artificial conditions of growth, namely the composition
of the heterotrophic media. Upon transfer to ex vitro, the higher irradiances,
compared to in vitro, can lead to oxidative stress symptoms, which can be
counteracted by CO2 concentrations above atmospheric levels. Here we analyse the
stability of Rubisco in in vitro grapevine plantlets, and after transfer to ex
vitro under four acclimatization treatments: low irradiance (LL, 150 micromol
m(-2)s(-1)) and high irradiance (HL, 300 micromol m(-2)s(-1)) in association
with CO2 concentrations of 350 (LCO2) and 700 (HCO2) microL L(-1). Proteins were
separated with SDS polyacrylamide gel electrophoresis and two-dimensional
electrophoresis and Rubisco degradation peptides were analysed by immunoblotting
with anti-LSU antibodies. These degradation products were present in the leaves
of plantlets under both in vitro and ex vitro treatments. Under LCO2 they were
maintained for almost all of the 28 days of the acclimatization period, while
becoming scarcely detected after 14 days under HCO2 and after 7 days when HCO2
was associated with HL. These results appear to confirm the counteraction of
HCO2 concentrations over the oxidative stress eventually caused by HL. The
patterns of soluble sugars in acclimatizing leaves under HLHCO2 also gave an
indication of a faster acquisition of autotrophic characteristics.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15900878 [PubMed - indexed for MEDLINE]

161: J Agric Food Chem. 2005 May 18;53(10):4019-25.

Influence of ethanol concentration on the extraction of color and phenolic
compounds from the skin and seeds of Tempranillo grapes at different stages of
ripening.

Canals R, Llaudy MC, Valls J, Canals JM, Zamora F.

Departament de Bioquimica i Biotecnologia, Facultat d'Enologia de Tarragona
(CeRTA), Universitat Rovira i Virgili, C/Ramon y Cajal, 70, 43005 Tarragona,
Spain.

The aim of this paper is to study how grape ripeness and ethanol concentration
affect the extraction of color and phenolic compounds from skins and seeds
during the maceration/fermentation process. Simulated maceration assays were
carried out with the grapes at three stages of berry development (vitis vinifera
cv. Tempranillo) and different percentages of ethanol in the maceration media.
Both ripeness and ethanol content have a considerable effect on the extraction
of color and phenolic compounds. Of these two factors, ripeness increases the
extractability most. The presence of ethanol in the medium facilitates
anthocyanin and especially proanthocyanidin extraction, but it also decreases
copigmentation phenomena, which can decrease the color intensity. The higher the
ethanol concentration is in the maceration media, the higher the astringency of
proanthocyanidins.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15884833 [PubMed - indexed for MEDLINE]

162: Int J Mol Med. 2005 Jun;15(6):1013-22.

Effects of plant phenolics and grape extracts from Greek varieties of Vitis
vinifera on Mitomycin C and topoisomerase I-induced nicking of DNA.

Stagos D, Kazantzoglou G, Magiatis P, Mitaku S, Anagnostopoulos K, Kouretas D.

Department of Biochemistry and Biotechnology, University of Thessaly, GR-41221
Larissa, Greece.

In recent years, a number of reports have shown the anticancer activity of grape
extracts and wine against various types of cancer such as breast, lung and
gastric cancer. This property is mainly attributed to the plant polyphenols
identified in grapes. The aim of the present study was to investigate the
mechanisms by which grape extracts and plant polyphenols found in them exert
their chemopreventive and antitumour activities. Thus, aqueous and methanolic
extracts from two Greek varieties of Vitis vinifera, fractions enriched in
polyphenols of these extracts and polyphenolics (caffeic acid, ferulic acid,
gallic acid, protocatechuic acid and rutin) found in grapes were screened using
two in vitro assays: i) the topoisomerase I relaxation assay and ii) the
mitomycin C-induced DNA strand breakage. The grape extracts, the polyphenol-rich
fractions and some of the polyphenolics (caffeic acid and protocatechuic acid)
were potent inhibitors of topoisomerase I, indicating that the inhibition of
this enzyme may be one of the mechanisms accounting for the anticancer activity
of these compounds. Moreover, the grape extracts inhibited the mitomycin
C-induced DNA strand breakage suggesting that they could prevent ROS-mediated
DNA damage. On the other hand, the polyphenol-rich fractions and the plant
polyphenols enhanced the mitomycin C-induced DNA strand breakage indicating
prooxidant activity. Thus, it is of interest that whole grape extracts act as
chemopreventive agents by inhibiting topo I and mitomycin C-induced DNA damage,
while polyphenol enriched fractions and plant polyphenolics exert prooxidant
activity leading to enhancement of DNA damage which may account for the
cytotoxic and apoptosis-inducing properties of plant polyphenols against cancer
cells.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 15870908 [PubMed - indexed for MEDLINE]

163: New Phytol. 2005 Jun;166(3):895-905.

Jasmonates and Na-orthovanadate promote resveratrol production in Vitis vinifera
cv. Barbera cell cultures.

Tassoni A, Fornale S, Franceschetti M, Musiani F, Michael AJ, Perry B, Bagni N.

Department of Biology e.s and Interdepartmental Centre for Biotechnology,
University of Bologna, Via Irnerio 42, 40126 Bologna, Italy.

Here the effect of jasmonic acid, methyljasmonate and Na-orthovanadate on the
production of resveratrol was studied in Vitis vinifera cv. Barbera cell
suspension cultures. Na-orthovanadate at 0.1 mm and 1 mm concentration was
efficient in promoting the production and/or accumulation and release in the
culture medium of cis-resveratrol while trans-resveratrol levels were not
affected by this treatment. Methyljasmonate was highly effective in stimulating
both trans- and cis-resveratrol endogenous accumulation, as well as their
release into the culture medium. Cis-resveratrol was absent or detected in very
low amounts in the controls. Jasmonic acid was less efficient than
methyljasmonate in promoting endogenous resveratrol accumulation, but it
stimulated the release in the culture medium especially of cis-resveratrol. Gel
analysis was performed on control and 10 microm MeJA treated cell suspensions.
Results showed an up-regulation of the stilbene synthase demonstrating that MeJA
stimulated the synthesis ex-novo of this protein.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15869650 [PubMed - indexed for MEDLINE]

164: Funct Integr Genomics. 2005 Oct;5(4):208-17. Epub 2005 Apr 22.

Comparative analysis of expressed sequence tags from different organs of Vitis
vinifera L.

Moser C, Segala C, Fontana P, Salakhudtinov I, Gatto P, Pindo M, Zyprian E,
Toepfer R, Grando MS, Velasco R.

Istituto Agrario San Michele all'Adige, S. Michele a/Adige, 38010 Trento, Italy.
[email protected]

Expressed sequence tags (ESTs) are providing a valuable approach to sampling
organism-expressed genomes, especially when studying large genomes such as those
of many plants. We report on the comparison of 8,647 ESTs generated from six
different grape (Vitis vinifera L.) organs: berry, root, leaf, bud, shoot and
inflorescence. Clustering and assembly of these ESTs resulted in 4,203 unique
sequences and revealed that at this level of EST sampling, each organ shares a
low percentage of transcripts with the others. To define organ relationships
based on EST counts, we calculated a distance matrix of pairwise correlation
coefficients between the libraries which indicated bud, inflorescence and shoot
as a group distinct from the other organs considered in this study. A putative
function was identified for about 85% of the unique sequences. By assigning them
to specific functional classes, we were able to highlight strong differences
between organs in the metabolism, protein biosynthesis and photosynthesis
categories. This grape EST collection has also proven to be a valuable source
for the development of 'functional' simple sequence repeats (SSRs) markers: a
total of 405 SSRs have been identified. EST sequences and annotation results
have been organised in the IASMA-grape database, freely available at the address
http://genomics.iasma.it.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 15856347 [PubMed - indexed for MEDLINE]

165: Mutagenesis. 2005 May;20(3):229-33. Epub 2005 Apr 20.

Evaluation of photo-mutagenicity and photo-cytotoxicity of food coloring agents.

Arimoto-Kobayashi S, Machida M, Okamoto K, Yamaguchi A.

Faculty of Pharmaceutical Sciences, Okayama University, 1-1-1 Tsushima, Okayama
700-8530, Japan. [email protected]

Pigments extracted from natural products are widely used for food coloration in
Japan. An investigation concerning the photo-mutagenicity and
photo-carcinogenicity of frequently used colorants in Japan was performed.
Colorants examined were from Laccifer lacca (lac-color), Coccus cacti
(cochineal-color), Carthamus tinctorius (carthamus yellow), Gardenia augusta
(gardenia yellow and gardenia blue), Monascus anka and Monascus purpureus
(monascus red), the skin of Vitis vinifera and Vitis labrusca (grape-skin
color), Tamarindus indica (tamarind brown) and Beta vulgaris (beet red). No
significant increase in bacterial mutation was found when Salmonella typhimurium
TA98, TA100 and TA102 were simultaneously treated with colorants and subjected
to UVA irradiation for 30 min. When colorant solutions were subjected to UVA
irradiation for 4 h, irradiated solutions containing lac-color became slightly
mutagenic toward S.typhimurium TA98 without metabolic activation. A decrease in
cell survival resulted when WTK-1 cells were subjected to UVA irradiation for 60
min in the presence of purpurin at 1 mg/ml. Delayed cytotoxicity was also
observed following 24 h incubation in fresh medium of samples that were
subjected to UVA irradiation for 60 min in the presence of colorant (carthamus
yellow, grape-skin color, gardenia blue, cochineal-color, monascus red or
purpurin).

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15843386 [PubMed - indexed for MEDLINE]

166: J Chem Ecol. 2005 Jan;31(1):77-87.

Antennal and behavioral responses of grapevine moth Lobesia botrana females to
volatiles from grapevine.

Tasin M, Anfora G, Ioriatti C, Carlin S, De Cristofaro A, Schmidt S, Bengtsson
M, Versini G, Witzgall P.

Agricultural Research Station 38 010 San Michele all'Adige, Trento, Italy.

Grapevine moth Lobesia botrana is the economically most important insect of
grapevine Vitis vinifera in Europe. Flower buds, flowers, and green berries of
Chardonnay grapevine are known to attract L. botrana for oviposition. The
volatile compounds collected from these phenological stages were studied by gas
chromatography-mass spectrometry, and the antennal response of L. botrana
females to these headspace collections was recorded by gas
chromatography-electroantennography. The compounds found in all phenological
stages, which consistently elicited a strong antennal response, were
pentadecane, nonanal, and alpha-farnesene. In a wind tunnel, gravid L. botrana
females flew upwind to green grapes, as well as to headspace collections from
these berries released by a piezoelectric sprayer release device. However, no
females landed at the source of headspace volatiles, possibly due to
inappropriate concentrations or biased ratios of compounds in the headspace
extracts.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15839481 [PubMed - indexed for MEDLINE]

167: Genome. 2005 Apr;48(2):312-20.

Isolation by improved thermal asymmetric interlaced PCR and characterization of
a seed-specific 2S albumin gene and its promoter from grape (Vitis vinifera L.).

Li ZT, Gray DJ.

Mid-Florida Research and Education Center, Institute of Food and Agricultural
Sciences, University of Florida, Apopka 32703-8504, USA.

A seed-specific 2S albumin gene and its promoter region of grape (Vitis vinifera
L.) were isolated using an improved thermal asymmetric interlaced PCR that
allowed efficient amplification of target sequence of up to 3 kbp in length
directly from genomic DNA. The 2S albumin VvAlb1 (for V. vinifera 2S albumin 1)
gene from different grape cultivars encompasses a coding region of 504-540
nucleotides corresponding to a deduced amino acid sequence of 167-179 residues.
This deduced protein contains up to 30% glutamine residues and eight cysteine
residues arranged in a pattern highly conserved among 2S albumins for disulfide
bond formation. DNA sequence alignment revealed that the same VvAlb1 gene among
different grape cultivars varied greatly, including an insertion of up to 36 bp
near the 3' end of the gene sequence isolated from 'Thompson Seedless'. DNA
sequence analysis indicated that several conserved seed-specific regulatory
motifs were clustered within a 0.6-kbp region 5' upstream of the transcription
start site. To further test promoter activity, the sequence of this region was
used to drive a bifunctional EGFP/NPTII fusion gene in Agrobacterium-mediated
transformation of grape somatic embryos and leaf discs of grape and tobacco
(Nicotiana tabacum L.). A high level of GFP expression, comparable with that
derived from an enhanced double CsVMV promoter, was observed in the cotyledonary
but not hypocotyl and vegetative tissues of grape and tobacco. These results
suggest that the VvAlb1 gene promoter isolated is capable of conferring
seed-specific gene expression.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15838554 [PubMed - indexed for MEDLINE]

168: Theor Appl Genet. 2005 May;110(8):1363-71. Epub 2005 Apr 16.

Construction and characterization of BAC libraries from major grapevine
cultivars.

Adam-Blondon AF, Bernole A, Faes G, Lamoureux D, Pateyron S, Grando MS, Caboche
M, Velasco R, Chalhoub B.

Unite Mixte de Recherches sur les Genomes des Vegetaux, INRA, 2 rue Gaston
Cremieux, 5708 91 057, Evry Cedex, France. [email protected]

Genome projects were initiated on grapevine (Vitis vinifera L., 2n=38, genome
size 475 Mb) through the successful construction of four bacterial artificial
chromosome (BAC) libraries from three major cultivars, Cabernet Sauvignon
(Cabernet S), Syrah and two different clones of Pinot Noir (Pinot N). Depending
on the library, the genome coverage represented 4.5-14.8 genome equivalents with
clones having a mean insert size of 93-158 kb. BAC pools suitable for PCR
screening were constructed for two of these BAC libraries [Cabernet S and Pinot
N clone (cl) 115] and subsequently used to confirm the genome coverage of both
libraries by PCR anchoring of 74 genetic markers sampled from the 19 linkage
groups. For ten of these markers, two bands on separate BAC pools were
differentiated that could correspond either to different alleles or to a
duplication of the locus being studied. Finally, a preliminary assessment of the
correspondence between genetic and physical distances was made through the
anchoring of all the markers mapped along linkage group 1 of the V. vinifera
genetic map. A pair of markers, 2.1 cM apart, anchored the same BAC clones,
which allowed us to estimate that 1 cM corresponded in this particular region to
a maximum length of 130 kb.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 15834699 [PubMed - indexed for MEDLINE]

169: J Plant Physiol. 2005 Mar;162(3):301-8.

Possible role of catalase in post-dormancy bud break in grapevines.

Perez FJ, Lira W.

Lab. de Bioquimica Vegetal, Facultad de Ciencias, Universidad de Chile, Casilla
653, Santiago, Chile. [email protected]

Changes in the activity of catalase (Cat) and in the levels of H2O2 were
followed throughout dormancy in buds of grapevines (Vitis vinifera L.). In
grapevines grown in the Elqui valley in Chile, a region with warm-winters, the
activity of Cat increased during the recess period of buds, reaching a maximum
and thereafter decreased to less than one third of its maximal activity. Three
isoforms of Cat were detected in extracts of buds by native PAGE analysis, and
the extracted activity was inhibited competitively by hydrogen cyanamide (HC), a
potent bud-break agent. Furthermore, HC applications to field-grown grapevines
in addition to the expected effect on advancing bud break, reduced the Cat
activity during bud dormancy. Similar reductions were observed during dormancy
in buds of grapevines grown in the Central valley in Chile, a region with
temperate winters, suggesting that HC and winter chilling inhibits the activity
of the main H2O2 degrading enzyme in grape buds. A transient rise in H2O2 levels
preceded the release of buds from endodormancy, moreover, the peak of H2O2 and
the onset of bud break occurred earlier in HC treated than in control
grapevines, suggesting the participation of H2O2 as a signal molecule in the
release of endodormancy in grape buds. The relationship between Cat inhibition,
rise in H2O2 levels and initiation of bud break are discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15832682 [PubMed - indexed for MEDLINE]

170: Plant Cell Rep. 2005 Sep;24(7):433-8. Epub 2005 Apr 6.

Transgenic plants of Vitis vinifera cv. Seyval blanc.

Bornhoff BA, Harst M, Zyprian E, Topfer R.

BAZ-Institute for Grapevine Breeding, Geilweilerhof, 76833 Siebeldingen,
Germany. [email protected]

Leaf discs of grapevine cv. Seyval blanc originating from in vitro cultures were
transformed with Agrobacterium tumefaciens strain LBA 4404 harbouring the vector
pGJ42 carrying genes for chitinase and RIP (ribosome-inactivating protein) in an
attempt to improve fungal resistance. The gene for neomycin phosphotransferase
II (nptII) was used as the selectable marker gene. The explants were
cocultivated for 2 days with recombinant Agrobacteria and then submitted to
selection on NN69 medium containing 100 mg/l kanamycin. Successful regeneration
and conversion of transgenic plantlets were obtained. Stable integration of
foreign DNA was confirmed by PCR and Southern blot analyses, and protein
expression was detected by Western blot. The regenerated transgenic plants were
adapted to the greenhouse and showed no evidence of phenotypical alterations.
The foreign genes introduced into the transformed plants did not effect the
expected improvement in fungal disease resistance under field conditions for the
major pests Uncinula necator and Plasmopara viticola.

PMID: 15812658 [PubMed - indexed for MEDLINE]

171: Appl Environ Microbiol. 2005 Apr;71(4):1685-93.

Endophytic colonization of Vitis vinifera L. by plant growth-promoting bacterium
Burkholderia sp. strain PsJN.

Compant S, Reiter B, Sessitsch A, Nowak J, Clement C, Ait Barka E.

Laboratoire de Stress, Defenses et Reproduction des Plantes, Unite de Recherche
Vignes et Vins de Champagne, UPRES EA 2069, UFR Sciences, Universite de Reims
Champagne-Ardenne, Reims, France.

Patterns of colonization of Vitis vinifera L. cv. Chardonnay plantlets by a
plant growth-promoting bacterium, Burkholderia sp. strain PsJN, were studied
under gnotobiotic conditions. Wild-type strain PsJN and genetically engineered
derivatives of this strain tagged with gfp (PsJN::gfp2x) or gusA (PsJN::gusA11)
genes were used to enumerate and visualize tissue colonization. The rhizospheres
of 4- to 5-week-old plantlets with five developed leaves were inoculated with
bacterial suspensions. Epiphytic and endophytic colonization patterns were then
monitored by dilution plating assays and microscopic observation of organ
sections. Bacteria were chronologically detected first on root surfaces, then in
root internal tissues, and finally in the fifth internode and the tissues of the
fifth leaf. Analysis of the PsJN colonization patterns showed that this strain
colonizes grapevine root surfaces, as well as cell walls and the whole surface
of some rhizodermal cells. Cells were also abundant at lateral root emergence
sites and root tips. Furthermore, cell wall-degrading endoglucanase and
endopolygalacturonase secreted by PsJN explained how the bacterium gains entry
into root internal tissues. Host defense reactions were observed in the
exodermis and in several cortical cell layers. Bacteria were not observed on
stem and leaf surfaces but were found in xylem vessels of the fifth internode
and the fifth leaf of plantlets. Moreover, bacteria were more abundant in the
fifth leaf than in the fifth internode and were found in substomatal chambers.
Thus, it seems that Burkholderia sp. strain PsJN induces a local host defense
reaction and systemically spreads to aerial parts through the transpiration
stream.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15811990 [PubMed - indexed for MEDLINE]

172: J Exp Bot. 2005 May;56(415):1409-18. Epub 2005 Apr 4.

VvHT1 encodes a monosaccharide transporter expressed in the conducting complex
of the grape berry phloem.

Vignault C, Vachaud M, Cakir B, Glissant D, Dedaldechamp F, Buttner M,
Atanassova R, Fleurat-Lessard P, Lemoine R, Delrot S.

Unite Mixte de Recherches Centre National de la Recherche Scientifique 6161,
Transport des Assimilats, Laboratoire de Physiologie, Biochimie et Biologie
Moleculaire Vegetales, Batiment Botanique, Universite de Poitiers, Poitiers,
France.

The accumulation of sugars in grape berries requires the co-ordinate expression
of sucrose transporters, invertases, and monosaccharide transporters. A
monosaccharide transporter homologue (VvHT1, Vitis vinifera hexose transporter
1) has previously been isolated from grape berries at the veraison stage, and
its expression was shown to be regulated by sugars and abscisic acid. The
present work investigates the function and localization of VvHT1. Heterologous
expression in yeast indicates that VvHT1 encodes a monosaccharide transporter
with maximal activity at acidic pH (pH 4.5) and high affinity for glucose
(K(m)=70 muM). Fructose, mannose, sorbitol, and mannitol are not transported by
VvHT1. In situ hybridization shows that VvHT1 transcripts are primarily found in
the phloem region of the conducting bundles. Immunofluorescence and immunogold
labelling experiments localized VvHT1 in the plasma membrane of the sieve
element/companion cell interface and of the flesh cells. The expression and
functional properties of VvHT1 suggests that it retrieves the monosaccharides
needed to provide the energy necessary for cell division and cell growth at an
early stage of berry development.

PMID: 15809282 [PubMed - indexed for MEDLINE]

173: J Agric Food Chem. 2005 Apr 6;53(7):2652-7.

Induction of de novo volatile terpene biosynthesis via cytosolic and plastidial
pathways by methyl jasmonate in foliage of Vitis vinifera L.

Hampel D, Mosandl A, Wust M.

Institut fur Lebensmittelchemie, Johann Wolfgang Goethe-Universitat,
Marie-Curie-Strasse 9, D-60439 Frankfurt/Main, Germany.

The terpene biosynthesis in leaves of Vitis vinifera L. cv. Morio Muskat was
studied using methyl jasmonate to induce defensive responses in vivo. The
experiments demonstrated the strong activation of the de novo biosynthesis of
terpenoids via the octadecanoid-signaling cascade and release of the compounds
to the gas phase. Feeding experiments with [5,5-(2)H(2)]-1-deoxy-d-xylulose and
[5,5-(2)H(2)]mevalonic acid lactone allowed the investigation of the dynamic
allocation of resources via the mevalonic acid and
1-deoxy-d-xylulose/2-C-methyl-d-erythritol 4-phosphate (DOXP/MEP) pathway under
induced conditions and after treatment with the specific inhibitors mevastatin
and fosmidomycin. The experiments reveal that monoterpenes are almost
exclusively synthesized via the DOXP/MEP pathway, whereas sesquiterpenes are
generated via both pathways at approximately equal rates. The biosynthesis of
the homoterpene (E)-4,8-dimethyl-1,3,7-nonatriene was not affected by
mevastatine or fosmidomycin.

PMID: 15796607 [PubMed - indexed for MEDLINE]

174: Chem Commun (Camb). 2005 Apr 14;(14):1901-3. Epub 2005 Feb 8.

Highly chemoselective reduction of aromatic nitro compounds to the corresponding
hydroxylamines catalysed by plant cells from a grape (Vitis vinifera L.).

Li F, Cui J, Qian X, Zhang R, Xiao Y.

State Key Laboratory of Fine Chemicals, Dalian University of Technology, Dalian
116012, China. [email protected]

Cells from a grape (Vitis vinifera L.) reduce aromatic nitro compounds under
mild conditions to the corresponding hydroxylamines with unprecedented
chemoselectivity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15795781 [PubMed - indexed for MEDLINE]

175: Oecologia. 2005 May;143(4):548-57. Epub 2005 Mar 25.

Relative performance of European grapevine moth (Lobesia botrana) on grapes and
other hosts.

Thiery D, Moreau J.

UMR 1065 INRA-ENITAB Sante vegetale, INRA Institut Superieur de la Vigne et du
Vin, B.P.81, 33883, Villenave d'Ornon Cedex, France. [email protected]

The European grapevine moth, Lobesia botrana is a major grapevine pest, but
despite the abundance of vineyards it is a generalist and uses either grapes or
alternative species. Given the abundance and predictability of grape, L. botrana
could be expected to have evolved towards monophagy. In order to understand why
this species remains polyphagous, we hypothesized that larvae reared on rare
wild host plants should have higher fitness than those reared on the more
abundant grape host. For this, we compared larval performance and several life
history traits on three alternative host plants (Daphne gnidium, Olea europaea,
Tanacetum vulgare) and three Vitaceae (Vitis vinifera), two cultivars and one
wild species (Ampelopsis brevipedunculata), and two control groups raised on
either a low or a high nutritive value medium. Alternative hosts are more
suitable than Vitaceae for the reproductive performance of L. botrana: larval
mortality and development time was reduced, while pupal weight, growth rate,
female longevity, female fecundity, duration of laying and mating success were
increased. High quality food ingested by larvae promotes higher adult body
weight and enhances female reproductive output. This suggests that alternative
hosts provide greater nutritional value for L. botrana than Vitaceae. The use of
alternative host plants could thus be maintained in the host range because they
offer L. botrana a better fitness than on the Vitaceae. This could typically
represent an advantage for moths behaving in plant diversity grape landscapes.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 15791428 [PubMed - indexed for MEDLINE]

176: Exp Appl Acarol. 2005;35(1-2):83-109.

Eriophyoid mite damage in Vitis vinifera (grapevine) in Australia:
Calepitrimerus vitis and Colomerus vitis (Acari: Eriophyidae) as the common
cause of the widespread 'Restricted Spring Growth' syndrome.

Bernard MB, Horne PA, Hoffmann AA.

Centre for Environmental Stress and Adaptation Research, La Trobe University,
Bundoora, Victoria 3086, Australia. [email protected]

Leaf and shoot distortions and retarded shoot growth in Vitis vinifera L.
prevalent in Australian vineyards in early spring, were investigated in
replicated field experiments over 3 yrs. Leaf distortion and retarded shoot
growth were identified as damage due to feeding of extremely high populations of
over-wintered deutogynes of Calepitrimerus vitis (Nalepa) (grape rust mite).
This damage was hitherto known in Australia as 'Restricted Spring Growth' (RSG),
a syndrome comprising several growth abnormality symptoms, none with a clearly
identified cause or a successful treatment. A successful treatment against C.
vitis was used to selectively eliminate RSG, while C. vitis numbers were
recorded using a validated trapping technique; intercepting deutogynes migrating
from winter shelters in the wooden vine structure, to emerging green tissues.
Severe leaf distortion was associated with > 400 C. vitis deutogynes per spur,
while > 1000 per spur had the added effect of severely retarding shoot growth. A
43.0-47.2% shoot length reduction was recorded for Cabernet Sauvignon,
27.1-32.8% for Sauvignon Blanc, when 4-6 leaves were separated. Symptoms were
most prominent up to 8-9 separated leaves, however 24.7-30.4% shoot length
reduction was still evident at flowering, and 12.8% circa fruit set. C. vitis
effect on vine fruitfulness, and yield parameters at fruit set, were also
studied. Once successfully treated to prevent C. vitis damage, poor bud burst
remained evident in some vineyards. Surveys of unburst buds from such vineyards
revealed presence of Colomerus vitis (Pagenstecher) (grape bud mite). When Col.
vitis numbers in unburst buds reached 100-500 per bud, apical meristems of
primary, and commonly also secondary buds were dead, preventing bud burst. The
remaining living scale tissue was distinctly scarred. Bud and associated shoot
damage were documented. Retarded shoot growth and leaf distortion, previously
attributed to RSG, are misdiagnosed C. vitis spring feeding damage. Clustered
high infestations of Col. vitis can cause bud-axis necrosis, bud burst failure,
shoots with short basal internodes, and short, thin, zigzagged shoots with
absent fruit clusters; all previously considered RSG.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15777003 [PubMed - indexed for MEDLINE]

177: J Econ Entomol. 2005 Feb;98(1):182-7.

Application of competitive enzyme-linked immunosorbent assay for the
quantification of imidacloprid titers in xylem fluid extracted from grapevines.

Byrne FJ, Castle SJ, Bi JL, Toscano NC.

Department of Entomology, University of California, Riverside, CA 92521, USA.

A competitive enzyme-linked immunosorbent assay (ELISA) technique was evaluated
for quantifying titers of imidacloprid in xylem fluid extracted from Vitis
vinifera L. grapevines that were treated with systemic applications of the
neonicotinoid insecticide Admire. Evidence of matrix effects, factors that
compromise the precision and accuracy of the ELISA, was present in assays with
undiluted xylem fluid. These effects could be eliminated by dilution of extracts
in water, resulting in a lower sensitivity of the assay of 4 microg liter(-1).
In a field trial conducted in a commercial vineyard, there was an excellent
correlation between Admire application rates and xylem fluid concentrations of
imidacloprid. At an Admire application rate of 1.17 liter ha(-1) (16 fl oz per
acre), uptake of imidacloprid into vines was rapid. Imidacloprid was
consistently detected in the xylem for up to 3 mo after application at
concentrations known to be effective at managing populations of the sharpshooter
Homalodisca coagulata Say, an important vector of Xylella fastidiosa Wells in
California vineyards. The ELISA is a sensitive technique that can be used to
study the behavior of systemic insecticides within crop systems and their impact
on pest populations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15765681 [PubMed - indexed for MEDLINE]

178: Ann Bot (Lond). 2005 May;95(6):943-8. Epub 2005 Mar 4.

Phenology of flowering and starch accumulation in grape (Vitis vinifera L.)
cuttings and vines.

Lebon G, Duchene E, Brun O, Clement C.

Laboratoire de Stress, Defenses et Reproduction des Plantes, URVVC UPRES EA
2069, Universite de Reims Champagne Ardenne, UFR Sciences, Moulin de la Housse,
BP 1039, F-51687 Reims Cedex 2, France.

BACKGROUND AND AIMS: A reliable protocol for flowering and fruiting in cuttings
was developed with the aim of (a) studying inflorescence and flower development
in grapevine cuttings and field plants, and (b) assisting haploid plant
production. METHODS: Inflorescence and flower development was studied in
'Gewurztraminer' (GW) and 'Pinot Noir' (PN) grape vines and cuttings grown in a
glasshouse, along with variations in starch in the flowers. As there is a strong
relationship between flower development and starch, the starch content of
reproductive structures was estimated. KEY RESULTS: Inflorescence and flower
development were similar in the vines and cuttings with consistent differences
between the two cultivars. Indeed, the ontogenesis of male and female organs is
not synchronous in GW and PN, with both female and male meiosis occurring
earlier in PN than in GW. Moreover, changes of starch reserves were similar in
the two plant types. CONCLUSIONS: Cuttings have a similar reproductive
physiology to vines, and can be used to study grape physiology and to develop
haploid plants.

PMID: 15749750 [PubMed - indexed for MEDLINE]

179: Tree Physiol. 2005 May;25(5):633-9.

Photosynthesis of the grapevine (Vitis vinifera) inflorescence.

Lebon G, Brun O, Magne C, Clement C.

Laboratoire de Stress Defenses et Reproduction des Plantes, URVVC UPRES EA 2069,
Universite de Reims Champagne Ardenne, UFR Sciences, Moulin de la Housse, BP
1039, 51687 Reims Cedex 2, France.

To further characterize carbohydrate physiology in grapevine flowers, we
examined inflorescence autotrophy in the 'Gewurztraminer' and 'Pinot noir'
cultivars, which differ in sensitivity to flower abscission. In both cultivars,
positive net photosynthesis occurred in inflorescences. The rate of
photosynthesis gradually decreased throughout flower development and there was
no net carbon assimilation at fruit set. The rate of photosynthesis was
positively correlated with chlorophyll concentration but not to stomatal
conductance. Throughout flower development, the internal CO2 concentration
increased in inflorescence tissues, suggesting that assimilates are also formed
through refixation of respiratory CO2 by the phosphoenolpyruvate carboxylase
(PEPC) pathway. Significant differences between the two cultivars were recorded
during meiosis, when photosynthesis was higher in 'Gewurztraminer'. We conclude
that the inflorescence of grapevine contributes to its own carbon nutrition by
photosynthesizing throughout flower development. Moreover, the differential
patterns of photosynthesis in the inflorescences of 'Gewurztraminer' and 'Pinot
noir' might account for their differing fertilization rates and sensitivity to
flower abscission.

PMID: 15741155 [PubMed - in process]

180: Curr Microbiol. 2005 Jan;50(1):1-7. Epub 2005 Jan 17.

Establishment of a genetically marked insect-derived symbiont in multiple host
plants.

Bextine B, Lampe D, Lauzon C, Jackson B, Miller TA.

Department of Entomology, University of California, Riverside, CA 92507, USA.
[email protected]

Alcaligenes xylosoxidans subsp. denitrificans, originally isolated from the
cibarial region of the foregut of the glassy-winged sharpshooter (Homalodisca
coagulata), was transformed using the Himar1 transposition system to express
EGFP. Seedlings of six potential host plants were inoculated with transformed
bacteria and 2 weeks later samples were taken 5 cm away and analyzed by
quantitative real-time PCR using primers designed to amplify the gene insert.
The largest colony of 3,591,427 cells/2 cm of A. xylosoxidans subsp.
denitrificans was found in Citrus limon, with almost all plants testing positive
in both trials. The amount of colonization decreased in the other plants tested
in the following order: orange (Citrus sinensis "sweet orange") > chrysanthemum
(Chrysanthemum grandiflora cv. "White Diamond") > periwinkle (Vinca rosea) >
crepe myrtle (Lagerstroemia indica) > grapevine (Vitis vinifera cv. Chardonnay).
The bacterium's preference for citrus paralleled the host insect's preference
for this same plant. Additional tests determined that A. xylosoxidans subsp.
denitrificans thrives as a nonpathogenic, xylem-associated endophyte.

Publication Types:
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 15723145 [PubMed - indexed for MEDLINE]

181: Eur J Dermatol. 2005 Jan-Feb;15(1):31-6.

A randomised, double-blind, vehicle-controlled study to evaluate the efficacy
and safety of MAS063D (Atopiclair) in the treatment of mild to moderate atopic
dermatitis.

Belloni G, Pinelli S, Veraldi S.

European Institute of Dermatology, Viale Puglie 15, 20137 Milan, Italy.

MAS063D (Atopiclair is a hydrolipidic cream that has been developed for the
management of atopic dermatitis (AD). The putative active ingredients of MAS063D
are hyaluronic acid, telmesteine, Vitis vinifera, glycyrrhetinic acid. A
five-week study in 30 adult patients with mild to moderate AD showed that
MAS063D offered significant benefits over a vehicle-only control. MAS063D
improved the total body area affected (17.2% --> 13.2%, p < 0.001), itch score
(2.7 --> 1.3 on a 10-point scale, p = 0.001) and EASI score (28.3 --> 24.3, p =
0.024) after 22 days treatment compared to baseline. The patients' opinion of
MAS063D (patient's view of itch control, and view of study substance) was rated
by participating patients as significantly better than control (p = 0.008, p =
0.042 respectively). Based on these preliminary results in a small scale study,
it is suggested that MAS063D is a possible new treatment option for improving
signs and symptoms in adults with mild to moderate AD.

Publication Types:
Clinical Trial
Randomized Controlled Trial
Research Support, Non-U.S. Gov't

PMID: 15701590 [PubMed - indexed for MEDLINE]

182: Phytochem Anal. 2005 Jan-Feb;16(1):17-23.

An analytical survey of the polyphenols of seeds of varieties of grape (Vitis
vinifera) cultivated in Greece: implications for exploitation as a source of
value-added phytochemicals.

Guendez R, Kallithraka S, Makris DP, Kefalas P.

Department of Food Quality Management, Mediterranean Agronomic Institute of
Chania, Chania, Greece.

Seed samples from 12 white and 25 red international and Hellenic native grape
varieties (Vitis vinifera) were screened for their polyphenolic composition. The
polyphenols determined were mainly of low molecular weight, including gallic
acid, catechin, epicatechin, epicatechin gallate, epigallocatechin,
epigallocatechin gallate and the procyanidins B1 and B2. Average values of total
content for white and red varieties (376 and 388 mg/100 g seeds, respectively)
were very similar. Comparable results were observed with respect to the
individual polyphenol content with seeds from red varieties being, in general,
slightly richer. The predominant flavanol monomer in white and red varieties was
catechin (which accounted for 50.5 and 49.3%, respectively, of the total
content), whilst gallic acid and epigallocatechin were the constituents showing
the lowest content, respectively. The data obtained are discussed with regard to
the exploitation of grape seeds as a low-cost source of value-added
phytochemicals.

PMID: 15688951 [PubMed - indexed for MEDLINE]

183: J Agric Food Chem. 2005 Feb 9;53(3):644-50.

Development of activated carbon using vine shoots (Vitis vinifera) and its use
for wine treatment.

Corcho-Corral B, Olivares-Marin M, Valdes-Sanchez E, Fernandez-Gonzalez C,
Macias-Garcia A, Gomez-Serrano V.

Departamento de Quimica Inorganica, Facultad de Ciencias, Universidad de
Extremadura, Avenida de Elvas s/n, 06071 Badajoz, Spain.

An abundant and low-cost agricultural waste such as vine shoots (Vitis vinifera)
(VS), which is generated by the annual pruning of vineyards, has been used as
raw material in the preparation of powder activated carbon (AC) with a view to
develop a new fining agent for white wines. A commercial activated carbon,
S5X-Agrovin, was used for comparison purposes. From VS size-reduced pieces, AC
was prepared using phosphoric acid as activating agent. The concentration of the
H(3)PO(4) solution, the impregnation temperature, and the carbonization
conditions were controlled. The carbons were texturally characterized by gas
adsorption (N(2), -196 degrees C), mercury porosimetry, and density
measurements. FT-IR spectroscopy was used in the analysis of the surface
functional groups and structures of the carbons. Three varieties of white wine
(i.e., cv. Cayetana, cv. Macabeo, and cv. Sauvignon Blanc) were treated with the
activated carbons. Color changes were monitored by UV-vis spectrometry.
Significant differences in the degree of uptake of polyphenols were observed
depending on the wine variety and on the method of preparation of activated
carbon. The carbon prepared by first impregnation of VS with the 60 vol%
H(3)PO(4) solution at 50 degrees C and by then carbonization of the resultant
product at 400 degrees C for 2 h presents a higher ability to discolor the white
wines. The action of this carbon is comparable to that shown by the commercial
product. Both carbons possess a well-developed porosity in the macropore range.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15686414 [PubMed - indexed for MEDLINE]

184: Phytochemistry. 2005 Feb;66(3):295-303.

Metabolism of geraniol in grape berry mesocarp of Vitis vinifera L. cv.
Scheurebe: demonstration of stereoselective reduction, E/Z-isomerization,
oxidation and glycosylation.

Luan F, Mosandl A, Munch A, Wust M.

Institut fur Lebensmittelchemie, Johann Wolfgang Goethe-Universitat,
Marie-Curie-Strasse 9, D-60439 Frankfurt (Main), Germany.

The metabolism of deuterium labeled geraniol in grape mesocarp of Vitis vinifera
L. cv. Scheurebe was studied by in vivo-feeding experiments. Stereoselective
reduction to (S)-citronellol, E/Z-isomerization to nerol, oxidation to
neral/geranial and glycosylation of the corresponding monoterpene alcohols could
be demonstrated. Time course studies including the determination of conversion
rates revealed that the activity of these secondary transformations of
monoterpenes is dependent on the ripening stage and can be distinguished from
the development of the primary monoterpene synthase activities by the sharp
increase at the end of the ripening period. The stereoselective biosynthesis of
the potent odorant cis-(2S,4R)-rose oxide from labeled geraniol in grape berry
mesocarp is demonstrated as well. Since (S)-citronellol is the precursor of
cis-(2S,4R)-rose oxide it can be concluded that especially the last part of the
ripening period is important for the generation of this potent odorant. This
finding confirms the conclusion that a higher concentration of flavor compounds
could be established in the berries by leaving the fruit on the vine for
extended periods.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15680986 [PubMed - indexed for MEDLINE]

185: FEMS Microbiol Lett. 2005 Feb 1;243(1):107-15.

De novo synthesis of monoterpenes by Saccharomyces cerevisiae wine yeasts.

Carrau FM, Medina K, Boido E, Farina L, Gaggero C, Dellacassa E, Versini G,
Henschke PA.

Seccion Enologia, Departmento de Ciencia y Tecnologia de Alimentos, Facultad de
Quimica, Universidad de la Republica, 11800 Montevideo, Uruguay.
[email protected]

This paper reports the production of monoterpenes, which elicit a floral aroma
in wine, by strains of the yeast Saccharomyces cerevisiae. Terpenes, which are
typical components of the essential oils of flowers and fruits, are also present
as free and glycosylated conjugates amongst the secondary metabolites of certain
wine grape varieties of Vitis vinifera. Hence, when these compounds are present
in wine they are considered to originate from grape and not fermentation.
However, the biosynthesis of monoterpenes by S. cerevisiae in the absence of
grape derived precursors is shown here to be of de novo origin in wine yeast
strains. Higher concentration of assimilable nitrogen increased accumulation of
linalool and citronellol. Microaerobic compared with anaerobic conditions
favored terpene accumulation in the ferment. The amount of linalool produced by
some strains of S. cerevisiae could be of sensory importance in wine production.
These unexpected results are discussed in relation to the known sterol
biosynthetic pathway and to an alternative pathway for terpene biosynthesis not
previously described in yeast.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15668008 [PubMed - indexed for MEDLINE]

186: BMC Complement Altern Med. 2005 Jan 19;5:1.

Adaptogenic and nootropic activities of aqueous extract of Vitis vinifera (grape
seed): an experimental study in rat model.

Sreemantula S, Nammi S, Kolanukonda R, Koppula S, Boini KM.

Pharmacology Division, Department of Pharmaceutical Sciences Andhra University,
Visakhapatnam-530 003, Andhra Pradesh, India.
[email protected]

BACKGROUND: The aerial parts of Vitis vinifera (common grape or European grape)
have been widely used in Ayurveda to treat a variety of common and stress
related disorders. In the present investigation, the seed extract of V. vinifera
was evaluated for antistress activity in normal and stress induced rats.
Furthermore, the extract was studied for nootropic activity in rats and in-vitro
antioxidant potential to correlate its antistress activity. METHODS: For the
evaluation of antistress activity, groups of rats (n = 6) were subjected to
forced swim stress one hour after daily treatment of V. vinifera extract.
Urinary vanillylmandelic acid (VMA) and ascorbic acid were selected as
non-invasive biomarkers to assess the antistress activity. The 24 h urinary
excretion of vanillylmandelic acid (VMA) and ascorbic acid were determined by
spectrophotometric methods in all groups under normal and stressed conditions.
The nootropic activity of the extract as determined from acquisition, retention
and retrieval in rats was studied by conditioned avoidance response using Cook's
pole climbing apparatus. The in vitro antioxidant activity was determined based
on the ability of V. vinifera to scavenge hydroxyl radicals. RESULTS: Daily
administration of V. vinifera at doses of 100, 200 and 300 mg/kg body weight one
hour prior to induction of stress inhibited the stress induced urinary
biochemical changes in a dose dependent manner. However, no change in the
urinary excretion of VMA and ascorbic acid was observed in normal animals at all
the doses studied. The cognition, as determined by the acquisition, retention
and recovery in rats was observed to be dose dependent. The extract also
produced significant inhibition of hydroxyl radicals in comparison to ascorbic
acid in a dose dependent manner. CONCLUSION: The present study provides
scientific support for the antistress (adaptogenic), antioxidant and nootropic
activities of V. vinifera seed extract and substantiate the traditional claims
for the usage of grape fruits and seeds in stress induced disorders.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15656916 [PubMed - indexed for MEDLINE]

187: Phytochemistry. 2005 Jan;66(2):165-73.

Cyanogenic glucosides in grapevine: polymorphism, identification and
developmental patterns.

Franks TK, Hayasaka Y, Choimes S, van Heeswijck R.

Cooperative Research Centre for Viticulture, PO Box 154, Glen Osmond, SA 5064,
Australia. [email protected]

Twelve grapevine (Vitis vinifera L.) cultivars were surveyed for 'cyanide
potential' (i.e. the total cyanide measured in beta-glucosidase-treated crude,
boiled tissue extract) in mature leaves. Two related cultivars (Carignan and
Ruby Cabernet) had mean cyanide potential (equivalent to 110 mgHCNkg-1fr.wt) ca.
25-fold greater than that of the other 10 cultivars, and so the trait is
polymorphic in the species. In boiled leaf extracts of Carignan and Ruby
Cabernet, free cyanide constituted a negligible fraction of the total cyanide
potential because beta-glucosidase treatment was required to liberate the major
cyanide fraction - which is therefore bound in glucosylated cyanogenic
compound(s) (or cyanogenic glucosides). In addition, cyanide was liberated from
ground leaf tissue of Ruby Cabernet but not Sultana (a cultivar with low cyanide
potential). Hence, the high cyanide potential in Ruby Cabernet leaves is coupled
with endogenous beta-glucosidase(s) activity and this cultivar may be considered
'cyanogenic'. A method was developed to detect and identify cyanogenic
glucosides using liquid chromatography combined with tandem mass spectrometry
(LC-MS/MS). Two putative cyanogenic glucosides were found in extracts from
leaves of Carignan and Ruby Cabernet and were identified as the epimers prunasin
and sambunigrin. Cyanide potential measured at three times over the growing
season in young and mature leaves, petioles, tendrils, flowers, berries, seeds
and roots of Ruby Cabernet was substantially higher in the leaves compared with
all other tissues. This characterisation of cyanogenic glucoside accumulation in
grapevine provides a basis for gauging the involvement of the trait in
interactions of the species with its pests and pathogens.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15652573 [PubMed - indexed for MEDLINE]

188: Genome. 2004 Dec;47(6):1211-9.

Genetic relationship among cultivated and wild grapevine accessions from
Tunisia.

Snoussi H, Slimane MH, Ruiz-Garcia L, Martinez-Zapater JM, Arroyo-Garcia R.

Laboratory of Biotechnology, Tunisian National Agricultural Research Institute
(INRAT), Rue Hedi Harry, 2049, Ariana, Tunisia.

We have used nuclear and chloroplast molecular markers to genotype cultivated
and wild accessions of Vitis vinifera L. from Tunisia and assess their genetic
relationships. Fifty-five distinct genotypes were identified among 80 cultivated
accessions, including 18 genotypic groups containing between 2 and 5 accessions
per group. They could represent a total of 60 distinct cultivars owing to berry
colour variation found within identical genotype groups. Most of the 55
genotypes represent unique table grape genotypes except for one of them that was
found identical to the genotype of table grape cultivar Rosseti. Hybridization
among cultivars as well as self pollinations seems to have played an important
role in their origin since several groups of closely related cultivars were
observed. Furthermore, a parentage analysis showed a high probability for a
parent hybrid relationship within two groups of three cultivars. No strong
genetic similarities were found between cultivated and wild samples indicating
that the cultivated accessions do not derive from local Vitis vinifera L.
populations but could have been introduced from other regions in historic times.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15644980 [PubMed - indexed for MEDLINE]

189: J Agric Food Chem. 2005 Jan 12;53(1):49-56.

Stability of copigmented anthocyanins and ascorbic acid in a grape juice model
system.

Brenes CH, Del Pozo-Insfran D, Talcott ST.

Department of Food Technology, ITESM-Campus Monterrey, E. Garza Sada 2501 Sur,
C.P. 64849, Monterrey, N.L., Mexico. [email protected]

The stability of red grape anthocyanins (Vitis vinifera) was evaluated in a
model juice system during normal (25 degrees C) and accelerated storage (35
degrees C) in the presence of ascorbic acid. Rosemary polyphenolic cofactors (0,
0.2, and 0.4% v/v) were evaluated as anthocyanin stabilizing agents. Cofactor
addition resulted in concentration-dependent hyperchromic (up to 178%) and
bathochromic (up to 23 nm) shifts, indicating a more intense red coloration of
the models. Anthocyanin and ascorbic acid degradation followed first-order
kinetics during storage. Results showed that copigmented treatments underwent a
lower conversion of L-ascorbic acid into dehydroascorbic acid during storage
when compared to the control, favorably impacting the vitamin retention of these
models. Copigmentation did not affect anthocyanin degradation in the absence of
ascorbic acid but in its presence aided to retain a higher anthocyanin content
than the control. This study indicated that the addition of anthocyanin
cofactors could be used to reduce the pigment and vitamin degradation while
masking detrimental color changes in anthocyanin containing products.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 15631508 [PubMed - indexed for MEDLINE]

190: Electrophoresis. 2005 Jan;26(1):60-3.

Anomalous electrophoretic behavior of a chitinase isoform from grape berries and
wine in glycol chitin-containing sodium dodecyl sulfate-polyacrylamide gel
electrophoresis gels.

Vincenzi S, Curioni A.

Dipartimento di Biotecnologie agrarie, Facolta di Agraria, Universita di Padova,
Padova, Italy.

An anomalous electrophoretic behavior of a chitinase isoform present in both
grape (Vitis vinifera L.) berries and wine was observed in glycol
chitin-containing sodium dodecyl sulfate-polyacrylamide gel electrophoresis
(SDS-PAGE) gels. A progressive shift of the relative molecular mass M(r) of the
enzyme (from approximately 30,500 up to approximately 57,700) with increasing
glycol chitin concentration in the gels up to 0.1% was revealed when samples
were electrophoresed under nonreducing conditions, whereas the presence of
glycol chitin had no effects when samples were reduced before SDS-PAGE
separation. The M(r) of other grape and wine chitinase isoforms as well as that
of the chitinase from pomegranate (Punica granatum L.) fruit was unaffected by
the presence of the substrate in the gel under both reducing and nonreducing
conditions. Since the enzymes were inactive during the electrophoretic
separation, it is likely that the retarding effect of glycol chitin observed
specifically for the unreduced chitinase band from grape and wine was due to an
interaction between the substrate and a chitin-binding domain different from the
catalytic site, such as that typical of class I and class IV chitinases.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 15624140 [PubMed - indexed for MEDLINE]

191: J Mass Spectrom. 2005 Jan;40(1):83-90.

Mass spectrometry in the study of anthocyanins and their derivatives:
differentiation of Vitis vinifera and hybrid grapes by liquid
chromatography/electrospray ionization mass spectrometry and tandem mass
spectrometry.

Mazzuca P, Ferranti P, Picariello G, Chianese L, Addeo F.

Dipartimento di Scienza degli Alimenti, Universita degli Studi di Napoli
Federico II, Via Universita, I-80055 Portici, Naples, Italy.

A mass spectrometric-based procedure for anthocyanin profiling was set up to
distinguish authentic Vitis vinifera from hybrid red grapevine cultivars.
3-O-Monoglucoside and the related acetyl-, p-coumaryl- and
caffeoyl-monoglucoside anthocyanins occurred only in Vitis vinifera, whereas
3,5-O-diglucoside and the substituted acetyl-, p-coumaryl-, feruloyl- and
caffeoyl-diglucoside anthocyanins were the additional pigments in hybrid
grapevines. The procedure was applied expressly to identify red grape cultivars
based on the anthocyanin chemo-type determination. In particular, a red grape
cultivar, having 3,5-O-diglucoside anthocyanins and a novel class of anthocyanin
monoglucosides, such as cyanidin-3-O-, cyanidin-3-O-(6-O-acetyl)- and
cyanidin-3-O-(6-O-p-coumaryl)pentoside, was classified as hybrid. A second vine
cultivar, characterized exclusively by 3-O-monoglucoside anthocyanins, was
included among the Vitis vinifera species. Anthocyanin profiling by mass
spectrometry could represent the core of a chemotaxonomic procedure for
distinguishing American and European grapevines based on the identification of
post-synthetic anthocyanidin modification.

Publication Types:
Comparative Study

PMID: 15619268 [PubMed - indexed for MEDLINE]

192: Zhi Wu Sheng Li Yu Fen Zi Sheng Wu Xue Xue Bao. 2004 Jun;30(3):277-83.

[Induction and activity changes of protein kinase in grape leaves under heat
stress]

[Article in Chinese]

Yu SL, Huang WD.

College of Food Science and Nutritional Engineering, China Agricultural
University, Beijing 100083, China.

The type and activity of protein kinase induced and activated by heat stress
were studied in leaves of "Jingxiu" grape (Vitis vinifera L. cv. Jingxiu). The
results showed that a 52-kD protein kinase was evidently activated by heat
stress in 10 to 60 min. Myelin basic protein (MBP) embedded in gel could be
phosphorylated by the 52 kD protein kinase. The phosphorylating activity of this
kinase was determined by using MBP as the substrate and measuring the products
of phosphorylation by autoradiography. The 52 kD protein kinase could not affect
histone-III embedded as the substrate in gel and the products of phosphorylation
by autoradiography was not displayed. High phosphorylation activity of this
protein kinase was found with MBP substrate in solution reaction system. After
60 min heat stress, the activity of this protein kinase reached its maximum
value, and then declined rapidly. Compared with control, the activity of protein
kinase had no difference when histone-III was used as substrate in reaction
system. The activity of this protein kinase was not enhanced by Ca(2+), showing
that it was not Ca(2+)-dependent. The tyrosine-specific protein phosphatase
(YOP) could significantly inactivate the phosphorylation activity of protein
kinase. All results demonstrate that the 52 kD protein kinase activated by heat
stress belong to the MAPK family.

Publication Types:
English Abstract
Research Support, Non-U.S. Gov't

PMID: 15599023 [PubMed - indexed for MEDLINE]

193: Int J Biometeorol. 2005 May;49(5):303-9. Epub 2004 Dec 9.

Climate change and shifts in spring phenology of three horticultural woody
perennials in northeastern USA.

Wolfe DW, Schwartz MD, Lakso AN, Otsuki Y, Pool RM, Shaulis NJ.

Department of Horticulture, Cornell University, 14853 Ithaca, NY, USA.
[email protected]

We evaluated spring phenology changes from 1965 to 2001 in northeastern USA
utilizing a unique data set from 72 locations with genetically identical lilac
plants (Syringa chinensis, clone "Red Rothomagensis"). We also utilized a
previously validated lilac-honeysuckle "spring index" model to reconstruct a
more complete record of first leaf date (FLD) and first flower date (FFD) for
the region from historical weather data. In addition, we examined mid-bloom
dates for apple (Malus domestica) and grape (Vitis vinifera) collected at
several sites in the region during approximately the same time period. Almost
all lilac sites with significant linear trends for FLD or FFD versus year had
negative slopes (advanced development). Regression analysis of pooled data for
the 72 sites indicated an advance of -0.092 day/year for FFD (P=0.003). The
slope for FLD was also negative (-0.048 day/year), but not significant
(P=0.234). The simulated data from the "spring index" model, which relies on
local daily temperature records, indicated highly significant (P<0.001) negative
slopes of -0.210 and -0.123 day/year for FLD and FFD, respectively. Data
collected for apple and grape also indicated advance spring development, with
slopes for mid-bloom date versus year of -0.20 day/year (P=0.01) and -0.146
(P=0.14), respectively. Collectively, these results indicate an advance in
spring phenology ranging from 2 to 8 days for these woody perennials in
northeastern USA for the period 1965 to 2001, qualitatively consistent with a
warming trend, and consistent with phenology shifts reported for other mid- and
high-latitude regions.

Publication Types:
Historical Article
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 15592880 [PubMed - indexed for MEDLINE]

194: Micron. 2005;36(1):73-80.

Ultrastructural analysis of Vitis vinifera leaf tissues showing atypical
symptoms of Plasmopara viticola.

Musetti R, Stringher L, Borselli S, Vecchione A, Zulini L, Pertot I.

Dipartimento di Biologia Applicata alla Difesa delle Piante, Universita di
Udine, via delle Scienze, 208, 33100 Udine, Italy. [email protected]

In an abandoned farm in Tuscany a year by year regression of downy mildew
disease on grapevines was observed and a decrease in virulence as well as vigor
and fertility of the causal fungus, Plasmopara viticola. Anomalous spots of the
fungus (i.e. atypical coloration of leaves or mosaic) on leaf tissues of a
sensitive Vitis vinifera grapevine were observed. The anomalous symptoms were
often associated with the typical 'oil spots' and were present under
environmental conditions favourable for a normal development of the disease. An
ultrastructural study was carried out on leaf tissues of grapevine plants aimed
at clarifying the cause of this phenomenon and detecting whether there were
alterations in P. viticola mycelium and endophytes present. ELISA was also
performed to check the presence of grapevine viruses in the plants. TEM results
demonstrated that characteristic P. viticola was present in leaf samples showing
oil spots, while, both the fungus and the host tissues showed cytological
alterations in leaves with mosaic symptoms. Finally, hyphae were absent in leaf
tissues without downy mildew spots, but showing severe ultrastructural
modifications. Several plant virus infections were found in these grapevines.
Literature reports that the development and sporulation of some phytopathogenic
fungi inside their hosts can be limited by virus infections. Further
experimental approaches are required to determine if resistance to P. viticola
can be induced by viral infections in grapevines.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15582481 [PubMed - indexed for MEDLINE]

195: Commun Agric Appl Biol Sci. 2004;69(2):231-4.

Thermostability of Victoria grape (Vitis vinifera ssp. sativa)
polyphenoloxidase.

Rapeanu G, Van Loey A, Smout C, Hendrickx M.

Department of Food Bioengineering, Faculty of Food Science and Engineering,
University Dunarea de Jos, Galati, Romania.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15560229 [PubMed - indexed for MEDLINE]

196: Yi Chuan Xue Bao. 2004 Oct;31(10):1136-41.

[Cloning of a MADS box protein gene (GhMADS1) from cotton (Gossypium hirsutum
L.)]

[Article in Chinese]

Zheng SY, Guo YL, Xiao YH, Luo M, Hou L, Luo XY, Pei Y.

Key Laboratory of Biotechnology and Crop Quality Improvement Ministry of
Agriculture, Biotechnology Research Center, Southwest Agricultural University,
Chongqing 400716, China.

As a kind of transcription factors, MADS-box protein plays an important role in
various cellular processes, especially in the development of floral organs.
Based on the contig analysis of the cotton ESTs, the coding region of a cotton
MADS-box protein (GhMADS1) was obtained by RT-PCR from floral buds of cotton (G.
hirsutum). The cloned fragment of 713 bp (GhMADS1, GenBank accession no.
AF538965) contains an open reading frame of 711 bp,coding a polypeptide of 236
amino acids. It was demonstrated that the deduced GhMADS1 protein was highly
homologous to the AGL2 group of MADS-box proteins from Vitis vinifera, Nicotiana
sylvestris, Petunia hybrida, Arabidopsis thaliana and Antirrhinum majus.
Phylogenetic analysis also indicated that GhMADS1 belongs to the AGL2 group of
MADS-box proteins. RT-PCR analysis showed that GhMADS1 gene expressed in petals,
stamens, ovules and fibers, but not in roots, stems, leaves, bracts and sepals.
The strongest expression of GhMADS1 gene was detected in petals. But in floral
buds of a cotton homeotic mutant (CHV1), whose floral organs are all converted
to bract leaf-like organs, the transcript of GhMADS1 gene was not detected. It
was proposed that GhMADS1 gene would be crucial to the development of cotton
floral organs.

Publication Types:
English Abstract
Research Support, Non-U.S. Gov't

PMID: 15552050 [PubMed - indexed for MEDLINE]

197: Int J Food Sci Nutr. 2004 Aug;55(5):351-62.

Protection capacity against low-density lipoprotein oxidation and antioxidant
potential of some organic and non-organic wines.

Kalkan Yildirim H, Delen Akcay Y, Guvenc U, Yildirim Sozmen E.

Department of Food Engineering, Ege University, 35100 Bornova, Izmir, Turkey.

Current research suggests that phenolics from wine may play a positive role
against oxidation of low-density lipoprotein (LDL), which is a key step in the
development of atherosclerosis. Considering the effects of different wine-making
techniques on phenols and the wine consumption preference influencing the
benefical effects of the product, organically and non-organically produced wines
were obtained from the grapes of Vitis vinifera origin var: Carignan, Cabernet
Sauvignon, Merlot, Grenache, Columbard and Semillon. Levels of total phenols
[mg/l gallic acid equivalents (GAE)], antioxidant activity (%) and inhibition of
LDL oxidation [%, inhibition of diene and malondialdehyde (MDA) formation] were
determined. Some phenolic acids (gallic acid, p-hydroxybenzoic acid, syringic
acid, 2,3-dihydroxybenzoic acid, ferulic acid, p-coumaric acid and vanillic
acid) were quantified by high-performance liquid chromatography equipped with an
electrochemical detection carried at +0.65 V (versus Ag/AgCl, 0.5 microA full
scale). The highest concentrations of gallic, syringic and ferulic acids were
found in organic Cabernet Sauvignon; 2,3-dihydroxybenzoic acid in organic
Carignan and p-coumaric and vanillic acids in non-organic Merlot wine. High
levels of antioxidant activity (AOA), inhibition of LDL oxidation and total
phenol levels were found in non-organic Merlot (101.950% AOA; 88.570% LDL-diene;
41.000% LDL-MDA; 4700.000 mg/l GAE total phenol) and non-organic Cabernet
Sauvignon (92.420% AOA; 91.430% LDL-diene; 67.000% LDL-MDA; 3500.000 mg/l GAE
total phenol) grape varieties. Concentrations of some individual phenolic
constituents (ferulic, p-coumaric, vanillic) are correlated with high
antioxidant activity and inhibition of LDL oxidation.The best r value for all
examined characteristics was determined for gallic acid, followed by
2,3-dihydroxybenzoic, syringic, ferulic and p-coumaric acids. Negative
correlation of vanillic with MDA and p-hydroxybenzoic acid with LDL were
confirmed by principal component analysis (PCA) analyses. Red wines display a
higher antioxidant activity (81.110% AOA) than white ones (19.512% AOA). The
average level of LDL inhibition capacity in red wine was determined as 87.072%
and for the white as 54.867%.

PMID: 15545043 [PubMed - indexed for MEDLINE]

198: Mycol Res. 2004 Oct;108(Pt 10):1195-204.

Identification and characterization of Eutypa leptoplaca, a new pathogen of
grapevine in Northern California.

Trouillas FP, Gubler WD.

Department of Plant Pathology, University of California, Davis, California
95616, USA. [email protected]

Although Eutypa lata is the main agent of Eutypa dieback of grapevine, another
species of Eutypa has been isolated from diseased grapevines in Northern
California. Stromata of this recently discovered Eutypa were also collected from
Acer macrophyllum, A. negundo, and Fraxinus latifolia in the vicinity of
vineyards, and appeared commonly on Umbellularia californica in some
mixed-evergreen forests of Napa and Sonoma counties. This second species of
Eutypa was distinguished from E. lata because of the sulcate ostiole of the
perithecium and smaller ascospores. A morphological comparison with type
specimens revealed identical features between the Californian isolates and E.
leptoplaca sensu Rappaz (1987). This identification was confirmed through
phylogenetic analyses of Eutypa spp. based on the complete sequence of the
internal transcribed spacer (ITS) of the rDNA and partial sequence of the
beta-tubulin gene. These analyses also separated collections of E. maura, E.
sparsa, E. lejoplaca, E. tetragona, E. leptoplaca and E. lata, confirming the
previously proposed species concepts. The pathogenicity of E. leptoplaca on
grapevine was established using isolates collected from Vitis vinifera, U.
californica, and A. macrophyllum. The importance of E. leptoplaca in relation to
Eutypa dieback and its role as a necrotrophic pathogen are discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15535070 [PubMed - indexed for MEDLINE]

199: J Plant Physiol. 2004 Sep;161(9):1031-40.

Partitioning and mobilization of starch and N reserves in grapevine (Vitis
vinifera L.).

Zapata C, Deleens E, Chaillou S, Magne C.

Laboratoire de Biologie et Physiologie Vegetales, Universite de Reims
Champagne-Ardenne, UFR Sciences, UPRES EA 2069, BP 1039, 51687 Reims Cedex 2,
France.

We followed C and N reserves of grapevines grown in trenches under
semi-controlled conditions over a 3-year period after planting. Temporal
mobilization of stored C and N and subsequent distribution of reserve materials
within the vines were described in parallel with 15N uptake, particularly during
the third growing season. Storage C in the perennial tissues (roots, trunk,
canes) was mainly made of starch, which accumulated in the ray parenchyma of the
wood. In the permanent tissues, starch and total nitrogen contents were found to
decrease early in the development (bleeding sap, budbreak) whereas, on a
concentration basis, they decreased only after stage 7 (first leaf fully
expanded). Starch started to accumulate again in the perennial tissues during
flowering. The same observation was made with total nitrogen, although N levels
were much lower than those of starch. The 15N study showed that N uptake by the
roots started at budbreak and increased with vine development, becoming
predominant over reserve mobilization only after the onset of flowering. Taken
together, these results indicate that the spring growth period can be divided
into three main phases: In the first (dormancy to budbreak), significant losses
of C and N proceed mainly via root necrosis. In the second period (first leaf to
the onset of bloom), a strong mobilization of starch (and, to a lower extent, of
N) occurred for supporting vegetative and reproductive growth. At that point,
most of the C and N reserves used on the spring flush were those of the roots,
rather than those of the old wood (trunk, canes). In the third period (bloom and
early berry development), the mobilization process became low and was relieved
by N uptake (and CO2 assimilation) supplying nutrients to the sink structures.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15499905 [PubMed - indexed for MEDLINE]

200: Funct Integr Genomics. 2005 Jan;5(1):40-58. Epub 2004 Oct 5.

cDNA microarray analysis of developing grape (Vitis vinifera cv. Shiraz) berry
skin.

Waters DL, Holton TA, Ablett EM, Lee LS, Henry RJ.

Centre for Plant Conservation Genetics, Southern Cross University, Lismore, NSW,
2480, Australia. [email protected]

Microarray analysis of Vitis vinifera cv. Shiraz developing berries has revealed
the expression patterns of several categories of genes. Microarray slides were
constructed from 4,608 PCR-amplified cDNA clones derived from a ripening grape
berry cDNA library. The mRNA expression levels of the genes represented by these
cDNAs were measured in flowers, week 2 post-flowering whole berries, week 5,
week 8, week 10 (veraison, green berries), week 12 and week 13 berry skin. In
addition, a comparison of RNA expression in pigmented and unpigmented berry skin
at veraison (week 10) was undertaken. Image and statistical analysis revealed
four sets of genes with distinctive and similar expression profiles over the
course of berry development. The first set was composed of genes which had
maximum RNA expression in flowers, followed by a steady decrease in expression.
The most prominent group within this set were genes which have a role in
photosynthesis. The second set of cDNAs was dominated by genes involved in
flavonoid biosynthesis and had a peak of expression week 2 post-flowering. The
data indicate co-ordinate regulation of flavonoid biosynthetic genes which code
for the enzymes 4-coumarate-CoA ligase, chalcone synthase, chalcone isomerase,
flavonone hydroxylase, anthocyanidin reductase and cytochrome b5. The third set
of cDNAs exhibited maximum expression week 5 post-flowering, midway between
flowering and veraison, a period of rapid berry growth. This set of cDNAs is
dominated by genes which code for structural cell wall proteins. The fourth set
of genes was dramatically up-regulated at veraison and remained up-regulated
until 13 weeks post-flowering. This set of genes was composed of a diverse range
of genes, a reflection of the complexity of ripening, most with no known
function.

PMID: 15480888 [PubMed - indexed for MEDLINE]

201: Phytochemistry. 2004 Oct;65(19):2649-59.

Vitis vinifera terpenoid cyclases: functional identification of two
sesquiterpene synthase cDNAs encoding (+)-valencene synthase and (-)-germacrene
D synthase and expression of mono- and sesquiterpene synthases in grapevine
flowers and berries.

Lucker J, Bowen P, Bohlmann J.

Biotechnology Laboratory, Department of Botany, University of British Columbia,
6174 University Boulevard, Vancouver, BC, V6T 1Z3, Canada.

Valencene is a volatile sesquiterpene emitted from flowers of grapevine, Vitis
vinifera L. A full-length cDNA from the cultivar Gewurztraminer was functionally
expressed in Escherichia coli and found to encode valencene synthase (VvVal).
The two major products formed by recombinant VvVal enzyme activity with farnesyl
diphosphate (FPP) as substrate are (+)-valencene and (-)-7-epi-alpha-selinene.
Grapevine valencene synthase is closely related to a second sesquiterpene
synthase from this species, (-)-germacrene D synthase (VvGerD). VvVal and VvGerD
cDNA probes revealed strong signals in Northern hybridizations with RNA isolated
from grapevine flower buds. Transcript levels were lower in open pre-anthesis
flowers, flowers after anthesis, or at early onset of fruit development. Similar
results were obtained using a third probe, (-)-alpha-terpineol synthase, a
monoterpenol synthase. Sesquiterpene synthase and monoterpene synthase
transcripts were not detected in the mesocarp and exocarp during early stages of
fruit development, but transcripts hybridizing with VvVal appeared during late
ripening of the berries. Sesquiterpene synthase transcripts were also detected
in young seeds.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15464152 [PubMed - indexed for MEDLINE]

202: J Agric Food Chem. 2004 Oct 6;52(20):6122-8.

Effects of commercial anthocyanin-rich extracts on colonic cancer and
nontumorigenic colonic cell growth.

Zhao C, Giusti MM, Malik M, Moyer MP, Magnuson BA.

Department of Nutrition and Food Science, 0112 Skinner Building, University of
Maryland, College Park, MD 20742-7521, USA.

Commercially prepared grape (Vitis vinifera), bilberry (Vaccinium myrtillus L.),
and chokeberry (Aronia meloncarpa E.) anthocyanin-rich extracts (AREs) were
investigated for their potential chemopreventive activity against colon cancer.
The growth of colon-cancer-derived HT-29 and nontumorigenic colonic NCM460 cells
exposed to semipurified AREs (10-75 microg of monomeric anthocyanin/mL) was
monitored for up to 72 h using a sulforhodamine B assay. All extracts inhibited
the growth of HT-29 cells, with chokeberry ARE being the most potent inhibitor.
HT-29 cell growth was inhibited approximately 50% after 48 h of exposure to 25
microg/mL chokeberry ARE. Most importantly, the growth of NCM460 cells was not
inhibited at lower concentrations of all three AREs, illustrating greater growth
inhibition of colon cancer, as compared to nontumorigenic colon cells. Extracts
were semipurified and characterized by high-pressure liquid chromatography,
spectrophotometry, and colorimetry. Grape anthocyanins were the glucosylated
derivatives of five different anthocyanidin molecules, with or without
p-coumaric acid acylation. Bilberry contained five different anthocyanidins
glycosylated with galactose, glucose, and arabinose. Chokeberry anthocyanins
were cyanidin derivatives, monoglycosylated mostly with galactose and arabinose.
The varying compositions and degrees of growth inhibition suggest that the
anthocyanin chemical structure may play an important role in the growth
inhibitory activity of commercially available AREs.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15453676 [PubMed - indexed for MEDLINE]

203: Transgenic Res. 2004 Jun;13(3):203-14.

Expression of the stilbene synthase (StSy) gene from grapevine in transgenic
white poplar results in high accumulation of the antioxidant resveratrol
glucosides.

Giorcelli A, Sparvoli F, Mattivi F, Tava A, Balestrazzi A, Vrhovsek U, Calligari
P, Bollini R, Confalonieri M.

Istituto di Sperimentazione per la Pioppicoltura-MiPAF, via di Frassineto 35,
1-15033 Casale Monferrato(AL), Italy.

When present, stilbene synthase leads to the production of resveratrol
compounds, which are major components of the phytoalexin response against fungal
pathogens of the plant and are highly bioactive substances of pharmaceutical
interest. White poplar (Populus alba L.) was transformed with a construct
containing a cDNA insert encoding stilbene synthase from grapevine (Vitis
vinifera L.), under the control of the cauliflower mosaic virus (CaMV) 35S
promoter, and a chimeric kanamycin resistance gene. Southern blot hybridization
analysis demonstrated the presence and integration of exogenous DNA sequences in
the poplar genome. Expression of the stilbene synthase-encoding gene in
different transgenic lines was confirmed by Western blot and Northern analyses.
Compared to the controls, in the transgenic plants two new compounds were
detected and were identified as the trans- and cis-isomers of
resveratrol-3-glucoside (piceid) by high-pressure liquid chromatography (HPLC),
UV spectrophotometry, electrospray mass spectrometry (HPLC-ESI-MS) and enzymatic
hydrolysis. Since poplar is a good biomass producer and piceids are accumulated
in substantial amounts (up to 615.2 microg/g leaf fresh weight), the transgenic
plants represent a potential alternative source for the production of these
compounds with high pharmacological value. Despite the presence of piceid, in
our experimental conditions no increased resistance against the pathogen
Melampsora pulcherrima, which causes rust disease, was observed when in vitro
bioassays were performed.

Publication Types:
Comparative Study

PMID: 15359598 [PubMed - indexed for MEDLINE]

204: Plant Physiol Biochem. 2004 Jul-Aug;42(7-8):609-15.

Cloning and characterisation of a basic IAA oxidase associated with root
induction in Vitis vinifera.

Vatulescu AD, Fortunato AS, Sa MC, Amancio S, Ricardo CP, Jackson PA.

Instituto de Tecnologia Quimica e Biologica, Apartado 127, 2781-901 Oeiras,
Portugal.

Changes in apoplastic peroxidases during auxin-induced in vitro rooting of
cultured grapevine (Vitis vinifera L. cv. Touriga) stems have been studied. The
largest increase in peroxidase activity (EC 1.11.1.7) was associated with the
early stages of root initiation and could be attributed to an increase in
activity of an apoplastic 36 kDa cationic peroxidase (PxB2). Relative to other
peroxidases, PxB2 demonstrated high indole-3-acetic acid (IAA) oxidase activity
and apparently contributed the majority of potential IAA oxidase activity in
rooting tissues. The distribution of this peroxidase in developing roots
additionally associates it with early phases of growth restriction. PxB2 was
purified from cell wall extracts prepared from the basal 1 cm of rooting stems.
Microsequencing and subsequent cloning of its corresponding 3' truncated cDNA
(encoding 255 amino acids of the mature protein) revealed it to have a typical
class III peroxidase structure. The results suggest that this class III
peroxidase with IAA oxidase activity is important for the control of IAA levels
during root initiation and development. Copyright 2004 Elsevier SAS

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15331089 [PubMed - indexed for MEDLINE]

205: J Agric Food Chem. 2004 Aug 25;52(17):5396-403.

Determination of piceid and resveratrol in Spanish wines deriving from
Monastrell (Vitis vinifera L.) grape variety.

Moreno-Labanda JF, Mallavia R, Perez-Fons L, Lizama V, Saura D, Micol V.

Instituto de Biologia Molecular y Celular, Universidad Miguel Hernandez,
03202-Elche, Alicante, Spain.

The presence of stilbenes in wine is becoming an important issue due to their
claimed relation to a low incidence in coronary diseases and their increasing
implication as cancer chemopreventive and neuroprotective agents. Total
resveratrol content, quantified as glucoside and aglycone forms of resveratrol,
has been determined in a survey of 45 Monastrell monovarietal Spanish red wine
types (around 135 wine samples), belonging to Alicante and Bullas appellations.
The average between ratio glucoside/aglycone forms of resveratrol in these wines
was considerably high, ranging from 82 to 91% of resveratrol in its glycosidic
form. This characteristic was observed in a high percentage of the studied
wines, which were made under different winemaking procedures, and from different
vintages (1995-2002). In addition, wines made using macerative fermentations
with double amount of solid parts ("doble pasta") reached the highest levels of
total stilbene content expressed as resveratrol equivalent, i.e., 30 mg/L
(average of 18.8 mg/L). It can be concluded that high resveratrol glucoside
concentration and low free isomer content can be considered characteristics of
the Monastrell variety, as it happens to red wines deriving from other varieties
grown at warm climates. This fact, also observed for other French and Portuguese
red varieties, might play an important role in food habits involving these types
of wines.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15315376 [PubMed - indexed for MEDLINE]

206: Planta. 2004 Dec;220(2):241-50. Epub 2004 Aug 12.

Expression of the grapevine stilbene synthase gene VST1 in papaya provides
increased resistance against diseases caused by Phytophthora palmivora.

Zhu YJ, Agbayani R, Jackson MC, Tang CS, Moore PH.

Hawaii Agriculture Research Center, 99-193 Aiea Heights Drive, Aiea, Hawaii, HI
96701, USA. [email protected]

The phytoalexin resveratrol (trans-3,5,4'-trihydroxy-stilbene), a natural
component of resistance to fungal diseases in many plants, is synthesized by the
enzyme trihydroxystilbene synthase (stilbene synthase, EC 2.3.1.95), which
appears to be deficient or lacking in susceptible plants. Earlier workers
isolated a stilbene synthase gene (Vst1) from grapevine (Vitis vinifera L.),
which has subsequently been introduced as a transgene into a range of species to
increase resistance of hosts to pathogens to which they were originally
susceptible. Papaya (Carica papaya L.) is susceptible to a variety of fungal
diseases, including root, stem, and fruit rot caused by the pathogen
Phytophthora palmivora. Since resveratrol at 1.0 mM inhibited mycelium growth of
P. palmivora in vitro, we hypothesized that papaya resistance to this pathogen
might be increased by transformation with the grapevine stilbene synthase
construct pVst1, containing the Vst1 gene and its pathogen-inducible promoter.
Multiple transformed lines were produced, clonally propagated, and evaluated
with a leaf disk bioassay and whole plant response to inoculation with P.
palmivora. RNA transcripts of stilbene synthase and resveratrol glycoside were
induced in plant lines transformed with the grapevine pVst1 construct shortly
after pathogen inoculation, and the transformed papaya lines exhibited increased
resistance to P. palmivora. The immature transformed plants appear normal and
will be advanced to field trials to evaluate their utility.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 15309535 [PubMed - indexed for MEDLINE]

207: Acta Pharmacol Sin. 2004 Aug;25(8):1083-9.

Procyanidins from grape seeds protect against phorbol ester-induced oxidative
cellular and genotoxic damage.

Lu Y, Zhao WZ, Chang Z, Chen WX, Li L.

Department of Pharmacology, Marine Drug Research Center, Nanjing University of
Traditional Chinese Medicine, Nanjing 210029, China. [email protected]

AIM: To evaluate the inhibitory effects of Vitis vinifera procyanidins (PAs) on
carcinogen-induced oxidative stress. METHODS: The single cell gel
electrophoresis technique (comet assay) was employed to detect DNA damage
induced by the carcinogen phorbol-12-myristate-13-acetate (PMA). The release of
hydrogen peroxidase from polymorphonuclear leukocytes (PMNs) was assayed by the
horseradish peroxidase-mediated oxidation of phenol red. The microplate assay
was used to detect the presence of oxidative products by means of
2',7'-dichlorofluorescin-diacetate (DCFH-DA). The superoxide dismutase (SOD)
activity of liver mitochondria was assayed, based on the ability of SOD to
inhibit the generation of superoxidate anions by the xanthine-xanthine oxidase
system. The malondialdehyde (MDA) level was determined by the thiobarbituric
acid (TBA) assay. RESULTS: DNA of NIH3T3 cells was significantly damaged after
addition of PMA. The length of the comet tail was observed,while in normal cells
the comet tail could not be observed. PAs showed significant protective effects
on carcinogen PMA-induced DNA damage. Through assessment of DCFH-DA oxidation,
PAs were shown to inhibit the PMA-induced release of hydrogen peroxide by PMNs,
and to inhibit respiratory burst activity in NIH3T3 mouse fibroblasts. Ex vivo
study showed that serum from rats administered with PAs displayed similar
effects in a dose-dependent manner. In addition, PAs suppressed liver
mitochondrial lipid peroxidation induced by PMA. PAs protected the activity of
SOD and decreased the level of MDA in liver mitochondria damaged by PMA.
CONCLUSION: Dietary PAs from grape seeds protect against carcinogen-induced
oxidative cellular and genotoxic damage.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15301744 [PubMed - indexed for MEDLINE]

208: Microbiology. 2004 Aug;150(Pt 8):2527-34.

Killer toxin of Pichia membranifaciens and its possible use as a biocontrol
agent against grey mould disease of grapevine.

Santos A, Marquina D.

Department of Microbiology, Biology Faculty, Complutense University of Madrid,
Madrid 28040, Spain.

The use of Pichia membranifaciens CYC 1106 killer toxin against Botrytis cinerea
was investigated. This strain exerted a broad-specificity killing action against
other yeasts and fungi. At pH 4, optimal killer activity was observed at
temperatures up to 20 degrees C. At 25 degrees C the toxic effect was reduced to
70%. The killer activity was higher in acidic medium. Above about pH 4.5
activity decreased sharply and was barely noticeable at pH 6. The killer toxin
protein from P. membranifaciens CYC 1106 was purified to electrophoretic
homogeneity. SDS-PAGE of the purified killer protein indicated an apparent
molecular mass of 18 kDa. Killer toxin production was stimulated in the presence
of non-ionic detergents. The toxin concentrations present in the supernatant
during optimal production conditions exerted a fungicidal effect on a strain of
B. cinerea. The symptoms of infection and grey mould observed in Vitis vinifera
plants treated with B. cinerea were prevented in the presence of purified P.
membranifaciens killer toxin. The results obtained suggest that P.
membranifaciens CYC 1106 killer toxin is of potential use in the biocontrol of
B. cinerea.

PMID: 15289549 [PubMed - indexed for MEDLINE]

209: J Agric Food Chem. 2004 Jul 28;52(15):4821-7.

Stir bar sorptive extraction applied to volatile constituents evolution during
Vitis vinifera ripening.

Salinas M, Zalacain A, Pardo F, Alonso GL.

Catedra de Quimica Agricola, ETSI Agronomos, Universidad Castilla-La Mancha,
Campus Universitario, 02071 Albacete, Spain. [email protected]

The stir bar sorptive extraction (SBSE) technique is used to determine volatile
constituents in Monastrell grapes. The method proposed involves a maceration
step lasting 2 h and a sorption time of 6 h. The coefficients of variation
obtained were <4% in the case of the volatile compounds and <2% for the internal
standard. The behavior of the volatiles during grape ripening was studied, and
34 compounds were determined, among which geranyl acetone, geranyl butyrate,
farnesol, and a bicycloterpene, tentatively identified as
6-methylene[3.1.0]byciclohexane, were recorded for the first time in this
variety. The highest terpene and norisoprenoid contents together with the lowest
values of C6 compounds were observed during the fifth week of ripening, meaning
that this would be the optimal time for harvesting in terms of volatile
compounds. Copyright 2004 American Chemical Society

PMID: 15264921 [PubMed - indexed for MEDLINE]

210: Occup Environ Med. 2004 Aug;61(8):709-11.

Asymptomatic sensitisation to grapes in a sample of workers in the wine
industry.

Kalogeromitros D, Rigopoulos D, Gregoriou S, Mousatou V, Lyris N, Papaioannou D,
Katsarou-Katsari A.

Department of Allergy, University of Athens, Andreas Sygros Hospital, Athens,
Greece. [email protected]

AIMS: To assess the prevalence of sensitisation to grapes (Vitis vinifera var.
agiorghitiko) in a population with repeated exposure to grape allergens through
direct cutaneous contact as well as through the gastrointestinal tract. METHODS:
One hundred and twenty subjects were enrolled in each of four groups: grape
harvesters, winery workers in selection of grapes, winery workers operating
de-stemming/crushing/pressing machines, and administrative personnel.
Sensitisation to grapes was examined by skin prick-to-prick tests with fresh
fruit and juice. RESULTS: Eight harvesters and five workers in grape selection
had positive reaction to the grapes tested. No machine operators or
administrative personnel had positive tests. The likelihood of sensitisation was
estimated at 3.7% per year of occupation by logistic regression analysis. None
of the employees reported symptoms associated with sensitisation to grapes.
CONCLUSION: Asymptomatic sensitisation to grapes was detected only in workers
handling the fruit, suggesting that sensitisation is more likely to occur
through cutaneous exposure and/or minor wounding than through the
gastrointestinal tract. Prevalence rates were high and the clinical impact needs
to be further investigated.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15258279 [PubMed - indexed for MEDLINE]

211: Plant Physiol. 2004 Jul;135(3):1491-501. Epub 2004 Jul 9.

Floral meristem identity genes are expressed during tendril development in
grapevine.

Calonje M, Cubas P, Martinez-Zapater JM, Carmona MJ.

Departamento de Biotecnologia, Escuela Tecnica Superior Ingenieros Agronomos,
Universidad Politecnica de Madrid, 28040 Madrid, Spain.

To study the early steps of flower initiation and development in grapevine
(Vitis vinifera), we have isolated two MADS-box genes, VFUL-L and VAP1, the
putative FUL-like and AP1 grapevine orthologs, and analyzed their expression
patterns during vegetative and reproductive development. Both genes are
expressed in lateral meristems that, in grapevine, can give rise to either
inflorescences or tendrils. They are also coexpressed in inflorescence and
flower meristems. During flower development, VFUL-L transcripts are restricted
to the central part of young flower meristems and, later, to the prospective
carpel-forming region, which is consistent with a role of this gene in floral
transition and carpel and fruit development. Expression pattern of VAP1 suggests
that it may play a role in flowering transition and flower development. However,
its lack of expression in sepal primordia, does not support its role as an
A-function gene in grapevine. Neither VFUL-L nor VAP1 expression was detected in
vegetative organs such as leaves or roots. In contrast, they are expressed
throughout tendril development. Transcription of both genes in tendrils of very
young plants that have not undergone flowering transition indicates that this
expression is independent of the flowering process. These unique expression
patterns of genes typically involved in reproductive development have
implications on our understanding of flower induction and initiation in
grapevine, on the origin of grapevine tendrils and on the functional roles of
AP1-and FUL-like genes in plant development. These results also provide
molecular support to the hypothesis that Vitis tendrils are modified
reproductive organs adapted to climb.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15247405 [PubMed - indexed for MEDLINE]

212: Plant Physiol. 2004 Jul;135(3):1849-62. Epub 2004 Jul 9.

Novel expression pattern of cytosolic Gln synthetase in nitrogen-fixing root
nodules of the actinorhizal host, Datisca glomerata.

Berry AM, Murphy TM, Okubara PA, Jacobsen KR, Swensen SM, Pawlowski K.

Department of Environmental Horticulture, University of California, Davis,
California 95616, USA. [email protected]

Gln synthetase (GS) is the key enzyme of primary ammonia assimilation in
nitrogen-fixing root nodules of legumes and actinorhizal (Frankia-nodulated)
plants. In root nodules of Datisca glomerata (Datiscaceae), transcripts
hybridizing to a conserved coding region of the abundant nodule isoform,
DgGS1-1, are abundant in uninfected nodule cortical tissue, but expression was
not detectable in the infected zone or in the nodule meristem. Similarly, the GS
holoprotein is immunolocalized exclusively to the uninfected nodule tissue.
Phylogenetic analysis of the full-length cDNA of DgGS1-1 indicates affinities
with cytosolic GS genes from legumes, the actinorhizal species Alnus glutinosa,
and nonnodulating species, Vitis vinifera and Hevea brasilensis. The D.
glomerata nodule GS expression pattern is a new variant among reported root
nodule symbioses and may reflect an unusual nitrogen transfer pathway from the
Frankia nodule microsymbiont to the plant infected tissue, coupled to a
distinctive nitrogen cycle in the uninfected cortical tissue. Arg, Gln, and Glu
are the major amino acids present in D. glomerata nodules, but Arg was not
detected at high levels in leaves or roots. Arg as a major nodule nitrogen
storage form is not found in other root nodule types except in the
phylogenetically related Coriaria. Catabolism of Arg through the urea cycle
could generate free ammonium in the uninfected tissue where GS is expressed.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 15247391 [PubMed - indexed for MEDLINE]

213: J Agric Food Chem. 2004 Jul 14;52(14):4360-7.

Polyphenol screening of pomace from red and white grape varieties (Vitis
vinifera L.) by HPLC-DAD-MS/MS.

Kammerer D, Claus A, Carle R, Schieber A.

Institute of Food Technology, Section Plant Foodstuff Technology, Hohenheim
University, August-von-Hartmann-Strasse 3, D-70599 Stuttgart, Germany.

Phenolic compounds of 14 pomace samples originating from red and white
winemaking were characterized by HPLC-MS. Up to 13 anthocyanins, 11
hydroxybenzoic and hydroxycinnamic acids, and 13 catechins and flavonols as well
as 2 stilbenes were identified and quantified in the skins and seeds by
HPLC-DAD. Large variabilities comprising all individual phenolic compounds were
observed, depending on cultivar and vintage. Grape skins proved to be rich
sources of anthocyanins, hydroxycinnamic acids, flavanols, and flavonol
glycosides, whereas flavanols were mainly present in the seeds. However, besides
the lack of anthocyanins in white grape pomace, no principal differences between
red and white grape varieties were observed. This is the first study presenting
comprehensive data on the contents of individual phenolic compounds comprising
all polyphenolic subclasses of grapes including a comparison of several red and
white pomaces from nine cultivars. The results obtained in the present study
confirm that both skins and seeds of most grape cultivars constitute a promising
source of polyphenolics.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15237937 [PubMed - indexed for MEDLINE]

214: Ultramicroscopy. 2004 Aug;100(3-4):171-8.

Cell wall modification in grapevine cells in response to UV stress investigated
by atomic force microscopy.

Lesniewska E, Adrian M, Klinguer A, Pugin A.

Physics Laboratory LPUB UMR CNRS 5027, University of Bourgogne, B.P. 400,
F-21078 Dijon Cedex, France. [email protected]u-bourgogne.fr

Despite cell wall reinforcement being a well-known defence mechanism of plants,
it remains poorly characterized from a physical point of view. The objective of
this work was to further describe this mechanism. Vitis vinifera cv Gamay cells
were treated with UV-light (254 nm), a well-known elicitor of defence mechanisms
in grapevines, and physical cell wall modifications were observed using the
atomic force microscopy (AFM) under native conditions. The grapevine cell
suspensions were continuously observed in their culture medium from 30 min to
24h after elicitation. In the beginning, cellulose fibrils covered by a matrix
surrounded the control and treated cells. After 3 h, the elicited cells
displayed sprouted expansions around the cell wall that correspond to pectin
chains. These expansions were not observed on untreated grapevine cells. The AFM
tip was used to determine the average surface elastic modulus of cell wall that
account for cell wall mechanical properties. The elasticity is diminished in
UV-treated cells. In a comparative study, grapevine cells showed the same
decrease in cell wall elasticity when treated with a fungal biotic elicitor of
defence response. These results demonstrate cell wall strengthening by UV
stress.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15231307 [PubMed - indexed for MEDLINE]

215: Genome. 2004 Jun;47(3):579-89.

Diversification within grapevine cultivars goes through chimeric states.

Hocquigny S, Pelsy F, Dumas V, Kindt S, Heloir MC, Merdinoglu D.

Laboratoire de Genetique et d'Amelioration des Plantes, Unite Mixte de Recherche
1131 Institut National de la Recherche Agronomique, Universite Louis Pasteur,
Colmar, France.

Vitis vinifera 'Pinot' clones were analysed at 50 microsatellite loci to assess
intravarietal genetic diversity. When analysing leaf tissue DNAs, polymorphism
mainly resulted from the appearance of a third allele when two were expected for
heterozygous loci in a diploid species. The sequencing of the three
microsatellite alleles at two loci has confirmed their simultaneous presence in
the leaf tissues. A hypothesis explaining the triallelic profiles at a locus is
the presence of a periclinal chimera meristem structure, in which genetically
different cell layers coexist. The periclinal chimeric state of two Vitis
vinifera 'Pinot gris' clones was confirmed by splitting and analysing the
genotypes resulting from L1 and L2 cell layers in progeny derived from
self-fertilization, in root tissues, and in plants regenerated from somatic
embryogenesis. Prevalence of chimerism in polymorphic clones observed in a
collection of 145 accessions belonging to 'Pinot gris', 'Pinot noir', Pinot
blanc', 'Pinot meunier', and 'Pinot moure' cultivars was demonstrated. The
accumulation of somatic mutations and cell layer rearrangements allowed us to
deduce the relationships between the various genotypes and to open a way for
understanding the diversification process and the phylogeny in the 'Pinot'
group.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15190375 [PubMed - indexed for MEDLINE]

216: Theor Appl Genet. 2004 Sep;109(5):1017-27. Epub 2004 Jun 4.

Mapping 245 SSR markers on the Vitis vinifera genome: a tool for grape genetics.

Adam-Blondon AF, Roux C, Claux D, Butterlin G, Merdinoglu D, This P.

URGV, INRA, 2 rue Gaston Cremieux, 91058, Evry 5708, France. [email protected]

The aim of the present work was to develop a microsatellite marker-based map of
the Vitis vinifera genome (n=19), useful for genetic studies in this perennial
heterozygous species, as SSR markers are highly transferable co-dominant
markers. A total of 346 primer pairs were tested on the two parents (Syrah and
Grenache) of a full sib population of 96 individuals (S x G population),
successfully amplifying 310 markers. Of these, 88.4% markers were heterozygous
for at least one of the two parents. A total of 292 primer pairs were then
tested on Riesling, the parent of the RS1 population derived from selfing (96
individuals), successfully amplifying 299 markers among which 207 (62.9%) were
heterozygous. Only 6.7% of the markers were homozygous in all three genotypes,
stressing the interest of such markers in grape genetics. Four maps were
constructed based on the segregation of 245 SSR markers in the two populations.
The Syrah map was constructed from the segregations of 177 markers that could be
ordered into 19 linkage groups (total length 1,172.2 cM). The Grenache map was
constructed with the segregations of 178 markers that could be ordered into 18
linkage groups (total length 1,360.6 cM). The consensus S x G map was
constructed with the segregations of 220 markers that were ordered into 19
linkage groups (total length 1,406.1 cM). One hundred and eleven markers were
scored on the RS1 population, among them 27 that were not mapped using the S x G
map. Out of these 111 markers, 110 allowed to us to construct a map of a total
length of 1,191.7 cM. Using these four maps, the genome length of V. vinifera
was estimated to be around 2,200 cM. The present work allowed us to map 123 new
SSR markers on the V. vinifera genome that had not been ordered in a previous
SSR-based map (Riaz et al. 2004), representing an average of 6.5 new markers per
linkage group. Any new SSR marker mapped is of great potential usefulness for
many applications such as the transfer of well-scattered markers to other maps
for QTL detection, the use of markers in specific regions for the fine mapping
of genes/QTL, or for the choice of markers for MAS.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 15184982 [PubMed - indexed for MEDLINE]

217: Phytochemistry. 2004 May;65(9):1223-9.

Identification of Vitis vinifera (-)-alpha-terpineol synthase by in silico
screening of full-length cDNA ESTs and functional characterization of
recombinant terpene synthase.

Martin DM, Bohlmann J.

Biotechnology Laboratory, University of British Columbia, 6174 University
Boulevard, Rm. 237, Vancouver, BC, Canada V6T 1Z3.

The flavour and aroma of certain Vitis vinifera grape varieties is dominated by
volatile terpenes and small volatile aldehydes. Monoterpenes contribute to the
final grape and wine aroma and flavour in form of free volatiles and as
glycoside conjugates of monoterpene alcohols. Typical monoterpenol components of
the cultivar Gewurztraminer and other aroma-rich grape varieties are linalool,
geraniol, nerol, citronellol, and alpha-terpineol. In a functional genomics
effort to identify genes for the formation of monoterpene alcohols in V.
vinifera, a database of full-length cDNA sequences was screened in silico and
yielded two clones for putative monoterpene synthases. The gene products were
functionally characterized by expression in Escherichia coli, in vitro enzyme
assay and gas chromatography-mass spectrometry (GC-MS) product identification as
multi-product (-)-alpha-terpineol synthases.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15184006 [PubMed - indexed for MEDLINE]

218: J Agric Food Chem. 2004 Jun 2;52(11):3492-7.

Influence of vine training and sunlight exposure on the
3-alkyl-2-methoxypyrazines content in musts and wines from the Vitis vinifera
variety cabernet sauvignon.

Sala C, Busto O, Guasch J, Zamora F.

Departament de Quimica Analitica i Quimica Organica, Facultat d' Enologia de
Tarragona, Universitat Rovira i Virgili, Avda. Ramon y Cajal, 70, E-43005
Tarragona, Spain. [email protected]

The influence of vine training and sunlight exposure on the
3-alkyl-2-methoxypyrazines contents in musts and wines was studied by means of
two previously reported methods based on headspace solid-phase micro-extraction.
Experimental samples were monitored throughout grape ripening and wine making.
3-Isobutyl-2-methoxypyrazine, 3-sec-butyl-2-methoxypyrazine and
3-isopropyl-2-methoxypyrazine were identified. The 3-isobutyl-2-methoxypyrazine
content decreased throughout grape ripening in all of the sample types studied.
After 1 day of maceration with the skins, there was an increase, but after
racking, no further increase was observed. No significant differences between
samples were found during grape ripening. Wines from goblet-trained vines,
however, contained significantly less 3-isobutyl-2-methoxypyrazine. Clusters
protected from sunlight since the beginning of the veraison resulted in wines
with a significantly lower content of this compound than the control samples.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15161221 [PubMed - indexed for MEDLINE]

219: J Agric Food Chem. 2004 Jun 2;52(11):3467-72.

Polyphenol glucosylating activity in cell suspensions of grape (Vitis vinifera).

Krasnow MN, Murphy TM.

Division of Biological Sciences, University of California, One Shields Ave,
Davis, CA, 95616, USA. [email protected]

Stilbenes are phenolic molecules that have antifungal effects in the plant and
antioxidant and anti-cancer effects when consumed in the human diet.
Glycosylation of stilbenes increases their solubility and may make them more
easily absorbed by the intestine. We have found an activity in extracts of
cultured cells of Vitis vinifera (cv. Gamay Freaux) that glucosylates the
stilbene resveratrol to form piceid. The Km for UDP-Glucose was 1.2 mM, and the
Km for resveratrol was 0.06 mM, values similar to those of other phenolic
glucosyltransferases. We investigated the resveratrol glucosylating activity of
the enzyme extracted from cells grown under different light treatments (dark,
visible light, light + ultraviolet (UVC) radiation) and found the activity to be
unaffected or slightly reduced. In contrast, UVC light strongly stimulated
extractable quercetin glucosyltransferase activity. These results, combined with
analysis of phenolic compounds extracted from the differently treated cells,
suggest that the resveratrol glucosyltransferase is distinct from the
glucosyltransferase(s) active on other phenolics.

PMID: 15161217 [PubMed - indexed for MEDLINE]

220: Physiol Plant. 2004 Jun;121(2):272-281.

Processes contributing to photoprotection of grapevine leaves illuminated at low
temperature.

Hendrickson L, Forster B, Furbank RT, Chow WS.

Research School of Biological Sciences, Australian National University,
Canberra, ACT 0200, Australia.

Photoinactivation of photosystem II (PSII) and energy dissipation at low leaf
temperatures were investigated in leaves of glasshouse-grown grapevine (Vitis
vinifera L. cv. Riesling). At low temperatures (< 15 degrees C), photosynthetic
rates of CO(2) assimilation were reduced. However, despite a significant
increase in the amount of light excessive to that required by photosynthesis,
grapevine leaves maintained high intrinsic quantum efficiencies of PSII
(F(v)/F(m)) and were highly resistant to photoinactivation compared to other
species. Non-photochemical energy dissipation involving xanthophylls and fast D1
repair were the main protective processes reducing the 'gross' rate of
photoinactivation and the 'net' rate of photoinactivation, respectively. We
developed an improved method of energy dissipation analysis that revealed up to
75% of absorbed light is dissipated thermally via pH- and xanthophyll-mediated
non-photochemical quenching at low temperatures (5-15 degrees C) and moderate
(800 micro mol quanta m(-2) s(-1)) light. Up to 20% of the energy flux
contributing to electron transport was dissipated via photorespiration when
taking into account temperature-dependent mesophyll conductance; however, this
flux used in photorespiration was only a relatively small amount of the total
absorbed light energy. Photoreduction of O(2) at photosystem I (PSI) and
subsequent superoxide detoxification (water-water cycle) was more sensitive to
inhibition by low temperature than photorespiration. Therefore the water-water
cycle represents a negligibly small energy sink below 15 degrees C, irrespective
of mesophyll conductance.

PMID: 15153195 [PubMed - as supplied by publisher]

221: Drugs Exp Clin Res. 2003;29(5-6):207-16.

Procyanidins from Vitis vinifera seeds display cardioprotection in an
experimental model of ischemia-reperfusion damage.

Berti F, Manfredi B, Mantegazza P, Rossoni G.

Department of Pharmacology, Chemotherapy and Medical Toxicology, University of
Milan, Italy. [email protected]

Since the early 1970s, increasing evidence has suggested that the consumption of
moderate amounts of alcohol is inversely correlated with mortality from
myocardial infarction. There is also some evidence that the protective effects
of wine might be more pronounced than those of other alcoholic beverages. These
observations prompted us to investigate the cardioprotective activity of Vitis
vinifera seeds in experimental ischemia-reperfusion injury. An isolated rabbit
heart preparation paced electrically was used to evaluate the effects of a
highly purified, high molecular weight fraction of oligomeric procyanidins
isolated from Vitis vinifera seeds on myocardial reperfusion injury after 40 min
of low-flow (1 ml/min) ischemia. Infusion of the heart with 100 or 200 microg/ml
procyanidins dose-dependently reduced left ventricular end-diastolic pressure
during ischemia, decreased coronary perfusion pressure, improved cardiac
mechanical performance upon reperfusion, increased the release of
6-Keto-prostaglandin F1alpha into the perfusate in both the preischemic and the
reperfusion periods and suppressed rhythm irregularity. Procyanidins
dose-dependently relaxed human internal mammary aortic (IMA) rings (with intact
endothelium) precontracted with norepinephrine. This effect was completely
abolished in IMA-rings without functional endothelium or when this vascular
tissue was pretreated with nitric oxide synthase inhibitor
(NG-monomethyl-L-arginine) or with guanylate cyclase inhibitor
(1H-[1,2,4]oxadiazolo[4,3-a]quinoxalin-1-one). In conclusion, these results
indicate that procyanidins could be of therapeutical potential in cardiovascular
diseases. However, further investigations are required for a better definition
of the mode of action of these oligomers.

Publication Types:
In Vitro

PMID: 15134376 [PubMed - indexed for MEDLINE]

222: J Exp Bot. 2004 Jun;55(401):1423-31. Epub 2004 May 7.

Combining thermal and visible imagery for estimating canopy temperature and
identifying plant stress.

Leinonen I, Jones HG.

University of Dundee at SCRI, Plant Science Research Group, Invergowrie, Dundee
DD2 5DA, Scotland, UK. [email protected]

Thermal imaging is a potential tool for estimating plant temperature, which can
be used as an indicator of stomatal closure and water deficit stress. In this
study, a new method for processing and analysing thermal images was developed.
By using remote sensing software, the information from thermal and visible
images was combined, the images were classified to identify leaf area and sunlit
and shaded parts of the canopy, and the temperature statistics for specific
canopy components were calculated. The method was applied to data from a
greenhouse water-stress experiment of Vicia faba L. and to field data for Vitis
vinifera L. Vaseline-covered and water-sprayed plants were used as dry and wet
references, respectively, and two thermal indices, based on temperature
differences between the canopy and reference surfaces, were calculated for
single Vicia faba plants. The thermal indices were compared with measured
stomatal conductance. The temperature distributions of sunlit and shaded leaf
area of Vitis vinifera canopies from natural rainfall and irrigation treatments
were compared. The present method provides two major improvements compared with
earlier methods for calculating thermal indices. First, it allows more accurate
estimation of the indices, which are consequently more closely related to
stomatal conductance. Second, it gives more accurate estimates of the
temperature distribution of the shaded and sunlit parts of canopy, and, unlike
the earlier methods, makes it possible to quantify the relationship between
temperature variation and stomatal conductance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15133055 [PubMed - indexed for MEDLINE]

223: J Agric Food Chem. 2004 Apr 21;52(8):2292-6.

Analysis of the oxidative degradation of proanthocyanidins under basic
conditions.

Jorgensen EM, Marin AB, Kennedy JA.

Department of Food Science and Technology, Oregon State University, Corvallis,
Oregon 97331, USA.

Proanthocyanidin isolates from grape (Vitis vinifera L. cv. Pinot noir) skin and
seed underwent oxidative degradation in solution (10 g/L) under basic conditions
while exposed to atmospheric oxygen. Degradation was monitored by reversed-phase
HPLC following acid-catalyzed cleavage in the presence of excess phloroglucinol
(phloroglucinolysis) and by high-performance gel permeation chromatography. All
isolates degraded under these conditions and followed second-order kinetics for
over 1 half-life, consistent with an oxidation reaction. The conversion of
proanthocyanidins to known subunits (conversion yield) when measured by
phloroglucinolysis showed a dramatic decline over the course of the reaction.
With the exception of (+)-catechin extension subunits, all individual subunits
decreased in concentration during the oxidation process, also following
second-order kinetics for over 1 half-life. Skin proanthocyanidins degraded the
fastest due to the presence of (-)-epigallocatechin extension subunits. Seed
procyanidins were degraded with and without flavan-3-ol monomers. Flavan-3-ol
monomers slowed the rate of seed procyanidin degradation. The mean degree of
polymerization (mDP) determined by phloroglucinolysis indicated a large decrease
in mDP as the reaction progressed; yet, by GPC, the size distribution of all
proanthocyanidins changed little in comparison. The conversion yield could be an
important parameter to follow when using phloroglucinolysis as a means for
determining proanthocyanidin mDP, and when monitoring the oxidative degradation
of proanthocyanidins.

PMID: 15080635 [PubMed - indexed for MEDLINE]

224: J Chem Ecol. 2004 Jan;30(1):69-89.

Qualitative and quantitative variation among volatile profiles induced by
Tetranychus urticae feeding on plants from various families.

van den Boom CE, van Beek TA, Posthumus MA, de Groot A, Dicke M.

Phytochemical Section, Laboratory of Organic Chemistry, Wageningen University,
Dreijenplein 8, 6703 HB Wageningen, The Netherlands. [email protected]

Many plant species are known to emit herbivore-induced volatiles in response to
herbivory. The spider mite Tetranychus urticae Koch is a generalist that can
feed on several hundreds of host plant species. Volatiles emitted by T.
urticae-infested plants of 11 species were compared: soybean (Glycine max),
golden chain (Laburnum anagyroides), black locust (Robinia pseudo-acacia),
cowpea (Vigna unguiculata), tobacco (Nicotiana tabacum), eggplant (Solanum
melalonga), thorn apple (Datura stramonium), sweet pepper (Capsicum annuum), hop
(Humulus lupulus), grapevine (Vitis vinifera), and ginkgo (Ginkgo biloba). The
degree to which the plant species produced novel compounds was analyzed when
compared to the odors of mechanically damaged leaves. Almost all of the
investigated plant species produced novel compounds that dominated the volatile
blend, such as methyl salicylate, terpenes, oximes, and nitriles. Only spider
mite-infested eggplant and tobacco emitted a blend that was merely
quantitatively different from the blend emitted by mechanically damaged or clean
leaves. We hypothesized that plant species with a low degree of direct defense
would produce more novel compounds. However, although plant species with a low
direct defense level do use indirect defense to defend themselves, they do not
always emit novel compounds. Plant species with a high level of direct defense
seem to invest in the production of novel compounds. When plant species of the
Fabaceae were compared to plant species of the Solanaceae, qualitative
differences in spider mite-induced volatile blends seemed to be more prominent
in the Fabaceae than in the Solanaceae.

Publication Types:
Comparative Study

PMID: 15074658 [PubMed - indexed for MEDLINE]

225: Tsitol Genet. 2003 Nov-Dec;37(6):31-8.

[Study of the intra-cultivar variability of grape DNA using RFLP and PCR
methods]

[Article in Russian]

Barysheva IA, Tulaeva MI, Chisnikov VS.

Evaluation of DNA polymorphism among Vitis vinifera varieties using RFLP and PCR
methods has been performed to choose a DNA technology for detection of grape
intracultivar variation. DNA polymorphism of clones of the varieties Riparia
Gluar, Riparia x Rupestris 101-14, Cabernet Sauvignon, Riestling reinskiy has
been studied using Southern hybridization and amplification techniques. It has
been shown that grape intracultivar variability of rDNA in Riparia x Rupestris
101-14 and Cabernet Sauvignon clones was caused by the modification in Alul
restriction sites of rDNA. DNA variability of the randomly amplified and
inter-SSR sequences of the Riparia Gluar, Riparia x Rupestris 101-14, Cabernet
Sauvignon clones was also detected. A set of molecular DNA loci which can be
used for grape clone identification has been obtained.

Publication Types:
English Abstract

PMID: 15067943 [PubMed - indexed for MEDLINE]

226: J Biotechnol. 2004 Apr 29;109(3):287-94.

Production of highly 13C-labeled polyphenols in Vitis vinifera cell bioreactor
cultures.

Aumont V, Larronde F, Richard T, Budzinski H, Decendit A, Deffieux G, Krisa S,
Merillon JM.

Groupe d'Etudes des Substances Vegetales a Activite Biologique, EA 3675,
Universite de Bordeaux 2, Faculte des Sciences Pharmaceutiques, 146 rue Leo
Saignat, 33076 Bordeaux Cedex, France.

The use of Vitis vinifera cells grown in a 2 l-stirred tank bioreactor for
producing isotopically 13C-labeled phenolic substances is presented. Several
culture parameters were optimized to achieve characteristics of growth and
polyphenol metabolism similar to that recorded in shake flasks. Administration
of [1-13C]L-phenylalanine (3 mM) to grape cell suspension cultures led to the
production of 13C-labeled stilbenes (trans- and cis-piceids), catechins
(catechin and epicatechin) and anthocyanins (delphinidin-, cyanidin-,
petunidin-, peonidin- and malvidin-3-O-beta-glucosides). Incorporation of
[1-13C]L-phenylalanine into polyphenols was measured by means of 13C satellites
in the proton NMR spectrum and EA-IRMS. The enrichment of labeling obtained for
all the compounds (between 40 and 65%) is sufficient to investigate their
absorption and metabolism in humans.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15066766 [PubMed - indexed for MEDLINE]

227: J Agric Food Chem. 2004 Apr 7;52(7):2036-41.

Enantioselective analysis of free and glycosidically bound monoterpene polyols
in Vitis vinifera L. cvs. Morio Muscat and Muscat Ottonel: evidence for an
oxidative monoterpene metabolism in grapes.

Luan F, Hampel D, Mosandl A, Wust M.

Institut fur Lebensmittelchemie, Johann Wolfgang Goethe-Universitat Frankfurt am
Main, Marie-Curie-Strasse 9, D-60439 Frankfurt (Main), Germany.

The enantiomeric ratios of various free and glycosidically bound monoterpene
polyols in musts of the aromatic grapes Vitis vinifera L. cvs. Morio Muscat and
Muscat Ottonel were determined by means of enantioselective multidimensional gas
chromatography-mass spectrometry. Reference compounds of defined stereochemistry
were synthesized and permitted the unequivocal determination of the elution
order of the target compounds on chiral columns with modified cyclodextrins as
stationary phases. It could be shown for the first time that the
linalool-derived polyol 3,7-dimethylocta-1,7-dien-3,6-diol occurs predominately
as the (3S,6S)-configured stereoisomer, providing evidence that this compound is
generated by an enzymatic process in grape berries. The involvement of a
cytochrome P450 monooxygenase in the oxidative metabolism of monoterpenes in
grapes is dicussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15053548 [PubMed - indexed for MEDLINE]

228: Ann Bot (Lond). 2004 May;93(5):581-90. Epub 2004 Mar 24.

Interaction between Vitis vinifera and grape phylloxera: changes in root tissue
during nodosity formation.

Kellow AV, Sedgley M, Van Heeswijck R.

The University of Adelaide, Department of Horticulture, Viticulture and
Oenology, PMB 1 Glen Osmond 5064, Australia. [email protected]

BacKGROUND AND AIMS: The interaction between the gall-forming grapevine
parasite, phylloxera, and the susceptible grapevine species Vitis vinifera was
investigated. METHODS: Phylloxera and grapevines were cocultivated using both
potted and micropropagated grapevines. Development of nodosities on primary
roots was studied by microscopy and histochemistry, and nodosities were analysed
for biochemical changes and changes in gene expression. KEY RESULTS: Within a
nodosity, phylloxera fed at a site in the root cortex. Nodosity development was
characterized by swelling of the root tissue distal to the feeding site, lack of
development of a suberized endodermis, and starch and amino acid accumulation,
and was eventually followed by root necrosis. No evidence of a defence response
was observed in pre-necrotic nodosities, but defence-type responses were
observed in tissue adjacent to necrotic regions. Changes in gene expression were
not detected by northern hybridization using DNA probes encoding a range of V.
vinifera transcripts. CONCLUSIONS: Nodosities on V. vinifera potentially
function as nutrient reservoirs, and defence responses to phylloxera attack were
not detected.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 15044213 [PubMed - indexed for MEDLINE]

229: Physiol Plant. 2004 Apr;120(4):657-666.

Polyamines and somatic embryogenesis in two Vitis vinifera cultivars.

Bertoldi D, Tassoni A, Martinelli L, Bagni N.

Department of Biology e.s. and Interdepartmental Centre for Biotechnology,
University of Bologna, Via Irnerio 42, 40126, Bologna, Italy.

Polyamine content and activities of enzymes of polyamine biosynthesis were
assayed during somatic embryogenesis in Vitis vinifera callus cultures of
Chardonnay and Brachetto 'a grappolo lungo' (Brachetto g.l.) cultivars. The
analyses were carried out on embryogenic callus samples, embryos at different
stages and developing plants. Polyamine content, both in the free and
PCA-soluble conjugated form, was higher in Brachetto g.l. than in Chardonnay,
and putrescine was present at higher concentrations than the other polyamines.
In all samples of both cultivars, ornithine decarboxylase activity (ODC, EC
4.1.1.17) was higher than arginine decarboxylase (ADC, EC 4.1.1.19), with a
maximum in developing plant roots. S-Adenosylmethionine decarboxylase (SAMDC, EC
4.1.1.50) activity displayed a similar trend. The activities of all three
enzymes were detected both in the supernatant and pellet fractions, indicating
for the first time the presence of SAMDC activity in the particulate fraction.
Particularly in the Chardonnay cultivar, an increase in the mRNAs expression
patterns of ODC and SAMDC during morphogenesis from small embryos to plantlets
was detected by northern blot, suggesting a direct correlation with enzymatic
activities.

PMID: 15032828 [PubMed - as supplied by publisher]

230: J Agric Food Chem. 2004 Mar 10;52(5):1300-6.

Occurrence of pyranoanthocyanins in sparkling wines manufactured with red grape
varieties.

Pozo-Bayon MA, Monagas M, Polo MC, Gomez-Cordoves C.

Instituto de Fermentaciones Industriales, CSIC, Juan de la Cierva 3, 28006
Madrid, Spain.

The anthocyanin pigments in rose (Vitis vinifera cv. Garnacha) and blanc de noir
(V. vinifera cv. Monastrell) base and sparkling wines were studied by
LC-DAD/ESI-MS. Anthocyanins of grape origin and pyranoanthocyanins resulting
from C-4/C-5 cycloaddition of the former ones with pyruvic acid, acetaldehyde,
4-vinylphenol, 4-vinylguaiacol, and 4-vinylcatechol were identified in the
different wines. Rose wines presented a higher total pigment content than blanc
de noir wines. Pyranoanthocyanins represented 68.9-76.0% of total pigment
content in rose wines and 49.4-60.7% in blanc de noir wines. Malvidin
3-glucoside-pyruvate was the most abundant pigment in both rose and blanc de
noir base wines. Important qualitative and quantitative changes were observed in
terms of the anthocyanin and pyranoanthocyanin pigments after the second
(bottle) fermentation and 9 months of aging on yeast lees, but not after a
further time (3-9 additional months) of aging on lees. Evaluation of the wine
color characteristics was consistent with a greater color stability for the rose
sparkling wines that could be associated with the high content, structural
diversity, and spectroscopic features of the pyranoanthocyanins present in these
wines.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14995137 [PubMed - indexed for MEDLINE]

231: J Agric Food Chem. 2004 Mar 10;52(5):1241-7.

Effect of fungicide residues on the aromatic composition of white wine
inoculated with three Saccharomyces cerevisiae strains.

Garcia MA, Oliva J, Barba A, Camara MA, Pardo F, Diaz-Plaza EM.

Departamento de Quimica Agricola, Geologia y Edafologia, Facultad de Quimica,
Universidad de Murcia, Campus de Espinardo, 30100 Murcia, Spain.

The effects of three fungicide residues (cyprodinil, fludioxonil, and
pyrimethanil) on the aromatic composition (acids, alcohols, and esters) of Vitis
vinifera white wines (var. Airen) inoculated with three Saccharomyces cerevisiae
strains (syn. bayanus, cerevisiae, and syn. uvarum) are studied. The aromatic
exponents were extracted and concentrated by adsorption-thermal desorption and
were determined by gas chromatography using a mass selective detector. The
addition of the three fungicides at different doses (1 and 5 mg/L) produces
significant differences in the acidic fraction of the aroma, especially in the
assays inoculated with S. cerevisiae, although the final contents do not exceed
the perception thresholds. The lower quality wines, according to isomeric
alcohol content [(Z)-3-hexen-1-ol and 3-(methylthio)propan-1-ol] are those
obtained by inoculation with S. cerevisiae(syn. bayanus) and addition of
cyprodinil. The addition of fungicides in the assays inoculated with S.
cerevisiae (syn. bayanus) produces an increase in the ethyl acetate and isoamyl
acetate contents, which causes a decrease in the sensorial quality of the wine
obtained.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14995128 [PubMed - indexed for MEDLINE]

232: J Agric Food Chem. 2004 Feb 25;52(4):891-7.

Effect of antioxidant protection of must on volatile compounds and aroma shelf
life of Falanghina (Vitis vinifera L.) wine.

Moio L, Ugliano M, Genovese A, Gambuti A, Pessina R, Piombino P.

Dipartimento di Scienze degli Alimenti, Universita degli Studi di Foggia, Via
Napoli 25, 71100 Foggia, Italy. [email protected]

Two vinification methods involving different degrees of antioxidant protection
of Falanghina must during prefermentative steps, and referred as HAMP (high
antioxidant must protection) and LAMP (low antioxidant must protection), were
compared in terms of fermentation performances of four different yeast strains,
composition of the volatile fraction of wines at the end of alcoholic
fermentation, and shelf life of wines during storage. The use of HAMP technology
resulted in wines with lower volatile acidity and higher concentrations of
medium-chain fatty acid ethyl esters, acetates, and volatile fatty acids. For
two of the four strains a lower concentration of isoamyl alcohol was also
observed. HAMP wines also revealed increased shelf life because of the higher
concentration of odor active esters at the end of storage and better
preservation of varietal aromas.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14969547 [PubMed - indexed for MEDLINE]

233: FEMS Microbiol Lett. 2004 Jan 30;230(2):283-90.

Colony aggregation and biofilm formation in xylem chemistry-based media for
Xylella fastidiosa.

Leite B, Andersen PC, Ishida ML.

North Florida Research and Education Center, Institute of Food and Agricultural
Sciences, University of Florida, 155 Research Road, Quincy, FL 32351, USA.
[email protected]

Two chemically defined media based on xylem fluid chemistry were developed for
Xylella fastidiosa. These media were tested and compared to chemically defined
media XDM2, XDM4 and XF-26. New media were evaluated for the Pierce's disease
(PD) strain UCLA-PD. Our media either was similar to the concentration of some
amino acids found in the xylem fluid of the PD-susceptible Vitis vinifera cv.
Chardonnay (medium CHARD2) or incorporated the tripeptide glutathione found in
xylem fluid composition (medium 3G10-R). CHARD2 and 3G10-R are among the
simplest chemically defined media available. Xylem fluid chemistry-based media
supported X. fastidiosa growth and especially stimulated aggregation and biofilm
formation.

Publication Types:
Evaluation Studies
Research Support, Non-U.S. Gov't

PMID: 14757251 [PubMed - indexed for MEDLINE]

234: J Agric Food Chem. 2004 Feb 11;52(3):498-504.

Investigations on anthocyanins in wines from Vitis vinifera cv. pinotage:
factors influencing the formation of pinotin A and its correlation with wine
age.

Schwarz M, Hofmann G, Winterhalter P.

Institute of Food Chemistry, Technical University of Braunschweig,
Schleinitzstrasse 20, 38106 Braunschweig, Germany.

Pinotage red wines were found to contain a reaction product of malvidin
3-glucoside and caffeic acid, the so-called pinotin A. A total of 50 Pinotage
wines from the vintages 1996-2002 were analyzed for the content of pinotin A,
malvidin 3-glucoside, caffeic acid, and caftaric acid. Statistical analyses were
performed to reveal variations in the content of these compounds and to
determine the factors that influence pinotin A formation during wine aging. An
exponential increase of the concentration of this aging product was observed
with prolonged storage time. The most rapid synthesis of pinotin A was observed
in 2.5-4 year old wines, although at this age malvidin 3-glucoside is already
degraded to a large extent. This phenomenon is explained by the increased ratio
of caffeic acid/malvidin 3-glucoside, which strongly favors the formation of
pinotin A and makes side reactions less likely. Pinotin A formation proceeds as
long as a certain level of malvidin 3-glucoside is maintained in the wines. In
wines >5-6 years old degradation or polymerization of pinotin A finally exceeds
the rate of its de novo synthesis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14759139 [PubMed - indexed for MEDLINE]

235: J Agric Food Chem. 2004 Feb 11;52(3):467-73.

Efficient one pot extraction and depolymerization of grape (Vitis vinifera)
pomace procyanidins for the preparation of antioxidant thio-conjugates.

Selga A, Sort X, Bobet R, Torres JL.

Institute for Chemical and Environmental Research (IIQAB-CSIC), Jordi Girona
18-26, 08034 Barcelona, Spain.

Antioxidant thio-conjugates were obtained from white grape pomace by a clean and
efficient one pot extraction and depolymerization method using water and
cysteamine hydrochloride. To evaluate the potential of grape pomaces of
different origins as sources of proanthocyanidins and conjugates, we conducted
varietal comparative studies of polyphenol content, antioxidant power, and
procyanidin composition. Xarel.lo proved to be the richest source of
polyphenols. The total conversion into the conjugates was as high as 8 g/kg of
Xarel.lo grape pomace, with a 50-fold excess of cysteamine, and 3 g/kg, with a
5-fold excess of cysteamine. After purification by preparative cation exchange
and reversed phase high-performance liquid chromatography, 17 g of 63% pure
4beta-(2-aminoethylthio)epicatechin (acetate salt) was obtained from 35 kg of
moist pomace. The procedure described here will make the antioxidant
thio-derivatives efficiently available directly from raw plant byproducts such
as grape pomace.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14759134 [PubMed - indexed for MEDLINE]

236: Ann Bot (Lond). 2004 Mar;93(3):263-74. Epub 2004 Jan 28.

Shoot development in grapevine (Vitis vinifera) is affected by the modular
branching pattern of the stem and intra- and inter-shoot trophic competition.

Lebon E, Pellegrino A, Tardieu F, Lecoeur J.

Institut National de la Recherche Agronomique - Ecole Nationale Superieure
Agronomique, Laboratoire d'Ecophysiologie des Plantes sous Stress
Environnementaux, UMR 759, INRA-ENSA.M, 2 place Viala, 34060 Montpellier Cedex
01, France. [email protected]

BACKGROUND AND AIMS: Shoot architecture variability in grapevine (Vitis
vinifera) was analysed using a generic modelling approach based on thermal time
developed for annual herbaceous species. The analysis of shoot architecture was
based on various levels of shoot organization, including pre-existing and newly
formed parts of the stem, and on the modular structure of the stem, which
consists of a repeated succession of three phytomers (P0-P1-P2). METHODS: Four
experiments were carried out using the cultivar 'Grenache N': two on potted
vines (one of which was carried out in a glasshouse) and two on mature vines in
a vineyard. These experiments resulted in a broad diversity of environmental
conditions, but none of the plants experienced soil water deficit. KEY RESULTS:
Development of the main axis was highly dependent on air temperature, being
linearly related to thermal time for all stages of leaf development from
budbreak to veraison. The stable progression of developmental stages along the
main stem resulted in a thermal-time based programme of leaf development. Leaf
expansion rate varied with trophic competition (shoot and cluster loads) and
environmental conditions (solar radiation, VPD), accounting for differences in
final leaf area. Branching pattern was highly variable. Classification of the
branches according to ternary modular structure increased the accuracy of the
quantitative analysis of branch development. The rate and duration of leaf
production were higher for branches derived from P0 phytomers than for branches
derived from P1 or P2 phytomers. Rates of leaf production, expressed as a
-function of thermal time, were not stable and depended on trophic competition
and environmental conditions such as solar radiation or VPD. CONCLUSIONS: The
application to grapevine of a generic model developed in annual plants made it
possible to identify constants in main stem development and to determine the
hierarchical structure of branches with respect to the modular structure of the
stem in response to intra- and inter-shoot trophic competition.

PMID: 14749253 [PubMed - indexed for MEDLINE]

237: J Agric Food Chem. 2004 Jan 28;52(2):255-60.

Major flavonoids in grape seeds and skins: antioxidant capacity of catechin,
epicatechin, and gallic acid.

Yilmaz Y, Toledo RT.

Department of Food Science and Technology, The University of Georgia, Athens,
Georgia 30602-7610, USA.

Grape seeds and skins are good sources of phytochemicals such as gallic acid,
catechin, and epicatechin and are suitable raw materials for the production of
antioxidative dietary supplements. The differences in levels of the major
monomeric flavanols and phenolic acids in seeds and skins from grapes of Vitis
vinifera varieties Merlot and Chardonnay and in seeds from grapes of Vitis
rotundifolia variety Muscadine were determined, and the antioxidant activities
of these components were assessed. The contribution of the major monomeric
flavonols and phenolic acid to the total antioxidant capacity of grape seeds and
skins was also determined. Gallic acid, monomeric catechin, and epicatechin
concentrations were 99, 12, and 96 mg/100 g of dry matter (dm) in Muscadine
seeds, 15, 358, and 421 mg/100 g of dm in Chardonnay seeds, and 10, 127, and 115
mg/100 g of dm in Merlot seeds, respectively. Concentrations of these three
compounds were lower in winery byproduct grape skins than in seeds. These three
major phenolic constituents of grape seeds contributed <26% to the antioxidant
capacity measured as ORAC on the basis of the corrected concentrations of gallic
acid, catechin, and epicatechin in grape byproducts. Peroxyl radical scavenging
activities of phenolics present in grape seeds or skins in decreasing order were
resveratrol > catechin > epicatechin = gallocatechin > gallic acid = ellagic
acid. The results indicated that dimeric, trimeric, oligomeric, or polymeric
procyanidins account for most of the superior antioxidant capacity of grape
seeds.

PMID: 14733505 [PubMed - indexed for MEDLINE]

238: J Agric Food Chem. 2003 Dec 31;51(27):8051-4.

Composition and cellular localization of tannins in grape seeds during
maturation.

Geny L, Saucier C, Bracco S, Daviaud F, Glories Y.

UMR OEnologie-Ampelologie 1219, Faculte d'oenologie-INRA, Universite Victor
Segalen Bordeaux 2, 33405 Talence, France.
[email protected]

Cell walls were isolated from seeds of grape berries (Vitis vinifera L.), and
proanthocyanidin composition was determined over the course of ripening for
different levels of vine water status. During the ripening period the tannins
from the cell walls were always more polymerized than those from the inner part
of the cell. At maturity this difference becomes more significant compared to
veraison, due to a significant increase in the mean degree of polymerization of
the cell wall tannins. The tannin composition was typical of grape seed tannins
and was quite similar in the two cell fractions studied, but the epicatechin
gallate proportion was significantly higher in the cell wall fraction. There
were no significant effects of water deficit on composition and polymerization
of seed tannins.

PMID: 14690395 [PubMed - indexed for MEDLINE]

239: Genome. 2003 Dec;46(6):1037-48.

Molecular genetic diversity of the French-American grapevine hybrids cultivated
in North America.

Pollefeys P, Bousquet J.

Chaire de recherche du Canada en genomique forestiere et environnementale,
Centre de recherche en biologie forestiere, Universite Laval, Sainte-Foy,
Canada.

French-American hybrid grapevines are most popular in eastern and mid-western
North America: they are hardy cultivars derived from crosses between the
European Vitis vinifera and American wild vines. The aim of this study was to
characterize their genetic background using 6 microsatellite (SSR) markers and a
set of 33 diagnostic RAPD markers. The latter were reproducible with different
PCR thermal cyclers. Two SSR loci were found to be synonymous, VrZAG47 and
VVMD27. The DNA profile frequencies estimated for each cultivar were much lower
with multi-locus SSR data than that obtained from multi-fragment RAPD data.
There was no significant correlation between the multi-locus DNA profile
frequencies derived from SSRs and those from RAPDs. Estimates of genetic
diversity derived from SSRs were generally higher and the average similarity
between cultivars was generally lower than values reported for subgroups of V.
vinifera, in accordance with expectations for hybrid cultivars. The phenetic
relationships depicted by UPGMA (unweighted pair-group method with arithmetic
averaging) and neighbor-joining analyses of microsatellite data were congruent
and, to a large extent, in agreement with the known pedigree or history of each
cultivar. A major dichotomy was observed between one group where the known
genetic background was dominated by the North American Vitis riparia and Vitis
labrusca, and another one where the genetic background was dominated by the
European V. vinifera. Two Kulhmann varieties thought to be synonymous were found
to be different, though closely related.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14663522 [PubMed - indexed for MEDLINE]

240: Appl Environ Microbiol. 2003 Dec;69(12):7319-27.

Use of a green fluorescent strain for analysis of Xylella fastidiosa
colonization of Vitis vinifera.

Newman KL, Almeida RP, Purcell AH, Lindow SE.

Department of Plant and Microbial Biology, University of California, Berkeley,
California 94720, USA.

Xylella fastidiosa causes Pierce's disease of grapevine as well as several other
major agricultural diseases but is a benign endophyte in most host plants. X.
fastidiosa colonizes the xylem vessels of host plants and is transmitted by
xylem sap-feeding insect vectors. To understand better the pattern of host
colonization and its relationship to disease, we engineered X. fastidiosa to
express a green fluorescent protein (Gfp) constitutively and performed confocal
laser-scanning microscopic analysis of colonization in a susceptible host, Vitis
vinifera. In symptomatic leaves, the fraction of vessels colonized by X.
fastidiosa was fivefold higher than in nearby asymptomatic leaves. The fraction
of vessels completely blocked by X. fastidiosa colonies increased 40-fold in
symptomatic leaves and was the feature of colonization most dramatically linked
to symptoms. Therefore, the extent of vessel blockage by bacterial colonization
is highly likely to be a crucial variable in symptom expression. Intriguingly, a
high proportion (>80%) of colonized vessels were not blocked in infected leaves
and instead had small colonies or solitary cells, suggesting that vessel
blockage is not a colonization strategy employed by the pathogen but, rather, a
by-product of endophytic colonization. We present evidence for X. fastidiosa
movement through bordered pits to neighboring vessels and propose that
vessel-to-vessel movement is a key colonization strategy whose failure results
in vessel plugging and disease.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 14660381 [PubMed - indexed for MEDLINE]

241: Mol Plant Microbe Interact. 2003 Dec;16(12):1118-28.

Laminarin elicits defense responses in grapevine and induces protection against
Botrytis cinerea and Plasmopara viticola.

Aziz A, Poinssot B, Daire X, Adrian M, Bezier A, Lambert B, Joubert JM, Pugin A.

Unite de Recherche Vignes et Vins de Champagne, URVVC-UPRES EA 2069, UFR
Sciences, Moulin de la Housse, Universite de Reims Champagne-Ardenne, BP 1039,
F-51687 Reims cedex 2 France.

Grapevine (Vitis vinifera L.) is susceptible to many pathogens, such as Botrytis
cinerea, Plasmopara viticola, Uncinula necator, and Eutypa lata. Phytochemicals
are used intensively in vineyards to limit pathogen infections, but the
appearance of pesticide-resistant pathogen strains and a desire to protect the
environment require that alternative strategies be found. In the present study,
the beta-1,3-glucan laminarin derived from the brown algae Laminaria digitata
was shown both to be an efficient elicitor of defense responses in grapevine
cells and plants and to effectively reduce B. cinerea and P. viticola
development on infected grapevine plants. Defense reactions elicited by
laminarin in grapevine cells include calcium influx, alkalinization of the
extracellular medium, an oxidative burst, activation of two mitogen-activated
protein kinases, expression of 10 defense-related genes with different kinetics
and intensities, increases in chitinase and beta-1,3-glucanase activities, and
the production of two phytoalexins (resveratrol and epsilon-viniferin). Several
of these effects were checked and confirmed in whole plants. Laminarin did not
induce cell death. When applied to grapevine plants, laminarin reduced infection
by B. cinerea and P. viticola by approximately 55 and 75%, respectively. Our
data describing a large set of defense reactions in grapevine indicate that the
activation of defense responses using elicitors could be a valuable strategy to
protect plants against pathogens.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14651345 [PubMed - indexed for MEDLINE]

242: Biosystems. 2003 Dec;72(3):253-61.

A study on the interplay between energy and matter transformation: the effect of
elevated temperatures on the leaf morphology of Vitis vinifera var. Merlot.

Maze J, Bowen P, Bogdanoff C.

Department of Botany, University of British Columbia, BC V6T 1Z4, Vancouver,
Canada. [email protected]

This investigation explores the relationship between increased energy levels and
leaf morphology. It tests the idea that the causal agent of development is the
dissipation of energy into transformed matter. The energy under which leaves
developed was modified by increasing temperatures in grape cordons through
wrapping them in clear plastic sleeves in the early spring. At the higher
temperatures, and energy levels, there was a small but statistically significant
decrease in leaf size and a change in organization the leaves. The decrease in
leaf size may be due to a reallocation of resources, either to greater shoot
growth as a previous study demonstrated or to the appearance of more vectors of
development in the leaves, i.e. the appearance of more developmental subsystems.
The leaves that grew under the higher temperature regime were more complex,
perhaps indicating that the grapes on those same vines may produce more complex
juice, another expression of more developmental subsystems. The change in
organization in these leaves that developed at higher temperature argues that
the causal agent in plant development is energy dissipation and the concomitant
transformation of matter, the latter expressed in the appearance of more growth
vectors.

PMID: 14643493 [PubMed - indexed for MEDLINE]

243: J Agric Food Chem. 2003 Dec 3;51(25):7402-9.

Pyruvic acid and acetaldehyde production by different strains of Saccharomyces
cerevisiae: relationship with Vitisin A and B formation in red wines.

Morata A, Gomez-Cordoves MC, Colomo B, Suarez JA.

Departamento de Tecnologia de Alimentos, Escuela Tecnica Superior de Ingenieros
Agronomos, Universidad Politecnica de Madrid, Ciudad Universitaria S/N, 28040
Madrid, Spain. [email protected]

The production of pyruvate and acetaldehyde by 10 strains of Saccharomyces
cerevisiae was monitored during the fermentation of Vitis vinifera L. variety
Tempranillo grape must to determine how these compounds might influence the
formation of the pyroanthocyanins vitisin A and B
(malvidin-3-O-glucoside-pyruvate acid and malvidin-3-O-glucoside-4 vinyl,
respectively). Pyruvate and acetaldehyde production patterns were determined for
each strain. Pyruvate production reached a maximum on day four of fermentation,
while acetaldehyde production was at its peak in the final stages. The
correlation between pyruvate production and vitisin A formation was especially
strong (R (2) = 0.80) on day 4, when the greatest quantity of pyruvate was found
in the medium. The correlation between acetaldehyde production and the formation
of vitisin B was strongest (R (2) = 0.81) at the end of fermentation when the
acetaldehyde content of the medium was at its highest. Identification and
quantification experiments were performed by HPLC-DAD. The identification of the
vitisins was confirmed by LC/ESI-MS.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 14640591 [PubMed - indexed for MEDLINE]

244: J Investig Allergol Clin Immunol. 2003;13(3):211-2.

Grape anaphylaxis.

Caiaffa MF, Tursi A, Macchia L.

Medical Faculty, University of Foggia, Italy.

Grape allergy is particularly rare in spite of the vast extension of Vitis
vinifera cultivation on all continents. We report on the case of a 28-year-old
woman who presented with allergic systemic reaction after eating white grapes
(Vitis vinifera). She complained of two severe episodes of anaphylaxis after
eating grapes, with generalized pruritus, acute generalized urticaria, facial
swelling, lip and oropharingeal angioedema, and dysphagia. Both the episodes
were treated at the Emergency Room level, with parenteral administration of
corticosteroids and antihistamines. Skin prick tests with commercial extract of
grapes provided a negative result, while prick by prick procedure performed with
white grapes and white grape juice yielded a positive result. Grape-specific
serum IgE were also detected. We conclude that in the diagnosis of grape allergy
the currently available commercial extracts might not be completely reliable and
the prick-by-prick procedure with fresh grapes should always be performed.

Publication Types:
Case Reports

PMID: 14635473 [PubMed - indexed for MEDLINE]

245: Planta. 2004 Mar;218(5):767-74. Epub 2003 Nov 14.

Oligogalacturonide signal transduction, induction of defense-related responses
and protection of grapevine against Botrytis cinerea.

Aziz A, Heyraud A, Lambert B.

Unite de Recherche Vignes et Vins de Champagne, UPRES EA 2069, Universite de
Reims Champagne-Ardenne, BP 1039, 51687, Reims cedex 2, France.
[email protected]

Grapevine ( Vitis vinifera L.) is vulnerable to a variety of pathogenic fungi,
among them Botrytis cinerea, the causal agent of grey mould, is responsible for
worldwide yield losses that would be even more important without a successful
control that relies mainly on fungicides. In the present work we investigated an
alternative way of using oligogalacturonides (OGA) to induce defense responses
in grapevine and protection against B. cinerea. Kinetic experiments with
grapevine cells showed that OGA induced a rapid and transient generation of
H(2)O(2), followed by differential expression of nine defense-related genes and
stimulation of chitinase and beta-1,3-glucanase activities. Inhibition of
OGA-induced oxidative burst by diphenylene iodonium (DPI), an inhibitor of NADPH
oxidase, lowered induction levels of six genes and chitinase activity.
Interestingly, the induction of three other genes and beta-1,3-glucanase
activity were inhibited by K252a, a protein kinase inhibitor, but not by DPI.
Treatment of grapevine leaves with OGA also reduced infection by B. cinerea by
about 55-65%. Accordingly, DPI or K252a with or without OGA increased the
susceptibility of grapevine leaves to B. cinerea. We suggest that treatment of
grapevine with OGA elicits different signalling pathways, which might act in
tandem with the oxidative burst to increase grapevine defense responses required
for protection against B. cinerea.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14618326 [PubMed - indexed for MEDLINE]

246: Medicina (Kaunas). 2003;39 Suppl 2:104-12.

[Polyphenols and anthocyanins in fruits, grapes juices and wines, and evaluation
of their antioxidant activity]

[Article in Lithuanian]

Briedis V, Povilaityte V, Kazlauskas S, Venskutonis PR.

Department of Drug Technology and Pharmaceutical Management, Kaunas University
of Medicine, A. Mickeviciaus 9, 3000 Kaunas, Lithuania.

The objective of this study was to produce grape (Vitis vinifera L.) extracts
using efficient solvents and to compare antioxidant activity, polyphenol and
anthocyanin content and composition in the produced extracts to the same
characteristics of grape juices and wines. Methanolic and aqueous extracts of
red and blue grapes were produced. Total amounts of polyphenols and anthocyanins
in grape extracts, juices and wines were determined. Total polyphenols were
spectrophotometrically determined measuring absorption after using
Folin-Ciocalteau reagent. Total anthocyanins were determined by pH-differential
method. Antioxidant activity of all those grape products was analyzed in
2,2-diphenyl-1-picrylhydrazil (DPPH) and
2,2'-azine-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) free radical
binding model systems. The results obtained by both antioxidant activity
determination methods were comparable and permitted to evaluate the ability to
inactivate free radicals. Individual polyphenols and anthocyanins were separated
by high performance liquid chromatography. Compounds of malvidine, delphinidine
and petunidine were identified as main anthocyanins present in grape products
demonstrating antioxidant activity. The determined amounts of polyphenols and
anthocyanins in red wines were relatively higher than in other investigated
objects. The wines possessed the highest antioxidant activity, which correlated
with the determined amounts of total polyphenols. Correlation between total
anthocyanins contents and antioxidant activity of appropriate objects was not
established.

PMID: 14617869 [PubMed - in process]

247: Theor Appl Genet. 2004 Mar;108(5):864-72. Epub 2003 Nov 6.

A microsatellite marker based framework linkage map of Vitis vinifera L.

Riaz S, Dangl GS, Edwards KJ, Meredith CP.

Department of Viticulture and Enology, University of California, One Shields
Avenue, Davis, CA 95616, USA.

We have constructed a framework linkage map based on microsatellite markers for
Vitis vinifera L., the European wine grape. The mapping population consisted of
153 progeny plants from a cross of Vitis vinifera cvs. Riesling x Cabernet
Sauvignon. One hundred fifty-two microsatellite markers and one polymorphic EST
marker have been mapped to 20 linkage groups (2 n=38). The map covers 1,728 cM
with an average distance between markers of 11.0 cM. Estimates of genome size,
expected genome coverage, and observed genome coverage were determined with
135-140 markers. Genome length estimates differed between paternal and maternal
data sets. Observed approximate genome coverage was 65% versus an expected
coverage of 90%. Meiotic recombination rates were not significantly different
between maternal and paternal parents. This map has been adopted as a reference
map for the International Grape Genome Program.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 14605808 [PubMed - indexed for MEDLINE]

248: Phytochemistry. 2003 Dec;64(7):1179-86.

Why are grape/fresh wine anthocyanins so simple and why is it that red wine
color lasts so long?

Brouillard R, Chassaing S, Fougerousse A.

Laboratoire de Chimie des Polyphenols, Universite Strasbourg I, UMR CNRS 7509,
Faculte de Chimie, 1 rue Blaise Pascal, 67 Strasbourg, France.
[email protected]

Vitis vinifera red berries are characterized by anthocyanins whose chemical
structures are among the simplest encountered in higher plants. On the contrary,
many plants, including orchids, petunias, red cabbage, elderberries, potatoes
for instance, have developed very complicated anthocyanins featuring side-chains
at the available positions of the aglycone skeleton. Such pigments were shown to
possess bio-physico-chemical properties not to be seen with the grape common
anthocyanins. Among beverages (water, tea, beer, wine, coffee, juices, milk),
red wine is the only one whose organoleptic properties improve with time and
this is called ageing. The grape/fresh red wine pigments, after a few months,
disappear from the wine giving birth to new pigments resulting from the wine
spontaneous chemistry allowing it to remain red for many years. What are the
wine pigments and why are they so stable is the purpose of this mini-review. The
structural simplicity of grape anthocyanins and the long lasting colour of red
wine is another French paradox; we call it French paradox II.

Publication Types:
Review

PMID: 14599515 [PubMed - indexed for MEDLINE]

249: Gig Sanit. 2003 Sep-Oct;(5):58-61.

[Regulation of the metabolism of ethyl alcohol in the body with oligomeric
proanthocyanides as a preventive measure against its toxic effect]

[Article in Russian]

Kushnerova NF, Sprygin VG, Rakhmanin IuA.

The study evaluated the effects of a complex of oligomer proanthocyanidines
derived from the crowns of the grapes Vitis vinifera on the oxidation of ethanol
and on the activity of alcohol dehydrogenase and aldehyde dehydrogenase in the
plasma of male volunteers given ethanol once, as well as in the rat liver during
long-term alcohol use. Oligomer proanthocyanides were shown to be actively
involved in the human and animal metabolism of ethyl alcohol in vivo. Under
their action, the reductase activity was preserved, which prevented acetaldehyde
from accumulating in the body since it reduces to less toxic ethanol. Oligomer
proanthocyanidines from the crowns of the grape may be used to prevent a
significant toxic effect of excess doses of ethyl alcohol on the body.

Publication Types:
Comparative Study
English Abstract

PMID: 14598757 [PubMed - indexed for MEDLINE]

250: Plant Cell Rep. 2004 Mar;22(8):608-14. Epub 2003 Nov 1.

Chitosan improves development, and protects Vitis vinifera L. against Botrytis
cinerea.

Ait Barka E, Eullaffroy P, Clement C, Vernet G.

UFR Sciences, URVVC, UPRES EA 2069, Laboratoire de Biologie et Physiologie
Vegetales, Universite de Reims Champagne-Ardenne, B.P. 1039, 51687 Reims Cedex
2, France. [email protected]

We evaluated the potential of chitosan both to stimulate plant development and
to induce protection from Botrytis cinerea in Vitis vinifera L. plantlets. The
presence of 1.75% (v/v) chitogel in the culture medium was the optimal
concentration for in vitro grapevine plantlet growth, as determined by
measurements on enhancement of root and shoot biomass. Photosynthesis and
related parameters were also stimulated in chitogel-treated plantlets. Chitogel
reduced the development of Botrytis cinerea and induced cytological alterations
to the pathogen. When challenged with the fungus, a significant decrease in
disease incidence was observed in plants growing on medium supplemented with
chitogel. Furthermore, exogenous foliar applications of chitogel to plantlets
growing on chitogel-free medium sensitized them so as to be protected against
Botrytis cinerea attack. Our results indicate that chitogel can be used in the
vineyard as a means to attain protection against Botrytis cinerea and that its
application may counteract the wide use of chemical pesticides.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14595516 [PubMed - indexed for MEDLINE]

251: Cryo Letters. 2003 Sep-Oct;24(5):315-22.

Genetic stability assessments of plantlets regenerated from cryopreserved in
vitro cultured grape and kiwi shoot-tips using RAPD.

Zhai Z, Wu Y, Engelmann F, Chen R, Zhao Y.

College of Life Science, Zhongshan University, Xinggang West Road 135,
Guangzhou, China.

In vitro cultured shoot-tips of four grape (Vitis vinifera) cultivars and one
kiwi (Actinidia deliciosa) cultivar were cryopreserved using the
encapsulation-dehydration method. Genomic DNA of plantlets regenerated directly
from cryopreserved shoot-tips was extracted and analyzed using the RAPD (Random
Amplified Polymorphic DNA) technique. The RAPD profiles obtained were highly
reproducible and no differences were found between the DNA patterns obtained
with plantlets regenerated from control and cryopreserved plantlets. The RAPD
technique therefore appears to be a fast, simple and efficient method for
evaluating genetic stability of cryopreserved material, which can be used
rapidly after the completion of a freezing experiment and will efficiently
complement other genetic stability evaluation methods.

Publication Types:
In Vitro

PMID: 14566391 [PubMed - indexed for MEDLINE]

252: Pest Manag Sci. 2003 Oct;59(10):1083-92.

Effects of fludioxonil and pyrimethanil, two fungicides used against Botrytis
cinerea, on carbohydrate physiology in Vitis vinifera L.

Saladin G, Magne C, Clement C.

Laboratoire de Biologie et Physiologie Vegetales, URWC UPRES EA 2069, Universite
de Reims Champagne Ardenne, UFR Sciences, Moulin de la Housse, BP 1039, F-51687
Reims 2, France.

In Vitis vinifera L, photosynthesis and photosynthate partitioning are affected
in the presence of fludioxonil and pyrimethanil, two fungicides commonly used in
vineyards against Botrytis cinerea Pers. However, the effects were found to be
different according to the model studied: plantlets (cv Chardonnay) grown in
vitro, fruiting cuttings (cv Chardonnay) and plants grown in vineyards (cvs
Chardonnay, Pinot noir and Pinot Meunier). In the plantlets grown in vitro, both
fungicides decreased gas exchanges, photosynthetic pigment and starch
concentrations in the leaves, whereas soluble carbohydrates transiently
accumulated, suggesting that plantlets mobilised starch in response to
photosynthesis inhibition caused by fungicides. In the fruiting cuttings, the
fungicides did not affect photosynthesis, although fludioxonil caused starch
decrease in parallel with sucrose accumulation, suggesting that the fungicide
effects were of lower intensity than in vitro. Conversely, in vineyard, the two
fungicides stimulated photosynthesis and increased pigment concentrations in the
three vine cultivars tested. In the meantime, glucose, fructose and starch
levels of the leaves declined after fungicide exposure, whereas sucrose
accumulated, indicating that sucrose synthesis increased in the leaves following
the fungicide treatment. Among the three varieties, Chardonnay was the most
sensitive to the fungicides as revealed by the intensity of the responses and
the longer period for recovery. In vineyard, the results suggested that the two
fungicides, in addition to inhibiting B cinerea development, had a beneficial
effect on vine physiology through the stimulation of leaf carbon nutrition,
which may further enable the plant to rapidly make use of its defence reactions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14561065 [PubMed - indexed for MEDLINE]

253: J Agric Food Chem. 2003 Oct 22;51(22):6475-81.

Monomeric, oligomeric, and polymeric flavan-3-ol composition of wines and grapes
from Vitis vinifera L. Cv. Graciano, Tempranillo, and Cabernet Sauvignon.

Monagas M, Gomez-Cordoves C, Bartolome B, Laureano O, Ricardo da Silva JM.

Instituto de Fermentaciones Industriales, CSIC, Juan de la Cierva 3, 28006
Madrid, Spain.

The monomeric, oligomeric, and polymeric flavan-3-ol composition of wines, grape
seeds, and skins from Vitis vinifera L. cv. Graciano, Tempranillo, and Cabernet
Sauvignon has been studied using (1) fractionation by polyamide column
chromatography followed by HPLC/ESI-MS analysis, (2) fractionation on C(18)
Sep-Pak cartridges followed by reaction with vanillin and acid-catalyzed
degradation in the presence of toluene-alpha-thiol (thiolysis). The content of
monomers ((+)-catechin and (-)-epicatechin), procyanidin dimers (B3, B1, B4, and
B2), trimers (T2 and C1), and dimer gallates (B2-3-O-gallate, B2-3'-O-gallate,
and B1-3-O-gallate) ranged from 76.93 to 133.18 mg/L in wines, from 2.30 to 8.21
mg/g in grape seeds, and from 0.14 to 0.38 mg/g in grape skins. In wines, the
polymeric fraction represented 77-84% of total flavan-3-ols and showed a mean
degree of polymerization (mDP) value of 6.3-13.0. In grapes, the polymeric
fraction represented 75-81% of total flavan-3-ols in seeds and 94-98% in skins
and showed mDP values of 6.4-7.3 in seeds and 33.8-85.7 in skins. All the
monomeric flavan-3-ols and oligomeric procyanidins found in wines were also
present in seeds, although differences in their relative abundances were seen.
The skin polymeric proanthocyanidins participated in the equilibration of the
wine polymeric proanthocyanidin fraction, especially contributing to the polymer
subunit composition and mDP.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14558765 [PubMed - indexed for MEDLINE]

254: FEMS Yeast Res. 2003 Oct;4(1):79-85.

Metabolic engineering of Saccharomyces cerevisiae for the synthesis of the
wine-related antioxidant resveratrol.

Becker JV, Armstrong GO, van der Merwe MJ, Lambrechts MG, Vivier MA, Pretorius
IS.

Institute for Wine Biotechnology, Department of Viticulture and Oenology,
Stellenbosch University, 7600 Stellenbosch, South Africa.

The stilbene resveratrol is a stress metabolite produced by Vitis vinifera
grapevines during fungal infection, wounding or UV radiation. Resveratrol is
synthesised particularly in the skins of grape berries and only trace amounts
are present in the fruit flesh. Red wine contains a much higher resveratrol
concentration than white wine, due to skin contact during fermentation. Apart
from its antifungal characteristics, resveratrol has also been shown to have
cancer chemopreventive activity and to reduce the risk of coronary heart
disease. It acts as an antioxidant and anti-mutagen and has the ability to
induce specific enzymes that metabolise carcinogenic substances. The objective
of this pilot study was to investigate the feasibility of developing wine yeasts
with the ability to produce resveratrol during fermentation in both red and
white wines, thereby increasing the wholesomeness of the product. To achieve
this goal, the phenylpropanoid pathway in Saccharomyces cerevisiae would have to
be introduced to produce p-coumaroyl-CoA, one of the substrates required for
resveratrol synthesis. The other substrate for resveratrol synthase,
malonyl-CoA, is already found in yeast and is involved in de novo fatty-acid
biosynthesis. We hypothesised that production of p-coumaroyl-CoA and resveratrol
can be achieved by co-expressing the coenzyme-A ligase-encoding gene (4CL216)
from a hybrid poplar and the grapevine resveratrol synthase gene (vst1) in
laboratory strains of S. cerevisiae. This yeast has the ability to metabolise
p-coumaric acid, a substance already present in grape must. This compound was
therefore added to the synthetic media used for the growth of laboratory
cultures. Transformants expressing both the 4CL216 and vst1 genes were obtained
and tested for production of resveratrol. Following beta-glucosidase treatment
of organic extracts for removal of glucose moieties that are typically bound to
resveratrol, the results showed that the yeast transformants had produced the
resveratrol beta-glucoside, piceid. This is the first report of the
reconstruction of a biochemical pathway in a heterologous host to produce
resveratrol.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14554199 [PubMed - indexed for MEDLINE]

255: J Agric Food Chem. 2003 Oct 8;51(21):6261-7.

Vitisin A content in Chilean wines from Vitis vinifera Cv. Cabernet Sauvignon
and contribution to the color of aged red wines.

Schwarz M, Quast P, von Baer D, Winterhalter P.

Institute of Food Chemistry, Technical University of Braunschweig,
Schleinitzstrasse 20, 38106 Braunschweig, Germany.

Vitisin A was prepared from malvidin 3-glucoside and pyruvic acid in model wine
medium, isolated by countercurrent chromatography, and purified by preparative
high-performance liquid chromatography (HPLC). The synthesized compound was used
as a reference standard to quantify vitisin A in Chilean wines from Vitis
vinifera cv. Cabernet Sauvignon, including a vertical row of wines from the same
vineyard over 16 years. Maximum vitisin A content was reached within the first
year of storage. Importantly, up to half of the initial amount of vitisin A in
young wines was still present in 15 year old wines. Although vitisin A was found
to be much more stable as compared to other monomeric C-4 underivatized
anthocyanins, it also slowly degrades after reaching its peak concentration. The
"color activity concept" was applied to vitisin A, malvidin 3-glucoside,
malvidin 3-(6' '-acetylglucoside), and polymeric pigments isolated by
countercurrent chromatography in order to estimate their contribution toward the
overall color expression of wines. It was found that vitisin A is only a minor
contributor to the visually perceived color of aged red wines (color
contribution approximately 5%). The major contributor is the polymeric fraction
(color contribution approximately 70-90%).

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14518953 [PubMed - indexed for MEDLINE]

256: J Agric Food Chem. 2003 Oct 8;51(21):6151-7.

Effect of ochratoxin A-producing Aspergilli on stilbenic phytoalexin synthesis
in grapes.

Bavaresco L, Vezzulli S, Battilani P, Giorni P, Pietri A, Bertuzzi T.

Istituto di Frutti-Viticoltura, Universita Cattolica del Sacro Cuore, 29100
Piacenza, Italy. [email protected]

Berries of Vitis vinifera L. cv. Barbera were infected, at veraison and during
ripening, by a conidial suspension of A. japonicus, A. ochraceus, A. fumigatus
and two isolates of A. carbonarius to control ochratoxin A production and
stilbene induced synthesis. The experimental design provided also for intact and
punctured berries and incubation temperature of 25 degrees C and 30 degrees C.
All the tested fungi, except A. fumigatus, significantly increased
trans-resveratrol synthesis over the control, while trans-piceid was not
affected; only A. ochraceus significantly elicited the berries to synthesize
piceatannol. The two isolates of A. carbonarius produced higher amounts of
ochratoxin A than did the other fungi. A positive correlation between ochratoxin
A and trans-resveratrol synthesis occurred. trans-Resveratrol and piceatannol
showed fungicidal activity against A. carbonarius, being able to completely
inhibit fungal growth at a concentration of 300 microg/g and 20 microg/g,
respectively.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 14518937 [PubMed - indexed for MEDLINE]

257: Life Sci. 2003 Oct 17;73(22):2883-98.

Procyanidins from grape seeds protect endothelial cells from peroxynitrite
damage and enhance endothelium-dependent relaxation in human artery: new
evidences for cardio-protection.

Aldini G, Carini M, Piccoli A, Rossoni G, Facino RM.

Istituto Chimico Farmaceutico Tossicologico, University of Milan, Viale Abruzzi
42-20131 Milan, Italy. [email protected]

The peroxynitrite scavenging ability of Procyanidins from Vitis vinifera L.
seeds was studied in homogeneous solution and in human umbilical endothelial
cells (EA.hy926 cell line) using 3-morpholinosydnonimine (SIN-1) as
peroxynitrite generator. In homogeneous phase procyanidins dose-dependently
inhibited 2',7'-dichloro-dihydrofluorescein (DCFH) oxidation induced by SIN-1
with an IC50 value of 0.28 microM. When endothelial cells (EC) were exposed to 5
mM SIN-1, marked morphological alterations indicating a necrotic cell death
(cell viability reduced to 16 +/- 2.5%) were observed. Cell damage was
suppressed by procyanidins, with a minimal effective concentration of 1 microM
(cell morphology and integrity completely recovered at 20 microM). Cellular
localization of procyanidins in EC was confirmed using a new staining procedure
and site-specific peroxyl radical inducers: AAPH and cumene hydroperoxide
(CuOOH). Endothelial cells (EC) pre-incubated with procyanidins (20 microM) and
exposed to FeCl3/K3Fe(CN)6 showed a characteristic blue staining, index of a
site-specific binding of procyanidins to EC. Procyanidins dose-dependently
inhibit the AAPH induced lipid oxidation and reverse the consequent loss of cell
viability, but were ineffective when oxidation was driven at intracellular level
(CuOOH). This demonstrates that the protective effect is due to their specific
binding to the outer surface of EC thus to quench exogenous harmful radicals.
Procyanidins dose-dependently relaxed human internal mammary aortic (IMA) rings
(with intact endothelium) pre-contracted with norepinephrine (NE), showing a
maximal vasorelaxant effect (85 +/- 9%) at 50 microM (catechin: 18 +/- 2%
relaxation at 50 microM). This effect was completely abolished when IMA-rings
were de-endothelized and when IMA-rings with intact endothelium were pretreated
with L-NMMA or with the soluble guanylate cyclase inhibitor, ODQ. Pre-incubation
with indomethacin reduces (by almost 50%) the vasodilating effect of
procyanidins, indicating the involvement also of a COX-dependent mechanism. This
was confirmed in another set of experiments, where procyanidins dose-dependently
stimulate the prostacyclin (PGI2) release, reaching a plateau between 25 and 50
microM. Finally, pre-incubation of IMA-rings with procyanidins (from 6.25 to 25
microM) resulted in a dose-dependent prevention of the endothelin-1 (ET-1)
vasoconstriction. The ability of procyanidins to prevent peroxynitrite attack to
vascular cells, by layering on the surface of coronary EC, and to enhance
endothelial NO-synthase-mediated relaxation in IMA rings provide further insight
into the molecular mechanisms through which they exert cardioprotective activity
in ischemia/reperfusion injury in vivo.

Publication Types:
In Vitro
Research Support, Non-U.S. Gov't

PMID: 14511773 [PubMed - indexed for MEDLINE]

258: Plant Cell Rep. 2003 Sep;22(2):141-9. Epub 2003 Jul 9.

Transformation of apple ( Malus domestica Borkh.) with the stilbene synthase
gene from grapevine ( Vitis vinifera L.) and a PGIP gene from kiwi ( Actinidia
deliciosa).

Szankowski I, Briviba K, Fleschhut J, Schonherr J, Jacobsen HJ, Kiesecker H.

Institut fur Gemuse- und Obstbau, Universitat Hannover, Am Steinberg 3, 31157
Sarstedt, Germany. [email protected]

The objective of the present research was to introduce genes with antifungal
potential into the commercially important apple cvs. Elstar and Holsteiner Cox
in order to establish resistance against fungal diseases. The gene encoding the
stilbene synthase (Vst1) from Vitis vinifera L., responsible for the synthesis
of the phytoalexin resveratrol in grapevine, and the gene for a
polygalacturonase-inhibiting protein (PGIP) from kiwi ( Actinidia deliciosa)
were transferred into Holsteiner Cox and Elstar via Agrobacterium
tumefaciens-mediated transformation. A total of nine transgenic Holsteiner Cox
clones and one transgenic E clone carrying the stilbene-synthase gene as well as
three transgenic Holsteiner Cox lines harbouring the
polygalacturonase-inhibiting protein from Kiwi were identified via polymerase
chain reaction and Southern blot analysis. High performance liquid
chromatography analysis revealed the accumulation of a resveratrol-derivate, a
glycoside, in transgenic Vst1 plants.

PMID: 14504909 [PubMed - indexed for MEDLINE]

259: Theor Appl Genet. 2004 Jan;108(2):231-7. Epub 2003 Sep 16.

Evidence on the evolution of polymorphism of microsatellite markers in varieties
of Vitis vinifera L.

Crespan M.

Istituto Sperimentale per la Viticoltura, Sezione Ampelografia e Miglioramento
Genetico, Via Casoni, 13/A-31058, Susegana (TV), Italy.
[email protected]

This paper contributes a description of four mutations in microsatellite
allele-length found within strains of each of the varieties known as Muscat
d'Alsace, Greco di Tufo, Primitivo and Corvina veronese: the comparison of
microsatellite sequences of wild and mutated strains showed in all cases an
increase of one or more repeats of the core GA sequence. Whereas studies on
animals reported mutations which originated in gametes, in grapevines somatic
cell-mutations take place and are eventually fixed and transmitted to new
individuals through vegetative propagation. The mutations detected in 1998 were
found again 3 years later in seven out of eight plants. A chimeric situation was
clearly noted and resolved in three of the four cases described. Considerations
about the genesis of microsatellite alleles and the reliability of
fingerprinting with these markers are provided.

PMID: 13679982 [PubMed - indexed for MEDLINE]

260: J Agric Food Chem. 2003 Sep 24;51(20):5967-71.

Carotenoid compounds in grapes and their relationship to plant water status.

Oliveira C, Silva Ferreira AC, Mendes Pinto M, Hogg T, Alves F, Guedes de Pinho
P.

Escola Superior de Biotecnologia, Universidade Catolica Portuguesa, R. Dr.
Antonio Bernardino de Almeida, 4200-072 Porto, Portugal.

The aim of this work was to study the relationship between carotenoid contents
in grapevine berries and plant water status. For this purpose, a black grapevine
variety, Vitis vinifera L. cv. Touriga Nacional, was studied. The experiments
were carried out in the same Douro vineyards, with plants of the same age, in
two different water retention soils. A higher water retention capacity soil,
soil A, and a lower water retention capacity soil, soil B, were both in a 1.2 m
deep silt-loam schist-derived soil. The training system was the double cordon
trained and spur pruned. A first range was nonirrigated (NI) and a second one
was irrigated (I), 60% of evapotranspiration (ET(0)). For soil B, a 30% of ET(0)
treatment was also applied. The plant water status was estimated by predawn leaf
water potential. The effects of plant water status on berry growth were studied
by measurement of the berry weight and total soluble solids (degrees Brix). The
carotenoid profile was quantitatively determined by high-performance liquid
chromatography/diode array. Carotenoids determined were beta-carotene, lutein,
neoxanthin, violaxanthin, and luteoxanthin. The comparison between irrigated and
nonirrigated grapes was followed from 2 weeks before veraison until the ripe
stage. Results showed that at harvest time, berries exposed to the NI had a
lower weight than those exposed to the irrigated treatment (60% of ET(0)), 0.89
vs 1.36 g/berry and 0.94 vs 1.34 g/berry, for soils A and B, respectively. The
irrigated treatment contributed to a higher sugar concentration in both soils.
However, depending on the soil water retention capacity, the carotenoid contents
were different in soils A and B. For soil A, the total carotenoid content was
similar for both NI and I treatments. However, with regard to soil B, in
irrigated treatment, levels of carotenoids were approximately 60% lower than
those found for the NI. It seems to be possible to produce higher weight berries
(with higher sugar levels) with similar carotenoid contents. On the other hand,
soil characteristics had a larger influence than irrigation on the concentration
of carotenoids in grapes, resulting in an important viticultural parameter to
take into account in aroma precursor formation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 13129303 [PubMed - indexed for MEDLINE]

261: Biotechnol Lett. 2003 Jul;25(14):1131-5.

Manipulating anthocyanin composition in Vitis vinifera suspension cultures by
elicitation with jasmonic acid and light irradiation.

Curtin C, Zhang W, Franco C.

Cooperative Research Centre for Bioproducts and Department of Medical
Biotechnology, School of Medicine, Flinders University, Bedford Park, SA 5042,
Adelaide, Australia.

Jasmonic acid altered the accumulation of major anthocyanins in Vitis vinifera
cell culture. Peonidin 3-glucoside content at day three was increased from 0.3
to 1.7 mg g(-1) dry cell wt while other major anthocyanins were increased by
smaller increments. By day 14, the content of methylated and acylated
anthocyanins (peonidin 3-p-coumaroylglucoside and malvidin
3-p-coumaroylglucoside) was 6.3 mg g(-1) DCW, in response to treatment with
jasmonic acid, and comprising approximately 45% (w/w) of total anthocyanins. In
comparison, the untreated control culture contained 1.2 mg g(-1) DCW which made
up approximately 32% (w/w) of total anthocyanins. Light further enhanced
anthocyanin accumulation induced by jasmonic acid elicitation. The content of
peonidin 3-glucoside at day 3 was 6.6 mg g(-1) DCW, 22-fold higher than control
cultures while the content in response to light irradiation alone was 0.6 mg
g(-1) DCW. When a highly pigmented cell line was elicited with jasmonic acid
total anthocyanins increased from 9.2 to 20.7 mg g(-1) DCW, but there was no
change in the anthocyanin composition.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12966999 [PubMed - indexed for MEDLINE]

262: J Plant Physiol. 2003 Aug;160(8):971-5.

Control of anthocyanin biosynthesis pathway gene expression by eutypine, a toxin
from Eutypa lata, in grape cell tissue cultures.

Afifi M, El-Kereamy A, Legrand V, Chervin C, Monje MC, Nepveu F, Roustan JP.

Institut National Polytechnique, Ecole Nationale Superieure Agronomique de
Toulouse, UMR INRA/INP-ENSAT 990, BP 107, F-31326 Castanet Tolosan cedex,
France.

Eutypine, 4-hydroxy-3-(3-methyl-3-butene-1-ynyl) benzaldehyde, is a toxin
produced by Eutypa lata, the causal agent of Eutypa dieback in grapevine. The
effect of the toxin on anthocyanin synthesis has been investigated in Vitis
vinifera cv. Gamay cell cultures. At concentrations higher than 200 micromol/L,
eutypine reduced anthocyanin accumulation in cells. The reduction in anthocyanin
accumulation was proportional to the eutypine concentrations and HPLC analysis
showed that eutypine affected the levels of all anthocyanins. The effect of
eutypine application on the expression of five genes of the anthocyanin
biosynthesis pathway, including chalcone synthase (CHS), flavonone-3-hydroxylase
(F3H), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin dioxygenase (LDOX),
and UDP glucose-flavonoid 3-O-glucosyl transferase (UFGT) was determined.
Expression of CHS, F3H, DFR and LDOXwas not affected by the addition of eutypine
to grapevine cell cultures. In contrast, expression of the UFGT gene was
dramatically inhibited by the toxin. These results suggest that in grapevine
cell cultures, eutypine strongly affects anthocyanin accumulation by inhibiting
UFGT gene expression. The mechanism of action of eutypine is discussed.

PMID: 12964874 [PubMed - indexed for MEDLINE]

263: Genet Res. 2003 Jun;81(3):179-92.

Genetic structure and differentiation in cultivated grape, Vitis vinifera L.

Aradhya MK, Dangl GS, Prins BH, Boursiquot JM, Walker MA, Meredith CP, Simon CJ.

National Clonal Germplasm Repository, US Department of Agriculture, One Shields
Avenue, University of California, Davis, CA 95616, USA.

222 cultivated (Vitis vinifera) and 22 wild (V. vinifera ssp. sylvestris) grape
accessions were analysed for genetic diversity and differentiation at eight
microsatellite loci. A total of 94 alleles were detected, with extensive
polymorphism among the accessions. Multivariate relationships among accessions
revealed 16 genetic groups structured into three clusters, supporting the
classical eco-geographic grouping of grape cultivars: occidentalis, pontica and
orientalis. French cultivars appeared to be distinct and showed close affinity
to the wild progenitor, ssp. sylvestris from south-western France (Pyrenees) and
Tunisia, probably reflecting the origin and domestication history of many of the
old wine cultivars from France. There was appreciable level of differentiation
between table and wine grape cultivars, and the Muscat types were somewhat
distinct within the wine grapes. Contingency chi2 analysis indicated significant
heterogeneity in allele frequencies among groups at all loci. The observed
heterozygosities for different groups ranged from 0.625 to 0.9 with an overall
average of 0.771. Genetic relationships among groups suggested hierarchical
differentiation within cultivated grape. The gene diversity analysis indicated
narrow divergence among groups and that most variation was found within groups
(approximately 85%). Partitioning of diversity suggested that the remaining
variation is somewhat structured hierarchically at different levels of
differentiation. The overall organization of genetic diversity suggests that the
germplasm of cultivated grape represents a single complex gene pool and that its
structure is determined by strong artificial selection and a vegetative mode of
reproduction.

PMID: 12929909 [PubMed - indexed for MEDLINE]

264: J Agric Food Chem. 2003 Aug 27;51(18):5408-13.

Aroma composition of Vitis vinifera Cv. tannat: the typical red wine from
Uruguay.

Boido E, Lloret A, Medina K, Farina L, Carrau F, Versini G, Dellacassa E.

Seccion Enologia and Catedra de Farmacognosia y Productos Naturales, Facultad de
Quimica, Universidad de la Republica, Avenida General Flores 2124, 11800
Montevideo, Uruguay.

The free volatiles, as well as those released from the glycosidically bound
forms by enzyme hydrolysis, have been analyzed to chracterize young Tannat wines
from two successive vintages. The Tannat wine showed some aroma profile
peculiarities detected in the free forms but, above all, in the bound fraction
for the level and profile of the norisoprenoidic fraction. Among the free
volatile compunds, a rather low content of C(6) alcohols with a prevalence of
cis-3-hexen-1-ol on the trans form and sometimes a remarkable level of
trans-2-hexen-1-ol seem to be typical for the variety. C(13)-norisoprenoidic and
monoterpenic volatiles made up approximately 42% of the total level of the
volatiles observed in the glycosidase enzyme-released fraction. The other
volatiles were C(6) alcohols (6%) and benzenoid compounds (51%). The dominating
monoterpene alcohols were the cis and trans isomers of
3,7-dimethyl-1,6-octadiene-3,8-diol (8-hydroxylinalool). The C(13)-norisoprenoid
pattern was composed by 3-hydroxy-beta-damascone, 3-oxo-alpha-ionol, vomifoliol,
4-oxo-beta-ionol, 3-oxo-7,8-dihydro-alpha-ionol, 4-oxo-7,8-dihydro-beta-ionol,
grasshopper ketone, and 7,8-dihydrovomifoliol.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12926890 [PubMed - indexed for MEDLINE]

265: J Agric Food Chem. 2003 Aug 27;51(18):5329-36.

Metabolism of fungicide diethofencarb in grape (Vitis vinifera L.): definitive
identification of thiolactic acid conjugated metabolites.

Fujisawa T, Ichise-Shibuya K, Katagi T, Ruzo LO, Takimoto Y.

Environmental Health Science Laboratory, Sumitomo Chemical Co., Ltd., 2-1,
Takatsukasa 4-Chome, Takarazuka 665-8555, Japan.
[email protected]

The metabolic fate of diethofencarb (isopropyl 3,4-diethoxycarbanilate)
separately labeled with (14)C at the phenyl ring and 2-position of the isopropyl
moiety was studied in grape (Vitis vinifera L.). The acetonitrile solution of
(14)C-diethofencarb at a rate of 500 g a.i. ha(-)(1) was once applied topically
to fruits or leaves at the maturity stage of fruits (PHI 35 days), and the
plants were grown in the greenhouse until harvest. In the grape plants,
diethofencarb was scarcely translocated to the untreated portion and was
degraded more in the fruit as compared to the leaf. For the fruit, diethofencarb
primary underwent O-deethylation at the 4-position of the phenyl ring to form
the phenolic derivative, isopropyl 3-ethoxy-4-hydroxycarbanilate (0.9% of the
total radioactive residue, TRR). This metabolite was successively transformed
via conjugation with glucose at the phenolic hydroxy group (8.1-18.1% TRR) or
with thiolactic acid at the 5-position of the phenyl ring (1.5-1.7% TRR). The
thiolactic acid conjugate was further metabolized mainly to two different types
of glucose conjugates at the 4-position of the phenyl ring (8.7-13.5% TRR) and
the hydroxy group in the thiolactic acid moiety (6.4-7.3% TRR), as evidenced by
(1)H NMR and atmospheric pressure chemical ionization-liquid chromatography-mass
spectrometry together with cochromatographies with synthetic standards.

PMID: 12926879 [PubMed - indexed for MEDLINE]

266: Chemosphere. 2003 Oct;53(3):199-206.

Stress reactions in Vitis vinifera L. following soil application of the
herbicide flumioxazin.

Saladin G, Magne C, Clement C.

Laboratoire de Biologie et Physiologie Vegetales, URVVC UPRES EA 2069,
Universite de Reims Champagne Ardenne, UFR Sciences, Moulin de la Housse, BP
1039, F-51687 Reims Cedex 2, France.

In order to evaluate the stress effects of flumioxazin (fmx) on grapevine, a
non-target plant (Vitis vinifera L.), physiological parameters such as
carbohydrate content, water status or nitrogenous metabolites were investigated
on fruiting cuttings and plants grown in vineyard. In the leaves of cuttings,
the soil-applied herbicide induced stress manifestations including a decrease of
the dry weight percentage and the soluble carbohydrate content during the first
week after treatment. Thereafter, a decrease of the osmotic potential was
observed, as well as a decrease of total protein content and a parallel
accumulation of free amino acids, including proline. Altogether, these results
suggest that soil-applied fmx induced a stress in grapevines, leading to leaf
proteolysis. However, this stress was partially recovered 3 weeks after
herbicide application, suggesting that the cuttings were capable to adapt to the
fmx exposure. In the vineyard, the flumioxazin effects were still significant 5
months after the treatment, particularly in the CH cv. They included a decrease
of the leaf dry weight percentage and soluble carbohydrate content, as well as
an increase of the osmotic potential. The decrease of leaf soluble carbohydrates
may have dramatic consequences for the berry growth and the reserve
constitution. Moreover, treated plants were characterized by a decrease of the
free amino acid content and an accumulation of ammonium, while the protein level
did not significantly increase, suggesting a degradation of amino acids. The
alteration of carbon and nitrogen status after herbicide treatment may affect
the grapevine vigour in a long term.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12919779 [PubMed - indexed for MEDLINE]

267: Plant Cell Rep. 2003 Nov;22(4):252-60. Epub 2003 Aug 8.

High-efficiency biolistic co-transformation and regeneration of 'Chardonnay'
(Vitis vinifera L.) containing npt-II and antimicrobial peptide genes.

Vidal JR, Kikkert JR, Wallace PG, Reisch BI.

Department of Horticultural Sciences, New York State Agricultural Experiment
Station, Cornell University, Geneva, NY 14456, USA.

A reliable and efficient system for transformation and regeneration of
'Chardonnay' (Vitis vinifera L.) plants via microprojectile bombardment was
developed. Improvements over the previous biolistic transformation system
included: (1) the use of gold particles for bombardment; (2) step-wise selection
at 10 then 15 mg/l kanamycin; and (3) embryo induction at 27 degrees C.
Embryogenic cell cultures were either bombarded with pBI426, which contains the
reporter gene gus (uidA) coding for beta-glucuronidase (GUS), or were
co-bombarded with pSAN237 carrying the npt-II (neomycin phosphotransferase II)
selectable marker gene, and a second plasmid with an antimicrobial peptide gene.
A large number of transient (7,883 +/- 1,928) and stable (46 +/- 32) blue spots
per plate at 2 and 95 days after bombardment, respectively, were obtained
according to GUS expression analyses. A total of 447 putative transgenic embryos
was harvested from 84 bombarded plates. From these embryos, 242 (54%) were
regenerated into plants within the first year of the experiment. Southern blot
analyses confirmed integration of the transgenes into the grape genome.
Co-transformation was tested with four separate antimicrobial constructs. The
co-transformation frequency of unlinked genes was 48% as measured by polymerase
chain reaction (PCR), and 56% as estimated by dot blot hybridization. Expression
of the gus gene, and PCR and Southern blot analyses of npt-II and antimicrobial
genes from regenerated plants document stable transformation of 'Chardonnay' and
establish the parameters for highly-efficient biolistic transformation in V.
vinifera.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 12908080 [PubMed - indexed for MEDLINE]

268: Theor Appl Genet. 2003 Oct;107(6):1043-7. Epub 2003 Jul 24.

AFLP analysis of genetic relationships among aromatic grapevines (Vitis
vinifera).

Fanizza G, Chaabane R, Lamaj F, Ricciardi L, Resta P.

DIBCA: Genetics and Plant Breeding Unit, University of Bari, Via Amendola 165/A,
70126 Bari, Italy. [email protected]

Genotypic diversity has been detected among aromatic grapevines (Vitis vinifera)
by molecular markers (AFLPs). The 22 primer-pairs generated a total of 1,331
bands of which 564 (40%) were polymorphic over all the genotypes. The bootstrap
analysis pointed out that a large number of polymorphic bands (200-400) has to
be used for a better estimation of the genetic distances among genotypes; 383
polymorphic AFLP bands were used for the cluster and the principal coordinate
analyses because they did not present missing data across all the genotypes. The
cluster analysis (UPGMA), based on polymorphic AFLP markers, revealed no
relationship between the Moscato and Malvasia grapevines. The Malvasias, unlike
the Moscatos distinguished by their distinct muscat aroma, have to be considered
a more complex group because it includes muscat and non-muscat grapevines. The
principal coordinate analysis (PCO) confirmed the pattern of the cluster
analysis only for those varieties which presented a low coefficient of
dissimilarity, while for the other varieties there was no correspondence between
the two analyses. The pattern of aggregation among aromatic grapevines in the
cluster and principal coordinate analyses does not support any classification
that might include an aromatic grapevine group in V. vinifera. Even though some
synonyms and homonyms are present among aromatic grapevines (V. vinifera),
genetic diversity exists among genotypes in AFLP markers.

PMID: 12898028 [PubMed - indexed for MEDLINE]

269: Biotechnol Lett. 2003 Jun;25(11):835-9.

Anthocyanic vacuolar inclusions (AVIs) selectively bind acylated anthocyanins in
Vitis vinifera L. (grapevine) suspension culture.

Conn S, Zhang W, Franco C.

Cooperative Research Centre for Bioproducts and Department of Medical
Biotechnology, School of Medicine, Flinders University, Bedford Park, SA 5042,
Adelaide, Australia.

Anthocyanic vacuolar inclusions (AVIs) appear as dark red-to-purple spheres of
various sizes in vacuoles of grapevine (Vitis vinifera L.) cell suspension
culture due to their interaction with anthocyanins. AVIs were purified and the
bound anthocyanins extracted and analysed by HPLC from two lines of V. vinifera
isolated from the same callus accumulating anthocyanin in the dark, yet varying
in their anthocyanin profiles and accumulation. An intermediate-pigmented line
(FU-1) with a 1.3:1 ratio of acylated:non-acylated anthocyanins, a colour value
of 0.84 units and cyanidin and peonidin as the dominant species was compared
with a high-pigmented line (FU-2) with a 1.2:1 ratio of acylated:non-acylated
anthocyanins, a colour value of 3.72 units and malvidin predominating. The
profile of AVI-bound anthocyanins showed an increase in acylated anthocyanins in
both lines of approx. 28-29%, with no apparent preference for anthocyanin
species. This resulted in a ratio of acylated:non-acylated anthocyanins of 6.2:1
for FU-1 and 4.9:1 for FU-2. The reasons for the selectivity of the AVIs for
acylated (specifically p-coumaroylated) species compared with the whole cell
profile are discussed.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 12889790 [PubMed - indexed for MEDLINE]

270: J Agric Food Chem. 2003 Jul 2;51(14):4046-53.

Environmental conditions during vegetative growth determine the major proteins
that accumulate in mature grapes.

Monteiro S, Picarra-Pereira MA, Teixeira AR, Loureiro VB, Ferreira RB.

Departamento de Botanica e Engenharia Biologica, Instituto Superior de
Agronomia, Universidade Tecnica de Lisboa, 1349-017 Lisboa, Portugal.

Despite recent reports in the literature that chitinases comprise approximately
50% of the soluble proteins present in mature Vitis vinifera L. (cv. Moscatel)
grapes, with the other major proteins being thaumatin-like proteins, a careful
inspection of the published data reveals inconsistencies as to which proteins
actually accumulate in mature grapes. Mature Moscatel grapes were harvested in
the same vineyard in two consecutive years, 1999 and 2000. The grapes exhibited
widely distinct polypeptide patterns when analyzed by either FPLC cation
exchange chromatography or two-dimensional electrophoresis: whrereas the 2000
grapes possessed a much higher protein content (1.96 versus 1.11 mg g(-)(1) of
fresh weight), the 1999 grapes contained a greater heterogeneity of
polypeptides. In addition, highly specific antibodies that recognize the
pathogenesis-related proteins present in the grapes demonstrated that the 2000
harvest grapes had a wider variety of pathogenesis-related polypeptides.
N-Terminal sequencing of the major polypeptides revealed differences in the
relative abundance and amino acid sequence of thaumatin-like and osmotin-like
polypeptides present in the 1999 and 2000 harvest Moscatel grapes and did not
detect the presence of chitinase. As a whole, the data indicate that the
expression and accumulation of the major proteins in grapes is essentially
determined by the environmental and/or pathological conditions prevailing during
grape development and maturation. The potential physiological and technological
implications are discussed. The results of the present work suggest that it is
not possible to base varietal differentiation of grapes on the profile of the
pathogenesis-related polypeptides present in the mature berries.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12822945 [PubMed - indexed for MEDLINE]

271: J Agric Food Chem. 2003 Jul 2;51(14):4017-22.

Effects of flumioxazin herbicide on carbon nutrition of Vitis vinifera L.

Saladin G, Magne C, Clement C.

Laboratoire de Biologie et Physiologie Vegetales, URVVC UPRES EA 2069,
Universite de Reims Champagne-Ardenne, Reims Cedex 2, France.

To evaluate the impact of the herbicide flumioxazin (fmx) on nontarget
grapevines, its effects were assessed on fruiting cuttings and field-grown
plants. The stress caused by the herbicide differed according to the grapevine
model. In cuttings, leaf gas exchange and photosynthetic pigment levels as well
as hexose contents decreased, whereas sucrose and starch accumulated, suggesting
an inhibition of photosynthesis and an increase of carbohydrate reserves as a
response to the fmx-induced stress. Paradoxically, in the field-grown grapevine
leaves, fmx caused a stimulation of photosynthesis, an accumulation of
photosynthetic pigments and monosaccharides, in parallel with a mobilization of
sucrose and starch. These results suggest that fmx reaches grapevine leaves via
root uptake and has prolonged effects. In cuttings, fmx generated a toxic effect
related to its target, whereas in field-grown plants, fmx had rather positive
physiological effects and acts as a signal further stimulating photosynthesis
and related parameters.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12822940 [PubMed - indexed for MEDLINE]

272: Plant Cell Rep. 2003 Aug;21(12):1221-7. Epub 2003 Jun 18.

Stress effects of flumioxazin herbicide on grapevine ( Vitis vinifera L.) grown
in vitro.

Saladin G, Clement C, Magne C.

UPRES EA 2069 URVVC, Laboratoire de Biologie et Physiologie Vegetales, UFR
Sciences, Universite de Reims Champagne-Ardenne, Moulin de la Housse, BP 1039,
51687, Reims Cedex 2, France.

Among the herbicides used in vineyards, the pre-emergence soil-applied
flumioxazin (FMX) is a recently synthesized molecule that inhibits chlorophyll
biosynthesis in weed species. The aim of this work was to characterize the
effects of FMX on non-target grapevine ( Vitis vinifera L. cv. Chardonnay)
plantlets grown in vitro. FMX treatment (from 1 to 100 micro M) represented a
stress, as revealed by measurement of several parameters. Stem and leaves
underwent dehydration and a decrease in both water- and osmotic-potential.
Treated plantlets exhibited concomitant accumulation of soluble carbohydrates in
all tissues and of free proline in stems and leaves. Moreover, FMX caused lipid
peroxidation and electrolyte leakage in leaf tissues. These results indicate
that the herbicide FMX is toxic for grapevine grown in vitro. In addition to
inhibiting protoporphyrinogen IX oxidase, it causes water stress and membrane
alteration in tissues and, as a consequence, generates the accumulation of
carbohydrates and free proline.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12819925 [PubMed - indexed for MEDLINE]

273: New Phytol. 2002 Oct;156(1):65-74.

Are xylem radial development and hydraulic conductivity in downwardly-growing
grapevine shoots influenced by perturbed auxin metabolism?

Lovisolo C, Schubert A, Sorce C.

Dip. Colture Arboree Universita Torino, Grugliasco, Italy.
[email protected]

Downwardly-growing grapevine shoots have smaller and more frequent vessels than
upwardly-growing ones and, as a consequence, a lower hydraulic conductivity.
Here, grapevine (Vitis vinifera L.) shoot growth orientation was manipulated to
test whether downward shoot orientation negatively affects vessel growth in the
apex via a shortage of water and nutrients. The orientation of the central vine
shoot portion was inverted by two consecutive 135 degrees bends, resulting in
double-bent N-shaped vines; the central downward shoot portion was of different
lengths in the experimental treatments to induce increasing reductions of shoot
conductivity. These treatments reduced shoot conductivity and water flow, but
had no effects on vessel development and frequency in the apex. In a second
experiment, auxin concentration was assessed in shoots of upwardly- and
downwardly-growing plants. IAA concentration at the apical internodes was higher
in downwardly-oriented shoots than in shoots growing upwards. In addition, a
higher density and a lower vessel diameter were observed in the lower, than the
upper side, of the downwardly-oriented shoot, suggesting increased accumulation
of auxin in the lower side. These results suggest that the downward orientation
induces accumulation of auxin in the apex, which in turn affects the density and
the size of the xylem vessels, causing reduction of hydraulic conductivity.

PMID: 12817565 [PubMed - indexed for MEDLINE]

274: Free Radic Res. 2003 May;37(5):573-84.

Grape and grape seed extract capacities at protecting LDL against oxidation
generated by Cu2+, AAPH or SIN-1 and at decreasing superoxide THP-1 cell
production. A comparison to other extracts or compounds.

Shafiee M, Carbonneau MA, Urban N, Descomps B, Leger CL.

Laboratoire de Nutrition Humaine et Atherogenese, UPRES-EA 2033, Faculte de
Medecine, Universite de Montpellier I, F-34060 Montpellier, France.

A large body of evidence supports the key role of oxidized low-density
lipoprotein in atherosclerosis. The aim of this study was to compare the
capacity of natural polyphenols (PP) from Vitis vinifera and Olea europea at
protecting LDL against oxidation brought about by Cu2+, oxygen-centered
radical-generating AAPH, or peroxynitrite-generating SIN-1 in vitro systems, or
at impairing superoxide production in promonocyte cells (THP-1) conveniently
differentiated into adherent macrophages. PP were either from the whole grape
(fraction A) containing mainly procyanidins, (epi)-catechin and anthocyanins, or
from grape seed extracts (fractions B and C) consisting of tannins and
procyanidin oligomers with a higher content in B than in C, or from a grape skin
extract (fraction D) consisting mainly of anthocyanins, or from a hydrosoluble
olive mill wastewater PP extract (fraction E) containing hydroxytyrosol and
oleuropein. Chlorogenic acid (F) and catechin (G) were taken as archetypes of PP
preventing oxidation partly as copper scavenger and as radical scavenger only,
respectively. All grape fractions were efficient towards Cu2+ system (equally or
more efficient than F), whereas they were rather poorly efficient towards AAPH
and SIN-1 (less efficient than G but as efficient as F). Among the PP fractions,
B was the most effective at protecting LDL in the SIN-1 system and at impairing
THP-1 superoxide production. Taken together, these data suggest that the PP
fraction from grape seed rich in procyanidins achieves the best compromise
between the direct and indirect (i.e. cell-mediated) types of action in
protecting LDL against oxidation, strengthening the need for improving the
knowledge of its bioavailability in humans.

Publication Types:
Comparative Study

PMID: 12797479 [PubMed - indexed for MEDLINE]

275: Mol Plant Microbe Interact. 2003 Jun;16(6):553-64.

The endopolygalacturonase 1 from Botrytis cinerea activates grapevine defense
reactions unrelated to its enzymatic activity.

Poinssot B, Vandelle E, Bentejac M, Adrian M, Levis C, Brygoo Y, Garin J,
Sicilia F, Coutos-Thevenot P, Pugin A.

Laboratoire de Biochimie, Biologie cellulaire et Ecologie des Interactions
Plantes/Micro-Organismes, UMR 1088 INRA/Universite de Bourgogne, 17 rue Sully,
BP 86510, 21065 Dijon cedex, France.

A purified glycoprotein from Botrytis cinerea (strain T4), identified as
endopolygalacturonase 1 (T4BcPG1) by mass spectrometry analysis, has been shown
to activate defense reactions in grapevine (Vitis vinifera cv. Gamay). These
reactions include calcium influx, production of active oxygen species,
activation of two mitogen-activated protein kinases, defense gene transcript
accumulation, and phytoalexin production. Most of these defense reactions were
also activated in grapevine in response to purified oligogalacturonides (OGA)
with a degree of polymerization of 9 to 20. In vivo, these active OGA might be a
part of the released products resulting from endopolygalacturonase activity on
plant cell walls. Nevertheless, the intensity and kinetics of events triggered
by OGA were very different when compared with T4BcPG1 effects. Moreover,
chemical treatments of T4BcPG1 and desensitization assays have allowed us to
discriminate enzymatic and elicitor activities, indicating that elicitor
activity was not due to released oligogalacturonides. Thus, BcPG1 should be
considered as both an avirulence and a virulence factor. The role of the
secreted BcPG1 in the pathogenicity of Botrytis cinerea is discussed.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 12795381 [PubMed - indexed for MEDLINE]

276: Plant Cell Rep. 2003 Apr;21(8):821-7. Epub 2003 Mar 6.

Impact of flumioxazin herbicide on growth and carbohydrate physiology in Vitis
vinifera L.

Saladin G, Magne C, Clement C.

Laboratoire de Biologie et Physiologie Vegetales, URVVC UPRES EA 2069,
Universite de Reims Champagne-Ardenne, UFR Sciences, Moulin de la Housse, BP
1039, 51687, Reims Cedex 2, France.

The impact of flumioxazin herbicide on in vitro-grown grapevine ( Vitis vinifera
L. cv. Chardonnay) was investigated. The herbicide treatments (1, 10 or 100
micro M flumioxazin in MM medium) had a negative impact on photosynthesis, as
revealed by a reduction in foliar chlorophyll and carotenoid contents, gas
exchanges and alteration in plastid structure and, consequently, resulted in a
strong inhibition of plantlet growth. Surprisingly, soluble sugars and starch
accumulated in all organs, suggesting a stimulation of sugar uptake from the
medium. Moreover, photosynthetic activity and starch content partially recovered
within 3 weeks of treatment at the weakest herbicide concentration. These
results provide new insights into the physiological responses of non-target
crops to flumioxazin, showing that flumioxazin is active in photosynthetic
tissues of the non-target grapevine via root uptake, which is contrary to what
is mentioned in the literature, and that the in vitro-grown plantlet is a good
model for investigating the physiological effects of pesticides on crop species.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12789529 [PubMed - indexed for MEDLINE]

277: J Agric Food Chem. 2003 Jun 4;51(12):3682-7.

Pathway leading to the formation of anthocyanin-vinylphenol adducts and related
pigments in red wines.

Schwarz M, Wabnitz TC, Winterhalter P.

Institute of Food Chemistry, Technical University of Braunschweig,
Schleinitzstrasse 20, Germany.

On the basis of observations from Vitis vinifera cv. Pinotage wines and
experiments performed in model wine medium, a new chemical pathway responsible
for the formation of anthocyanin-vinylphenol adducts in red wines is described.
Until now, these pigments have been considered to be reaction products of
anthocyanins and vinylphenols, the latter being generated during fermentation by
enzymatic decarboxylation of the respective cinnamic acids. The mechanism of the
novel pathway, involving intact hydroxycinnamic acid and anthocyanin, is
explained. Only cinnamic acids with electron-donating substituents on the
aromatic ring, such as coumaric acid, ferulic acid, caffeic acid, and sinapic
acid, undergo this conversion, as they stabilize an intermediately formed
carbenium ion. Decarboxylation and oxidation of the pyran moieties are the final
steps in the generation of the corresponding 4-vinylphenol, 4-vinylguaiacol,
4-vinylcatechol, and 4-vinylsyringol adducts of anthocyanins in red wine.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12769545 [PubMed - indexed for MEDLINE]

278: Bull Entomol Res. 2003 Jun;93(3):193-201.

Evidence for host-associated clones of grape phylloxera Daktulosphaira
vitifoliae (Hemiptera: Phylloxeridae) in Australia.

Corrie AM, van Heeswijck R, Hoffmann AA.

Centre for Environmental Stress and Adaptation Research, La Trobe University,
Bundoora, Victoria 3086, Australia. [email protected]

Grape phylloxera, Daktulosphaira vitifoliae Fitch, is an important pest of
grapevines (Vitis vinifera L.) (Vitaceae). Using microsatellite DNA markers it
was demonstrated strong associations can exist between D. vitifoliae asexual
lineages and vine host type within a vineyard. Also, in excised root bioassays,
D. vitifoliae collected from three regions where different genotypic classes
predominated showed host-specific differences in life table parameters of
reproductive rate and intrinsic rate of increase. Lastly, comparisons of
mitochondrial DNA (cytochrome oxidase I) sequences revealed that D. vitifoliae
in Australia have paraphyletic origins and fall into two clades partially
related to vine host usage. These findings indicate introduction of separate
lineages of D. vitifoliae which have close host associations and as such, have
important implications for management of this pest in Australia.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12762861 [PubMed - indexed for MEDLINE]

279: J Plant Physiol. 2003 Mar;160(3):239-45.

Crystallochemical characterization of calcium oxalate crystals isolated from
seed coats of Phaseolus vulgaris and leaves of Vitis vinifera.

Jauregui-Zuniga D, Reyes-Grajeda JP, Sepulveda-Sanchez JD, Whitaker JR, Moreno
A.

Instituto de Quimica, UNAM, Circuito Exterior, C.U. Mexico, D.F. 04510, Mexico.

Calcium oxalate crystals are a major biomineralization product in higher plants.
Their biological function and use are not well understood. In this work, we
focus on the isolation and crystallochemical characterization of calcium oxalate
crystals from seed coats of Phaseolus vulgaris (prisms) and leaves of Vitis
vinifera (raphides and druses) using ultrastructural methods. A proposal based
on crystal growth theory was used for explaining the existence of different
morphologies shown by these crystals grown inside specialized cells in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12749080 [PubMed - indexed for MEDLINE]

280: Theor Appl Genet. 2003 May;106(7):1213-24. Epub 2002 Dec 10.

Molecular linkage maps of Vitis vinifera L. and Vitis riparia Mchx.

Grando MS, Bellin D, Edwards KJ, Pozzi C, Stefanini M, Velasco R.

Istituto Agrario di San Michele all'Adige, Via Mach 1 - I38010 San Michele
all'Adige, Trento, Italy. [email protected]

Two linkage maps for grape (Vitis spp.) have been developed based on 81 F(1)
plants derived from an interspecific cross between the wine cultivar Moscato
bianco (Vitis vinifera L.) and a Vitis riparia Mchx. accession, a donor of
pathogen resistance traits. The double pseudotest-cross mapping strategy was
applied using three types of molecular markers. The efficiency of SSRs to anchor
homologous linkage groups from different Vitis maps and the usefulness of AFLPs
in saturating molecular linkage maps were evaluated. Moreover, the SSCP
technique was developed based on sequence information in public databases
concerning genes involved in flavonoid and stilbene biosynthesis. For the
maternal genetic map a total of 338 markers were assembled in 20 linkage groups
covering 1,639 cM, whereas 429 loci defined the 19 linkage groups of the
paternal map which covers 1,518 cM. The identification of 14 linkage groups
common to both maps was possible based on 21 SSR and 19 AFLP loci. The position
of SSR loci in the maps presented here was consistent with other published
mapping experiments in Vitis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12748772 [PubMed - indexed for MEDLINE]

281: Mol Plant Microbe Interact. 2003 May;16(5):456-64.

Nonspecific lipid-transfer protein genes expression in grape (Vitis sp.) cells
in response to fungal elicitor treatments.

Gomes E, Sagot E, Gaillard C, Laquitaine L, Poinssot B, Sanejouand YH, Delrot S,
Coutos-Thevenot P.

Laboratoire de Physiologie et Biochimie Vegetale, CNRS UMR 6161, Universite de
Poitiers, 40 avenue du Recteur Pineau, 86022 Poitiers, France.
[email protected]

Nonspecific lipid transfer proteins (nsLTPs) are small, basic cystein-rich
proteins believed to be involved in plant defense mechanisms. Three cDNAs coding
nsLTPs from grape (Vitis vinifera sp.) were cloned by
reverse-transcriptase-polymerase chain reaction (RT-PCR) and PCR. The expression
of nsLTP genes was investigated in 41B-rootstock grape cell suspension, in
response to various defense-related signal molecules. Ergosterol (a
fungi-specific sterol) and a proteinaceous elicitor purified from Botrytis
cinerea strongly and rapidly induced the accumulation of nsLTP mRNAs. Jasmonic
acid, cholesterol, and sitosterol also promoted nsLTPs mRNA accumulation,
although to a lesser extent, whereas salicylic acid had no effect. High
performance liquid chromatography analysis indicated that the amounts of three
LTP isoforms (previously named P1, P2, and P4) were increased by ergosterol.
None of the four isoforms displayed any significant antifungal properties, with
the exception of the P4 isoform, which reduced Botrytis mycelium growth in
vitro, but only in calcium-free medium. The results are discussed in the context
of plant-pathogen interactions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12744517 [PubMed - indexed for MEDLINE]

282: Theor Appl Genet. 2003 Aug;107(3):448-54. Epub 2003 May 7.

Microsatellite analysis of ancient alpine grape cultivars: pedigree
reconstruction of Vitis vinifera L. 'Cornalin du Valais'.

Vouillamoz J, Maigre D, Meredith CP.

Department of Viticulture and Enology, University of California, One Shields
Avenue, Davis 95616 CA, USA. [email protected]

Ancient and closely related grape cultivars from the Alps were analyzed with 50
microsatellite markers: 'Cornalin', 'Humagne Rouge' and 'Goron' from Valais
(Switzerland); 'Cornalin', 'Petit Rouge' and 'Mayolet' from the Aosta Valley
(Italy). Our results confirmed previous studies showing that the 'Cornalin'
cultivars from Switzerland and Italy are distinct, and that 'Humagne Rouge' is
identical to 'Cornalin' from the Aosta Valley. We propose the nomenclature
'Cornalin du Valais' and 'Cornalin d'Aoste' in order to prevent further
confusion. At each locus, 'Goron', 'Petit Rouge', 'Mayolet' and 'Cornalin
d'Aoste' all share at least one allele with 'Cornalin du Valais', strongly
suggesting parent/offspring relationships. Alleles at 49 out of 50
microsatellite loci are consistent with 'Cornalin du Valais' being the progeny
of 'Petit Rouge' and 'Mayolet'. The exception is a 10-base pair discrepancy at
one locus, most likely the result of somatic mutation in one of the parents,
since this parentage is supported by high likelihood ratios and historical data.
We hypothesize that 'Cornalin du Valais' originated in the Aosta Valley through
a natural cross and was then introduced into Valais centuries ago, probably via
the Great St. Bernard Pass. Furthermore, 'Cornalin du Valais' is likely to be
one of the parents of both 'Goron' and 'Cornalin d'Aoste', the respective second
parents remaining unknown. This pedigree provides a convincing explanation for
the allele-sharing patterns and is strongly supported by historical data. The
present work is the first grapevine parentage study to deal with a multiple
repeat unit discrepancy at a microsatellite locus. We suggest that the use of
increasingly large numbers of loci in making parentage determinations leads to a
corresponding increase in the probability of encountering a locus with
intra-cultivar variability during the analysis. We therefore assume that a sole
multiple repeat unit discrepancy is not sufficient to discard a parentage
hypothesis.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 12734654 [PubMed - indexed for MEDLINE]

283: Chemosphere. 2003 Jun;51(9):913-23.

Background ozone monitoring and phytodetection in the greater rural area of
Corinth--Greece.

Saitanis CJ.

Laboratory of Ecology and Environmental Sciences, Agricultural University of
Athens, Iera Odos 75, Greece. [email protected]

Natural background ozone levels were monitored in three places within the
greater rural area of Corinth, namely Bogdani Hill, Astronomical Observatory of
Krionerion, and Kiato, and compared with ambient ozone monitored in the
metropolitan area of Athens. Measurements were made sequentially, for a few
weeks at each place, during the summer of 2000. In addition, ozone
phytodetection, using tobacco (Nicotiana tabacum L.) plants of the Bel-W3 and
Zichnomirodata varieties, was conducted in 12 places (the above included).
Moreover, stomatal conductance was measured in the Bel-W3 plants, as well as in
leaves of cultivated grape-vines (Vitis vinifera L.) and in needles of Aleppo
pine (Pinus halepensis Mill.) trees and compared with the diurnal pattern of
ozone concentrations.The 24 and 12 (08:00-20:00) hourly averages of ozone
concentrations were high in Athens (37; 51 ppb), at Bogdani Hill (53; 56 ppb)
and at the Astronomical Observatory (56; 55 ppb), but relatively low in Kiato
(30; 34 ppb). Furthermore, the average daily AOT40 (accumulated exposure over 40
ppb for the daylight hours) (ppbh) was 193 in Athens, 212 at Bogdani Hill, 192
at the Astronomical Observatory and 47 in Kiato. Ozone concentrations exhibited
the usual diurnal pattern in Athens (altitude 50 m), where they were maximum
during midday and early afternoon hours, as well as at Bogdani Hill (300 m) and
in Kiato (5 m) where, however, they were maximal 1-3 h later. At the
Astronomical Observatory (altitude 920 m) ozone remained constant during both
daylight and night hours. The differences in diurnal patterns are consistent
with those in places of different elevation, reported elsewhere.The Bel-W3
plants were injured at all 12 places; Zichnomirodata plants exhibited lower
injury and only in some of the places; probable ozone symptoms were also
observed on vine plants and pine trees. The greatest injury was observed at the
high altitude places of Astronomical Observatory and Mougostos. Stomatal
conductance, in all three species, peaked during morning and early midday hours
when ozone levels were higher in the high altitude, and lower in the low
altitude, places.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12697182 [PubMed - indexed for MEDLINE]

284: Appl Environ Microbiol. 2003 Apr;69(4):1904-12.

Colonization of Vitis vinifera by a green fluorescence protein-labeled,
gfp-marked strain of Xylophilus ampelinus, the causal agent of bacterial
necrosis of grapevine.

Grall S, Manceau C.

UMR Pathologie Vegetale, INRA-INH-Universite d'Angers, Institut National de la
Recherche Agronomique, Centre d'Angers, F-49071 Beaucouze, France.

The dynamics of Xylophilus ampelinus were studied in Vitis vinifera cv. Ugni
blanc using gfp-marked bacterial strains to evaluate the relative importance of
epiphytic and endophytic phases of plant colonization in disease development.
Currently, bacterial necrosis of grapevine is of economic importance in
vineyards in three regions in France: the Cognac, Armagnac, and Die areas. This
disease is responsible for progressive destruction of vine shoots, leading to
their death. We constructed gfp-marked strains of the CFBP2098 strain of X.
ampelinus for histological studies. We studied the colonization of young plants
of V. vinifera cv. Ugni blanc by X. ampelinus after three types of artificial
contamination in a growth chamber and in a greenhouse. (i) After wounding of the
stem and inoculation, the bacteria progressed down to the crown through the
xylem vessels, where they organized into biofilms. (ii) When the bacteria were
forced into woody cuttings, they rarely colonized the emerging plantlets. Xylem
vessels could play a key role in the multiplication and conservation of the
bacteria, rather than being a route for plant colonization. (iii) When bacterial
suspensions were sprayed onto the plants, bacteria progressed in two directions:
both in emerging organs and down to the crown, thus displaying the importance of
epiphytic colonization in disease development.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12676663 [PubMed - indexed for MEDLINE]

285: J Exp Bot. 2003 Apr;54(385):1193-204.

Expression of a putative grapevine hexose transporter in tobacco alters
morphogenesis and assimilate partitioning.

Leterrier M, Atanassova R, Laquitaine L, Gaillard C, Coutos-Thevenot P, Delrot
S.

UMR CNRS 6161, Transport des Assimilats, Laboratoire de Physiologie, Biochimie
et Biologie Moleculaire Vegetales, Batiment Botanique, UFR Sciences, 40 Avenue
du Recteur Pineau, F-86022 Poitiers Cedex, France.

Tobacco plants were transformed by leaf disc regeneration with the VvHT1 (Vitis
vinifera hexose transporter 1) cDNA under the control of the constitutive CaMV
35S promoter in a sense or antisense orientation. Among the 20 sense plants and
10 antisense plants obtained, two sense plants showed a mutant phenotype when
grown in vitro, with stunted growth and an increase in the (leaves+stem)/roots
dry weight ratio. The rate of [(3)H]-glucose uptake in leaf discs from these
plants was decreased to 25% of the value measured in control plants. The amount
of VvHT1 transgene and of host monosaccharide transporter MST transcripts in the
leaves were studied by RNA gel blot analysis. The VvHT1 transcripts were usually
present, but the amount of MST transcripts was the lowest in the plants that
exhibited the most marked phenotype. Although the phenotype was lost when the
plants were transferred from in vitro to greenhouse conditions, it was found
again in vitro in the progeny obtained by self-pollination or by back-cross. The
data show that VvHT1 sense expression resulted in unidirectional
post-transcriptional gene inactivation of MST in some of the transformants, with
dramatic effects on growth. They provide the first example of plants modified
for hexose transport by post-transcriptional gene silencing. Some of the
antisense plants also showed reduced expression of MST, and decreased growth.
These results indicate that, like the sucrose transporters, hexose transporters
play an important role in assimilate transport and in morphogenesis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12654870 [PubMed - indexed for MEDLINE]

286: Theor Appl Genet. 2003 Mar;106(5):912-22. Epub 2002 Dec 3.

Development and transferability of apricot and grape EST microsatellite markers
across taxa.

Decroocq V, Fave MG, Hagen L, Bordenave L, Decroocq S.

INRA Centre de Bordeaux, U.R.E.F.V, 71 Avenue E. Bourleaux, 33883 Villenave
d'Ornon France. [email protected]

EST microsatellite markers were developed in apricot (Prunus armeniaca L.) and
grape (Vitis vinifera L.). cDNA libraries from either apricot leaves or grape
roots were used in an enrichment procedure for GA and CA repeats. The
transferability of EST simple sequence repeat (SSR) markers from apricot and
grapevine to other related and unrelated species was examined. Overall, grape
primers amplified products in most of the Vitaceae accessions while the apricot
primers amplified polymorphic alleles only in closely related species of the
Rosaceae. In this taxonomic family, ten EST SSR loci were tested, and one single
primer pair, PacB22, was amplified across species and sections in the Prunoideae
and Maloideae. Sequencing of EST SSR loci in other species and genera confirmed
a higher level of conservation in the microsatellite motif and flanking regions
in the Vitaceae compared to the Rosaceae. Two distinct fragments of the PacB22
locus amplified across the Malus and Pyrus genera; however, while the coding
region was highly conserved, the microsatellite repeat motif was no longer
present. The banding pattern was explained by base substitution and
insertion/deletion events in the intronic region of PacB22. This study includes
the determination of the degree of polymorphism detected among species and
genera in two unrelated taxonomic families and the evaluation of the information
provided by the microsatellite repeats and the flanking regions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12647067 [PubMed - indexed for MEDLINE]

287: J Agric Food Chem. 2003 Mar 26;51(7):1839-44.

Characterization of anthocyanins in grape juices by ion trap liquid
chromatography-mass spectrometry.

Wang H, Race EJ, Shrikhande AJ.

Research and Development, Canandaigua Wine Company, 12667 Road 24, Madera,
California 93639, USA. [email protected]

A reverse phase HPLC and electrospray interface with ion trap mass spectrometer
method was developed for the characterization of anthocyanins in Concord,
Rubired, and Salvador grape juices. Rubired and Salvador grapes are hybrids from
Vitis vinifera and Vitis rupestris. Concord grape is a grape from the native
American cultivar Vitis labrusca. Individual anthocyanins in these three
varieties were identified on the basis of UV-vis and MS spectra and further
elucidated by MS/MS spectra. Anthocyanins in Salvador and Concord grapes were
3-O-glucosides, 3-O-(6' '-O-p-coumaroyl)glucosides, 3-O-(6'
'-O-p-acetyl)glucosides, 3,5-O-diglucosides, and 3-O-(6'
'-O-p-coumaroyl)-5-O-diglucosides of delphinidin, cyanidin, petunidin, peonidin,
and malvidin. Vitisin B was detected in Salvador grape juice. Anthocyanins in
Rubired grape juice were primarily anthocyanin diglucosides: peonidin
3,5-O-diglucoside, malvidin 3,5-O-diglucoside, peonidin 3-O-(6'
'-O-p-coumaroyl)-5-O-diglucoside, and malvidin 3-O-(6'
'-O-p-coumaroyl)-5-O-diglucoside are the four major anthocyanins. The presence
of pelargonidin 3-O-glucoside, not previously reported, has been established for
the first time in all three juices.

PMID: 12643639 [PubMed - indexed for MEDLINE]

288: J Med Food. 2001 Summer;4(2):93-105.

Grape Extract, Resveratrol, and Its Analogs: A Review.

Sovak M.

Biophysica Foundation and Radiology Research, University of California Medical
School, San Diego, 3333 N. Torrey Pines Ct., Ste. 100, La Jolla, CA 92037.

The recent and essential reports on the biological activity of the principal
phytophenols of Vitis vinifera and wine, with special attention to resveratrol,
are reviewed. The phytophenols are arbitrarily divisible into single-ring
phenolic acids, bisphenols including stilbenes, tricyclic phenols (flavonoids)
and their subclasses, and oligomeric and polymeric species, the
proanthocyanidins and anthocyanidins. Their precursors and the stilbenes,
including resveratrol with its analogs and conjugates, appear to be of
preventative and possibly therapeutic value in atherosclerosis and certain
neoplastic and inflammatory afflictions. The probable mechanisms are free
radical scavenging and selective interference with a multitude of factors
affecting the division cycle of rapidly and abnormally proliferating mammalian
cells. Reviewed are studies of natural occurrence, extraction methods,
bioavailability, analytical detection, and metabolism of resveratrol, as well as
its effects on cancer and inflammation, atherosclerosis, and neurons. Because
grape extracts are a convenient alimentary source of salutary phytochemicals to
supplement currently prevalent occidental food and resveratrol appears to be
especially useful, it could conveniently be added in biosignificant amounts to
the grape extracts provided that their extraction, contents, and quality
controls are instituted.

PMID: 12639418 [PubMed - as supplied by publisher]

289: J Agric Food Chem. 2003 Mar 12;51(6):1543-7.

Analysis of wine components in Cynthiana and Syrah wines.

Walker T, Morris J, Threlfall R, Main G.

Department of Food Science and Institute of Food Science and Engineering,
University of Arkansas, 2650 North Young Avenue, Fayetteville 72704, USA.

Red wine is composed of a complex matrix of compounds that can interfere with
analysis. A high-performance liquid chromatography (HPLC) procedure was
developed to efficiently analyze organic acids, sugars, glycerol, and ethanol in
Cynthiana (Vitis aestivalis) wine. Standard laboratory procedures (pH,
titratable acidity, and color attributes) and HPLC were found reproducible for
Cynthiana wine. HPLC recovery efficiency was determined by analysis of spiked
and unspiked samples (model, Cynthiana, and Syrah (Vitis vinifera) wines).
Although recovery of components was greater in the model wine, recovery in
Cynthiana and Syrah wine was comparable. The HPLC procedure was further compared
to commercial rapid enzyme analysis tests using model, Cynthiana, and Syrah
wines. HPLC analyses were more accurate than enzymatic tests for determining
components in the model, Cynthiana, and Syrah wines. Considering the complexity
of the wines analyzed, reproducibility and recovery of the HPLC procedure was
demonstrated and showed improvement and precision when compared to existing
methods.

PMID: 12617581 [PubMed - indexed for MEDLINE]

290: J Agric Food Chem. 2003 Feb 26;51(5):1373-6.

Contribution of benzenemethanethiol to smoky aroma of certain Vitis vinifera L.
wines.

Tominaga T, Guimbertau G, Dubourdieu D.

Faculte d'Oenologie, Universite de Victor Segalen Bordeaux 2, 351 cours de la
Liberation, 33405 Talence, France. [email protected]

Benzenemethanethiol, a volatile thiol with a strong empyreumatic aroma
reminiscent of smoke, has been identified in boxwood (Buxus sempervirens L.) as
well as in red and white Vitis vinifera L. wines. The perception threshold in a
model hydroalcoholic solution is approximately 0.3 ng/L. All of the wines
analyzed for this study contained this compound in concentrations of several
dozen nanograms per liter. The Chardonnay wines had 30-40 ng/L. Sensory
discrimination between a wine containing 7 ng/L benzenemethanethiol and the same
wine with an additional 4 ng/L is very significant; the difference in smell is
described as "empyreumatic". This compound can therefore significantly
contribute to the aroma of certain wines (Sauvignon Blanc, Semillon, Chardonnay,
etc.) containing concentrations as high as 30-100 times higher than their
perception threshold.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12590483 [PubMed - indexed for MEDLINE]

291: Theor Appl Genet. 2002 Mar;104(4):610-618.

Identification of resistance gene analogs linked to a powdery mildew resistance
locus in grapevine.

Donald TM, Pellerone F, Adam-Blondon AF, Bouquet A, Thomas MR, Dry IB.

CSIRO Plant Industry, Horticulture Unit, PO Box 350, Glen Osmond SA 5064,
Australia. [email protected]

Oligonucleotide primers, designed to conserved regions of nucleotide binding
site (NBS) motifs within previously cloned pathogen resistance genes, were used
to amplify resistance gene analogs (RGAs) from grapevine. Twenty eight unique
grapevine RGA sequences were identified and subdivided into 22 groups on the
basis of nucleic acid sequence-identity of approximately 70% or greater.
Representatives from each group were used in a bulked segregant analysis
strategy to screen for restriction fragment length polymorphisms linked to the
powdery mildew resistance locus, Run1, introgressed into Vitis vinifera L. from
the wild grape species Muscadinia rotundifolia. Three RGA markers were found to
be tightly linked to the Run1 locus. Of these markers, two (GLP1-12 and MHD145)
cosegregated with the resistance phenotype in 167 progeny tested, whereas the
third marker (MHD98) was mapped to a position 2.4 cM from the Run1 locus. The
results demonstrate the usefulness of RGA sequences, when used in combination
with bulked segregant analysis, to rapidly generate markers tightly linked to
resistance loci in crop species.

PMID: 12582665 [PubMed - as supplied by publisher]

292: Theor Appl Genet. 2002 Sep;105(4):614-621. Epub 2002 Jun 21.

Complete sequence of Tvv1, a family of Ty 1 copia-like retrotransposons of Vitis
vinifera L., reconstituted by chromosome walking.

Pelsy F, Merdinoglu D.

Laboratoire de Genetique et Amelioration des Plantes, UMR Vigne et Vins
d'Alsace, INRA-Colmar, 28 rue de Herrlisheim, 68000 Colmar. France,
[email protected]

A chromosome-walking strategy was used to sequence and characterize
retrotransposons in the grapevine genome. The reconstitution of a family of
retroelements, named Tvv1, was achieved by six successive steps. These elements
share a single, highly conserved open reading frame 4,153 nucleotides-long,
putatively encoding the gag, pro, int, rt and rh proteins. Comparison of the
Tvv1 open reading frame coding potential with those of drosophila copia and
tobacco Tnt1, revealed that Tvv1 is closely related to Ty 1 copia-like
retrotransposons. A highly variable untranslated leader region, upstream of the
open reading frame, allowed us to differentiate Tvv1 variants, which represent a
family of at least 28 copies, in varying sizes. This internal region is flanked
by two long terminal repeats in direct orientation, sized between 149 and 157
bp. Among elements theoretically sized from 4,970 to 5,550 bp, we describe the
full-length sequence of a reference element Tvv1-1, 5,343 nucleotides-long. The
full-length sequence of Tvv1-1 compared to pea PDR1 shows a 53.3% identity. In
addition, both elements contain long terminal repeats of nearly the same size in
which the U5 region could be entirely absent. Therefore, we assume that Tvv1 and
PDR1 could constitute a particular class of short LTRs retroelements.

PMID: 12582512 [PubMed - as supplied by publisher]

293: Theor Appl Genet. 2002 Oct;105(5):780-795. Epub 2002 May 25.

Genetic mapping of grapevine ( Vitis vinifera L.) applied to the detection of
QTLs for seedlessness and berry weight.

Doligez A, Bouquet A, Danglot Y, Lahogue F, Riaz S, Meredith P, Edwards J, This
P.

INRA, UMR Diversite et Genomes des Plantes Cultivees, equipe Genetique Vigne,
Bat 6, 2 place Viala, F-34060 Montpellier Cedex 1, France, [email protected]

Parental and consensus genetic maps of Vitis vinifera L. (2n = 38) were
constructed using a F(1) progeny of 139 individuals from a cross between two
partially seedless genotypes. The consensus map contained 301 markers [250
amplification fragment length polymorphisms (AFLPs), 44 simple sequence repeats
(SSRs), three isozymes, two random amplified polymorphic DNAs (RAPDs), one
sequence-characterized amplified region (SCAR), and one phenotypic marker, berry
color] mapped onto 20 linkage groups, and covered 1,002 cM. The maternal map
consisted of 157 markers covering 767 cM (22 groups). The paternal map consisted
of 144 markers covering 816 cM (23 groups). Differences in recombination rates
between these maps and another unpublished map are discussed. The major gene for
berry color was mapped on both the paternal and consensus maps. Quantitative
trait loci (QTLs) for several quantitative subtraits of seedlessness in 3
successive years were searched for, based on parental maps: berry weight, seed
number, seed total fresh and dry weights, seed percent dry matter, and seed mean
fresh and dry weights. QTLs with large effects (R(2) up to 51%) were detected
for all traits and years at the same location on one linkage group, with some
evidence for the existence of a second linked major QTL for some of them. For
these major QTLs, differences in relative parental effects were observed between
traits. Three QTLs with small effects (R(2) from 6% to 11%) were also found on
three other linkage groups, for berry weight and seed number in a single year,
and for seed dry matter in 2 different years.

PMID: 12582493 [PubMed - as supplied by publisher]

294: Theor Appl Genet. 2002 Jan;104(1):61-6.

Evaluating the potential of SSR flanking regions for examining taxonomic
relationships in the Vitaceae.

Rossetto M, McNally J, Henry RJ.

Centre for Plant Conservation Genetics, Southern Cross University, PO Box 157
Lismore NSW, Australia 2480. [email protected]

Three EST-derived microsatellite loci from Vitis vinifera were amplified and
sequenced across eight species of Vitaceae from four different genera.
Phylogenetic analysis of the microsatellite's flanking regions produced
informative results in congruence with previous studies. Generic relationships
were respected and the data produced sufficient inter-specific variation to
distinguish between Cayratia acris and Cayratia saponaria, two very closely
related species. Overall, the sequence alignments showed that priming sites were
conserved, whereas microsatellite repeats were present in most cases but
structurally variable. The sequence data provided information on the
evolutionary patterns of various microsatellite repeats and their correlation to
evolutionary relationships among taxa.

PMID: 12579429 [PubMed - as supplied by publisher]

295: Ying Yong Sheng Tai Xue Bao. 2002 Oct;13(10):1323-38.

[Responses of agricultural crops of free-air CO2 enrichment]

[Article in Chinese]

Kimball BA, Zhu J, Cheng L, Kobayashi K, Bindi M.

U. S. Water Conservation Laboratory, USDA, Agricultural Research Service
Phoenix, Arizona 85040, USA. [email protected]

Over the past decade, free-air CO2 enrichment (FACE) experiments have been
conducted on several agricultural crops: wheat(Triticum aestivum L.), perennial
ryegrass (Lolium perenne), and rice(Oryza sativa L.) which are C3 grasses;
sorghum (Sorghum bicolor (L.) Moench), a C4 grass; white clover (Trifolium
repens), a C3 legume; potato (Solanum tuberosum L.), a C3 forb with tuber
storage; and cotton (Gossypium hirsutum L.) and grape (Vitis vinifera L.) which
are C3 woody perennials. Using reports from these experiments, the relative
responses of these crops was discussed with regard to photosynthesis, stomatal
conductance, canopy temperature, water use, water potential, leaf area index,
shoot and root biomass accumulation, agricultural yield, radiation use
efficiency, specific leaf area, tissue nitrogen concentration, nitrogen yield,
carbohydrate concentration, phenology, soil microbiology, soil respiration,
trace gas emissions, and soil carbon sequestration. Generally, the magnitude of
these responses varied with the functional type of plant and with the soil
nitrogen and water status. As expected, the elevated CO2 increased
photosynthesis and biomass production and yield substantially in C3 species, but
little in C4, and it decreased stomatal conductance and transpiration in both C3
and C4 species and greatly improved water-use efficiency in all the crops.
Growth stimulations were as large or larger under water-stress compared to
well-watered conditions. Growth stimulations of non-legumes were reduced at low
soil nitrogen, whereas elevated CO2 strongly stimulated the growth of the clover
legume both at ample and under low N conditions. Roots were generally stimulated
more than shoots. Woody perennials had larger growth responses to elevated CO2,
while at the same time, their reductions in stomatal conductance were smaller.
Tissue nitrogen concentrations went down while carbohydrate and some other
carbon-based compounds went up due to elevated CO2, with leaves and foliage
affected more than other organs. Phenology was accelerated slightly in most but
not all species. Elevated CO2 affected some soil microbes greatly but not
others, yet overall activity appears to be stimulated. Detection of
statistically significant changes in soil organic carbon in any one study was
impossible, yet combining results from several sites and years, it appears that
elevated CO2 did increase sequestration of soil carbon. Whenever possible,
comparisons were made between the FACE results and those from prior
chamber-based experiments reviewed in the literature. Over all the data and
parameters considered in this review, there are only two parameters for which
the FACE- and chamber-based data appear to be inconsistent. One is that elevated
CO2 from FACE appears to reduce stomatal conductance about one and a half times
more than observed in prior chamber experiments. Similarly, elevated CO2 appears
to have stimulated root growth relatively more than shoot growth under FACE
conditions compared to chamber conditions. Nevertheless, for the most part, the
FACE- and chamber-based results have been consistent, which gives confidence
that conclusions drawn from both types of data are accurate. However, the more
realistic FACE environment and the larger plot size have enabled more extensive
robust multidisciplinary data sets to be obtained under conditions
representative of open fields in the future high-CO2 world.

Publication Types:
English Abstract

PMID: 12557686 [PubMed - indexed for MEDLINE]

296: Plant Physiol. 2003 Jan;131(1):326-34.

Sugar-regulated expression of a putative hexose transport gene in grape.

Atanassova R, Leterrier M, Gaillard C, Agasse A, Sagot E, Coutos-Thevenot P,
Delrot S.

Unite Mixte de Recherche-Centre National de la Recherche Scientifique 6161,
Transport des Assimilats, Laboratoire de Physiologie, Biochimie et Biologie
Moleculaires Vegetales, Poitiers, France. [email protected]

Different lengths of the promoter of grape (Vitis vinifera) VvHT1 (Hexose
Transporter 1) gene, which encodes a putative hexose transporter expressed
during the ripening of grape, have been transcriptionally fused to the
beta-glucuronidase reporter gene. In transgenic tobacco (Nicotiana tabacum)
transformed with these constructs, VvHT1 promoters were clearly responsible for
the sink organ preferential expression. The potential sugar effectors of VvHT1
promoter were studied in tobacco cv Bright-Yellow 2 cells transformed with
chimeric constructs. Glucose (56 mM), sucrose (Suc; 58 mM), and the
non-transported Suc isomer palatinose doubled the beta-glucuronidase activity
conferred by the VvHT1 promoter, whereas fructose did not affect it. These
effects were the strongest with the 2.4-kb promoter, which contains all putative
sugar-responsive elements (activating and repressing), but they were also
significant with the 0.3-kb promoter, which contains only activating sugar
boxes. The induction of VvHT1 expression by both Suc and palatinose was
confirmed in the homologous grape berry cell culture. The data provide the first
example of a putative sugar transporter, which is induced by both glucose and
Suc in higher plants. Although induction of VvHT1 expression by Suc does not
require transport, the presence of glucosyl moiety is necessary for Suc sensing.
These results provide new insights into sugar sensing and signaling in plants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12529540 [PubMed - indexed for MEDLINE]

297: Plant Physiol. 2003 Jan;131(1):129-38.

trans-Resveratrol and grape disease resistance. A dynamical study by
high-resolution laser-based techniques.

Montero C, Cristescu SM, Jimenez JB, Orea JM, te Lintel Hekkert S, Harren FJ,
Gonzalez Urena A.

Unidad de Laseres y Haces Moleculares Instituto Pluridisciplinar, Universidad
Complutense de Madrid Po Juan XXIII, 1. 28040 Madrid, Spain.

Two modern laser-based techniques were synchronously applied to study the
dynamics of the trans-resveratrol activity in Botrytis cinerea-infected grapes.
Direct analysis of trans-resveratrol in both infected and noninfected grapes
(Vitis vinifera, Aledo variety) was performed by using an analytical technique
incorporating laser desorption coupled with laser resonant ionization and
time-of-flight mass spectrometry. On the other hand, one of the most sensitive
on-line methods for trace gas detection, laser photoacoustic spectroscopy, was
used to investigate the involvement of the plant hormone ethylene (C(2)H(4)) in
the B. cinerea grapes interaction and its temporal relationship with the
trans-resveratrol content upon infection. The trans-resveratrol content and the
ethylene released by noninfected grapes showed an opposite behavior. In this
case, a high trans-resveratrol content corresponds to a low ethylene emission.
For the B. cinerea-infected grapes, ethylene emission rises up after 48 h when
the analogous content of trans-resveratrol started to decrease irreversibly.
Moreover, the activity of trans-resveratrol as natural pesticide has been
investigated by exogenous application on grapes. A short submerge (5 s) of the
grapes in 1.6 x 10(-4) M solution of trans-resveratrol delays the increase of
C(2)H(4) emission with about 48 h and produces a decrease of the C(2)H(4)
concentration and its emission rate. The treatment has positive effects on fruit
conservation during storage; it doubled the normal shelf-life of grapes at room
temperature, maintaining their post-harvest quality within 10 d.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12529521 [PubMed - indexed for MEDLINE]

298: Genome. 2002 Dec;45(6):1142-9.

Chloroplast microsatellite polymorphisms in Vitis species.

Arroyo-Garcia R, Lefort F, de Andres MT, Ibanaez J, Borrego J, Jouve N, Cabello
F, Martinez-Zapater JM.

Departamento de Genetica Molecular de Plantas, Centro Nacional de
Biotecnologia-CSIC, Campus de la Universidad Autonoma de Madrid, Cantoblanco,
Spain.

The use of consensus chloroplast microsatellites primers for dicotyledonous
chloroplast genomes revealed the existence of intra and interspecific length
variation within the genus Vitis. Three chloroplast microsatellite loci were
found to be polymorphic in samples of Vitis vinifera, Vitis berlandieri, Vitis
riparia, and Vitis rupestris out of a total of 10 consensus primer pairs tested.
These polymorphisms were always due to a variable number of mononucleotide
residues within A and (or) T stretches in the amplified regions. Chloroplast
microsatellite polymorphisms were used to demonstrate the maternal inheritance
of chloroplast in V. vinifera and to characterise the chloroplast haplotypes
present in wine grape cultivars of this species grown in Spain and Greece. The
different distribution of haplotype frequencies in the two ends of the
Mediterranean growth area suggests the existence of independent domestication
events for grapevine.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12502260 [PubMed - indexed for MEDLINE]

299: J Exp Bot. 2003 Jan;54(381):355-63.

Spermidine and related-metabolic inhibitors modulate sugar and amino acid levels
in Vitis vinifera L.: possible relationships with initial fruitlet abscission.

Aziz A.

Unite de Recherche Vignes et Vins de Champagne, UPRES EA 2069, UFR Sciences,
Universite de Reims Champagne-Ardenne, BP 1039, F-51687 REIMS cedex 2, France.
[email protected]

The relationships between free polyamines (PAs), sugar and amino acid status
were investigated in cuttings from two cultivars of Vitis vinifera L., Pinot
noir (PN), a low abscising cultivar and Merlot (MRT), a high abscising one. In
both cultivars free PAs decreased in inflorescences, but more drastically in MRT
plants. Upon anthesis, this was associated with a decreased sugar content,
especially sucrose, and an increase in total free amino acids. Thereafter,
sucrose and amino acids showed opposite trends. In addition, darkening the PN
plants at full flowering resulted in a dramatic decrease of PAs and sugars in
inflorescences, but an increase in amino acid content, followed by high
abscission. The concept that initial fruitlet abscission might be regulated by
free PAs through changes in primary metabolites was hypothesized. Hence, the
application of exogenous spermidine (Spd), but not putrescine (Put), prior to
flowering markedly inhibits abscission. The Spd treatment also increased soluble
sugar content but reduced amino acids in both leaves and inflorescences, while
Put had no significant effect. By contrast, cyclohexylamine and
beta-hydroxyethylhydrazine, as potent inhibitors of Spd synthase and
PA-oxidases, respectively, exerted inverse effects on sugar, amino acid and
abscission levels. Sucrose and free proline seemed to be highly sensitive to
these treatments. This study suggests that Spd could regulate fruitlet
abscission in grapevine by modulating, in a reverse way, the levels of sugars
and amino acids in inflorescences.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12493864 [PubMed - indexed for MEDLINE]

300: J Med Food. 2002 Summer;5(2):69-78.

Synergistic antioxidative properties of phenolics from natural origin toward
low-density lipoproteins depend on the oxidation system.

Shafiee M, Carbonneau MA, d'Huart JB, Descomps B, Leger CL.

Laboratoire de Nutrition Humaine et Atherogenese, UPRES-EA 2033, Faculte de
Medecine, Universite de Montpellier I, F-34060, Montpellier, France.

Using an approach in line with that of a previous report, we assessed the
antioxidant activity of several natural, polyphenol- or tocotrienol-rich
mixtures: extracts from Elaesis Guineensis oil (A) and Vitis vinifera (B), a
Coffea robusta powder (C), and extracts from Olea europea mill wastewaters (D),
Solanum melongena (E), and Lycopersicon esculentum (F). The copper- and
2-2'-azobis(2-amidinopropane) hydrochloride (AAPH)-oxidation systems were used
in the presence of low-density lipoprotein. For comparison, antioxidant
activities of chlorogenic acid and catechin, as archetypes of molecules highly
efficient with the copper- and the AAPH-oxidation system, respectively, were
assessed. The aim was to establish the occurrence of synergistic antioxidant
actions among some of these natural mixtures. On a molar basis, the highest
specific antioxidant activities (SAA) were found for B, chlorogenic acid, and C
in the copper system, and for A, catechin, and B in the AAPH system. On a mass
basis, the highest SAA were found, respectively, for chlorogenic acid, B, and
catechin, and for catechin, chlorogenic acid, and B. These results show that
large discrepancies take place in the evaluations between the two systems. B and
C exhibited a synergistic antioxidant efficiency, in the presence or absence of
A, but only with the copper system. This was also true for the two types of
A+B+C mixture that were tested. It is thought that this association might
provide an ideal combination, incorporating both the radical scavenger and the
transition-metal ion chelation properties of B and C.

Publication Types:
Comparative Study
In Vitro

PMID: 12487753 [PubMed - indexed for MEDLINE]

301: Rapid Commun Mass Spectrom. 2003;17(1):9-16.

Characterization of proanthocyanidins in grape seeds using electrospray mass
spectrometry.

Hayasaka Y, Waters EJ, Cheynier V, Herderich MJ, Vidal S.

The Australian Wine Research Institute, PO Box 197, Glen Osmond, SA, 5064,
Australia. [email protected]

Two proanthocyanidin (PA) fractions, one (Sdp3) with the mean degree of
polymerization (mDP) of 3 and the other (Sdp9) with mDP of 9, were obtained from
a Vitis vinifera cv. Shiraz grape seed extract. The PA fractions were directly
analyzed by electrospray ionization mass spectrometry (ES-MS) and negative ion
mass spectra were recorded. The mass spectrum of Sdp3 exhibited only singly
charged ions corresponding to the molecular mass of PA with a degree of
polymerization (DP) up to 9 (nonamers). In contrast, Sdp9 yielded rather complex
mass spectra featuring ions with single [M - H](-), double [M - 2H](2-) and
triple [M - 3H](3-) charge representing the molecular masses of PAs up to a DP
of 28. In addition, the degree of galloylation per procyanidin (DG) was observed
to be up to 5 (pentagallates) in Sdp3 and 8 (octagallates) in the Sdp9. This is
the first evidence obtained by mass spectrometry for the distribution of grape
seed PAs with such a high degree of polymerization and a broad diversity of
galloylation. ES-MS data together with the complementary information provided by
acid hydrolysis provides a detailed picture of the composition of grape seed
PAs. Copyright 2002 John Wiley & Sons, Ltd.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12478550 [PubMed - indexed for MEDLINE]

302: J Agric Food Chem. 2002 Dec 18;50(26):7548-55.

Valorization of grape (Vitis vinifera) byproducts. Antioxidant and biological
properties of polyphenolic fractions differing in procyanidin composition and
flavonol content.

Torres JL, Varela B, Garcia MT, Carilla J, Matito C, Centelles JJ, Cascante M,
Sort X, Bobet R.

Department of Peptide and Protein Chemistry, Department of Surfactant
Technology, and Thermal Analysis Laboratory, Institute for Chemical and
Environmental Research (IIQAB-CSIC), Jordi Girona 18-26, 08034 Barcelona, Spain.
[email protected]

Many byproducts and wastes generated by agroindustries contain polyphenols with
potential application as food antioxidants and preventive agents against skin
cancer and other diseases. The performance of polyphenolic fractions from
Parellada grape (Vitis vinifera) pomace as antioxidants in different
physicochemical environments was tested. Fractions containing oligomers with
mean degree of polymerization between 3 and 4 and percentage galloylation ca.
30% were the most potent free radical scavengers and efficient antioxidants in
an oil-in-water emulsion. A fraction including glycosylated flavonols was also
efficient in the emulsion. All the fractions showed low aquatic toxicity and
weak influence on proliferation of human melanoma cells.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 12475269 [PubMed - indexed for MEDLINE]

303: Pharmacol Res. 2002 Dec;46(6):557-64.

Chemopreventive effect of Vitis vinifera extract on
12-O-tetradecanoyl-13-phorbol acetate-induced cutaneous oxidative stress and
tumor promotion in murine skin.

Alam A, Khan N, Sharma S, Saleem M, Sultana S.

Department of Medical Elementology and Toxicology, Faculty of Science, Jamia
Hamdard University, New Delhi 110062, India.

Vitis vinifera (grapes) is used as a fruit worldwide and known for its
pharmacological properties. The present paper assesses the chemopreventive
potential of Vitis vinifera against 12-O-tetradecanoyl-13-phorbol acetate
(TPA)-mediated tumor promotion in 7,12-dimethyl-benz[a]anthracene (DMBA)
initiated mice skin. Skin tumor initiation was achieved by a single topical
application of DMBA (40 microg/animal/0.20 ml acetone) to mice. Two weeks after
the initiation, promoting agent, TPA (5.0 microg/animal/0.2 ml acetone) was
applied two times a week for 20 weeks. Pretreatment of Vitis vinifera 1h prior
to each application of TPA resulted in protection against cutaneous
tumorigenesis in dose-dependent manner. This inhibition was evident when tumor
data was considered as the percentage of mice with tumor and the number of
tumors per mouse. We have shown that typical application of Vitis vinifera prior
to that of TPA resulted in significant inhibition against TPA-caused induction
of epidermal ODC activity (P<0.001) and DNA synthesis. Application of Vitis
vinifera at a dose level of 5.0 mg and 10.0 mg kg(-1) body weight in acetone
prior to that of TPA treatment resulted in partial significant inhibition of
oxidative stress in dose-dependent manner. The concomitant increase in the
microsomal lipid peroxidation and xanthine oxidase activities were significantly
reduced (P<0.001). In addition, the depleted level of glutathione and inhibited
activities of antioxidant enzymes were recovered to the partial significant
level. Hence, it can be suggested that Vitis vinifera can be used as a
chemopreventive agent against oxidative stress and carcinogenesis.

PMID: 12457631 [PubMed - indexed for MEDLINE]

304: Planta. 2002 Dec;216(2):245-54. Epub 2002 Aug 27.

Cellular and subcellular localisation of glutamine synthetase and glutamate
dehydrogenase in grapes gives new insights on the regulation of carbon and
nitrogen metabolism.

Paczek V, Dubois F, Sangwan R, Morot-Gaudry JF, Roubelakis-Angelakis KA, Hirel
B.

Unite de Nutrition Azotee des Plantes, INRA de Versailles, Route de Saint-Cyr,
78026 Versailles Cedex, France.

The subcellular localisation of glutamine synthetase (GS) and glutamate
dehydrogenase (GDH) in grapevine (Vitis vinifera L.) leaves and flowers was
investigated using immunogold-labelling experiments. In mature leaf tissue or
fully developed flowers, GS was visualised both in the cytosol and in the
chloroplasts, a high proportion of the protein being present in the phloem
companion cells. GDH was preferentially located in the mitochondria of the
phloem companion cells in both leaves and flowers. This observation suggests
that, in conjunction with GS, GDH plays a major role in controlling the
translocation of organic carbon and nitrogen metabolites in both vegetative and
reproductive organs. Significant amounts of GDH protein were also visualised in
multivesicular bodies within the flower receptacle. Although the function of
such organelles is still unknown, its is possible that the presence of GDH in
such cellular structures is important for the recycling of carbon and nitrogen
molecules in senescing tissues in which the enzyme is generally induced.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12447538 [PubMed - indexed for MEDLINE]

305: Plant Physiol. 2002 Nov;130(3):1172-80.

A novel nucleus-targeted protein is expressed in barley leaves during senescence
and pathogen infection.

Krupinska K, Haussuhl K, Schafer A, van der Kooij TA, Leckband G, Lorz H, Falk
J.

Institute of Botany, University of Kiel, Olshausenstrasse 40, D-24098 Kiel,
Germany. [email protected]

The barley (Hordeum vulgare) cDNA HvS40 represents a gene with enhanced mRNA
level during leaf senescence. Biolistic transformation of onion (Allium cepa)
epidermal cell layers with a glucuronidase fusion protein construct provided
evidence that the 15.4-kD protein encoded by HvS40 is localized to the nucleus.
Expression of the gene is induced by jasmonate and salicylic acid; both are
known to act as signaling compounds during senescence and defense toward
pathogens. Transcript levels of HvS40 were observed to be particularly high in
leaf sectors that undergo necrosis and chlorosis after infection with
Pyrenophora teres. This pathogen-related expression is, in contrast, clearly
reduced in transgenic barley plants expressing a stilbene synthase from grape
(Vitis vinifera), whereas the mRNA level of a gene encoding the pathogen-related
protein HvPR1 is unaffected. In situ hybridization with HvS40 antisense RNA
revealed that during leaf senescence, the HvS40 transcript predominantly
accumulates in the mesophyll. Taken together, the findings suggest a connection
between the nuclear protein encoded by HvS40 and the degeneration of
chloroplasts occurring during senescence and during infection of barley
wild-type plants with P. teres.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12427984 [PubMed - indexed for MEDLINE]

306: Trends Biotechnol. 2002 Nov;20(11):472-8.

Genetically tailored grapevines for the wine industry.

Vivier MA, Pretorius IS.

Institute for Wine Biotechnology, Stellenbosch University, South Africa.

Grapevine biotechnology is one of the most promising developments in the global
wine industry, which is increasingly faced with conflicting demands from
markets, consumers and environmentalists. In the grapevine industries, this
technology and its supporting disciplines entail the establishment of stress
tolerant and disease resistant varieties of Vitis vinifera, with increased
productivity, efficiency, sustainability and environmental friendliness,
especially regarding improved pest and disease control, water use efficiency and
grape quality. The implementation and successful commercialisation of
genetically improved grapevine varieties will only be realized if an array of
hurdles, both scientific and otherwise, can be overcome.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 12413822 [PubMed - indexed for MEDLINE]

307: Plant Mol Biol. 2002 Oct;50(3):497-509.

The vascular expression pattern directed by the Eucalyptus gunnii cinnamyl
alcohol dehydrogenase EgCAD2 promoter is conserved among woody and herbaceous
plant species.

Lauvergeat V, Rech P, Jauneau A, Guez C, Coutos-Thevenot P, Grima-Pettenati J.

IFR40, UMR CNRS-UPS 5546, Signaux et Messages Cellulaires chez les Vegetaux,
Pjle de Biotechnologie Vegetale, Castanet Tolosan, France.

Cinnamyl alcohol dehydrogenase (CAD; EC 1.1.1.195) catalyses the last step in
the synthesis of the monomeric precursors of lignin. Here, we demonstrate that
the vascular expression pattern conferred by the Eucalyptus gunnii EgCAD2
promoter in transgenic poplar (Populus tremula x Populus alba) is conserved in
another perennial woody angiosperm of economic interest (Vitis vinifera L.), as
well as in a model herbaceous plant (Nicotiana tabacum L.). Furthermore,
promoter deletion analysis performed in both tobacco and poplar allowed us to
identify the proximal region [-340/-124] as essential for vascular
cambium/xylem-specific expression whereas the [-124/+117] region was shown to
contain cis element-driving activity in phloem fibres. Interestingly, the
[-340/-124] fragment contains an AC-rich cis-acting element present in numerous
genes of the phenylpropanoid pathway expressed in xylem tissues, and known as a
consensus Myb transcription factor binding site, suggesting that common Myb
sites may provide a mechanism by which different steps of phenylpropanoid
metabolism are coordinately regulated and expressed in vascular tissues. We have
also shown in both tobacco and poplar that the EgCAD2 promoter is inducible by
wounding and the cis-elements responsible for wounding responsiveness are
located in the distal promoter region. Taken together, our data suggest that the
mechanisms controlling developmental and wounding inducible expression of the
EgCAD2 promoter are conserved among perennial woody and annual herbaceous plant
species enabling us now to investigate in depth the transcriptional regulation
of the EgCAD2 promoter in tobacco.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 12369625 [PubMed - indexed for MEDLINE]

308: J Agric Food Chem. 2002 Oct 9;50(21):6217-21.

Procyanidins from Vitis vinifera seeds: in vivo effects on oxidative stress.

Simonetti P, Ciappellano S, Gardana C, Bramati L, Pietta P.

Department of Food Science and Microbiology, Division of Human Nutrition,
University of Milan, Via Celoria 2, 20133 Milan, Italy. [email protected]

The purpose of this study was to evaluate the effect of supplementation with
procyanidins from Vitis vinifera on markers of oxidative stress. Ten healthy
volunteers received a daily dose of 110 mg of procyanidins for 30 days. Fasting
venous blood samples were taken before and at the end of the supplementation
period and after 7 days of wash-out. The total antioxidant activity and the
plasma concentrations of alpha-tocopherol were not modified. Conversely, the
levels of alpha-tocopherol in red blood cell membranes increased significantly
from 1.8 +/- 0.1 to 2.8 +/- 0.2 mg/g. Similarly, the lymphocyte oxidized DNA
[8-oxo-7,8-dihydro-2'-deoxyguanosine/2'-deoxyguanosine ratio] was reduced from
7.23 +/- 2.47 to 2.34 +/- 0.51, and the red blood cell membrane fatty acid
composition shifted to a higher level of polyunsaturated fatty acids. On the
basis of these results, it may be suggested that dietary procyanidins exert
their antioxidant protection in vivo by sparing liposoluble vitamin E and
reducing DNA oxidative damage.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12358505 [PubMed - indexed for MEDLINE]

309: J Agric Food Chem. 2002 Oct 9;50(21):6090-6.

Application of a DNA analysis method for the cultivar identification of grape
musts and experimental and commercial wines of Vitis vinifera L. using
microsatellite markers.

Garcia-Beneytez E, Moreno-Arribas MV, Borrego J, Polo MC, Ibanez J.

Instituto Madrileno de Investigacion Agraria y Alimentaria, Finca El Encin,
Apdo. 127, Alcala de Henares, 28800 Madrid, Spain.

A DNA-based method has been applied to the identification of several musts and
wines using microsatellite markers. DNA was extracted from the solid phases of
sixteen monovarietal and five multivarietal musts (mixtures of two musts down to
a 4:1 proportion) and they were genotyped at seven microsatellites through a
multiplex PCR reaction and automated fluorescent detection. PCR multiplexing was
successful in monovarietal musts, but should be used with caution with at least
some markers and in multivarietal musts. The same extraction and detection
methods were unsuccessfully applied to the solid and liquid phases of five
monovarietal commercial wines, even after using different concentration
procedures. Nucleic acids presence was then studied in a recent must, during the
fermentation process, and during the subsequent steps of winemaking. Genotyping
was possible in the resulting experimental wine until decanting, when the
particles in suspension were removed. These results suggest that wine
authentication through DNA analysis is not possible in commercial wines, in the
tested conditions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12358485 [PubMed - indexed for MEDLINE]

310: J Agric Food Chem. 2002 Oct 9;50(21):5909-14.

Antioxidant activities of grape (Vitis vinifera) pomace extracts.

Chidambara Murthy KN, Singh RP, Jayaprakasha GK.

Government College of Pharmacy, Bangalore 560 027, India.

Antioxidant-rich fractions were extracted from grape (Vitis vinifera) pomace
using ethyl acetate, methanol, and water. The extracts were screened for their
potential as antioxidants in different models. The ethyl acetate, methanol, and
water extracts showed 76, 87.1, and 21.7% antioxidant activities at 100 ppm,
respectively, using the 1,1-diphenyl-2-picrylhydrazyl model system. As the
methanol extract of grape pomace showed maximum antioxidant activity among all
of the extracts, it was selected to determine its effect on lipid peroxidation,
hydroxyl radical scavenging activity, and human low-density lipoprotein (LDL)
oxidation. The methanol extract showed 71.7, 73.6, and 91.2% inhibition using
the thiobarbituric acid method, hydroxyl radical scavenging activity, and LDL
oxidation, respectively, at 200 ppm. Treatment of albino rats of the Wistar
strain with a single dose of CCl(4) at 1.25 mL/kg of body weight decreases the
activities of catalase, superoxide dismutase (SOD), and peroxidase by 81, 49,
and 89%, respectively, whereas the lipid peroxidation value increased nearly
3-fold. Pretreatment of the rats with the methanolic extract of grape pomace at
50 mg/kg (in terms of catechin equivalents) followed by CCl(4) treatment causes
restoration of catalase, SOD, and peroxidase by 43.6, 73.2, and 54%,
respectively, as compared with control, whereas lipid peroxidation was restored
to values comparable with the control. Histopathological studies of the liver of
different groups also support the protective effects exhibited by the methanol
extract of grape pomace by restoring the normal hepatic architecture. Owing to
this property, the studies on grape pomace can be further extended to exploit
its possible application for the preservation of food products as well as a
health supplement and neutraceutical.

PMID: 12358458 [PubMed - indexed for MEDLINE]

311: BMC Biotechnol. 2002 Sep 27;2:18.

Genetic transformation of Vitis vinifera via organogenesis.

Mezzetti B, Pandolfini T, Navacchi O, Landi L.

Dipartimento Biotecnologie Agrarie e Ambientali, University of Ancona, Via
Brecce Bianche 60100-Ancona Italy. [email protected]

BACKGROUND: Efficient transformation and regeneration methods are a priority for
successful application of genetic engineering to vegetative propagated plants
such as grape. The current methods for the production of transgenic grape plants
are based on Agrobacterium-mediated transformation followed by regeneration from
embryogenic callus. However, grape embryogenic calli are laborious to establish
and the phenotype of the regenerated plants can be altered. RESULTS: Transgenic
grape plants (V. vinifera, table-grape cultivars Silcora and Thompson Seedless)
were produced using a method based on regeneration via organogenesis. In vitro
proliferating shoots were cultured in the presence of increasing concentrations
of N6-benzyl adenine. The apical dome of the shoot was removed at each
transplantation which, after three months, produced meristematic bulk tissue
characterized by a strong capacity to differentiate adventitious shoots. Slices
prepared from the meristematic bulk were used for Agrobacterium-mediated
transformation of grape plants with the gene DefH9-iaaM. After rooting on
kanamycin containing media and greenhouse acclimatization, transgenic plants
were transferred to the field. At the end of the first year of field
cultivation, DefH9-iaaM grape plants were phenotypically homogeneous and did not
show any morphological alterations in vegetative growth. The expression of
DefH9-iaaM gene was detected in transgenic flower buds of both cultivars.
CONCLUSIONS: The phenotypic homogeneity of the regenerated plants highlights the
validity of this method for both propagation and genetic transformation of table
grape cultivars. Expression of the DefH9-iaaM gene takes place in young flower
buds of transgenic plants from both grape cultivars.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12354328 [PubMed - indexed for MEDLINE]

312: Biocell. 2002 Aug;26(2):263-6.

Improved in vitro embryo development of stenospermic grape by putrescine.

Ponce MT, Guinazu ME, Tizio R.

Departamento de Biologia, Catedra de Fisiologia Vegetal, Facultad de Ciencias
Agrarias, Universidad Nacional de Cuyo, Mendoza, Argentina.
[email protected]

The goal of this study was to determine the effect of putrescine, added to the
culture medium, on the in vitro development of stenospermic grape (Vitis
vinifera L) embryos. The cross breedings of Perlon x G.C88552 and Perlon x
Argentina were used. 0 (control), 2 and 4 mM of putrescine were added to the
immature seed's culture medium. In Perlon x Argentina, 2 mM of putrescine
statistically increased the percentage of total embryos, direct germination,
polyembryos and normal plants. In Perlon x G.C88552, only 2 mM of putrescine
increased all the variables considered, eventually tripling the percentage of
normal plants obtained. The results suggest that the endogenous concentration of
putrescine may be a growth limiting factor. Adding putrescine to the culture
medium of immature grape seeds is a legitimate resource to significantly
increase the results of this technique.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12240561 [PubMed - indexed for MEDLINE]

313: Plant Physiol. 2002 Sep;130(1):68-77.

VFL, the grapevine FLORICAULA/LEAFY ortholog, is expressed in meristematic
regions independently of their fate.

Carmona MJ, Cubas P, Martinez-Zapater JM.

Departmento de Biotecnologia, Escuela Tecnics Suoerior Ingenieros Agronomos,
Universidad Politecnica de Madrid, Spain.

The flowering process in grapevine (Vitis vinifera) takes place in buds and
extends for two consecutive growing seasons. To understand the genetic and
molecular mechanisms underlying this process, we have characterized grapevine
bud development, cloned the grapevine FLORICAULA/LEAFY (FLO/LFY) ortholog, VFL,
and analyzed its expression patterns during vegetative and reproductive
development. Flowering induction takes place during the first season. Upon
induction, the shoot apical meristem begins to produce lateral meristems that
will give rise to either inflorescences or tendrils. During the second season,
after a winter dormancy period, buds reactivate and inflorescence meristems give
rise to flower meristems. VFL is expressed in lateral meristems that give rise
to inflorescence and flower meristems, consistent with a role in reproductive
development. Furthermore, VFL is also detected in other meristematic regions
such as the vegetative shoot apical meristem and the lateral meristems that will
give rise to tendrils. VFL is also expressed in leaf primordia and in growing
leaf margins until later stages of development. Accumulation of VFL transcripts
in cell-proliferating regions suggests a role for VFL not only in flower
meristem specification, but also in the maintenance of indeterminacy before the
differentiation of derivatives of the apical meristem: flowers, leaves, or
tendrils.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12226487 [PubMed - indexed for MEDLINE]

314: Plant Physiol. 1997 Nov;115(3):1155-1161.

Treatment of Grape Berries, a Nonclimacteric Fruit with a Synthetic Auxin,
Retards Ripening and Alters the Expression of Developmentally Regulated Genes.

Davies C, Boss PK, Robinson SP.

Cooperative Research Centre for Viticulture, P.O. Box 145, Glen Osmond, South
Australia 5064, Australia (C.D., P.K.B., S.P.B.).

Treatment of grape (Vitis vinifera L.) berries with the synthetic auxin-like
compound benzothiazole-2-oxyacetic acid (BTOA) caused a delay in the onset of
ripening of approximately 2 weeks. This was manifested as a retardation of the
increases in berry weight, color, deformability, and hexose concentration. BTOA
treatment also delayed by 2 weeks the increase in abscisic acid level that
normally accompanies ripening and altered the expression of a number of
developmentally regulated genes. A putative vacuolar invertase, which is
normally expressed from berry set until ripening and turned off after ripening
commences, remained expressed throughout development in BTOA-treated grape
berries. This elevated expression resulted in increased levels of invertase
activity. In contrast, the up-regulation of four other genes normally switched
on at the time of ripening was delayed in BTOA-treated fruit. These included
chalcone synthase and UDP-glucose-flavonoid 3-O-glucosyl transferase, both of
which are involved in anthocyanin synthesis, a chitinase, and a ripening-related
gene of an unknown function. These observations support the view that auxins
(perhaps in conjunction with abscisic acid) may have a role in the control of
grape berry ripening by affecting the expression of genes involved in the
ripening process.

PMID: 12223864 [PubMed - as supplied by publisher]

315: J Chromatogr A. 2002 Jul 26;964(1-2):123-8.

Preparative isolation of polyphenolic compounds from Vitis vinifera by
centrifugal partition chromatography.

Delaunay JC, Castagnino C, Cheze C, Vercauteren J.

Laboratoire de Pharmacognosie, EA no. 491, Faculte des Sciences Pharmaceutiques,
Universite Victor Segalen Bordeaux 2, 146 Rue Leo-Saignat, 33076 Bordeaux,
France.

This study deals with a centrifugal partition chromatography developed for the
separation of phenolic compounds from Vitis vinifera. EtOAc grape seed extracts
were separated using the solvent system hexane-ethyl acetate-ethanol-water
(1:8:2:7; v/v) in two fractions: one containing about 75% of flavanol monomers
(catechin and epicatechin) corresponding to 18% of crude extract and another
fraction B-type dimers (22% of crude extract). From the stalk extracts, we could
separate stilbenoid compounds (resveratrol and its oligomers; 12% of crude
extract) which were eluted in less than 30 min from flavanols (which required a
few hours of additional elution). Using the same solvent system but in different
ratios (4:5:3:3; v/v), we isolated the trans-resveratrol ([email protected]; 90% purity).

PMID: 12198840 [PubMed - indexed for MEDLINE]

316: Planta. 2002 Aug;215(4):630-8. Epub 2002 May 21.

Immunocharacterization of Vitis vinifera L. ferredoxin-dependent glutamate
synthase, and its spatial and temporal changes during leaf development.

Loulakakis KA, Primikirios NI, Nikolantonakis MA, Roubelakis-Angelakis KA.

Department of Horticulture, Technological Educational Institute of Crete, 71 500
Heraklion, Crete, Greece.

The grapevine (Vitis vinifera L.) partial fragment of cDNA clone pGOGAT1
[Loulakakis and Roubelakis-Angelakis (1997) Physiol Plant 101:220-228], encoding
the ferredoxin-dependent glutamate synthase (Fd-GOGAT; EC 1.4.7.1), was
overexpressed in Escherichia coli cells. A hybrid between the Fd-GOGAT fragment
and maltose-binding protein was purified and used to raise a polyclonal antibody
in a rabbit. The prepared antibody appeared to be specific towards Fd-GOGAT; it
recognized a protein band of approximately 160 kDa on nitrocellulose blots after
SDS-PAGE of total proteins from leaves, internodes, roots and calluses, and
precipitated most of the enzyme activity present in grapevine protein extracts.
The quantity of Fd-GOGAT protein was substantially higher in leaves than in
other grapevine tissues tested, coincident with a similar distribution of the
enzyme specific activity. Intracellular localization studies revealed that both
the enzyme activity and the 160-kDa immunoreactive protein were associated with
the chloroplastic fraction. Furthermore, the accumulation of Fd-GOGAT, glutamine
synthetase (GS) and glutamate dehydrogenase (GDH), at the activity and protein
levels, was monitored during leaf development of field-grown plants, from the
stage of the newly expanding leaf to the senescing old leaf. Both the specific
activity and quantity of the 160-kDa polypeptide of Fd-GOGAT were higher in the
mature, full sized leaves and substantially lower in young and senescing leaves.
GS specific activity and immunoreactive protein followed the same trend as
Fd-GOGAT, while GDH showed opposite developmental patterns of accumulation. The
biological significance of the presence of Fd-GOGAT in the various grapevine
tissues and its physiological role during early development and natural
senescence of the leaves are discussed.

PMID: 12172846 [PubMed - indexed for MEDLINE]

317: Phytochemistry. 2002 Aug;60(8):795-8.

Galloylated catechins and stilbene diglucosides in Vitis vinifera cell
suspension cultures.

Decendit A, Waffo-Teguo P, Richard T, Krisa S, Vercauteren J, Monti JP, Deffieux
G, Merillon JM.

Laboratoire de Mycologie et Biotechnologie Vegetale, EA 491, Faculte des
Sciences Pharmaceutiques, Universite de Bordeaux II, 3 ter, Place de la
Victoire, F-33076 Bordeaux Cedex, France.

Suspension cultures of Vitis vinifera were found to produce catechins and
stilbenes. When cells were grown in a medium inducing polyphenol synthesis,
(-)-epicatechin-3-O-gallate, dimeric procyanidin B-2 3'-O-gallate and two
resveratrol diglucosides were isolated, together with a new natural compound
that was identified as cis-resveratrol-3,4'-O-beta-diglucoside by
spectroscopical methods.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12150801 [PubMed - indexed for MEDLINE]

318: Planta. 2002 Jul;215(3):387-93. Epub 2002 Apr 10.

The host guides morphogenesis and stomatal targeting in the grapevine pathogen
Plasmopara viticola.

Kiefer B, Riemann M, Buche C, Kassemeyer HH, Nick P.

Staatliches Weinbauinstitut, Merzhauserstrasse 119, 79100 Freiburg, Germany.

The oomycete grape downy mildew (Plasmopara viticola Berk. & Curt. Ex de Bary)
is a serious pathogen of grapevine and spreads by extremely efficient cycles of
asexual propagation. The high efficiency must involve efficient sensing of the
host. We therefore analyzed the time course and morphology of the early
development of this pathogen in a host system, by infection of leaf discs of
grapevine (Vitis vinifera L. cv. Muller-Thurgau), and in a host-free system.
Host factors were demonstrated to influence pathogen development in the
following ways: (i) the release of zoospores from mature sporangia was
accelerated, (ii) the morphogenesis of the germ tube was coordinated, and (iii)
the zoospores were targeted to the stomata by factors that depended on stomata
closure. The findings show that the early development of P. viticola is
regulated, specifically and coordinately, by factors originating from the host
plant.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12111219 [PubMed - indexed for MEDLINE]

319: J Agric Food Chem. 2002 Jul 17;50(15):4303-9.

"Untypical aging off-flavor" in wine: synthesis of potential degradation
compounds of indole-3-acetic acid and kynurenine and their evaluation as
precursors of 2-aminoacetophenone.

Hoenicke K, Borchert O, Gruning K, Simat TJ.

Institute of Biochemistry and Food Chemistry, University of Hamburg,
Grindelallee 117, D 20146 Hamburg, Germany.

Kynurenine (1) and indole-3-acetic acid (2) are considered as potential
precursors of 2-aminoacetophenone (3), which is regarded to be the aroma impact
compound causing an "untypical aging off-flavor" (UTA) in Vitis vinifera wines.
The mechanism of the formation of 3 was studied using model fermentation and
model sulfuration media spiked with 1 or 2 as potential precursors. Possible
degradation products such as kynurenamine (4) and kynurenic acid (5), or skatole
(6), 2-oxoskatole (7), 2-formamidoacetophenone (8), 2-oxindole-3-acetic acid
(9), and 3-(2-formylaminophenyl)-3-oxopropionic acid (10) were evaluated by
HPLC-UV of the fermentation and sulfuration media and comparison with
synthesized 7, 8, 9, and 10. The synthesis of the possible precursor
4-(2-aminophenyl)-2,4-dioxobutanoic acid (11), a proposed metabolite of 1 failed
because a spontaneous cyclization yields 5 and N-oxo-kynurenic acid (12), but
not 11. It could be shown that the formation of 3 is triggered by an oxidative
degradation of 2 after sulfuration with potassium bisulfite via the
intermediates 10 and 8. However, no formation of 3 occurred during sulfuration
of a model wine spiked with 1 or during fermentation of a model must spiked with
1 or 2.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12105962 [PubMed - indexed for MEDLINE]

320: Ann Bot (Lond). 2002 Apr;89(4):401-8.

Allometric relationships to estimate seasonal above-ground vegetative and
reproductive biomass of Vitis vinifera L.

Castelan-Estrada M, Vivin P, Gaudillere JP.

Ecophysiology and Agronomy of Grapevine (ECAV), Department of Agronomy, INRA
Bordeaux, Villenave d'Ornon, France.

A procedure is described for obtaining allometric regression equations to
estimate non-destructively and in a cost-effective manner the current year's
above-ground vegetative and reproductive biomass of Vitis vinifera L. Merlot'
throughout the growing season. Significant relationships were obtained over a
3-year period (1998-2000) between the dimensions of an individual shoot per vine
(i.e. diameter and length) and dry weights of its primary stem, primary leaves
and lateral growth. The dry mass of a grape was best estimated from measurements
of the basal diameter of the bunch peduncle. Introducing cumulative degree-days
as an additional explanatory variable in the equations allowed them to be used
irrespective of year and growth stage. Multi-year regressions were used to
quantify in detail the seasonal evolution of mature grapevine biomass under the
climatic conditions of the Bordeaux area, France, and for differing levels of
soil nitrogen.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12096800 [PubMed - indexed for MEDLINE]

321: J Agric Food Chem. 2002 Jul 3;50(14):4076-9.

Sulfur aroma precursor present in S-glutathione conjugate form: identification
of S-3-(hexan-1-ol)-glutathione in must from Vitis vinifera L. cv. Sauvignon
blanc.

Peyrot Des Gachons C, Tominaga T, Dubourdieu D.

Faculte d'oenologie, Universite Victor Segalen Bordeaux 2, 351 cours de la
Liberation, 33405 Talence Cedex France.

When Sauvignon blanc or Gros Manseng grape must was percolated through an
immobilized gamma-glutamyltranspeptidase column, there was a significant
increase in the concentration of S-3-(hexan-1-ol)-L-cysteine, the precursor of
3-mercaptohexan-1-ol, a compound that contributes to the varietal aroma of wines
made from these grapes. Low- and high-resolution liquid secondary ion mass
sepectrometry (LSIMS) analyses established the presence of
S-3-(hexan-1-ol)-glutathione in Sauvignon blanc must. The identification of this
compound suggests that the S-3-(hexan-1-ol)-L-cysteine in grapes is produced by
the catabolism of S-3-(hexan-1-ol)-glutathione. As is the case in other plant or
animal organisms, S-glutathione conjugates may be involved in certain
detoxification systems in vines.

PMID: 12083886 [PubMed - indexed for MEDLINE]

322: J Ind Microbiol Biotechnol. 2002 Jul;29(1):28-33.

Influence of Saccharomyces cerevisiae strains on fermentation and flavor
compounds of white wines made from cv. Emir grown in Central Anatolia, Turkey.

Nurgel C, Erten H, Canbas A, Cabaroglu T, Selli S.

Department of Food Engineering, Faculty of Agriculture, University of Cukurova,
Adana 01300, Turkey.

The effect of inoculation with selected Saccharomyces cerevisiae strains was
studied on fermentation and flavor compounds of wines made from Vitis vinifera
L. cv. Emir grown in Central Anatolia, Turkey. Flavor compounds were analysed
and identified by GC-FID and GC-MS, respectively. The total concentrations of
flavor compounds did not increase with the addition of indigenous and commercial
wine yeasts, but differences were noted in individual volatile compounds.
Cluster and factor analyses of flavor compounds also showed that wines produced
were different depending on the wine strain used. Wines were completely
fermented to less than 1.4 g/l residual sugar. Yeasts other than S. cerevisiae
survived longer than previously reported. Inoculation with selected strains
increased the ethanol level.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12080424 [PubMed - indexed for MEDLINE]

323: J Agric Food Chem. 2002 Jun 19;50(13):3822-7.

Analysis of grape Vitis vinifera L. DNA in must mixtures and experimental mixed
wines using microsatellite markers.

Siret R, Gigaud O, Rosec JP, This P.

Unite Mixte de Recherches 1097, Diversite et Genomes des Plantes Cultivees
(ENSA.M, CIRAD, INRA, IRD), Equipe Genetique Vigne, ENSA.M-INRA, 2 Place P.
Viala, 34060 Montpellier, France. [email protected]

Because wine quality highly relies on the varietal composition of the must, the
development of methods allowing the authentication of varieties in musts and
wines would be of great value as a guarantee of quality. Microsatellite markers
have already been applied to the authentication of grape juices (Faria, M. A.;
Magalhaes, R.; Ferreira, M. A.; Meredith, C. P.; Ferreira Monteiro, F. J. Agric.
Food Chem. 2000, 48, 1096-1100) and to the analysis of experimental wines
(Siret, R.; Boursiquot, J. M.; Merle, M. H.; Cabanis, J. C.; This, P. J. Agric
Food Chem. 2000, 48, 5035-5040). In the present paper, we accessed the
usefulness of this technology for the analysis of must and wine mixtures. The
detection limit of DNA mixtures was first estimated on DNA extracted from
leaves: 4% of a foreign DNA can be detected. Analysis of must and wine mixtures
(Chardonnay B/Clairette B and Syrah N/Grenache N) was performed on experimental
fermentations. DNA was extracted along the fermentation process and analyzed
using five microsatellite loci. The 70:30 (v/v) mixtures were successfully
analyzed until the end of the fermentation. The applications of these results to
commercial purposes are discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12059166 [PubMed - indexed for MEDLINE]

324: J Chromatogr B Analyt Technol Biomed Life Sci. 2002 Apr
25;770(1-2):297-301.

Influence of storage conditions on the stability of monomeric anthocyanins
studied by reversed-phase high-performance liquid chromatography.

Morais H, Ramos C, Forgacs E, Cserhati T, Oliviera J.

National Agronomical Station, Quinta do Marques, Oeiras, Portugal.

The effect of light, storage time and temperature on the decomposition rate of
monomeric anthocyanin pigments extracted from skins of grape (Vitis vinifera
var. Red globe) was determined by reversed-phase high-performance liquid
chromatography (RP-HPLC). The impact of various storage conditions on the
pigment stability was assessed by stepwise regression analysis. RP-HPLC
separated well the five anthocyanins identified and proved the presence of other
unidentified pigments at lower concentrations. Stepwise regression analysis
confirmed that the overall decomposition rate of monomeric anthocyanins,
peonidin-3-glucoside and malvidin-3-glucoside significantly depended on the time
and temperature of storage, the effect of storage time being the most important.
The presence or absence of light exerted a negligible impact on the
decomposition rate.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 12013239 [PubMed - indexed for MEDLINE]

325: Nutr Cancer. 2001;40(2):173-9.

Potential cancer-chemopreventive activities of wine stilbenoids and flavans
extracted from grape (Vitis vinifera) cell cultures.

Waffo-Teguo P, Hawthorne ME, Cuendet M, Merillon JM, Kinghorn AD, Pezzuto JM,
Mehta RG.

Program for Collaborative Research in the Pharmaceutical Sciences, Department of
Medicinal Chemistry and Pharmacognosy, College of Pharmacy, University of
Illinois at Chicago, Chicago, IL 60612, USA.

Moderate consumption of wine is associated with a reduced risk of cancer. Grape
plant cell cultures were used to purify 12 phenols: the stilbenoids
trans-astringin, trans-piceid (2), trans-resveratroloside, trans-resveratrol,
trans-piceatannol, cis-resveratroloside, cis-piceid, and cis-resveratrol; the
flavans (+)-catechin, (-)-epicatechin, and epicatechin 3-O-gallate; and the
flavan dimer procyanidin B2 3'-O-gallate. These compounds were evaluated for
potential to inhibit cyclooxygenases and preneoplastic lesion formation in
carcinogen-treated mouse mammary glands in organ culture. At 10 micrograms/ml,
trans-astringin and trans-piceatannol inhibited development of
7,12-dimethylbenz[a]anthracene-induced preneoplastic lesions in mouse mammary
glands with 68.8% and 76.9% inhibition, respectively, compared with untreated
glands. The latter compound was the most potent of the 12 compounds tested in
this assay, with the exception of trans-resveratrol (87.5% inhibition). In the
cyclooxygenase (COX)-1 assay, trans isomers of the stilbenoids appear to be more
active than cis isomers: trans-resveratrol [50% inhibitory concentration (IC50)
= 14.9 microM, 96%] vs. cis-resveratrol (IC50 = 55.4 microM). In the COX-2
assay, among the compounds tested, only trans- and cis-resveratrol exhibited
significant inhibitory activity (IC50 = 32.2 and 50.2 microM, respectively).
This is the first report showing the potential cancer-chemopreventive activity
of trans-astringin, a plant stilbenoid recently found in wine. trans-Astringin
and its aglycone trans-piceatannol were active in the mouse mammary gland organ
culture assay but did not exhibit activity in COX-1 and COX-2 assays.
trans-Resveratrol was active in all three of the bioassays used in this
investigation. These findings suggest that trans-astringin and trans-piceatannol
may function as potential cancer-chemopreventive agents by a mechanism different
from that of trans-resveratrol.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

PMID: 11962253 [PubMed - indexed for MEDLINE]

326: Physiol Plant. 2002 Feb;114(2):231-240.

Steady-state chlorophyll fluorescence (Fs) measurements as a tool to follow
variations of net CO2 assimilation and stomatal conductance during water-stress
in C3 plants.

Flexas J, Escalona JM, Evain S, Gulias J, Moya I, Osmond CB, Medrano H.

Laboratori de Fisiologia Vegetal, Departament de Biologia, Universitat de les
Illes Balears-Institut Mediterrani d'Estudis Avancats (UIB-CSIC). Carretera de
Valldemossa Km. 7.5, E-07071 Palma de Mallorca. Balears, Spain Laboratoire pour
l'Utilization du Rayonnement Electromagnetique (LURE), Centre Universitaire
Paris-Sud, B.P. 34; F-91898 Orsay Cedex, France Research School of Biological
Sciences, Institute of Advanced Studies, Australian National University, Box
475, Canberra, ACT 2601, Australia.

Water stress experiments were performed with grapevines (Vitis vinifera L.) and
other C3 plants in the field, in potted plants in the laboratory, and with
detached leaves. It was found that, in all cases, the ratio of steady state
chlorophyll fluorescence (Fs) normalized to dark-adapted intrinsic fluorescence
(Fo) inversely correlated with non-photochemical quenching (NPQ). Also, at high
irradiance, the ratio Fs/Fo was positively correlated with CO2 assimilation in
air, with electron transport rate calculated from fluorescence, and with
stomatal conductance, but no clear correlation was observed with qP. The
significance of these relationships is discussed. The ratio Fs/Fo, measured with
a portable instrument (PAM-2000) or with a remote sensing FIPAM system, provides
a good method for the early detection of water stress, and may become a useful
guide to irrigation requirements.

PMID: 11903970 [PubMed - as supplied by publisher]

327: J Biotechnol. 2002 Apr 25;95(1):49-56.

Carbon-14 biolabelling of wine polyphenols in Vitis vinifera cell suspension
cultures.

Vitrac X, Krisa S, Decendit A, Vercauteren J, Nuhrich A, Monti JP, Deffieux G,
Merillon JM.

GESNIT EA 491, Universite de Bordeaux 2, Faculte des Sciences Pharmaceutiques 3
ter, Place de la Victoire, 33076 Bordeaux Cedex, France.

14C-L-phenylalanine is incorporated into a range of polyphenolic compounds when
fed to grape cell cultures. Optimisation of several parameters such as the
quantity of precursor applied and the duration of metabolism led to
incorporation yields of 15% and to specific activities of 875 mu Ci g(-1) in
stilbenes. Purification of the products by several chromatographic steps is
reported. Both trans- and cis-resveratrols were easily obtained by enzymatic
hydrolysis of their corresponding glucosides, with specific activity of
1200-1400 mu Ci g(-1). The specific radioactivity obtained for all the compounds
is suitable for in vivo feeding trials to trace their metabolic fate when
consumed by animals and for in vitro activity mechanism studies. Indeed, these
polyphenols seem to be implicated in the health benefits associated with regular
and moderate wine consumption but little is known about their pharmacokinetics
and cellular uptake.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11879711 [PubMed - indexed for MEDLINE]

328: J Agric Food Chem. 2002 Feb 27;50(5):1213-7.

Antioxidant properties of trans-epsilon-viniferin as compared to stilbene
derivatives in aqueous and nonaqueous media.

Privat C, Telo JP, Bernardes-Genisson V, Vieira A, Souchard JP, Nepveu F.

Laboratoire Pharmacophores Redox, Phytochimie et Radiobiologie, EA-3030,
Universite Paul Sabatier, 35 Chemin des Maraichers, 31062 Toulouse Cedex 4,
France.

trans-Epsilon-viniferin, the dimer of resveratrol, extracted from Vitis
vinifera, has been evaluated for its antioxidant capacity. Its properties have
been compared to those of resveratrol and synthetic stilbenic derivatives
(4-hydroxystilbene, 4,4'-dihydroxystilbene, 3,5-dihydroxystilbene, and
trimethylresveratrol), in regard to their liposolubility using two media with
different polarity. The bleaching of beta-carotene by lipoperoxyl (LOO.)
radicals in an oil/water (O/W) emulsion and the scavenging of superoxide anions
(O(-)(2) in dimethyl sulfoxide (DMSO) using 5,5-dimethyl-1-pyrroline-N-oxide as
a spin trap were followed using UV-visible and electron paramagnetic resonance,
respectively. Epsilon-viniferin exhibits the best antioxidant capacity in the
DMSO/O(-)(2) polar system (IC(50) = 0.14 mM) while 4,4'-dihydroxystilbene
presents the highest antioxidant capacity in the O/W/LOO. system (inhibition of
beta-carotene bleaching, 82%). Partition coefficients and kinetics of partition
between 1-octanol and water were measured to discuss the antioxidant efficiency
of the compounds in relation with their chemical structure.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 11853506 [PubMed - indexed for MEDLINE]

329: Plant Physiol. 2002 Feb;128(2):564-77.

A grapevine gene encoding a guard cell K(+) channel displays developmental
regulation in the grapevine berry.

Pratelli R, Lacombe B, Torregrosa L, Gaymard F, Romieu C, Thibaud JB, Sentenac
H.

Biologie du Developpement des Plantes Perennes Cultivees, Unite Mixte de
Recherche 1098 Agro-M/Centre de Cooperation Internationale en Recherche
Agronomique pour le Developpement, 2 place Viala, F-34060 Montpellier cedex 1,
France.

SIRK is a K(+) channel identified in grapevine (Vitis vinifera), belonging to
the so-called Shaker family. The highest sequence similarities it shares with
the members of this family are found with channels of the KAT type, although
SIRK displays a small ankyrin domain. This atypical feature provides a key to
understand the evolution of the plant Shaker family. Expression in Xenopus
laevis oocytes indicated that SIRK is an inwardly rectifying channel displaying
functional properties very similar to those of KAT2. The activity of SIRK
promoter region fused to the GUS reporter gene was analyzed in both grapevine
and Arabidopsis. Like other KAT-like channels, SIRK is expressed in guard cells.
In Arabidopsis, the construct is also expressed in xylem parenchyma.
Semiquantitative reverse transcriptase-polymerase chain reaction experiments
indicated that SIRK transcript was present at low levels in the berry, during
the first stages of berry growth. After veraison, the period of berry
development that corresponds to the inception of ripening and that is associated
with large biochemical and structural modifications, such as evolution of
stomata in nonfunctional lenticels and degeneration of xylem vasculature, the
transcript was no longer detected. The whole set of data suggests that in the
berries SIRK is expressed in guard cells and, possibly, in xylem tissues. The
encoded channel polypeptide could therefore play a role in the regulation of
transpiration and water fluxes in grapevine fruits.

PMID: 11842160 [PubMed - indexed for MEDLINE]

330: Planta. 2001 Dec;214(2):257-64.

Expression patterns of cell wall-modifying enzymes during grape berry
development.

Nunan KJ, Davies C, Robinson SP, Fincher GB.

Department of Plant Science, University of Adelaide, Glen Osmond, South
Australia, Australia.

During ripening of grape (Vitis vinifera L.) berries, softening occurs
concomitantly with the second growth phase of the fruit and involves significant
changes in the properties of cell wall polysaccharides. Here, the activities of
enzymes that might participate in cell wall modification have been monitored
throughout berry development. Alpha-galactosidase (EC 3.2.1.22),
beta-galactosidase (EC 3.2.1.23) and pectin methylesterase (EC 3.1.1.11)
activities were present, but no polygalacturonase (EC 3.2.1.15), cellulase (EC
3.2.1.4), xyloglucanase (xyloglucan-specific cellulase EC 3.2.1.4) or
galactanase (EC 3.2.1.89) could be detected. The accumulation of mRNAs encoding
wall-modifying enzymes was examined by northern hybridization analysis.
Transcripts for beta-galactosidase, pectin methylesterase, polygalacturonase,
pectate lyase (EC 4.2.2.2) and xyloglucan endotransglycosylase (EC 2.4.1.207)
were present during ripening, although polygalacturonase activity had not been
detected in berry extracts. Cellulases could not be detected in ripening
berries, either at the enzyme or mRNA levels. The increase in beta-galactosidase
activity and mRNA is consistent with the observed decrease in type-I
arabinogalactan content of the walls during ripening, and the detection of
polygalacturonase and pectate lyase mRNAs might explain the increased solubility
of galacturonan in walls of ripening grapes. Thus, the modification of cell wall
polysaccharides during softening of grape berries is a complex process involving
subtle changes to different components of the wall, and in many cases only small
amounts of enzyme activity are required to effect these changes.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11800390 [PubMed - indexed for MEDLINE]

331: Cryo Letters. 2001 Sep-Oct;22(5):321-8.

Cryopreservation of axillary buds of grape (Vitis vinifera) in vitro plantlets.

Zhao C, Wu Y, Engelmann F, Zhou M.

Changli Institute of Pomology, Hebei Academy of Agricultural and Forestry
Sciences, Hebei 066600, Changli Town, China. [email protected]

Axillary buds sampled from in vitro plants of four grape varieties could
withstand cryopreservation using the encapsulation-dehydration technique.
Regrowth percentages ranged between 15 and 40%. Buds were encapsulated in
alginate beads containing 0.5M sucrose, precultured at 5 degrees C on media
containing daily increasing sucrose concentrations from 0.1 to 1.0M, then
desiccated to 21% moisture content (fresh weight basis) and frozen slowly
(pre-freezing to -40 degrees C at 0.2 degrees C/min followed by immersion in
liquid nitrogen). The physiological state of the in vitro mother-plants and the
freezing procedure employed dramatically influenced the results. Regrowth after
cryopreservation was achieved only when buds were sampled from mother-plants
that had been kept without subculture for 3 to 4 months. An additional one-month
period of cold-acclimation of mother-plants at 5 degrees C and a two-step
freezing procedure improved growth recovery. The multiplication rate of shoots
produced from cryopreserved buds was lower than that of controls during the
first subcultures after thawing. However, the rooting ability of control and
cryopreserved plantlets became similar from the fourth subculture onwards.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11788874 [PubMed - indexed for MEDLINE]

332: Radiats Biol Radioecol. 2001 Nov-Dec;41(6):664-70.

[Photoprotective properties of melanins from grape (Vitis vinifera) and black
tea (Thea sinensis)]

[Article in Russian]

Novikov DA, Kurchenko VP, Azarko II.

Department of Biochemistry, Biological Faculty, Belarus State University, Minsk,
220050, Belarus. [email protected]

Dark pigments of melanin type were extracted from the rind of ripe grapes Vitis
vinifera (ring "Alfa") and pack tea (Thea sinensis). The study of
photoprotection activity of the extracted pigments have shown that they
intensively absorb radiation in the UV and visible. The increase of the
photoprotection effect correlates with the concentration of paramagnetic centers
in the melanins. The pigments reduce the number of damaged molecules of plasmid
DNA pBR-322 and inhibition the IUV-induced lipid peroxidation.

Publication Types:
English Abstract

PMID: 11785308 [PubMed - indexed for MEDLINE]

333: J Chromatogr A. 2001 Nov 30;936(1-2):145-57.

Monoterpenic and norisoprenoidic glycoconjugates of Vitis vinifera L. cv. Melon
B. as precursors of odorants in Muscadet wines.

Schneider R, Razungles A, Augier C, Baumes R.

Unite Mixte de Recherche - Sciences pour l'OEnologie, 2 place Viala, 34060
Montpellier cedex 1, France. [email protected]

The volatile monoterpenic and norisoprenoidic compounds released by glycosidase
enzyme hydrolysis of C18 reversed-phase isolates from the juice of Vitis
vinifera L. cv. Melon B. have been qualitatively and quantitatively determined
using GC-MS and GC-FID. The components analyzed were broadly similar to those
previously reported for other varieties but the level of bound
p-menth-1-en-7,8-diol was higher in this cultivar. Then the monoterpenic and
norisoprenoidic volatiles released from the same glycosidic extracts under mild
acid conditions, mimicking wine aging conditions, have been analyzed using
GC-Olfactometry and GC-MS. The most odorous compounds detected were p-cymene,
terpinen-4-ol, cis- and trans-vitispiranes,
1,6,6-trimethyl-1,2-dihydronaphtalene (TDN), beta-damascenone and riesling
acetal. To assess their potential levels in corresponding wines after ageing,
most of these odorants were generated by harsh acid hydrolysis from the
precursors extracts and quantitatively determined using SPME and GC-MS/MS. For
the development and application of this analysis, the odorants not commercialy
available were synthesized. The total amounts of norisoprenoidic odorants
generated by acid hydrolysis of the glycosidic extracts were shown to be
proportional to the total amounts of these precursors.

PMID: 11760996 [PubMed - indexed for MEDLINE]

334: Biosci Biotechnol Biochem. 2001 Oct;65(10):2213-9.

Purification and characterization of a O-methyltransferase capable of
methylating 2-hydroxy-3-alkylpyrazine from Vitis vinifera L. (cv. Cabernet
Sauvignon).

Hashizume K, Tozawa K, Hiraga Y, Aramaki I.

National Research Institute of Brewing, Higashihiroshima, Japan.
[email protected]

An S-adenosyl-L-methionine-dependent O-methyltransferase capable of methylating
2-hydroxy-3-alkylpyrazine (HP) was purified 7,300-fold to apparent homogeneity
with an 8.2% overall recovery from Vitis vinifera L. (cv. Cabernet Sauvignon)
through a purification procedure including column chromatography on
DEAE-Sepharose FF, Ether-5PW, hydroxyapatite, G2000SW(XL), and DEAE-5PW. The
relative molecular mass of the native enzyme estimated on gel permeation
chromatography was 85 kDa, and the subunit molecular mass was estimated to be 41
kDa on SDS-polyacrylamide gel electrophoresis. The enzyme also methylates
caffeic acid. The Vmax for IBHP and caffeic acid were 0.73 and 175 pkatals/mg,
respectively, and the respective Km for IBHP and caffeic acid were 0.30 and
0.032 mm. The optimum pH for IBHP (8.5) was different from that for caffeic acid
(7.5).

PMID: 11758912 [PubMed - indexed for MEDLINE]

335: Planta Med. 2001 Nov;67(8):714-7.

Procyanidins from Vitis vinifera seeds inhibit the respiratory burst of
activated human neutrophils and lysosomal enzyme release.

Carini M, Stefani R, Aldini G, Ozioli M, Facino RM.

Istituto Chimico Farmaceutico Tossicologico, University of Milan, Italy.
[email protected]

The inhibitory properties of procyanidins (a standardized oligomeric catechin
fraction) from Vitis vinifera L. seeds on the respiratory burst and on the
release of granule components myeloperoxidase, beta-glucuronidase and elastase
were studied in activated human neutrophils. Procyanidins strongly inhibit
superoxide generation with an IC(50) of 7.2 microM, through a direct scavenging
of superoxide and prevent the release from calcium ionophore activated
neutrophils of beta-glucuronidase (IC(50) = 13.9 microM), myeloperoxidase
(IC(50) = 7.2 microM) and elastase (IC(50) = 5.4 microM). In addition they
dose-dependently inhibit the activity of myeloperoxidase released from calcium
ionophore-stimulated cells with an IC(50) value of 2 microM. The monomeric
constitutive unit (+)-catechin was far less active than procyanidins in all the
models tested. These results evidence that procyanidins efficiently restrain the
inflammatory response of activated neutrophils in vitro and whenever absorbed in
vivo can prevent their oxidative discharge at the site(s) of their adhesion.

PMID: 11731911 [PubMed - indexed for MEDLINE]

336: J Agric Food Chem. 2001 Nov;49(11):5531-6.

Stilbene compounds and stilbene synthase expression during ripening, wilting,
and UV treatment in grape cv. Corvina.

Versari A, Parpinello GP, Tornielli GB, Ferrarini R, Giulivo C.

Corso di Laurea in Scienze e Tecnologie Alimentari, Universita di Bologna, via
Ravennate 1020, Cesena (FC) 47023 Italy.

The content of selected phenolic compounds including resveratrol
(trans-3,5,4'-trihydroxystilbene) and its cis-isomer and glucosides (piceides)
were monitored in grape (Vitis vinifera L. cv. Corvina) berry skin during
ripening in the vineyard and in response to the post-harvest drying process
(wilting). Four wilting conditions were compared (traditional, mixed,
low-temperature, and high-temperature) to verify the eliciting effect of drying
on resveratrol production. During fruit ripening the cis-piceid was the major
stilbene found in berry skins, and a weak accumulation of stilbene synthase
(STS) mRNA was observed, whereas UV-light irradiation greatly stimulated STS
transcript of unripe berries. A time-course experiment showed the highest STS
mRNA accumulation and resveratrol content (34 microg/g fresh weight at 58 days)
occurring in berry skins in a mixed wilting process.

PMID: 11714355 [PubMed - indexed for MEDLINE]

337: J Agric Food Chem. 2001 Nov;49(11):5494-501.

Determination of free and conjugated indole-3-acetic acid, tryptophan, and
tryptophan metabolites in grape must and wine.

Hoenicke K, Simat TJ, Steinhart H, Kohler HJ, Schwab A.

University of Hamburg, Institute of Biochemistry and Food Chemistry,
Grindelallee 117, D-20146 Hamburg, Germany.

Tryptophan (Trp) and its metabolites, especially indole-3-acetic acid (IAA), are
considered to be potential precursors of 2-aminoacetophenone (AAP), an aroma
compound that causes an "untypical aging off-flavor" (UTA) in Vitis vinifera
wines. In this study, RP-HPLC with fluorescence detection was used for the
qualitative and quantitative analysis of Trp and Trp metabolites in grape musts
and wines to which different viticultural measures had been applied (time of
harvest, soil treatment, leaf plucking, vine prune). An alkaline hydrolysis was
developed to release bound IAA and Trp. A sensitive and selective determination
of different Trp metabolites was achieved after solid phase extraction using a
strong anion exchange material. In the examined grape musts, more than 95% of
the total IAA was bound either as ester conjugate or as amide conjugate. Free
IAA and other Trp metabolites were below the detection limit (<3 microg/L) or
could be determined only in traces. Their amounts increased significantly during
fermentation, whereas the amount of Trp decreased. It could be shown that the
different viticultural measures applied (except the vine prune) as well as the
climatic conditions of the vintage exhibited significant influences on the
amounts of Trp and Trp metabolites in grape musts or wines.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11714350 [PubMed - indexed for MEDLINE]

338: J Agric Food Chem. 2001 Nov;49(11):5484-8.

Determination of carotenoid profiles in grapes, musts, and fortified wines from
Douro varieties of Vitis vinifera.

Guedes De Pinho P, Silva Ferreira AC, Mendes Pinto M, Benitez JG, Hogg TA.

Escola Superior de Biotecnologia, Universidade Catolica Portuguesa, R. Dr.
Antonio Bernardino de Almeida 4200-072 Porto, Portugal. [email protected]

beta-Carotene and six xanthophylls (lutein, neoxanthin, violaxanthin,
luteoxanthin, cryptoxanthin, and echinenone) have been identified and
semiquantitatively or quantitatively determined in musts and port wines for the
first time. An HPLC method was developed and compared with that of one based on
thin layer cromatography with scanning densitometry. The most abundant
carotenoids present in red grape varieties are beta-carotene and lutein. In
wines, significant quantities of violaxanthin, luteoxanthin, and neoxanthin were
found. This study was done with berries (skin and pulp), musts, and fortified
wines. Some experiments were performed to follow carotenoid content from grapes
to wines. Although the levels of beta-carotene and lutein found in fortified
wines were lower than those found in musts, other xanthophylls, such as
neoxanthin, violaxanthin, and luteoxanthin, exist in appreciable amounts in
young ports.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11714348 [PubMed - indexed for MEDLINE]

339: J Agric Food Chem. 2001 Nov;49(11):5348-55.

Composition of grape skin proanthocyanidins at different stages of berry
development.

Kennedy JA, Hayasaka Y, Vidal S, Waters EJ, Jones GP.

Department of Horticulture, Viticulture and Oenology, Adelaide University, Waite
Campus, PMB 1, Glen Osmond, South Australia 5064, Australia.
[email protected]

The composition of grape (Vitis vinifera L. cv. Shiraz) skin proanthocyanidins
has been determined at different stages of berry development. Beginning
approximately 3 weeks after fruit set and concluding at commercial ripeness, the
composition of isolated skin proanthocyanidins was determined using the
following analytical techniques: elemental analysis, UV-Vis absorption
spectroscopy, reversed-phase HPLC after acid-catalysis in the presence of excess
phloroglucinol, gel permeation chromatography, electrospray ionization mass
spectrometry (ESI-MS), and (13)C NMR. On the basis of these analyses, berry
development was correlated with an increase in proanthocyanidin mean degree of
polymerization, an increase in the proportion of (-)-epigallocatechin extension
subunits, and increases in the level of anthocyanins associated with the
proanthocyanidin fraction. Additionally, data acquired from ESI-MS of the
isolates following acid-catalysis in the presence of excess phloroglucinol is
consistent with pectin-bound proanthocyanidins.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11714327 [PubMed - indexed for MEDLINE]

340: J Exp Bot. 2001 Dec;52(365):2313-22.

Sink feedback regulation of photosynthesis in vines: measurements and a model.

Quereix A, Dewar RC, Gaudillere JP, Dayau S, Valancogne C.

Unite de Bioclimatologie, INRA, Centre de Bordeaux, BP81, 33883 Villenave
d'Ornon CEDEX, France.

An experimental and modelling study of source-sink interactions in Vitis
vinifera L., cv. Cabernet Sauvignon, rooted cuttings under non-limiting
environmental conditions with a 12 h photoperiod is presented here. After 4 h,
measured photosynthesis, stomatal conductance and leaf carbohydrate content
reached maximum values. Over the remainder of the photoperiod, photosynthesis
and stomatal conductance decreased continuously, whereas leaf carbohydrate
content remained relatively constant. Because the experiment took place in a
non-limiting environment, the results suggest that stomatal regulation of
photosynthesis was mediated by an internal factor, possibly related to sink
activity. A simple 1-source, 2-sink model was developed to examine the extent to
which the data could be explained by a hypothetical sink-to-source feedback
mechanism mediated by carbohydrate levels in either the mesophyll, the source
phloem or the phloem of one of the two sinks. Model simulations reproduced the
data well under the hypothesis of a phloem-based feedback signal, although the
data were insufficient to elucidate the detailed nature of such a signal. In a
sensitivity analysis, the steady-state response of photosynthesis to sink
activity was explored and predictions made for the partitioning of photosynthate
between the two sinks. The analysis highlights the effectiveness of a
phloem-based feedback signal in regulating the balance between source and sink
activities. However, other mechanisms for the observed decline in
photosynthesis, such as photoinhibition, endogenous circadian rhythms or
hydraulic signals in the leaf cannot be excluded. Nevertheless, it is concluded
that the phloem-based feedback model developed here may provide a useful working
hypothesis for incorporation into plant growth models and for further
development and testing.

PMID: 11709581 [PubMed - indexed for MEDLINE]

341: Plant Physiol. 2001 Nov;127(3):1065-76.

Rapid deposition of extensin during the elicitation of grapevine callus cultures
is specifically catalyzed by a 40-kilodalton peroxidase.

Jackson PA, Galinha CI, Pereira CS, Fortunato A, Soares NC, Amancio SB, Pinto
Ricardo CP.

Instituto de Tecnologia Quimica e Biologica, Apartado 127, 2781-901 Oeiras,
Portugal.

Elicitation or peroxide stimulation of grape (Vitis vinifera L. cv Touriga) vine
callus cultures results in the rapid and selective in situ insolubilization of
an abundant and ionically bound cell wall protein-denominated GvP1.
Surface-enhanced laser desorption/ionization/time of flight-mass spectrometry
analysis, the amino acid composition, and the N-terminal sequence of purified
GvP1 identified it as an 89.9-kD extensin. Analysis of cell walls following the
in situ insolubilization of GvP1 indicates large and specific increases in the
major amino acids of GvP1 as compared with the amino acids present in
salt-eluted cell walls. We calculate that following deposition, covalently bound
GvP1 contributes up to 4% to 5% of the cell wall dry weight. The deposition of
GvP1 in situ requires peroxide and endogenous peroxidase activity. Isoelectric
focusing of saline eluates of callus revealed only a few basic peroxidases that
were all isolated or purified to electrophoretic homogeneity. In vitro and in
situ assays of extensin cross-linking activity using GvP1 and peroxidases showed
that a 40-kD peroxidase cross-linked GvP1 within minutes, whereas other
grapevine peroxidases had no significant activity with GvP1. Internal peptide
sequences indicated this extensin peroxidase (EP) is a member of the class III
peroxidases. We conclude that we have identified and purified an EP from
grapevine callus that is responsible for the catalysis of GvP1 deposition in
situ during elicitation. Our results suggest that GvP1 and this EP play an
important combined role in grapevine cell wall defense.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11706187 [PubMed - indexed for MEDLINE]

342: Plant Physiol. 2001 Nov;127(3):863-75.

Effects of natural intensities of visible and ultraviolet radiation on epidermal
ultraviolet screening and photosynthesis in grape leaves.

Kolb CA, Kaser MA, Kopecky J, Zotz G, Riederer M, Pfundel EE.

Lehrstuhl fur Botanik II, Universitat Wurzburg, Julius-von-Sachs-Platz 3,
D-97082 Wurzburg, Germany.

Grape (Vitis vinifera cv Silvaner) vine plants were cultivated under shaded
conditions in the absence of ultraviolet (UV) radiation in a greenhouse, and
subsequently placed outdoors under three different light regimes for 7 d.
Different light regimes were produced by filters transmitting natural radiation,
or screening out the UV-B (280-315 nm), or screening out the UV-A (315-400 nm)
and the UV-B spectral range. During exposure, synthesis of UV-screening
phenolics in leaves was quantified using HPLC: All treatments increased
concentrations of hydroxycinnamic acids but the rise was highest, reaching 230%
of the initial value, when UV radiation was absent. In contrast, UV-B radiation
specifically increased flavonoid concentrations resulting in more than a 10-fold
increase. Transmittance in the UV of all extracted phenolics was lower than
epidermal UV transmittance determined fluorimetrically, and the two parameters
were curvilinearly related. It is suggested that curvilinearity results from
different absorption properties of the homogeneously dissolved phenolics in
extracts and of the non-homogeneous distribution of phenolics in the epidermis.
UV-B-dependent inhibition of maximum photochemical yield of photosystem II
(PSII), measured as variable fluorescence of dark-adapted leaves, recovered in
parallel to the buildup of epidermal screening for UV-B radiation, suggesting
that PSII is protected against UV-B damage by epidermal screening. However, UV-B
inhibition of CO(2) assimilation rates was not diminished by efficient UV-B
screening. We propose that protection of UV-B inactivation of PSII is observed
because preceding damage is efficiently repaired while those factors determining
UV-B inhibition of CO(2) assimilation recover more slowly.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11706169 [PubMed - indexed for MEDLINE]

343: Phytochemistry. 2001 Nov;58(5):693-701.

Involvement of pectin methyl-esterase during the ripening of grape berries:
partial cDNA isolation, transcript expression and changes in the degree of
methyl-esterification of cell wall pectins.

Barnavon L, Doco T, Terrier N, Ageorges A, Romieu C, Pellerin P.

Unite de Recherches Biopolymeres et Aromes, IPV-INRA, 2 Place Viala, 34 060,
Montpellier, France. [email protected]

Grape berries (Vitis vinifera L., cv Ugni blanc) were harvested at 12 different
weeks of development in 1996 and 1997. Ripening was induced at veraison, the
crucial stage of berry softening, and was followed by a rapid accumulation of
glucose and fructose and an increase of pH. Total RNAs, crude proteins and cell
wall material were isolated from each developmental stage. A partial length cDNA
(pme1, accession number AF159122, GenBank) encoding a pectin methyl-esterase
(PME, EC 3.1.1.11) was cloned by RT-PCR with degenerate primers. Northern blots
revealed that mRNAs coding for PME accumulate from one week before the onset of
ripening until complete maturity, indicating that this transcript represents an
early marker of veraison and could be involved in berry softening. However, PME
activity was detected during all developmental stages. Total activity per berry
increased, whereas "specific" activity, on a fresh weight basis, decreased
during development. The amount of cell wall material (per berry and per g of
berry) followed the same pattern as that of PME activity (total and "specific"
respectively), indicating they were tightly correlated and that PME levels
varied very little in the cell walls. Nevertheless, the degree of
methyl-esterification of insoluble pectins decreased throughout the development
from 68% in green stages to less than 20% for the ripe berries, and this
observation is consistent with the induction of PME mRNAs during ripening.
Relations between transcript expression, PME activity, the DE of insoluble
pectic polysaccharides and their involvement in grape berry ripening are
discussed.

PMID: 11672733 [PubMed - indexed for MEDLINE]

344: J Agric Food Chem. 2001 Oct;49(10):4627-34.

New flavanol derivatives from grape (Vitis vinifera) byproducts. Antioxidant
aminoethylthio-flavan-3-ol conjugates from a polymeric waste fraction used as a
source of flavanols.

Torres JL, Bobet R.

Department of Peptide and Protein Chemistry, Institute for Chemical and
Environmental Research (IIQAB-CSIC), Jordi Girona 18-26, 08034 Barcelona, Spain.
[email protected]

A new family of antioxidants has been obtained from a residual fraction of
polymeric polyphenols of grape origin. The integral exploitation of resources is
important in any sustainable production scheme. Many byproducts and residues
generated by the agroindustries contain polyphenols with potential application
as preventative agents against cancer and cardiovascular diseases. Among these
polyphenols oligomeric proanthocyanidins are particularly significant. The
polymeric forms, considered of less interest because of their astringent
properties, constitute the largest portion of the biologically active plant
proanthocyanidins. The new compounds described here result from the breakdown of
polymeric flavanols in the presence of cysteamine and bear an amino function,
which facilitates their isolation from complex mixtures by cation-exchange gels
or resins. In this way, valuable antioxidant molecules can be efficiently
obtained from otherwise wasted polymers. The new molecules appear to be as
effective as their underivatized counterparts (flavan-3-ols) as antioxidants.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11599999 [PubMed - indexed for MEDLINE]

345: J Environ Qual. 2001 Sep-Oct;30(5):1844-7.

Pesticide applications of copper on perennial crops in California, 1993 to 1998.

Epstein L, Bassein S.

Department of Plant Pathology, Univ. of California, Davis 95616-8680, USA.
[email protected]

Inorganic copper is used as a broad-spectrum fungicide and bacteriocide on a
variety of agricultural crops. After application, the copper residue typically
accumulates in the upper 15 cm of soil. Data from the California Pesticide Use
Reports were used to estimate the augmentation of copper in the soil that
resulted from pesticide applications for the six years from 1993 to 1998 on 12
crops that are grown without rotation. The estimated mean mg Cu kg(-1) soil
added to the upper 15 cm during the six years was the following: walnut (Juglans
regia L.), 28; peach [Prunus persica (L.) Batsch var. persica], 22; nectarine
[Prunuspersica (L.) Batsch var. nucipersica (Suckow) C.K. Schneid], 19; cherry
(Pseudolmedia oxyphyllaria Donn. Sm.), 18; rice (Orvza sativa L.), 16; apricot
(Prunus armeniaca L.), 11; orange [Citrus sinensis (L.) Osbeck) and plum (Prunus
domestica L. subsp. domestica ), 9; lemon [Citrus limon (L.) Burm.f.] and almond
[Prunus dulcis (Mill.) D.A. Webb], 6; pear (Pyrus communis L.), 4; and grape
(Vitis vinifera L.), 3. In addition, for the first five of these crops, we
estimated the area that was treated with each level of kg Cu ha(-1). For
example, for walnut orchards, we estimated that 12 500 ha, or 17% of the planted
area, was treated with a quantity of Cu that would increase the total
concentration of Cu in the upper 15 cm of soil by at least 50 mg Cu kg(-1) soil.
A comparison of the amount of Cu per unit planted area that was applied in the
first and second half of the study indicated that the intensity of copper use is
either relatively constant or increasing, depending on the crop. The findings
are discussed in relation to the potential effect of continued long-term use of
Cu pesticides on soil sustainability.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11577894 [PubMed - indexed for MEDLINE]

346: J Nat Prod. 2001 Sep;64(9):1174-8.

A hemicellulose B fraction from grape skin (Vitis vinifera, Palomino variety).

Igartuburu JM, Pando E, Rodriguez Luis F, Gil-Serrano A.

Departamento de Quimica Organica, Facultad de Ciencias, Universidad de Cadiz,
Apartado 40, 11510-Puerto Real, Cadiz, Spain. [email protected]

The structure of a hemicellulose B fraction (B-1) isolated from grape skins
(Vitis vinifera) of the Palomino variety has been studied by methylation
analysis, (1)H NMR and (13)C NMR spectroscopy, and partial acid hydrolysis.
Hemicellulose B-1 appeared to be homogeneous by gel filtration with a
weight-average molecular weight of 22 600. This polysaccharide is a linear
xyloglucan chain composed of xylopyranosyl and glucopyranosyl residues linked by
beta-(1-->4) glycosidic bonds. Attached to this backbone,
4-O-methyl-D-glucuronopyranosyl acid, D-glucopyranosyl, and L-fucopyranosyl
residues occur at position 2 in a ratio of one residue for every five units of
xylose in the main chain, with D-xylopyranosyl residues attached at position 6
of glucose units in a ratio of one residue for every two glucose-derived
moieties in the main chain.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11575951 [PubMed - indexed for MEDLINE]

347: J Agric Food Chem. 2001 Sep;49(9):4183-7.

Determination of anthocyanidins in berries and red wine by high-performance
liquid chromatography.

Nyman NA, Kumpulainen JT.

Food Research, L-Building, MTT Agrifood Research Finland, FIN-31600 Jokioinen,
Finland.

A high-performance liquid chromatographic (HPLC) method for the determination of
anthocyanidins from berries and red wine is described. Delphinidin, cyanidin,
petunidin, pelargonidin, peonidin, and malvidin contents of bilberry (Vaccinium
myrtillus), black currant (Ribes nigrum), strawberry (Fragaria ananassa cv.
Jonsok), and a Cabernet sauvignon (Vitis vinifera) red wine were determined. The
aglycon forms of the anthocyanins present in the samples were revealed by acid
hydrolysis. A reversed phase analytical column was employed to separate the
anthocyanidins before identification by diode array detection. The suitability
of the method was tested by determining the recovery (95-102% as aglycons and
69-104% from glycosides) for each anthocyanidin. Method repeatability was tested
by charting the total aglycon content of two samples over a period of 14
analyses and determining the coefficients of variation (1.41% for bilberry and
2.56% for in-house reference material). The method developed proved thus to be
effective for reliable determination of anthocyanidins from freeze-dried berry
samples and red wine. The total anthocyanidin content of the tested samples was
as follows: in-house reference material, 447 +/- 8 mg/100 g; strawberry, 23.8
+/- 0.4 mg/100 g; black currant, 135 +/- 3 mg/100 g; bilberry, 360 +/- 3 mg/100
g; and Cabernet sauvignon red wine, 26.1 +/- 0.1 mg/100 mL.

PMID: 11559107 [PubMed - indexed for MEDLINE]

348: Plant Cell Environ. 1999 Feb;22(2):197-204.

Shoot orientation affects vessel size, shoot hydraulic conductivity and shoot
growth rate in Vitis vinifera L.

Schubert A, Lovisolo C, Peterlunger E.

Centro di Studio per il Miglioramento genetico e la Biologia della Vite, CNR.
[email protected]

Vitis vinifera L. plants were grown in containers and each plant's single shoot
was orientated upwards or downwards. Some plants were trained first upwards,
then downwards, then again upwards (N-shaped plants). Vegetative growth was
reduced in plants trained downwards compared to that in upward and N-shaped
plants. Shoot growth rate slowed in downward shoot portions, but only after the
apex had grown downwards for at least 10 internodes. Shoot hydraulic
conductivity kh, measured after elimination of xylem embolisms, was lower in
downward than in upward plants. In N-shaped plants kh was higher in the
upward-growing shoot portions, and lower in the central, downward-growing
portion. Shoot- and leaf-specific conductivities were also lower in downward
than in upward shoot portions. Xylem cross-sectional area and xylem structure
(number of wedges, number of vessels per unit xylem area) differed little in the
three orientations. In contrast, vessel diameter and the sum of vessel
cross-sectional areas were significantly smaller in downward than in upward
shoot portions. These differences could explain the reduction in conductivity
observed in the downward-orientated shoot portions. The measurements taken on
N-shaped plants showed that the decreases in kh and in vessel size were a result
of shoot orientation, not shoot bending.

PMID: 11542249 [PubMed - indexed for MEDLINE]

349: Planta. 2001 May;213(1):20-8.

Changes in acidity and in proton transport at the tonoplast of grape berries
during development.

Terrier N, Sauvage FX, Ageorges A, Romieu C.

Unite de Recherche de Biochimie Metabolique et Technologie, Institut des
Produits de la Vigne, INRA, Montpellier, France. [email protected]

As in many fruits, the induction of grape berry (Vitis vinifera L.) ripening
results in intense breakdown of malic acid. Using membrane fractions, we tested
the hypothesis that changes in acidity resulted from malate vacuolar
decompartmentation. The hydrolytic activities of the two primary vacuolar pumps
inorganic pyrophosphatase (V-PPase; EC 3.6.1.1) and vacuolar ATPase (V-ATPase;
EC 3.6.1.3) increased throughout development with an acceleration during
ripening, as confirmed by Western blotting and analysis of transcript
expression. The ratio of V-PPase activity to V-ATPase activity was always in
favour of V-PPase and reached its maximum value at veraison. The rate of anion
transport strongly increased during ripening. Before ripening, tonoplast passive
permeability was low, but rose during ripening. Our data indicate that tonoplast
leakage dramatically increased during ripening. This leakage is probably the
prime cause of malate decompartmentation, amplified by the incapacity of
oxidative phosphorylation to face increased energy demand.

PMID: 11523652 [PubMed - indexed for MEDLINE]

350: Mutat Res. 2001 Sep 1;480-481:243-68.

Molecular mechanisms underlying chemopreventive activities of anti-inflammatory
phytochemicals: down-regulation of COX-2 and iNOS through suppression of
NF-kappa B activation.

Surh YJ, Chun KS, Cha HH, Han SS, Keum YS, Park KK, Lee SS.

College of Pharmacy, Seoul National University, Shinlim-dong, Kwanak-ku, Seoul
151-742, South Korea. [email protected]

A wide array of phenolic substances, particularly those present in edible and
medicinal plants, have been reported to possess substantial anticarcinogenic and
antimutagenic activities. The majority of naturally occurring phenolics retain
antioxidative and anti-inflammatory properties which appear to contribute to
their chemopreventive or chemoprotective activity. Cyclooxygenase-2 (COX-2)
inducible and nitric oxide synthase (iNOS) are important enzymes that mediate
inflammatory processes. Improper up-regulation of COX-2 and/or iNOS has been
associated with pathophysiology of certain types of human cancers as well as
inflammatory disorders. Since inflammation is closely linked to tumor promotion,
substances with potent anti-inflammatory activities are anticipated to exert
chemopreventive effects on carcinogenesis, particularly in the promotion stage.
Examples are curcumin, a yellow pigment of turmeric (Curcuma longa L.,
Zingiberaceae), the green tea polyphenol epigallocatechin gallate (EGCG), and
resveratrol from grapes (Vitis vinifera, Vitaceae) that strongly suppress tumor
promotion. Recent studies have demonstrated that eukaryotic transcription factor
nuclear factor-kappa B (NF-kappa B) is involved in regulation of COX-2 and iNOS
expression. Several chemopreventive phytochemicals have been shown to inhibit
COX-2 and iNOS expression by blocking improper NF-kappa B activation. Multiple
lines of compelling evidence indicate that extracellular-regulated protein
kinase and p38 mitogen-activated protein kinase are key elements of the
intracellular signaling cascades responsible for NF-kappa B activation in
response to a wide array of external stimuli. Curcumin, EGCG and resveratrol
have been shown to suppress activation of NF-kappa B. One of the plausible
mechanisms underlying inhibition of NF-kappa B activation by aforementioned
phytochemicals involves repression of degradation of the inhibitory unit I kappa
B alpha, which hampers subsequent nuclear translocation of the functionally
active subunit of NF-kappa B.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 11506818 [PubMed - indexed for MEDLINE]

351: J Agric Food Chem. 2001 Jul;49(7):3321-7.

Free radical scavenging activities measured by electron spin resonance
spectroscopy and B16 cell antiproliferative behaviors of seven plants.

Calliste CA, Trouillas P, Allais DP, Simon A, Duroux JL.

UPRES EA 1085, Biomolecules et Cibles Cellulaires Tumorales, Laboratoire de
Biophysique, Faculte de Pharmacie, 2 rue du Dr. Marcland, 87025 Limoges Cedex,
France.

In an effort to discover new antioxidant natural compounds, seven plants that
grow in France (most of them in the Limousin countryside) were screened. Among
these plants, was the extensively studied Vitis vinifera as reference. For each
plant, sequential percolation was realized with five solvents of increasing
polarities (hexane, chloroform, ethyl acetate, methanol, and water). Free
radical scavenging activities were examined in different systems using electron
spin resonance (ESR) spectroscopy. These assays were based on the stable free
radical 1,1-diphenyl-2-picrylhydrazyl (DPPH), the hydroxyl radicals generated by
a Fenton reaction, and the superoxide radicals generated by the X/XO system.
Antiproliferative behavior was studied on B16 melanoma cells. ESR results showed
that three plants (Castanea sativa, Filipendula ulmaria, and Betula pendula)
possessed, for the most polar fractions (presence of phenolic compounds), high
antioxidant activities in comparison with the Vitis vinifera reference. Gentiana
lutea was the only one that presented a hydroxyl scavenging activity for the
ethyl acetate and chloroform fractions. The antiproliferative test results
showed that the same three plants are the most effective, but for the apolar
fractions (chloroform and hexane).

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11453770 [PubMed - indexed for MEDLINE]

352: J Agric Food Chem. 2001 Jul;49(7):3310-5.

Changes in nitrogen compounds in must and wine during fermentation and
biological aging by flor yeasts.

Mauricio JC, Valero E, Millan C, Ortega JM.

Departamento de Microbiologia, Universidad de Cordoba, Campus Universitario de
Rabanales, Edificio C-6, 14014 Cordoba, Spain. [email protected]

Urea, ammonium, and free amino acid contents were quantified in a must from
Vitis vinifera cv. Pedro Ximenez grapes and in fermented wine and after a short
aging of this wine by Saccharomyces cerevisiae race capensis yeast under
variable oxygen availability conditions. The previous compounds were also
determined in a wine in which the nitrogen source was depleted by the same race
of flor yeast (old wine) and also following the addition of ammonium ion,
L-glutamic acid, and L-proline. Under specific conditions such as low oxygen
level and the absence of some nutrients, the yeasts release some amino acids
including L-threonine, L-tryptophan, L-cysteine, and L-methionine to the medium.
These amino acids must originate primarily in a de novo synthesis from ethanol
that regenerates NAD(P)+. On the basis of these results, the yeasts may be able
to use amino acids not only as nitrogen sources but also as redox agents to
balance the oxidation-reduction potential under conditions of restricted oxygen,
when electron transport along the respiratory chain may be hindered or limited.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11453768 [PubMed - indexed for MEDLINE]

353: Plant Mol Biol. 2001 Apr;45(6):669-78.

Indications for post-translational regulation of Vitis vinifera L. arginine
decarboxylase.

Primikirios NI, Roubelakis-Angelakis KA.

Department of Biology, University of Crete, Heraklion, Greece.

In order to study arginine decarboxylase regulation, we produced an antiserum
against a hybrid of a 615 amino acid residue fragment of grapevine arginine
decarboxylase cDNA with maltose-binding protein. The antiserum generated
recognized mainly a protein band of ca. 80 kDa in extracts from grapevine
tissues. Extracts from leaves and internodes in different developmental stages
showed differences in the quantity of the 80 kDa band recognized by the
antiserum. However, these differences did not correspond with changes in
arginine decarboxylase specific activity. Furthermore, western blot analysis of
extracts from cell cultures, where enzyme-specific activity was induced or
repressed, did not reveal respective changes in the quantity of the 80 kDa
protein band. Digestion of the hybrid by the specific protease factor Xa
resulted in a polypeptide of 90 kDa instead of the expected two polypeptides of
43 and 66 kDa. Finally, western blot analysis of shoot extract incubated with
factor Xa or the hybrid protein previously digested by factor Xa revealed that
factor Xa-digested hybrid protein cleaved the 80 kDa band, resulting in two
bands of ca. 38 and 40 kDa, whereas factor Xa alone did not affect it. These
results suggest that arginine decarboxylase protein levels and/or activity is
post-translationally regulated, as has been shown for other enzymes of polyamine
biosynthesis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11430429 [PubMed - indexed for MEDLINE]

354: Plant Mol Biol. 2001 Mar;45(5):541-53.

A cDNA from grapevine (Vitis vinifera L.), which shows homology to AGAMOUS and
SHATTERPROOF, is not only expressed in flowers but also throughout berry
development.

Boss PK, Vivier M, Matsumoto S, Dry IB, Thomas MR.

CSIRO Plant Industry, Horticulture Unit, Glen Osmond, SA, Australia.
[email protected]

An AGAMOUS/SHATTERPROOF homologue (Vvmads1) was isolated from grapevine by
differential display between berry and leaf mRNA. The predicted protein sequence
of the full-length clone shows a high degree of homology to PLENA (77% identity)
and to SHP1 and SHP2 (75% and 74% identity respectively), and is grouped with
AGAMOUS/PLENA homologues when the conserved MADS and K domains are compared.
Vvmads1 is expressed only in the later stages of flower development and
throughout berry development, although expression is reduced after ripening
commenced. When Vvmads1 was over-expressed in tobacco, the resulting plants
display altered morphologies in the outer two floral whorls. In the most extreme
cases, the inner whorls were surrounded by a carpelloid structure created by the
modified sepals. Within these sepals were petals which had been split into
sections and which were attached at the base of the flower by structures with
the appearance of filaments. The results of this study suggest that Vvmads1 has
a regulatory role in flower development before fertilisation and a role in fruit
development after fertilisation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11414613 [PubMed - indexed for MEDLINE]

355: J Agric Food Chem. 2001 Jun;49(6):2917-23.

Volatile compounds released by enzymatic hydrolysis of glycoconjugates of leaves
and grape berries from Vitis vinifera Muscat of Alexandria and Shiraz cultivars.

Wirth J, Guo W, Baumes R, Gunata Z.

Unite de recherches Biopolymeres et Aromes, IPV-ENSAM-INRA, 2 place viala, 34060
Montpellier Cedex 1, France.

Glycoconjugates from Muscat of Alexandria and Shiraz leaves and grape berries
were isolated by adsorption on Amberlite XAD-2 resin, and enzymatically released
aglycons were analyzed by GC-FID and GC-MS. About 120 aglycons were fully or
tentatively identified. Compositional differences were observed between leaves
and berries of the two varieties in five aglycon chemical groups: C6 alcohols,
aliphatic alcohols, monoterpenes, shikimates, and C(13)-norisoprenoids, which
were much more abundant in the leaves than in the berries. The differences
observed for C(13)-norisoprenoids were in agreement with their hypothetical
independent biosynthesis in leaves and berries. Thus, 3-hydroxy-beta-damascone,
an important norisoprenoid aglycon of grape berries, was not detected in leaves,
whereas its oxidized derivative, 3-oxo-alpha-damascone, was absent in berries.
Compositional differences were also observed between Muscat and Shiraz leaves.
3-Oxo-alpha-ionol was not detected in Shiraz leaves, and its retro derivatives
were less abundant than in Muscat of Alexandria leaves. Conversely, in Shiraz
leaves the levels of 7,8-dihydroionone derivatives, such as megastigman-3,9-diol
and 3-oxo-7,8-dihydro-alpha-ionol, were higher than in Muscat of Alexandria
leaves.

PMID: 11409987 [PubMed - indexed for MEDLINE]

356: J Gen Virol. 2001 Jul;82(Pt 7):1791-5.

Quasispecies nature of the genome of Grapevine fanleaf virus.

Naraghi-Arani P, Daubert S, Rowhani A.

Department of Plant Pathology, University of California, 1 Shields Avenue,
Davis, CA 95616, USA.

Genetic diversity was characterized in 14 isolates of Grapevine fanleaf virus
(GFLV) recovered from grapevine (Vitis vinifera). Virions were collected by
immunocapture, and a 1557 bp fragment containing part of the viral coat protein
gene and part of the untranslated region to its 3' side was amplified by RT-PCR.
Sequence variation among isolates was characterized by restriction fragment
length polymorphism (RFLP) analysis and by sequencing. The AvaII-generated RFLP
patterns from the various isolates were highly variable. The isolates were
passaged in Chenopodium quinoa. The RFLP patterns altered with passage through
the alternate host, but the variation stabilized after a number of passages.
Individual genomes were recovered by cloning. The subcloned sequences were found
to vary from each other by as much as 13%, and the encoded amino acid sequences
by as much as 9%. The data suggest that the GFLV genome consists of quasispecies
populations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11413392 [PubMed - indexed for MEDLINE]

357: Biol Pharm Bull. 2001 May;24(5):592-3.

Effects of leucocyanidines on activities of metabolizing enzymes and antioxidant
enzymes.

Seo K, Jung S, Park M, Song Y, Choung S.

Toxicology Department, National Institute of Toxicological Research, Korea Food
and Drug Administration, Eunpyunggu, Seoul, Korea. [email protected]

Procyanidolic oligomers (leucocyanidines, LCs) extracted from grape seeds (Vitis
vinifera) are known to have antioxidant and antimutagenic activities, and a
protective effect against cardiovascular disease. In the present study we
examined the influence of LCs on the activities of phase 1 enzymes and
conjugation enzymes and on antioxidant enzymes such as superoxide dismutase,
catalase, and glutathione peroxidase. Administration of LCs (25, 50, and 100
mg/kg. p.o. for 7 d) markedly decreased the activities of NADPH-cytochrome P450
reductase, P4501A1, P4501A2, and P4503A4, but significantly increased the
activities of glutathione S-transferase and phenolsulfotransferase in rat liver.
However, the activities of antioxidant enzymes were not affected by LC
administration. The inhibition of P450s and increases in phase II enzyme
activities indicate a role for LCs as a chemopreventive agent against toxic or
carcinogenic metabolites of P450 isozymes.

PMID: 11379789 [PubMed - indexed for MEDLINE]

358: Plant Physiol. 2001 May;126(1):434-44.

Reduced activity of antioxidant machinery is correlated with suppression of
totipotency in plant protoplasts.

Papadakis AK, Siminis CI, Roubelakis-Angelakis KA.

Department of Biology, University of Crete, P.O. Box 2208, 71409 Heraklio,
Greece.

We previously showed that during protoplast isolation, an oxidative burst
occurred and the generation of active oxygen species was differentially mediated
in tobacco (Nicotiana tabacum) and grapevine (Vitis vinifera), accompanied by
significant quantitative differences (A.K. Papadakis, K.A. Roubelakis-Angelakis
[1999] Plant Physiol 127: 197-205). We have now further tested if the expression
of totipotency in protoplasts is related to the activity of cellular antioxidant
machinery during protoplast culture. Totipotent (T) tobacco protoplasts had
2-fold lower contents of intracellular O2*- and H2O2 and 7-fold lower levels of
O2*- and H2O2 in the culture medium, compared with non-totipotent (NT) tobacco
protoplasts. Addition of alkaline dimethylsulfoxide, known to generate O2*-,
resulted in isolation of tobacco protoplasts with reduced viability and cell
division potential during subsequent culture. Active oxygen species levels
decreased in tobacco and grapevine protoplasts during culturing, although higher
contents of O2*- and H2O2 were still found in NT- compared with T-tobacco
protoplasts, after 8 d in culture. In T-tobacco protoplasts, the reduced forms
of ascorbate and glutathione predominated, whereas in NT-tobacco and grapevine
protoplasts, the oxidized forms predominated. In addition, T-tobacco protoplasts
exhibited severalfold lower lipid peroxidation than NT-tobacco and grapevine
protoplasts. Furthermore, several antioxidant enzyme activities were increased
in T-tobacco protoplasts. Superoxide dismutase activity increased in tobacco,
but not in grapevine protoplasts during culturing due to the increased
expression of cytoplasmic Cu/Zn-superoxide dismutase. The increase was only
sustained in T-tobacco protoplasts for d 8. Together, these results suggest that
suppressed expression of totipotency in protoplasts is correlated with reduced
activity of the cellular antioxidant machinery.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11351105 [PubMed - indexed for MEDLINE]

359: Plant Physiol. 2001 May;126(1):27-31.

In vivo observation of cavitation and embolism repair using magnetic resonance
imaging.

Holbrook NM, Ahrens ET, Burns MJ, Zwieniecki MA.

Department of Organismic and Evolutionary Biology, Harvard University,
Cambridge, Massachusetts 02138, USA. [email protected]

Magnetic resonance imaging (MRI) was used to noninvasively monitor the status of
individual xylem vessels in the stem of an intact, transpiring grape (Vitis
vinifera) plant over a period of approximately 40 h. Proton density-weighted MRI
was used to visualize the distribution of mobile water in the stem and
individual xylem vessels were scored as either water or gas filled (i.e.
embolized). The number of water-filled vessels decreased during the first 24 h
of the experiment, indicating that approximately 10 vessels had cavitated during
this time. Leaf water potentials decreased from -1.25 to -2.1 MPa during the
same period. Watering increased leaf water potentials to -0.25 MPa and prevented
any further cavitation. Refilling of xylem vessels occurred as soon as the
lights were switched off, with the majority of vessels becoming refilled with
water during the first 2 to 3 h in darkness. These measurements demonstrate that
MRI can be used to monitor the functional status of individual xylem vessels,
providing the first method to study the process of cavitation and embolism
repair in intact plants.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.
Research Support, U.S. Gov't, P.H.S.

PMID: 11351066 [PubMed - indexed for MEDLINE]

360: J Agric Food Chem. 2000 Dec;48(12):6103-5.

Stilbene content of mature Vitis vinifera berries in response to UV-C
elicitation.

Adrian M, Jeandet P, Douillet-Breuil AC, Tesson L, Bessis R.

IUVV, Universite de Bourgogne, B.P. 27877, 21078 Dijon Cedex, France.
[email protected]

A method using HPLC analysis has been used to compare the level of resveratrol
and its derivatives, piceid, pterostilbene and epsilon-viniferin, in grapevine
berries of three Vitis vinifera varieties. The concentration of these compounds
has been evaluated in healthy and Botrytis cinerea infected grape clusters, both
in natural vineyard conditions and in response to UV elicitation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11312782 [PubMed - indexed for MEDLINE]

361: J Agric Food Chem. 2000 Dec;48(12):5903-12.

Genetic fingerprinting of grape plant (Vitis vinifera) using random amplified
polymorphic DNA (RAPD) analysis and dynamic size-sieving capillary
electrophoresis.

Siles BA, O'Neil KA, Fox MA, Anderson DE, Kuntz AF, Ranganath SC, Morris AC.

The College of William and Mary, Department of Chemistry, P.O. Box 8795,
Williamsburg, Virginia 23187-8795, USA. [email protected]

Dynamic size-sieving capillary electrophoresis with laser-induced fluorescence
detection (DSCE-LIF) was combined with random amplified polymorphic DNA (RAPD)
analysis to demonstrate the feasibility of the genetic analysis of grape plant
varieties and clones within a variety. Parameters of the genomic DNA extraction
process, as well as those of the RAPD analysis, were optimized specifically for
this application. Polymorphic DNA fragments were generated for four different
grape plant varieties including Cabernet Franc, Cabernet Sauvignon, Merlot, and
Chardonnay. Relative to slab gel electrophoresis (SGE) with ethidium bromide
staining, DSCE-LIF provided superior separation efficiency and detection limits
in the analysis of DNA polymorphic bands. Optimal DSCE-LIF analyses were
achieved using a 10-fold RAPD sample dilution, hydrodynamic sample injection,
and 100 ng/mL of YO-PRO-1 DNA intercalator in the dynamic size-sieving buffer
solution. In addition, the reproducibility of both the DSCE-LIF and RAPD
analyses were demonstrated.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11312766 [PubMed - indexed for MEDLINE]

362: J Agric Food Chem. 2001 Apr;49(4):1830-9.

Use of electrospray mass spectrometry for mass determination of grape (Vitis
vinifera) juice pathogenesis-related proteins: a potential tool for varietal
differentiation.

Hayasaka Y, Adams KS, Pocock KF, Baldock GA, Waters EJ, Hoj PB.

The Australian Wine Research Institute, P.O. Box 197, Glen Osmond, South
Australia, Australia 5064. [email protected]

Methods based on liquid chromatography-mass spectrometry (LC-MS) and protein
trap mass spectrometry (trap-MS) were developed to determine the complement of
pathogenesis-related (PR) proteins in grape juice. Trap-MS was superior to LC-MS
in terms of simplicity, rapidity, and sensitivity. Proteins with a wide range of
masses (13--33 kDa) were found in the juices of 19 different varieties of grape
(Vitis vinifera) and were identified as mostly PR-5 type (thaumatin-like) and
PR-3 type (chitinases) proteins. Although the PR proteins in juices of grapes
are highly conserved, small consistent differences in molecular masses were
noted when otherwise identical proteins were compared from different varieties.
These differences persisted through different harvest years and in fruits grown
in different Australian locations. With the definition of four different masses
for PR-5 proteins (range = 21,239--21,272 Da) and nine different masses of PR-3
proteins (range = 25,330--25,631 Da) and using statistical analysis, the methods
developed could be used for varietal differentiation of grapes grown in several
South Australian locations on the basis of the PR protein composition of the
juice. It remains to be seen whether this technology can be extended to grapes
grown worldwide and to wine and other fruit-derived products to assist with
label integrity to the benefit of consumers.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11308333 [PubMed - indexed for MEDLINE]

363: J Agric Food Chem. 2001 Apr;49(4):1740-6.

Analysis of proanthocyanidin cleavage products following acid-catalysis in the
presence of excess phloroglucinol.

Kennedy JA, Jones GP.

Department of Horticulture, Viticulture and Oenology, University of Adelaide,
Waite Campus, PMB 1, Glen Osmond, South Australia 5064, Australia.
[email protected]

The analysis of proanthocyanidin cleavage products after acid-catalysis in the
presence of excess phloroglucinol was investigated. In the developed analytical
method, a solution of 0.1 N HCl in methanol, containing 50 g/L phloroglucinol
and 10 g/L ascorbic acid was prepared. The proanthocyanidin of interest was
reacted in this solution (5 g/L) at 50 degrees C for 20 min, and afterward
combined with 5 volumes of 40 mM aqueous sodium acetate before analysis by
reversed-phase HPLC using an aqueous acetic acid and methanol gradient. This
procedure was used to investigate the composition of proanthocyanidins isolated
from the seed and skin tissue of Vitis vinifera L. berries. The results compared
favorably to results obtained when benzyl mercaptan was used as the trapping
nucleophile, indicating that phloroglucinol is an effective reagent for this
analysis.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11308320 [PubMed - indexed for MEDLINE]

364: Transgenic Res. 2001 Apr;10(2):105-12.

Genetic transformation of major wine grape cultivars of Vitis vinifera L.

Iocco P, Franks T, Thomas MR.

CSIRO Plant Industry, Urrbrae, SA, Australia.

We have developed an Agrobacterium-mediated transformation system for a number
of important grapevine cultivars used in wine production. Transgenic plants were
obtained for the seven cultivars: Cabernet Sauvignon, Shiraz, Chardonnay,
Riesling, Sauvignon Blanc, Chenin Blanc and Muscat Gordo Blanco. Embryogenic
callus was initiated from anther filaments and genotypic differences were
observed for initiation and subsequent proliferation with Chardonnay responding
most favourably to culture conditions. The transformation system allowed the
recovery of germinating transgenic embryos 10-12 weeks after Agrobacterium
inoculation and plants within 18 weeks. Examination of the expression patterns
of the green fluorescent protein gene under the control of the CAMV35S promoter
in leaf tissue of transgenic plants showed that for up to 35% of plants the
pattern was not uniform. The successful transformation of a genetically diverse
group of wine grape cultivars indicates that the transformation system may have
general application to an even wider range of Vitis vinifera cultivars.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11305357 [PubMed - indexed for MEDLINE]

365: Plant Sci. 2001 Apr;160(5):877-887.

Expression of a bifunctional green fluorescent protein (GFP) fusion marker under
the control of three constitutive promoters and enhanced derivatives in
transgenic grape (Vitis vinifera).

Li Z, Jayasankar S, Gray DJ.

Mid-Florida Research and Education Center, Institute of Food and Agricultural
Sciences, University of Florida, 2725 Binion Road, 32703, Apopka, FL, USA

Activity of three constitutive promoters and enhanced derivatives in transgenic
grape (Vitis vinifera L. cv. Thompson Seedless) was characterized using a
bifunctional fusion marker containing the enhanced green fluorescent protein
(EGFP) and neomycin phosphotransferase (NPTII) genes. Relative differences in
transient GFP expression and stable transformation efficiencies were used to
compare promoter activity. Expression patterns in transformed somatic embryos
revealed that the ACT2 promoter from Arabidopsis thaliana, previously shown to
be a strong constitutive promoter in A. thaliana and other species, failed to
promote strong expression in grape. In contrast, a promoter isolated from
cassava vein mosaic virus (CsVMV) supported high levels of transgene expression
equivalent to those achieved using an enhanced double cauliflower mosaic virus
(CaMV) 35S promoter. Duplication of the 5'-upstream enhancer region of the CsVMV
promoter further enhanced its ability to increase transgene expression. However,
the pattern of transgene expression driven by these two viral promoters was
significantly different at the whole plant level. The enhanced double CaMV 35S
promoter was highly active in most tissues and organs including roots, mature
leaves, shoot apices and lateral buds. In contrast, the CsVMV promoter and its
double enhancer derivative induced relatively weak expression in these tissues.
Our results suggest that activity of the CsVMV promoter, in contrast to the CaMV
35S promoter, was under developmental regulation in transgenic grape plants as
compared with the CaMV 35S promoter.

PMID: 11297784 [PubMed - as supplied by publisher]

366: J Econ Entomol. 2001 Feb;94(1):138-44.

Seasonal capacity of attached and detached vineyard roots to support grape
phylloxera (Homoptera: Phylloxeridae).

Granett J, Omer AD, Walker MA.

Department of Entomology, University of California, Davis 95616, USA.

Field experiments were conducted to evaluate population densities and survival,
developmental rate, and fecundity of grape phylloxera, Daktulosphaira vitifoliae
(Fitch), as influenced by root attachment or detachment from mature,
field-grown, Vitis vinifera L. grapevines through the growing season.
Experiments were performed using artificial infestations of California biotype A
grape phylloxera. Thirty-day bioassays on attached- and detached-roots were
repeated monthly from May to September in 1997 (cultivar 'Carignane') and April
to September in 1998 (cultivar 'Thompson Seedless'). The bioassays showed that
attached roots had lower population densities than detached roots in all months
of both years. Densities varied by month, tending to be higher in spring than in
summer. Of the population parameters studied, survival was most influenced by
attachment condition, being higher on detached than on attached roots by up to
25-fold. These results imply the importance of vine-related mortality factors to
grape phylloxera population density. Influence of root attachment condition on
developmental rate and fecundity was not uniform across bioassay months for
either year; however, in the four out of 21 assays where there was a significant
difference it favored detached roots by twofold. Fruit harvest resulted in
higher survival in the July assay but not for assays in August and September;
however, neither developmental rate nor fecundity was affected by harvest in any
ofthe assays. We conclude that mortality rather than nutritional factors are
most limiting for field populations on susceptible vines. This work suggests
that detachment of roots as occurs with root girdling by root pathogens may
increase grape phylloxera populations on infested, susceptible vines. These
results imply that excised root bioassays over-estimate grape phylloxera
virulence and underestimate rootstock resistance.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 11233103 [PubMed - indexed for MEDLINE]

367: Biochem Soc Trans. 2000 Dec;28(6):883-5.

Influence of UV-B irradiation on the carotenoid content of Vitis vinifera
tissues.

Steel CC, Keller M.

National Wine and Grape Industry Centre, School of Wine & Food Sciences, Charles
Sturt University, P.O. Box 588, Wagga Wagga NSW 2678, Australia.
[email protected]

The carotenoid content was examined in leaf and berry tissues of grapevines (cv.
Cabernet Sauvignon) grown either under ambient conditions or under a polyester
film to reduce UV light by 98%. Total carotenoids in leaves were less in vines
grown under the UV screen. Levels of beta-carotene decreased with berry
development around veraison. This effect was more pronounced in vines grown
under reduced UV light. The lutein content of berries appeared to remain
relatively constant with berry development, but levels were decreased under the
UV screen. These observations are important for the wine industry because of the
biosynthetic link between carotenoids and wine flavour and aroma compounds.

PMID: 11171244 [PubMed - indexed for MEDLINE]

368: J Nat Prod. 2001 Jan;64(1):136-8.

Two new stilbene dimer glucosides from grape (Vitis vinifera) cell cultures.

Waffo-Teguo P, Lee D, Cuendet M, Merillon J, Pezzuto JM, Kinghorn AD.

Program for Collaborative Research in the Pharmaceutical Sciences and Department
of Medicinal Chemistry and Pharmacognosy, College of Pharmacy, University of
Illinois at Chicago, Chicago, Illinois 60612, USA.

Two new stilbene dimer glucosides, resveratrol (E)-dehydrodimer
11-O-beta-D-glucopyranoside (1) and resveratrol (E)-dehydrodimer
11'-O-beta-D-glucopyranoside (2), were isolated together with the known
resveratrol (E)-dehydrodimer (3) and pallidol (4) from Vitis vinifera cell
cultures. The structures and stereochemistry of the new compounds were
determined on the basis of spectroscopic data analysis. Compounds 1 and 2 are
dimers that belong to a new type of oligostilbene formed from a resveratrol unit
and a resveratrol glucoside unit. Compounds 1 and 3 exhibited nonspecific
inhibitory activity against cyclooxygenase-1 and -2, with IC(50) values in the
range of 5 microM, whereas compound 4 was approximately 10-fold less active.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, P.H.S.

PMID: 11170689 [PubMed - indexed for MEDLINE]

369: J Agric Food Chem. 2001 Jan;49(1):26-31.

Quantitative analysis of polymeric procyanidins (Tannins) from grape (Vitis
vinifera) seeds by reverse phase high-performance liquid chromatography.

Peng Z, Hayasaka Y, Iland PG, Sefton M, Hoj P, Waters EJ.

Cooperative Research Centre for Viticulture, P.O. Box 154, Glen Osmond, South
Australia 5064, Australia. [email protected]

A reverse phase C(18) HPLC method with potential for high automated throughput
has been developed for the quantitative analysis of polymeric procyanidins
(tannins) in grape seed extracts. Chromatography gave rise to 13 distinct
UV-absorbing peaks with good baseline separation. The UV-absorbing peak eluting
last is distinct and therefore easily quantified. Biochemical analyses including
ultrafiltration, protein precipitation, and Sephadex LH20 chromatography
combined with electrospray mass spectrometric analyses establish that this peak
predominantly contains polymeric procyanidins. The polymers, which appear to be
galloylated to various degrees and seem to fragment in a characteristic manner
during electrospray mass spectrometry, are well separated from catechins and
procyanidin oligomers of up to 4 units. The recovery of polymeric grape seed
tannins with this HPLC method was 86%, which is similar to the 89% recovery
achieved with commercial quebracho tannins. The concentration of tannins in
seeds from ripe Vitis vinifera cv. Shiraz grapes ranged from 1360 to 2830 mg/kg
of berries.

PMID: 11170555 [PubMed - indexed for MEDLINE]

370: Plant Sci. 2001 Feb 5;160(3):543-550.

Comparison of UDP-glucose:flavonoid 3-O-glucosyltransferase (UFGT) gene
sequences between white grapes (Vitis vinifera) and their sports with red skin.

Kobayashi S, Ishimaru M, Ding CK, Yakushiji H, Goto N.

Persimmon and Grape Research Center, National Institute of Fruit Tree Science,
Akitsu, 729-2494, Hiroshima, Japan

The expression of the UDP-glucose:flavonoid 3-O-glucosyltransferase (UFGT) gene
has been shown to be critical for anthocyanin biosynthesis in the grape berry.
Using white cultivars and bud sports with red skin, we examined the expression
of seven anthocyanin biosynthetic genes including the UFGT gene and compared the
coding/promoter sequences of the UFGT gene. Northern blot analysis showed that
the seven anthocyanin biosynthetic genes were expressed coordinately at higher
levels in the red-skin sports than in the white-skin progenitors of the sports.
It was especially notable that UFGT gene expression was detected only in the
red-skin sports and Kyoho. However, there were no differences in either coding
or promoter sequences between Italia (Vitis vinifera) and its red-skin sport
Ruby Okuyama, or between Muscat of Alexandria (V. vinifera) and the red-skin
sport Flame Muscat. From these findings, the phenotypic change from white to red
in the sports is thought to be the result of a mutation in a regulatory gene
controlling the expression of UFGT.

PMID: 11166442 [PubMed - as supplied by publisher]

371: Plant Sci. 2000 Dec 7;160(1):159-163.

Effects of copper exposure upon nitrogen metabolism in tissue cultured Vitis
vinifera.

Llorens N, Arola L, Blade C, Mas A.

Unitat d'Enologia - Centre de Referencia en Tecnologia dels Aliments (UE-CeRTA),
Departament de Bioquimica i Biotecnologia, Facultat d'Enologia de Tarragona,
Universitat Rovira i Virgili, Ramon y Cajal, 70, 43005, Tarragona, Spain

The present study analyses the effects of copper treatment on nitrogen
metabolism in a closed system. Sauvignon grapevines were cultured in agar and
exposed to copper levels ranging from 0.07 (control) to 5 microg Cu g(-1)
medium. Ammonium, nitrate, nitrite, individual and total amino acids and protein
contents were determined in root, and leaves. Enzyme activities of nitrate and
nitrite reductases, glutamine synthetase, glutamate synthase and glutamate
dehydrogenase were also determined. Copper exposure produces a dramatic change
in nitrogen metabolism, with a reduction of total nitrogen, which reflects the
reduction on nitrate and free amino acid contents in both root and leaves. The
assimilation of nitrate the main nitrogen source in the medium, requires nitrate
reductase, which is reduced to negligible activity as response to copper
exposure. Primary nitrogen metabolism is also reduced in leaves, although to a
lesser extent than in roots, which may explain the differences between the two
organs in response to copper exposure. An alternative system for assimilation of
nitrogen through glutamate dehydrogenase in roots is proposed, while higher
levels of ammonium and glutamine may fullfil the needs of organic nitrogen in
the leaves.

PMID: 11164588 [PubMed - as supplied by publisher]

372: J Agric Food Chem. 1998 Jan 19;46(1):42-48.

Relation between Fatty Acid Content and Its Evolution during Fermentation and
Utilization of Free Amino Acids in Vacuum-Filtered Viura Must.

Ayestaran B, Garrido J, Ancin C.

Departamento de Quimica Aplicada, Universidad Publica de Navarra, Campus de
Arrosadia s/n, 31006 Pamplona, Spain.

The activity of proteins that transport amino acids changes depending on the
unsaturated residue which is introduced into the phospholipids of
Saccharomycescerevisiae membrane. The aim of this study was to observe the
influence of fatty acid content and its evolution during fermentation on the
utilization of free amino acids in Vitis vinifera var. Viura must clarified by
vacuum filtration. The results demonstrate that filtration of the must did not
reduce the concentration of free amino acids, but it did reduce fatty acids,
especially the unsaturateds. In the first half of fermentation (up to 50% sugar
consumption), fatty acid utilization was different in each sample without
involving an alteration in the uptake of amino acids. In the second half of
fermentation (from 50% sugar consumption until the end of fermentation), the
clarified sample had greater consumption of saturated fatty acids than
unsaturated, in contrast to the control; this, along with other factors,
produced a greater liberation of free amino acids in the filtered sample.

PMID: 10554194 [PubMed - as supplied by publisher]

373: J Agric Food Chem. 2000 Dec;48(12):6178-82.

Glycosides and phenylpropanoid glycerol in vitis vinifera cv. Gewurztraminer
wine.

Baltenweck-Guyot R, Trendel JM, Albrecht P, Schaeffer A.

Laboratoire de Chimie Organique des Substances Naturelles, UMR 7509
CNRS/Universite Louis Pasteur, Institut de Chimie, 1 rue Blaise Pascal, 67000
Strasbourg, Francece.

Eight glycosides and a phenylpropanoid glycerol were isolated from Vitis
vinifera cv. Gewurztraminer wine, and their structures were elucidated by MS and
NMR spectroscopies. cis-1-(5-Ethenyl-5-methyltetrahydrofuran-2-yl)-1-methylethyl
O-beta-D-apiofuranosyl-(1-->6)-O-beta-D-glucopyranoside, (E)-3,6,
9-trihydroxymegastigm-7-ene 9-O-beta-D-glucopyranoside, 2-phenylethyl
O-beta-D-apiofuranosyl-(1-->6)-O-beta-D-glucopyranoside, and
2-[4-(3-hydroxypropyl)-2-methoxyphenoxy]propane-1,3-diol are reported for the
first time as wine components.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11141277 [PubMed - indexed for MEDLINE]

374: Annu Rev Entomol. 2001;46:387-412.

Biology and management of grape phylloxera.

Granett J, Walker MA, Kocsis L, Omer AD.

Department of Entomology; University of California Davis, CA 95616, USA.
[email protected]

Grape phylloxera, Daktulosphaira vitifoliae (Homoptera: Phylloxeridae), is a
worldwide pest of grapevines. Its life cycle has sexual and asexual portions
with forms that feed from leaf and root galls. Not all forms occur throughout
the insect's range. Root forms predominate on Vitis vinifera cultivars; leaf
forms predominate on other Vitis species characteristic of the American native
range. Other conditions influence expression of the life cycle. Hosts and
conditions similarly affect life table performance. Damage to grapevines is by
secondary soilborne pathogens attacking the feeding site and by physiological
interaction of the insect with the grapevine, though the latter has not been
well studied. Resistant rootstocks derived from native American Vitis are the
primary control tool. The insect varies genetically and relative to performance
on hosts. Use of insecticides is limited in effect, and other control methods
are not proven. More research on the biology, ecology, and management of
phylloxera is needed.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 11112174 [PubMed - indexed for MEDLINE]

375: J Agric Food Chem. 2000 Nov;48(11):5369-75.

Phenolic and volatile composition of wines made from Vitis vinifera cv. Muscat
lefko grapes from the island of Samos.

Karagiannis S, Economou A, Lanaridis P.

The Athens Wine Institute, National Agricultural Research Foundation, 1 Sofokli
Venizelou Street, Athens 141 23, Greece.

A study of the phenolic and volatile composition of wines produced from the
white cultivar Muscat lefko from the island of Samos was conducted. Dry,
fortified, naturally sweet wines and mistelles (aged and nonaged) have been
studied. The phenolic components (flavan-3-ols, hydroxycinnamates, and
flavonols) were measured by high-performance liquid chromatography after solid
phase extraction (SPE). The terpenes (free and glycosidically linked) were
determined by the use of gas chromatography-mass spectrometry (GC-MS) after SPE.
The fermentation aroma components were analyzed by GC-MS after liquid-liquid
extraction. It was found that the dry wines contained lower amounts of most of
the phenolics and higher quantities of terpenes and fermentation aroma compounds
than the sweet wines. The aged mistelle wines contained lower levels of coutaric
and caftaric acids, higher concentrations of the free acids, and markedly fewer
free and bound terpenes and fermentation aroma components compared to the other
sweet wines. The naturally sweet wine contained relatively increased amounts of
phenolics, 2,3-butanediol, and glycosidically linked terpenes.

PMID: 11087487 [PubMed - indexed for MEDLINE]

376: J Agric Food Chem. 2000 Oct;48(10):5035-40.

Toward the authentication of varietal wines by the analysis of grape (Vitis
vinifera L.) residual DNA in must and wine using microsatellite markers.

Siret R, Boursiquot JM, Merle MH, Cabanis JC, This P.

Laboratoire Interregional de la Direction Generale de la Concurrence, de la
Consommation et de la Repression des Fraudes (DGCCRF), 205 Rue de la croix
verte, parc Euromedecine, 34196 Montpellier, France. [email protected]

In an attempt to develop a technique for the identification of grape cultivars
in commercial wines, a method for the extraction of DNA from must and
experimental wines was adapted and optimal PCR conditions for the amplification
of this DNA were established. DNA was analyzed during the fermentation process
for six cultivars (Chardonnay, Clairette blanche, Grenache noir, Merlot, Muscat
blanc a petits grains, and Syrah). The extractions were performed on solid parts
in suspension as well as on the aqueous fraction. Expected profiles for these
cultivars were obtained with DNA extracted from the solid parts during all of
the fermentation process and for the wine. The analysis of DNA extracted from
aqueous fractions was less reproducible, and microsatellite amplifications were
obtained only in the case of Clairette blanche, Merlot, and Syrah wines. Results
demonstrate that the purification process is adequate for the analysis but that
the DNA concentration represents the main limiting factor. Technical
improvements of the method are discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11052774 [PubMed - indexed for MEDLINE]

377: Phytochemistry. 2000 Sep;55(1):77-85.

Changes in grape seed polyphenols during fruit ripening.

Kennedy JA, Matthews MA, Waterhouse AL.

Department of Viticulture and Enology, University of California, Davis
95616-8749, USA.

The quantity and characterization of extracted flavan-3-ol monomers and
procyanidins was determined in seeds from Vitis vinifera cv. Cabernet Sauvignon
berries, over the course of ripening and at different levels of vine water
status. The per berry extractive yield of all polyphenols decreased with
maturity, and followed second-order kinetics. The flavan-3-ol monomers decreased
most rapidly, followed by the procyanidin extension units and finally, the
terminal units. The relative proportion of procyanidin extension units did not
vary with maturity. During fruit ripening, the mean degree of polymerization of
extracted procyanidins is unchanged when analyzed intact by HPLC, but decreases
by thiolytic degradation. The proportion of extracted procyanidins resistant to
acid catalyzed thiolysis increased with maturity. Changes in vine water status
affected polyphenol amounts, indicating that cultural practices can be used to
influence composition. Oxidation of the seed polyphenols during fruit ripening,
could explain these observations.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 11021647 [PubMed - indexed for MEDLINE]

378: Plant Sci. 2000 Oct 16;159(1):87-95.

Analysis of grape ESTs: global gene expression patterns in leaf and berry.

Ablett E, Seaton G, Scott K, Shelton D, Graham MW, Baverstock P, Lee LS, Henry
R.

Centre for Plant Conservation Genetics, Southern Cross University, P.O. Box 157,
NSW 2480, Lismore, Australia

Analysis of 2479 ESTs from Vitis vinifera berry tissue and 2438 from leaf
revealed that 1% of the ESTs match to known Vitis proteins, 72% to plant
proteins, 11% to non-plant, and 16% had no match (P[N]>0.5). The levels of
redundancy were similar in the leaf and berry libraries. Only 12% of the genes
matched by the ESTs were common to both libraries indicating marked differences
in the genes expressed in the two tissues. The abundance of transcripts with
predicted cellular roles in leaf and berry were estimated by classifying the
primary BLAST matches to known proteins (score >80) into functional categories.
Thirty-six percent of the leaf transcripts were involved in photosynthesis,
compared to 3% in the berry. This is a much higher proportion of transcripts
involved with a function limited to specialized cells, than was found when
transcripts of 33 human tissues were compared using a similar approach,
suggesting plant cells may involve their cellular machinery to a greater extent
in specialized activities than animal cells. Relatively enhanced expression of
specific transcription factors, and genes involved in defense, detoxification,
stress response, proteolysis, trafficing, and signal transduction, suggests
berry tissue is actively engaged in responding to environmental stimuli.

PMID: 11011096 [PubMed - as supplied by publisher]

379: J Agric Food Chem. 2000 Aug;48(8):3387-91.

Measuring the aromatic potential of Vitis vinifera L. Cv. Sauvignon blanc grapes
by assaying S-cysteine conjugates, precursors of the volatile thiols responsible
for their varietal aroma.

Peyrot Des Gachons C, Tominaga T, Dubourdieu D.

Faculte d'OEnologie, Universite Victor Segalen Bordeaux 2, Talence Cedex,
France. [email protected]

The method presented for measuring the aromatic potential of Sauvignon blanc
must is based on an assay of the S-cysteine conjugate precursors of three
volatile thiols involved in the characteristic aroma of wines made from this
grape variety: 4-mercapto-4-methylpentan-2-one, 4-mercapto-4-methylpentan-2-ol,
and 3-mercaptohexan-1-ol. These compounds were released enzymatically from their
precursors by percolating the must through an immobilized tryptophanase column
(EC 4.1.99.1), catalyzing an alpha, beta-elimination reaction on the S-cysteine
conjugate. The volatile thiols were analyzed by GC-MS, as were the deuterated
analogues that had been released from synthesized deuterated precursors and were
added as internal standards. The quantities of volatile thiols released under
these conditions were proportional to the S-cysteine conjugate content of the
must.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10956121 [PubMed - indexed for MEDLINE]

380: Planta. 2000 Jul;211(2):200-8.

In-vitro selection of Vitis vinifera 'Chardonnay' with Elsinoe ampelina culture
filtrate is accompanied by fungal resistance and enhanced secretion of
chitinase.

Jayasankar S, Li Z, Gray DJ.

Mid-Florida Research and Education Center, Institute of Food and Agricultural
Sciences, University of Florida, Apopka 32703, USA. [email protected]

Proembryogenic masses of grapevine (Vitis vinifera L.) 'Chardonnay' (clone 02Ch)
were exposed to the culture filtrate of Elsinoe ampelina (deBary) Shear, the
causal agent of anthracnose disease. After four or five cycles of recurrent
in-vitro selection with medium containing 40% fungal culture filtrate, putative
resistant lines RC1 and RC2 respectively, were established. The selected lines
inhibited the growth of E. ampelina and Fusarium oxysporium (Schlecht.)
(isolated from watermelon) in a dual-culture assay and reduced the growth of
mycelium on a conditioned-medium test, thus suggesting the involvement of
extracellular compounds in resistance. Sodium dodecyl sulfate-polyacrylamide
(SDS-PAGE) gel electrophoresis of extracellular proteins from spent
suspension-culture medium showed enhanced secretion of new proteins by selected
lines. A 36-kDa protein was immunodetected by a chitinase antiserum. This
chitinase continued to express constitutively in differentiated somatic embryos
and also in the intercellular fluids of plants regenerated from the selected
lines. Somatic embryos from selected lines grew uninhibitedly in a medium
containing 40% fungal culture filtrate, whereas non-selected (control) somatic
embryos became necrotic and died within a few days. Plants regenerated from
selected lines exhibited resistance to infection by E. ampelina in both
greenhouse tests and detached leaf bioassays. Results suggest that embryogenic
cells can be selected for resistance following in-vitro selection, resulting in
resistant plants. Whether or not resistant cells pre-existed in the original
embryogenic culture or were induced by the selection pressure could not be
determined.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10945214 [PubMed - indexed for MEDLINE]

381: Plant Sci. 2000 Aug 8;157(1):77-88.

Molecular cloning and expression of cDNAs encoding alcohol dehydrogenases from
Vitis vinifera L. during berry development.

Tesniere C, Verries C.

Unite de Recherche de Biochimie Metabolique et Technologie, INRA, ISVV M, IPV,
2, Place Viala, cedex 1, Montpellier, France

Three full-length cDNAs (VvAdh1, VvAdh2, and VvAdh3) encoding alcohol
dehydrogenases (EC 1.1.1.1) were obtained from grape berries (Vitis vinifera L.)
by means of PCR and RACE. Pairwise comparisons at the nucleotide level showed
that the three cDNAs displayed strong homology in the coding region, but were
highly divergent in the 5' and 3' untranslated regions. VvAdh1 and VvAdh2
corresponded to the two previously characterised Adh genes from grapevine, but
VvAdh3 was unrelated to known grapevine Adh sequences. The two first cDNAs
presented a single ORF of 380 amino acids, whereas the last one has two
additional residues. Moreover, the three encoded polypeptides possessed the 22
residues strictly conserved between Adh from different kingdoms. Expression
pattern of the individual isogenes was investigated during fruit development.
Specific primers were designed, and quantitative RT-PCR experiments were
performed to increase the sensitivity of detecting isogenes with a low
expression level. Results presented here revealed different developmental
regulation of the three Adh isogenes during fruit ripening. VvAdh1 and VvAdh3
transcripts were temporarily accumulated in young, developing berry, whereas
VvAdh2 was overexpressed later in fruit development, from the onset of ripening
(veraison). Expression analysis also indicated that VvAdh2 accounted for most of
the Adh mRNAs present in berries during development. The increased ADH activity
detected in berries correlated with the expression pattern of VvAdh2
transcripts. The VvAdh2 and VvAdh3 encoded enzymes were purified from
overexpressing E. coli cells. Comparison of kinetic properties of the two ADH
enzymes showed a difference in affinity with either ethanol or acetaldehyde as
substrates. Significance of multiple Adh expressed in berries is discussed.

PMID: 10940471 [PubMed - as supplied by publisher]

382: J Exp Bot. 2000 Apr;51(345):675-83.

An immunohistochemical study of the compartmentation of metabolism during the
development of grape (Vitis vinifera L.) berries.

Famiani F, Walker RP, Tecsi L, Chen ZH, Proietti P, Leegood RC.

Dipartimento di Arboricoltura e Protezione delle Piante, Universita degli Studi
di Perugia, Italy.

The compartmentation of key processes in sugar, organic acid and amino acid
metabolism was studied during the development of the flesh and seeds of grape
(Vitis vinifera L.) berries. Antibodies specific for enzymes involved in sugar
(cell wall and vacuolar invertases, pyrophosphate: fructose 6-phosphate
phosphotransferase, aldolase, NADP-glyceraldehyde-P dehydrogenase, cytosolic
fructose 1,6-bisphosphatase), photosynthesis (Rubisco, fructose
1,6-bisphosphatase, sedoheptulose 1,7-bisphosphatase), amino acid metabolism
(cytosolic and mitochondrial aspartate aminotransferases, alanine
aminotransferase, glutamate dehydrogenase, glutamine synthetase), organic acid
metabolism (phosphoenolpyruvate carboxylase, NAD- and NADP-dependent malic
enzyme, ascorbate peroxidase), and lipid metabolism (acetyl CoA carboxylase,
isocitrate lyase) were used to determine how their abundance changed during
development. There were marked changes in the abundance of many of these enzymes
in both the flesh and seeds. The intercellular location of some enzymes was
investigated using immunohistochemistry. Several enzymes (e.g.
phosphoenolpyruvate carboxylase and those involved in amino acid metabolism)
were associated with tissues likely to function in the transport of imported
assimilates, such as the vasculature. Although other enzymes (e.g. NADP-malic
enzyme and soluble acid invertase, involved in the metabolism of sugars and
organic acids) were largely present in the parenchyma cells of the flesh, their
distribution was extremely heterogeneous. This study shows that when considering
the metabolism of complex structures such as fruit, it is essential to consider
how metabolism is compartmentalized between and within different tissues, even
when they are apparently structurally homogeneous.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10938859 [PubMed - indexed for MEDLINE]

383: J Econ Entomol. 2000 Jun;93(3):1031-4.

Response of omnivorous leafroller (Lepidoptera: Tortricidae) and onion thrips
(Thysanoptera: Thripidae) to low-temperature storage.

Yokoyama VY, Miller GT.

Horticultural Crops Research Laboratory, USDA-ARS, Fresno, CA 93727, USA.

Eggs and first-fifth instars of omnivorous leafroller, Platynota stultana
Walshingham, had a mean percentage survival to the adult stage of 60.7-95.2% for
nonexposed immatures and 14.5-54.3% for immatures exposed to 1 wk at 0-1 degree
C. Exposures of 2-5 wk resulted in 0-6.7% survival, and a 6-wk exposure resulted
in < 1% survival of all stages tested. A significant reduction in survival of
all larval stages occurred between exposures of 0 and 1 wk and between 1 wk and
2-6 wk. Survival of eggs after exposures of 0 and 1 wk was significantly
different than survival after exposures of 2-6 wk. The second instar was the
stage least susceptible to low-temperature storage. Adults that were exposed to
low temperature for 1 wk in the third through fifth instars laid a mean of
120-289 eggs per female, and the mean percentage viability of the eggs ranged
from 56.2 to 71.4%. Mean percentage survival of adults and nymphs of onion
thrips, Thrips tabaci Lindeman, was inversely related to the duration of
exposure from 1 through 3-6 wk at 0-1 and 5 degrees C and was lower at 0-1
(0.2-52.5%) than at 5 degrees C (17.6-66.6%). Exposure to 0-1 degree C for 4 wk
attained 91.2% control, which increased to 99.8% after 6 wk. Low-temperature
storage has potential to control omnivorous leafroller in table grapes, Vitis
vinifera L., and onion thrips in onions, Allium cepa L.

PMID: 10902368 [PubMed - indexed for MEDLINE]

384: Planta. 2000 May;210(6):1006-13.

Cloning and expression of UDP-glucose: flavonoid 7-O-glucosyltransferase from
hairy root cultures of Scutellaria baicalensis.

Hirotani M, Kuroda R, Suzuki H, Yoshikawa T.

School of Pharmaceutical Sciences, Kitasato University, Tokyo, Japan.
[email protected]

A cDNA encoding UDP-glucose: baicalein 7-O-glucosyltransferase (UBGT) was
isolated from a cDNA library from hairy root cultures of Scutellaria baicalensis
Georgi probed with a partial-length cDNA clone of a UDP-glucose: flavonoid
3-O-glucosyltransferase (UFGT) from grape (Vitis vinifera L.). The heterologous
probe contained a glucosyltransferase consensus amino acid sequence which was
also present in the Scutellaria cDNA clones. The complete nucleotide sequence of
the 1688-bp cDNA insert was determined and the deduced amino acid sequences are
presented. The nucleotide sequence analysis of UBGT revealed an open reading
frame encoding a polypeptide of 476 amino acids with a calculated molecular mass
of 53,094 Da. The reaction product for baicalein and UDP-glucose catalyzed by
recombinant UBGT in Escherichia coli was identified as authentic baicalein
7-O-glucoside using high-performance liquid chromatography and proton nuclear
magnetic resonance spectroscopy. The enzyme activities of recombinant UBGT
expressed in E. coli were also detected towards flavonoids such as baicalein,
wogonin, apigenin, scutellarein, 7,4'-dihydroxyflavone and kaempferol, and
phenolic compounds. The accumulation of UBGT mRNA in hairy roots was in response
to wounding or salicylic acid treatments.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10872235 [PubMed - indexed for MEDLINE]

385: J Agric Food Chem. 2000 May;48(5):1799-802.

A powerful aromatic volatile thiol, 2-furanmethanethiol, exhibiting roast coffee
aroma in wines made from several Vitis vinifera grape varieties.

Tominaga T, Blanchard L, Darriet P, Dubourdieu D.

Faculte d'OEnologie, Universite Victor Segalen Bordeaux 2, 351 Cours de la
Liberation, 33405 Talence, France. [email protected]

The chemical compound 2-furanmethanethiol (2FM), with a strong roast coffee
aroma, has been identified in sweet white wines made from the Petit manseng
grape variety, and in certain red Bordeaux wines (made from the Merlot, Cabernet
franc, and Cabernet sauvignon grape varieties). This was done by extracting
specific volatile thiols using p-hydroxymercuribenzoate. The 2FM has also been
found in toasted oak used in barrel-making. All the Petit manseng sweet white
wines and some of the red Bordeaux wines analyzed contained between a few ng/L
and several dozen ng/L of 2FM. Taking into account its very low perception
threshold (0.4 ng/L in a model hydro alcoholic environment), 2FM could therefore
contribute to the roast coffee aroma of certain wines.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10820097 [PubMed - indexed for MEDLINE]

386: J Agric Food Chem. 2000 May;48(5):1738-45.

Antioxidant activity and radioprotective effects against chromosomal damage
induced in vivo by X-rays of flavan-3-ols (Procyanidins) from grape seeds (Vitis
vinifera): comparative study versus other phenolic and organic compounds.

Castillo J, Benavente-Garcia O, Lorente J, Alcaraz M, Redondo A, Ortuno A, Del
Rio JA.

Research and Development Department of Furfural Espanol S.A., Camino Viejo de
Pliego s/n, 80320 Alcantarilla, Murcia, Spain. [email protected]

The quantitative distribution of several flavan-3-ols was determined using HPLC
in a grape (Vitis vinifera) seed extract (GSE) of four cultivars grown in the
region of Murcia. Polymer >/= C(4) units made up the largest group of
procyanidins in the GSE (90.92%, expressed as HPLC % area). The antioxidant
activity of GSE and other reference compounds was investigated by measuring
their ability to scavenge the ABTS(*)(+) radical cation (TEAC). The most
effective compounds were, in order: GSE > rutin > (+)-catechin > diosmin >/=
ascorbic acid. The radioprotective effects of GSE and other reference compounds
were determined by using the micronucleus test for anticlastogenic activity, any
reduction of the frequency of micronucleated polychromatic erythrocytes (MnPCEs)
being evaluated in the bone marrow of mouse exposed to X-rays. The most
effective compounds were, in order: GSE > rutin > dimethyl sulfoxide (DMSO) >
ascorbic acid > 6-n-propyl-2-thiouracil-6c (PTU) > diosmin. The higher
ABTS(*)(+) scavenging capacity and anticlastogenic activity of GSE can be
explained, structurally, by the high number of conjugated structures between the
catechol groups in the B-rings and the 3-OH free groups of the polymeric
polyphenolic skeleton and, in addition, by the stability of the aroxyl flavonoid
radical generated in the above processes.

Publication Types:
Comparative Study

PMID: 10820088 [PubMed - indexed for MEDLINE]

387: J Agric Food Chem. 2000 May;48(5):1637-43.

Thaumatin-like proteins and chitinases, the haze-forming proteins of wine,
accumulate during ripening of grape (Vitis vinifera) berries and drought stress
does not affect the final levels per berry at maturity.

Pocock KF, Hayasaka Y, McCarthy MG, Waters EJ.

The Australian Wine Research Institute, PO Box 197, Glen Osmond, SA 5064,
Australia.

Thaumatin-like proteins and chitinases, which are pathogenesis-related (PR)
proteins, were the major soluble protein components of grapes from five
cultivars of Vitis vinifera. This dominance of PR proteins was apparent at berry
softening (veraison) and then throughout berry development for the Muscat of
Alexandria, Sultana, and Shiraz cultivars and in the berries of the Sauvignon
Blanc and Pinot Noir cultivars examined at commercial maturity. The M(r) of the
major thaumatin-like protein from Muscat of Alexandria grapes was 21 272, and
those of the three major chitinases from this cultivar, ChitB, ChitC, and ChitD,
were 25 588, 25 410, and 25 457, respectively. The vines in the study were
irrigated and showed no obvious signs of disease. Shiraz vines that had not been
irrigated throughout the season were clearly water stressed, but had levels of
PR proteins in the berry similar to vines that had been fully irrigated. It
appears that the production of PR proteins that cause protein instability in
wines by grapes may be little influenced by environmental conditions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10820071 [PubMed - indexed for MEDLINE]

388: J Nutr. 2000 May;130(5):1091-4.

Wine modifies the effects of alcohol on immune cells of mice.

Percival SS, Sims CA.

Food Science and Human Nutrition Department, University of Florida, Gainesville,
FL 32611, USA.

Ethanol may be detrimental to immune cells due to the generation of free
radicals during detoxification. If this is true, then alcoholic beverages that
contain antioxidants, like red wine, should be protective against immune cell
damage. We investigated this by giving mice either a red muscadine wine (Vitis
rotundifolia), a cabernet sauvignon (Vitis vinifera), ethanol (all at 6%
alcohol) or water in the water bottles as the sole fluid for 8 wk. Plasma
antioxidant capacity was measured with alphaalpha-diphenyl-beta-picrylhydrazyl
and was more than doubled in the mice that consumed wine compared to control
mice that consumed water or ethanol. Cytochrome P450-2E1 levels and
glutathione-S-transferase activity were modified in such a way as to be
interpreted as protective. An immune response was elicited by an intraperitoneal
injection of lipopolysaccharide. Later (24 h), natural killer cells and
T-lymphocytes derived from the circulation were quantitated in the leukocyte
fraction by flow cytometry. Ethanol consumption, as ethanol, significantly
suppressed baseline cell numbers relative to the other groups. However, the mice
that consumed the same amount of alcohol as wine had baseline cell numbers not
different from the water-consuming controls. The lymphocyte response to
lipopolysaccharide challenge was inhibited in the mice that consumed ethanol,
but was normal in those that consumed the same amount of alcohol in the form of
wine. We conclude that there are phytochemicals acting as antioxidants and
impacting on the detoxification pathway in the wine that offset the detrimental
effects of ethanol on immunity.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10801903 [PubMed - indexed for MEDLINE]

389: J Agric Food Chem. 1999 Jun;47(6):2193-7.

Isolation of isolectins from Vitis vinifera L. Cv. Chardonnay grape berries.

Berthier L, Marchal R, Debray H, Bonnet E, Jeandet P, Maujean A.

Laboratoire d'oenologie, URVVC EA 2069, Universite de Reims, France.

A lectin fraction from Chardonnay grape juice has been isolated by affinity
chromatography on a column of p-aminophenyl beta-D-glucoside-derivatized
agarose. The lectin fractions agglutinate rabbit and human erythrocytes without
serological specificity. None of the usual monosaccharides, glycosides, or
glycoproteins inhibit the hemagglutinating activity. Erythroagglutination is
only inhibited by nitrophenyl glycosides, p-nitrophenyl beta-D-glucoside being
the strongest inhibitor. In SDS-PAGE in the presence of 2-mercaptoethanol and
gel filtration HPLC, the lectin fraction gave a single band or peak
corresponding to M(r) 13.2-11.9 kDa, thus indicating it to be a monomer. Three
bands were observed by isoelectric focusing with pI values of 4.1, 4. 4, and
4.9. The isolectins seem to be glycoproteins since they are bound on a
concanavalin A-Sepharose column.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10794608 [PubMed - indexed for MEDLINE]

390: Genome. 2000 Apr;43(2):366-76.

Cloning and characterization of Vine-1, a LTR-retrotransposon-like element in
Vitis vinifera L., and other Vitis species.

Verries C, Bes C, This P, Tesniere C.

UR Biochimie Metabolique et Technologie, INRA, IPV, Montpellier, France.

We report the organization of a grapevine chimeric gene Adhr-Vine-1, composed by
an Adhr gene, into which a retroelement, Vine-1, was inserted. Sequence analysis
revealed that Adhr is a member of the Adh multigene family, but does not
correspond to any other grapevine Adh described to date. Vine-1, albeit
defective, is the most complete LTR (long terminal repeat)-retrotransposon-like
element described in Vitis vinifera L. It is 2392 bp long, with two almost
identical LTRs (287 bp) in the same orientation, and flanked by direct repeats
of a 5 bp host DNA. This element presents other features, characteristic of
retroviruses and retrotransposons including inverted repeats, a primer binding
site, and a polypurine tract. It has a single open reading frame (ORF) of 581
amino acids, potentially encoding for a gag protein and parts of the protease
and integrase proteins. Vine-1 is most likely related to the copia-like type
family, but with no significant similarity to any previously described plant
retrotransposon or inserted element, nor to any eukaryotic element described to
date. Vine-1 element has been found in Adhr at the same location in different V.
vinifera cultivars, but not in some other analyzed Vitis species. These data
suggest that Vine-1 insertion in Adhr is specific to V. vinifera, and has
occurred after the Adh isogene separation, but prior to cultivar development.
Sequences related to Vine-1 were revealed in multiple copies in the V. vinifera
genome and, to a lesser extent, in other analyzed Vitis species. The
polymorphism observed prompts us to question the role played by transposition in
the evolution of the Vitis genus.

PMID: 10791826 [PubMed - indexed for MEDLINE]

391: Yeast. 2000 Apr;16(6):499-506.

Biotransformation of monoterpene alcohols by Saccharomyces cerevisiae,
Torulaspora delbrueckii and Kluyveromyces lactis.

King A, Richard Dickinson J.

Cardiff School of Biosciences, Cardiff University, PO Box 915, Cardiff CF10 3TL,
UK. [email protected]

Monoterpenoids are important flavour compounds produced by many plant species,
including grapes (Vitis vinifera) and hops (Humulus lupulus). Biotransformation
reactions involving monoterpenoids have been characterized in filamentous fungi,
but few examples have been observed in yeasts. As monoterpenoids are in contact
with yeasts during beer and wine production, biotransformation reactions may
occur during the fermentation of these beverages. This paper describes the
biotransformation of monoterpene alcohols, of significance in the alcoholic
beverage industries, by three yeast species. All three species analysed had the
ability to convert monoterpenoids. Saccharomyces cerevisiae and Kluyveromyces
lactis reduced geraniol into citronellol, whilst all three yeasts produced
linalool from both geraniol and nerol. Monocyclic alpha-terpineol was formed
from both linalool and nerol, by all three yeasts. alpha-Terpineol was then
converted into the diol cis-terpin hydrate. K. lactis and Torulaspora
delbrueckii also had the ability to form geraniol from nerol. Finally, the
stereospecificity of terpenoid formation was analysed. Both (+) and (-)
enantiomers of linalool and alpha-terpineol were formed in roughly equal
quantities, from either geraniol or nerol. Copyright 2000 John Wiley & Sons,
Ltd.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10790686 [PubMed - indexed for MEDLINE]

392: FEMS Microbiol Lett. 2000 May 1;186(1):91-5.

Enhancement of in vitro growth and resistance to gray mould of Vitis vinifera
co-cultured with plant growth-promoting rhizobacteria.

Barka EA, Belarbi A, Hachet C, Nowak J, Audran JC.

Universite de Reims, UFR Sciences, P.O. Box 1039, Laboratoire de Biologie et
Physiologie Vegetales, 51687, Reims, France. [email protected]

The potential of a plant growth-promoting rhizobacterium, Pseudomonas sp.
(strain PsJN), to stimulate the growth and enhancement of the resistance of
grapevine (Vitis vinifera L.) transplants to gray mould caused by Botrytis
cinerea has been investigated. In vitro inoculation of grapevine plantlets
induced a significant plant growth promotion which made them more hardy and
vigorous when compared to non-inoculated plantlets. This ability increased upon
transplanting. When grown together with B. cinerea, the causal agent of gray
mould, significant differences of aggressiveness were observed between the
inoculated and non-inoculated plants. The presence of bacteria was accompanied
by an induction of plant resistance to the pathogen. The beneficial effect from
this plant-microbe association is being postulated.

PMID: 10779718 [PubMed - indexed for MEDLINE]

393: J Agric Food Chem. 2000 Apr;48(4):1290-7.

Effects of vine or bunch shading on the glycosylated flavor precursors in grapes
of Vitis vinifera L. Cv. syrah.

Bureau SM, Baumes RL, Razungles AJ.

Institut Superieur de la Vigne et du Vin, IPV-ENSA-INRA, UFR de
Technologie-Oenologie, Laboratoire des Aromes et Substances Naturelles 2,
Montepellier, France.

Effects of the modification of vine or bunch environment on glycoconjugates were
studied in Syrah berries over two years. Vines were shaded from berry set to
maturity, with black polyethylene nets of different mesh size to obtain 30 and
50% of the direct sunlight. Bunches were naturally shaded by the leaves or
artificially with 90% shade bags. Sun-exposed berries were chosen as control
berries. A quantitative decrease in levels of glycoconjugates was observed in
shaded bunches, particularly for phenolic and C(13)-norisoprenoidic glycosides.
In the same way, vine shading caused a decrease in the contents of glycosides of
terpenols, phenols, and C(13)-norisoprenoids in berries, but the grape
environment (microclimate) affected the berry composition more than the vine
environment. A cluster thinning experiment confirmed the independence of grapes
with regard to the plant for the biosynthesis of the C(13)-norisoprenoid
glycosides.

PMID: 10775388 [PubMed - indexed for MEDLINE]

394: J Agric Food Chem. 2000 Apr;48(4):1096-100.

Vitis vinifera must varietal authentication using microsatellite DNA analysis
(SSR).

Faria MA, Magalhaes R, Ferreira MA, Meredith CP, Monteiro FF.

CEQUP/Servico de Bromatologia, Faculdade de Farmacia, Universidade do Porto, Rua
de Anibal Cunha, 4050-047 Porto, Portugal. [email protected]

A microsatellite DNA-based method for Vitis vinifera grape must authentication
is presented. Five of the most important port wine producing grape cultivars
(Tinta Roriz, Tinto Cao, Touriga Francesa, Touriga Nacional, and Tinta Barroca)
were typed at four microsatellite loci described by Bowers et al. (Genome 1996,
39, 628-633) and Thomas and Scott (Theor. Appl. Genet. 1993, 86, 985-990). The
corresponding 5 varietal musts and 26 must mixtures that result from the
combination of the five varieties were also typed at the four loci. There were
no differences between the corresponding leaf and varietal must profiles. All
must combinations showed the expected band profiles corresponding to the sum of
the varietal band profile components. Among the 26 must mixtures, 8 could be
discriminated using the four loci.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10775355 [PubMed - indexed for MEDLINE]

395: Phytochemistry. 2000 Mar;53(6):659-65.

Sugar sensing and Ca2+-calmodulin requirement in Vitis vinifera cells producing
anthocyanins.

Vitrac X, Larronde F, Krisa S, Decendit A, Deffieux G, Merillon JM.

Laboratoire de Mycologie et Biotechnologie Vegetale, Faculte de Pharmacie,
Universite de Bordeaux II, France.

We have previously reported that sucrose modulates anthocyanin biosynthesis in
cell suspension cultures of Vitis vinifera L. The main role of sugar in this
response does not seem to be that of general carbohydrate source for the supply
of energy. In the present work, a number of pharmacological agents were used to
further investigate the components of the signal transduction pathway involved
in the induction of anthocyanin biosynthesis by sugar. We found that the
phosphorylation of hexose by hexokinase, but not its transport, has to be taken
into account for the sucrose signal transduction leading to anthocyanin
accumulation. Indeed, 3-O-methylglucose, a glucose analog transported into cells
but not phosphorylated by hexokinase, has no effect on anthocyanin production.
Mannose mimics the effect of sucrose in grape cells, and mannoheptulose, a
specific inhibitor of hexokinase, reduces the accumulation of anthocyanins in
response to sucrose. The results with the two latter analogs are discussed. Ca2+
channel blockers, verapamil and LaCl3, which were used to investigate the role
of extracellular Ca2+, all inhibited the sugar response. Ca2+ depletion by
pretreatment with ethylene glycol bis
(beta-aminoethylether)-N,N,N',N'-tetraacetic acid (EGTA) also blocked the sugar
response, which was partially recovered when Ca2+ was added exogenously after
Ca2+ depletion. The use of two potent calmodulin antagonists,
N-(6-aminohexyl)-5-chloro-1-naphtalenesulphonamide (W7) and chlorpromazine,
showed that calmodulin is involved in the sugar signal transduction. A protein
kinase inhibitor, 6-dimethylaminopurine (6-DMAP), and the protein phosphatase
inhibitors, endothall and cantharidin, also inhibited the sugar response. The
results of the present study suggest the involvement of several components of
general signal transduction pathways such as Ca2+, calmodulin, and protein
kinases phosphatases in the induction of anthocyanin biosynthesis by sugar.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10746878 [PubMed - indexed for MEDLINE]

396: Adv Exp Med Biol. 1999;467:671-7.

Determination of tryptophan and tryptophan metabolites in grape must and wine.

Hoenicke K, Simat TJ, Steinhart H, Christoph N, Kohler HJ, Schwab A.

Universitat Hamburg, Institut fur Biochemie und Lebensmittelchemie, Hamburg.
[email protected]

Tryptophan (Trp) and its metabolites, especially indole-3-acetic acid (IAA), are
considered as potential precursors of 2-aminoacetophenone (AAP), an aroma
compound which causes the "untypical aging off-flavor" (UTA) in Vitis vinifera
white wines. In this study RP-HPLC with fluorescence detection was used for the
qualitative and quantitative analysis of Trp and Trp-metabolites in 39 grapes,
22 grape musts and 16 wines, to which different viticultural conditions
(ripeness, pruning, strip of leaves, soil condition) have been applied. A
sensitive and selective determination was achieved after solid phase extraction
using an anion exchange material. Only traces of Trp-metabolites could be
determined in the examined grapes and grape musts, but their amounts increased
significantly during fermentation, whereas the amount of Trp decreased.
Different viticultural measures, besides the time of grape harvest, showed no
significant influences on the amount of Trp and Trp-metabolites.

PMID: 10721117 [PubMed - indexed for MEDLINE]

397: Plant Physiol. 2000 Mar;122(3):803-12.

Differential screening indicates a dramatic change in mRNA profiles during grape
berry ripening. Cloning and characterization of cDNAs encoding putative cell
wall and stress response proteins.

Davies C, Robinson SP.

Cooperative Research Centre for Viticulture, P.O. Box 145, Glen Osmond, South
Australia 5064, Australia. [email protected]

We used differential screening to isolate ripening-associated cDNAs from a
Shiraz grape (Vitis vinifera L.) berry cDNA library. A rapid increase in the
mRNA levels of a number of cDNAs not present in unripe fruit occurred in grape
berries at the onset of ripening. The putative translation products of some of
these clones had homologs in other species that are involved in cell wall
structure. These included four proline-rich proteins, a small protein that is
similar to the non-catalytic, N-terminal domain of some pectin methylesterases,
and two other glutamate-rich proteins. The remainder of the clones encoded
putative stress response proteins. These included two thaumatin-like proteins, a
metallothionein, a transcription factor, a cytochrome P450 enzyme, and proteins
induced by water, sugar, and/or cold stress in other species. Many of the
homologs of the grape cDNAs thought to be involved in cell wall structure or
stress-related responses also accumulate in a developmental manner in other
plants. This may indicate that the grape mRNAs accumulate in response to
stresses such as the storage of high concentrations of sugars and rapid cell
expansion, or they may accumulate as part of the ripening developmental program.

PMID: 10712544 [PubMed - indexed for MEDLINE]

398: J Ethnopharmacol. 1999 Dec 15;68(1-3):71-6.

Occurrence of resveratrol and pterostilbene in age-old darakchasava, an
ayurvedic medicine from India.

Paul B, Masih I, Deopujari J, Charpentier C.

Laboratoire des Sciences de la Vigne, Institut Jules Guyot, Universite de
Bourgogne, Dijon, France. [email protected]

'Darakchasava' is a well known Indian herbal preparation of which the main
ingredient is Vitis vinifera L. This 'ayurvedic' medicine is prescribed as a
cardiotonic and also given for other disorders. HPLC analysis of this age old
formulation revealed the presence of polyphenols like resveratrol and
pterostilbene. These phenolic compounds are now known as antioxidants, cancer
chemopreventive agents, and also known to reduce mortality from coronary heart
disease by increasing high density lipoproteins like cholesterol and inhibiting
platelet aggregation (Soleas, J.S., Diamandis, E.P., Goldberg, D.M., 1997.
Resveratrol: a molecule whose time has come? and gone? Clin. Biochem. 30 (2),
91-113). The study of darakchasava becomes interesting in the light of these
findings. A brief introduction of this medicinal preparation, its formulation,
its analysis by HPLC, and some of its properties are discussed in this article.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10624864 [PubMed - indexed for MEDLINE]

399: Clin Ter. 1999 Jul-Aug;150(4):275-8.

[Leukocyanidines and collagenases: in vitro enzyme inhibition activity]

[Article in Italian]

Barracchini A, Franceschini N, Filippello M, Pantaleoni P, Di Giulio A,
Amicosante G, Pantaleoni G.

Facolta di Medicina e Chirurgia, Universita degli Studi di L'Aquila, Italia.

OBJECTIVE: The matrix metalloproteinases (MMPs) play a key role in the
extracellular matrix turnover. This protein family has been involved in some
ocular pathologies such as glaucoma, diabetic retinopathy, macular degeneration,
vitreous degeneration and corneal stroma ulceration cleaving all the matrix
components. In the present study we evaluated the action of leucocyanidin from
Vitis vinifera seeds as non toxic inhibitor of these proteinases. MATERIALS AND
METHODS: To this purpose we used a fluorimetric method to evaluate the effect of
this substance on the collagenase activity. We evaluated "in vitro" the
inhibitory potency of the tested drug on type III collagenase activity, and the
recover of the metalloprotease activity upon removal by dialysis of the
inhibitor. RESULTS: The leucocyanidines extract (minimum procyanidines value of
95.0) resulted to be a good collagenase activity inhibitor showing an inhibition
constant value, Ki, of 82 microM, evident index of affinity between the extract
and the enzyme. Furthermore, the dialysis experiments demonstrated that the
inhibitory effect persisted 24 h later, probably because the extract forms a
stable complex with the enzyme. CONCLUSIONS: These results should be related to
the pharmacokinetic profile of leucoanthocyanins, a family of natural
polyphenols belonging to the class of bioflavonoids of grape seds extract (Vitis
vinifera L.).

Publication Types:
English Abstract
In Vitro

PMID: 10605164 [PubMed - indexed for MEDLINE]

400: J Agric Food Chem. 1999 Jul;47(7):2830-6.

Effect of pesticide residues on the aromatic composition of red wines.

Oliva J, Navarro S, Barba A, Navarro G, Salinas MR.

Departamento de Quimica Agricola, Geologia y Edafologia, Facultad de Quimica,
Universidad de Murcia, Campus Universitario de Espinardo, 30100 Murcia, Spain.

The influence of pesticide residues on the aromatic composition (major and minor
volatiles) of red wines made from Vitis vinifera (Monastrell var.) was studied
by comparing the concentration of aromas in wines made from grapes subjected (or
not) to phytosanitary treatment with chlorpyrifos, fenarimol, mancozeb,
metalaxyl, penconazole, and vinclozolin, according to the agricultural practice
of the area. The analytical determination of the major volatiles was made by gas
chromatography using a flame ionization detector, while the minor volatiles were
determined by adsorption-thermal desorption gas chromatography using a mass
selective detector. There were significant differences between the ethyl
acetate, methanol, isobutanol, and diethylacetal levels of the control wine and
that containing chlorpyrifos residues, although only the ethyl acetate exceeded
the olfactory threshold. With regard to the minor volatiles, significant
differences were detected in the concentrations of some esters, aldehydes, and
acids. However, only isoamyl acetate exceeded the olfactory threshold in wines
containing residues of chlorpyrifos, fenarimol, and vinclozolin.

PMID: 10552572 [PubMed - indexed for MEDLINE]

401: J Agric Food Chem. 1999 Jul;47(7):2645-52.

Regional origin assignment of red wines from Valencia (Spain) by (2)H NMR and
(13)C IRMS stable isotope analysis of fermentative ethanol.

Gimenez-Miralles JE, Salazar DM, Solana I.

Lehrstuhl fur Biologische Chemie, Technische Universitat Munchen, Vottinger
Strasse 40, 85354 Freising-Weihenstephan, Germany.
[email protected]

The use of the stable hydrogen and carbon isotope ratios of fermentative ethanol
as suitable environmental fingerprints for the regional origin identification of
red wines from Valencia (Spain) has been explored. Monovarietal Vitis vinifera
L. cvs. Bobal, Tempranillo, and Monastrell wines have been investigated by (2)H
NMR and (13)C IRMS for the natural ranges of site-specific (2)H/(1)H ratios and
global delta(13)C values of ethanol over three vintage years. Statistically
significant interregional and interannual (2)H and (13)C abundance differences
have been noticed, which are interpreted in terms of environmental and
ecophysiological factors of isotope content variation. Multivariate discriminant
analysis is shown to provide a convenient means for integration of the
classifying information, high discriminating abilities being demonstrated for
the (2)H and (13)C fingerprints of ethanol. Reasonable differentiation results
are achieved at a microregional scale in terms of geographic provenance and even
grapevine genotypic features.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10552539 [PubMed - indexed for MEDLINE]

402: J Agric Food Chem. 1999 Jul;47(7):2519-22.

Effects of copper exposure in tissue cultured Vitis vinifera.

Romeu-Moreno A, Mas A.

Unitat d'Enologia-Centre de Referencia en Tecnologia dels Aliments (UE-CeRTA),
Departament de Bioquimica i Biotecnologia, Facultat d'Enologia de Tarragona,
Universitat Rovira i Virgili, Ramon y Cajal 70, 43005 Tarragona, Spain.

The present study determined the effects of copper treatment on some biochemical
parameters in a closed system. Sauvignon grapevines were cultured in agar and
exposed to copper levels ranging from 0.07 to 10 microg Cu/g medium.
Chlorophylls, carotenoids, lipids, sucrose, soluble sugars, starch, cellulose,
and minerals in root, leaves, and sap were determined. Copper levels over 5
microg Cu/g inhibit root and plantlet development. Copper-exposed plants have
higher levels of chlorophylls and carotenoids as well as total lipids. Soluble
sugars decrease without changes in starch or sucrose. Copper concentration
increases dramatically in roots and leaves. Iron concentrations are lower in
leaves, although they increase in roots, with respect to control plants. Sap
flow and translocation of essential elements are reduced. Reduction of K
translocation can be related to limited use of water by the plant and, thus,
reduction in growth and physiological activity.

PMID: 10552520 [PubMed - indexed for MEDLINE]

403: Planta. 1999 Nov;210(1):9-18.

Phosphoenolpyruvate carboxykinase plays a role in interactions of carbon and
nitrogen metabolism during grape seed development

Walker RP, Chen ZH, Tecsi LI, Famiani F, Lea PJ, Leegood RC.

Robert Hill Institute and Department of Animal and Plant Sciences, University of
Sheffield, Sheffield, S10 2TN, UK.

Phosphoenolpyruvate carboxykinase (PEPCK) was shown to be present in a range of
developing seeds by measurement of its activity and by immunoblotting. Its
function was investigated during grape (Vitis vinifera L.) seed development. The
maximum abundance of PEPCK coincided with the deposition of storage reserves. At
this stage of development, immunohistochemistry showed that PEPCK was very
abundant in a layer of cells located at the boundary of developing storage
tissues and in the chalaza (close to the termination of the vascular supply to
the seed) and was also present in the palisade layer of the seed coat (the inner
layer of the outer integument). Earlier in development PEPCK was also present in
the developing palisade layer and in the inner region of the nucellus which
surrounds the developing endosperm. At later stages of development, PEPCK was
located in the outer region of the endosperm. However, PEPCK was present in the
phloem of the seed at all stages of development. Feeding of asparagine to
developing grape seeds led to a strong induction of PEPCK. We suggest that, in
developing grape seeds, both the chalaza and palisade tissue may distribute
imported assimilates from the vasculature to the developing storage tissues and
that PEPCK may play a role in the metabolism of nitrogenous assimilates during
their delivery from the vasculature to the storage tissues.

PMID: 10592027 [PubMed - as supplied by publisher]

404: J Agric Food Chem. 1999 Oct;47(10):4456-61.

Changes in the phytoalexin content of various Vitis spp. in response to
ultraviolet C elicitation.

Douillet-Breuil AC, Jeandet P, Adrian M, Bessis R.

Laboratoire des Sciences de la Vigne, Institut Universitaire de la Vigne et du
Vin, Universite de Bourgogne, B.P. 27-877, 21078 Dijon Cedex, France.
[email protected]

The phytoalexin production potential of three American Vitis species and that of
three cultivars of Vitis vinifera were evaluated in response to UV-C
irradiation. Time course changes in resveratrol, piceid, epsilon-viniferin, and
pterostilbene contents were studied within 3 days after a short UV-C
irradiation. Results show that the two major stilbenes accumulated as a response
to UV-C elicitation are resveratrol and epsilon-viniferin, a resveratrol
dehydrodimer, the concentration of both compounds usually reaching quantities
>100 microgram/g of fresh weight. In contrast, piceid and pterostilbene were
constantly produced in low quantities. Owing to the results obtained, the role
of stilbene phytoalexins in the resistance of grapevines to diseases is
discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10552833 [PubMed - indexed for MEDLINE]

405: Microbios. 1999;100(395):27-40.

Indigenous yeasts associated with two Vitis vinifera grape varieties cultured in
southern Spain.

De La Torre MJ, Millan MC, Perez-Juan P, Morales J, Ortega JM.

Department of Microbiology, Faculty of Science, University of Cordoba, Spain.

The yeast microbiota present on the surface of grapes of two Vitis vinifera
varieties, Pedro Ximenez and Tempranillo de Rioja, grown in the Montilla-Moriles
region of southern Spain was identified. The changes between veraison and the
physiological ripeness time during 3 years were monitored. Overall, the yeast
microbiota isolated was of oxidative metabolism. Sporobolomyces roseus and
Cryptococcus albidus species occurred at all physiological stages, in the two
Vitis vinifera varieties, and the three seasons studied. On the other hand,
Kloeckera apiculata was never detected and Saccharomyces cerevisiae was scarcely
isolated, it was only present, testimonial, in Tempranillo de Rioja grapes
during the 1992 vintage. The widest variety of yeast species was observed in the
1992 season, and in contrast, the lowest number of species in both varieties of
Vitis was detected in the 1994 season.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10582378 [PubMed - indexed for MEDLINE]

406: J Agric Food Chem. 1999 Jan;47(1):114-20.

Assessment of the native electrophoretic analysis of total grape must proteins
for the characterization of Vitis vinifera L. cultivars.

Moreno-Arribas MV, Cabello F, Polo MC, Martin-Alvarez PJ, Pueyo E.

Instituto de Fermentaciones Industriales (CSIC), Juan de la Cierva 3, 28006
Madrid, Spain.

The findings of an ampelographic analysis of vines belonging to a Germoplasm
Bank were compared to the results of native electrophoresis of the total
proteins in their musts. Cluster analysis of the data from the morphological
description produced correct groupings, in terms of variety, for all samples.
When cluster analysis was performed on the electrophoretic data, 10 of the 11
musts studied were grouped correctly. Electrophoresis was also performed on 30
musts made from a mixture of grapes from large vineyards. In the cluster
analysis of the electrophoretic data on the proteins of the 41 musts studied,
all the musts are grouped correctly in terms of variety. Electrophoretic
analysis of proteins is a simple technique that can be used routinely, provides
complementary information to morphological analysis for varietal
characterization of vines, and in the majority of cases, makes it possible to
ascertain the grape variety from which musts originate.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10563858 [PubMed - indexed for MEDLINE]

407: Plant Physiol. 1999 Oct;121(2):675-84.

Analysis of the relative increase in photosynthetic O(2) uptake when
photosynthesis in grapevine leaves is inhibited following low night temperatures
and/or water stress

Flexas J, Badger M, Chow WS, Medrano H, Osmond CB.

Molecular Plant Physiology and Photobioenergetics Groups, Research School of
Biological Sciences, Institute of Advanced Studies, Australian National
University, Box 475, Canberra, Australian Capital Territory 2601, Australia.

We found similarities between the effects of low night temperatures (5 degrees
C-10 degrees C) and slowly imposed water stress on photosynthesis in grapevine
(Vitis vinifera L.) leaves. Exposure of plants growing outdoors to successive
chilling nights caused light- and CO(2)-saturated photosynthetic O(2) evolution
to decline to zero within 5 d. Plants recovered after four warm nights. These
photosynthetic responses were confirmed in potted plants, even when roots were
heated. The inhibitory effects of chilling were greater after a period of
illumination, probably because transpiration induced higher water deficit.
Stomatal closure only accounted for part of the inhibition of photosynthesis.
Fluorescence measurements showed no evidence of photoinhibition, but
nonphotochemical quenching increased in stressed plants. The most characteristic
response to both stresses was an increase in the ratio of electron transport to
net O(2) evolution, even at high external CO(2) concentrations. Oxygen isotope
exchange revealed that this imbalance was due to increased O(2) uptake, which
probably has two components: photorespiration and the Mehler reaction. Chilling-
and drought-induced water stress enhanced both O(2) uptake processes, and both
processes maintained relatively high rates of electron flow as CO(2) exchange
approached zero in stressed leaves. Presumably, high electron transport
associated with O(2) uptake processes also maintained a high DeltapH, thus
affording photoprotection.

PMID: 10517860 [PubMed - as supplied by publisher]

408: Plant Physiol. 1999 Sep;121(1):197-206.

The generation of active oxygen species differs in tobacco and grapevine
mesophyll protoplasts.

Papadakis AK, Roubelakis-Angelakis KA.

Department of Biology, University of Crete, P.O. Box 2280, 71 409 Heraklio,
Greece.

Our previous results have shown that oxidative stress may reduce the
regeneration potential of protoplasts, but only protoplasts that are able to
supply extracellularly H(2)O(2) can actually divide (C.I. Siminis, A.K.
Kanellis, K.A. Roubelakis-Angelakis [1993] Physiol Plant 87: 263-270; C.I.
Siminis, A.K. Kanellis, K.A. Roubelakis-Angelakis [1994] Plant Physiol 1105:
1375-1383; A. de Marco, K.A. Roubelakis-Angelakis [1996a] Plant Physiol 110:
137-145; A. de Marco, K.A. Roubelakis-Angelakis [1996b] J Plant Physiol 149:
109-114). In the present study we have attempted to break down the oxidative
burst response into the individual active oxygen species (AOS) superoxide
(O(2)(*-)) and H(2)O(2), and into individual AOS-generating systems during the
isolation of regenerating tobacco (Nicotiana tabacum L.) and non-regenerating
grape (Vitis vinifera L. ) mesophyll protoplasts. Wounding leaf tissue or
applying purified cellulase did not elicit AOS production. However, the
application of non-purified cellulase during maceration induced a burst of
O(2)(*-) and H(2)O(2) accumulation in tobacco leaf, while in grape significantly
lower levels of both AOS accumulated. AOS were also generated when protoplasts
isolated with purified cellulase were treated with non-purified cellulase. The
response was rapid: after 5 min, AOS began to accumulate in the culture medium,
with significant quantitative differences between the two species. In tobacco
protoplasts and plasma membrane vesicles, two different AOS synthase activities
were revealed, one that showed specificity to NADPH and sensitivity to
diphenyleneiodonium (DPI) and was responsible for O(2)(*-) production, and a
second NAD(P)H activity that was sensitive to KCN and NaN(3), contributing to
the production of both AOS. The first activity probably corresponds to a
mammalian-like NADPH oxidase and the second to a NAD(P)H oxidase-peroxidase. In
grape, only one AOS-generating activity was detected, which corresponded to a
NAD(P)H oxidase-peroxidase responsible for the generation of both AOS.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 10482675 [PubMed - indexed for MEDLINE]

409: Science. 1999 Sep 3;285(5433):1562-1565.

Historical Genetics: The Parentage of Chardonnay, Gamay, and Other Wine Grapes
of Northeastern France.

Bowers J, Boursiquot JM, This P, Chu K, Johansson H, Meredith C.

Department of Viticulture and Enology, University of California, Davis, CA
95616, USA. Unite de Formation et de Recherche de Viticulture, Ecole Nationale
Superieure Agronomique, 34060 Montpellier, France. Unite de Recherches de
Genetique et d'Amelioration des Plantes-Viticulture, Institut National de la
Recherche Agronomique, 34060 Montpellier, France.

The origins of the classic European wine grapes (Vitis vinifera) have been the
subject of much speculation. In a search for parental relationships,
microsatellite loci were analyzed in more than 300 grape cultivars. Sixteen wine
grapes that have long been grown in northeastern France, including 'Chardonnay',
'Gamay noir', 'Aligote', and 'Melon', have microsatellite genotypes consistent
with their being the progeny of a single pair of parents, 'Pinot' and 'Gouais
blanc', both of which were widespread in this region in the Middle Ages.
Parentage analysis at 32 microsatellite loci provides statistical support for
these relationships.

PMID: 10477519 [PubMed - as supplied by publisher]

410: Plant J. 1999 Jul;19(2):229-236.

Technical Advance: Confocal measurement of the three-dimensional size and shape
of plant parenchyma cells in a developing fruit tissue.

Gray JD, Kolesik P, Hoj PB, Coombe BG.

Parenchyma cells from the inner mesocarp of a grape berry (Vitis vinifera L. cv.
Chardonnay) were visualised in three-dimensions within a whole mount of cleared,
stained tissue using confocal laser scanning microscopy and digital image
reconstruction. The whole berry was fixed, bisected longitudinally, cleared in
methyl salicylate, stained with safranin O and mounted in methyl salicylate.
Optical slices were collected at 1.0 microm intervals to a depth of 150 microm.
Neighbouring z-series were joined post-collection to double the field-of-view.
Attenuation at depth of the fluorescent signal from cell walls was quantified
and corrected. Axial distortion due to refractive index mismatch between the
immersion and mounting media was calibrated using yellow-green fluorescent
microspheres and corrected. Transmission electron microscopy was used to correct
fluorescent measurements of cell wall thickness. Digital image reconstructions
of wall-enclosed spaces enabled cells to be rendered as geometric solids of
measurable surface area and volume. Cell volumes within the inner mesocarp
tissue of a single grape berry exhibited a 14-fold range, with polysigmoidal
distribution and groupings around specific size classes. Cell shape was
irregular and the planes of contact were rarely flat or simple. Variability in
cell shape was indicated by the range in surface area to volume ratios, from
0.080 to 0.198 microm-1. Structural detail at the internal surface of the cell
wall was apparent. The technique is applicable to a wide range of morphometric
analyses in plant cell biology, particularly developmental studies, and reveals
details of cell size and shape that were previously unattainable.

PMID: 10476070 [PubMed - as supplied by publisher]

411: Plant Physiol. 1999 Aug;120(4):1083-94.

Cloning and expression of a hexose transporter gene expressed during the
ripening of grape berry.

Fillion L, Ageorges A, Picaud S, Coutos-Thevenot P, Lemoine R, Romieu C, Delrot
S.

Laboratoire de Physiologie et Biochimie Vegetales, Equipe Scientifique Associee
Centre National de la Recherche Scientifique 6161, Unite de Formation et de
Recherches Sciences, 40 Avenue du Recteur Pineau, Universite de Poitiers,
France.

The ripening of grape (Vitis vinifera L.) is characterized by massive sugar
import into the berries. The events triggering this process and the pathways of
assimilate transport are still poorly known. A genomic clone Vvht1 (Vitis
vinifera hexose transporter1) and the corresponding cDNA encoding a hexose
transporter whose expression is induced during berry ripening have been
isolated. Vvht1 is expressed mainly in the berries, with a first peak of
expression at anthesis, and a second peak about 5 weeks after veraison (a
viniculture term for the inception of ripening). Vvht is strictly conserved
between two grape cultivars (Pinot Noir and Ugni-Blanc). The organization of the
Vvht1 genomic sequence is homologous to that of the Arabidopsis hexose
transporter, but differs strongly from that of the Chlorella kessleri hexose
transporter genes. The Vvht1 promoter sequence contains several potential
regulating cis elements, including ethylene-, abscisic acid-, and
sugar-responsive boxes. Comparison of the Vvht1 promoter with the promoter of
grape alcohol dehydrogenase, which is expressed at the same time during
ripening, also allowed the identification of a 15-bp consensus sequence, which
suggests a possible co-regulation of the expression of these genes. The
expression of Vvht1 during ripening indicates that sucrose is at least partially
cleaved before uptake into the flesh cells.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 10444092 [PubMed - indexed for MEDLINE]

412: Ann Allergy Asthma Immunol. 1999 Jul;83(1):25-6.

Comment in:
Ann Allergy Asthma Immunol. 2000 Feb;84(2):265.
Ann Allergy Asthma Immunol. 2000 Feb;84(2):265-6.

Adult onset grape hypersensitivity causing life threatening anaphylaxis.

Vaswani SK, Chang BW, Carey RN, Hamilton RG.

Mid-Atlantic Permanente Medical Group & Johns Hopkins University School of
Medicine, Baltimore, Maryland 21244, USA.

BACKGROUND: Adverse reactions to foods are encountered much less frequently in
adults than in the children. Adult onset hypersensitivity to grapes has not been
previously reported. OBJECTIVE: Evaluation of a case of anaphylaxis that
occurred as a result of the consumption of white grapes (Vitis vinifera).
METHODS AND RESULTS: A 28-year-old woman experienced generalized urticaria,
facial/oropharyngeal angioedema, and dizziness after eating a bunch of white
grapes. She was treated in an emergency room for anaphylaxis. Previously, she
had experienced two similar episodes after eating white grapes. The grape prick
skin tests were strongly positive forming a pseudopod type reaction. The total
serum IgE was 1918 ng/mL. The grape-specific serum IgE was weakly positive by
the modified RAST and negative in the Pharmacia-Upjohn Cap System. CONCLUSION:
Hypersensitivity to a commonly consumed fruit such as grapes can develop late in
life causing a near-fatal anaphylaxis.

Publication Types:
Case Reports

PMID: 10437812 [PubMed - indexed for MEDLINE]

413: Planta. 1999 Jun;208(4):574-82.

Cloning and expression of an arginine decarboxylase cDNA from Vitis vinifera L.
cell-suspension cultures.

Primikirios NI, Roubelakis-Angelakis KA.

Department of Biology, University of Crete, Heraklion, Greece.

Arginine decarboxylase (ADC; EC 4.1.1.19) is a key enzyme in one of the two
pathways to putrescine. We present the first ADC cDNA from a woody perennial
plant species, the grapevine (Vitis vinifera L.), which exhibits 70-80% homology
with other dicot ADCs. The effects of ammonium, nitrate, and putrescine on ADC
specific activity, soluble polyamine levels, ADC-mRNA, endogeneous arginine and
ornithine, and arginase specific activity were investigated in suspension
cultures of grapevine cells. The addition of NH4+ to cells cultured in
NH4(+)-free medium, resulted in a 4-fold increase in ADC activity and
concomitantly in a 4-fold increase in putrescine and a 3-fold decrease in
arginine. During this period ornithine increased and arginase activity followed
a reverse pattern of changes compared with ADC. In contrast, the addition of
NO3- did not markedly affect ADC activity, putrescine, arginine and ornithine,
but transiently increased arginase activity. The addition of putrescine caused a
4-fold decrease in ADC activity and increased arginine, ornithine and arginase
activity. The changes in ADC specific activity were not accompanied by analogous
changes in the ADC transcript levels. These results further support the view
that ADC regulation is not exhibited, at least for the factors considered in
this work, at the transcriptional level.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10420650 [PubMed - indexed for MEDLINE]

414: Plant Physiol. 1999 Jul;120(3):923.

Proline accumulation in developing grapevine fruit occurs independently of
changes in the levels of delta1-pyrroline-5-carboxylate synthetase mRNA or
protein.

Stines AP, Naylor DJ, Hoj PB, van Heeswijck R.

Department of Horticulture, Viticulture, and Oenology, University of Adelaide,
Waite Campus, PMB 1, Glen Osmond, SA 5064, Australia.

Mature fruit of grapevine (Vitis vinifera) contains unusually high levels of
free proline (Pro; up to 24 micromol or 2.8 mg/g fresh weight). Pro accumulation
does not occur uniformly throughout berry development but only during the last 4
to 6 weeks of ripening when both berry growth and net protein accumulation have
ceased. In contrast, the steady-state levels of both the mRNA encoding V.
vinifera Delta1-pyrroline-5-carboxylate synthetase (VVP5CS), a key regulatory
enzyme in Pro biosynthesis, and its protein product remain relatively uniform
throughout fruit development. In addition, the steady-state protein levels of
Pro dehydrogenase, the first enzyme in Pro degradation, increased throughout
early fruit development but thereafter remained relatively constant. The
developmental accumulation of free Pro late in grape berry ripening is thus
clearly distinct from the osmotic stress-induced accumulation of Pro in plants.
It is not associated with either sustained increases in steady-state levels of
P5CS mRNA or protein or a decrease in steady-state levels of Pro dehydrogenase
protein, suggesting that other physiological factors are important for its
regulation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10398729 [PubMed - indexed for MEDLINE]

415: Phytochemistry. 1999 Jul;51(5):651-6.

Production of 13C-labelled anthocyanins by Vitis vinifera cell suspension
cultures.

Krisa S, Teguo PW, Decendit A, Deffieux G, Vercauteren J, Merillon JM.

Groupe d'Etude des Substances Naturelles a Interet Therapeutique, Faculte de
Pharmacie, Universite de Bordeaux II, France.

The use of plant cell cultures for producing isotopically (13C) labelled
phenolic substances is reported. Vitis vinifera cells synthesize high levels of
anthocyanins when they are cultured in a polyphenol synthesis-inducing medium.
Three major anthocyanin monoglucosides found in red wine were identified in
grape cells: cyanidin-3-O-beta-glucoside, peonidin-3-O-beta-glucoside, and
malvidin-3-O-beta-glucoside. Kinetic study of the intracellular level of
phenylalanine and its metabolites showed that it is preferable to add this
precursor to grape cell suspensions after the 5th day of culture, i.e. at the
beginning of the exponential growth phase. After adding phenylalanine to the
culture medium, its uptake was complete and the accumulation of anthocyanins in
grape cells was stimulated. Incorporation of [1-13C]-phenylalanine into
anthocyanins was measured by means of 13C satellites in the proton NMR spectrum.
The maximal rate of 13C enrichment anthocyanins obtained with this technique
reached 65%. The production of 13C labelled phenolic compounds was undertaken in
order to investigate their absorption and metabolism in humans.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 10392469 [PubMed - indexed for MEDLINE]

416: Drugs Exp Clin Res. 1999;25(2-3):57-63.

Stilbene compounds: from the grapevine to wine.

Bavaresco L, Fregoni C, Cantu E, Trevisan M.

Institute of Viticulture, Universita Cattolica S. Cuore, Piacenza, Italy.

Stilbenes are natural compounds occurring in a number of plant families,
including Vitaceae and (within this family) Vitis vinifera L., which is the most
important species grown worldwide for grape and wine production. Stilbenes
(resveratrol and viniferins) are present in grapevine as constitutive compounds
of the woody organs (roots, canes, stems) and as induced substances (in leaves
and fruit) acting as phytoalexins in the mechanisms of grape resistance against
certain pathogens. Resveratrol (3, 5, 4'-trihydroxystilbene) was also detected
in wine and it was thought to be the active principle of red wines that were
shown to reduce heart diseases. This paper reviews data, obtained by the
Viticulture Institute of the Catholic University at Piacenza and taken from the
literature, on some aspects of stilbene physiology in grapevine and on their
relation to resveratrol wine levels. Constitutive stilbene contents of woody
organs are reported, as well as the possible role of cluster stems as a source
of resveratrol for wine. The accumulation of stilbenes in grape berries infected
by grey mould (Botrytis cinerea Pers.) has been investigated and the effects of
environmental factors on resveratrol grape and wine levels will be discussed. An
unidentified new hydroxystilbene was detected in wine.

Publication Types:
Research Support, Non-U.S. Gov't
Review

PMID: 10370866 [PubMed - indexed for MEDLINE]

417: Minerva Cardioangiol. 1999 Jan-Feb;47(1-2):39-46.

[Clinical and capillaroscopic evaluation of chronic uncomplicated venous
insufficiency with procyanidins extracted from vitis vinifera]

[Article in Italian]

Costantini A, De Bernardi T, Gotti A.

Istituto di Chirurgia Vascolare e Angiologia, Universita degli Studi, Milano.

BACKGROUND: The pharmacological treatment of non-complicated chronic venous
insufficiency is a current and well-debated topic. The introduction of new
products with action on the venous system, improved knowledge on the
physiopathology of venous insufficiency and the possibility provided by new
analytical instruments, have given new impulse to the consolidation of the
clinical value of phlebotonics in this indication. METHODS: In light of this, 24
patients with non-complicated chronic venous insufficiency were treated with
oral administration of Oligomeric Proanthocyanidins (Pycnogenols-OPC) 100
mg/day. To evaluate the therapeutic efficacy of the treatment, an instrumental
evaluation by optical probe capillaroscope was employed in addition to the
traditional subjective clinical parameters: swelling, itching, heaviness and
pain. The videocapillaroscope examination was performed at the lower third of
the leg and the first toe. Edema in the capillaroscopic field, the number of
observable capillaries and the capillary dilatation were the parameter chosen to
evaluate the efficacy of treatment. All patients completed the study with no
reports of adverse events during the period of observation. RESULTS: The results
obtained show a positive clinical response (improved or absent symptoms) in over
80% of patients, with significant improvement of symptoms already evident after
the first 10 days of treatment. The mechanism of action of the OPCs explains the
rapid reduction of the swelling of the lower limbs and correlated with this are
the other evaluable symptoms: heaviness and itching. Particularly striking
results were observed for itching and pain which completely disappeared during
the course of therapy in 80% and 53% of the patients respectively. Noteworthy is
the good correlation between the clinical and instrumental data, with
improvement in a total of 70% of patients. CONCLUSIONS: The results obtained in
the course of this clinical experience, with evident improvement already during
the first weeks of treatment, the absence of adverse events added to the benefit
of a once-a-day administration, justify the use of OPC in the treatment of
non-complicated chronic venous insufficiency.

Publication Types:
English Abstract

PMID: 10356940 [PubMed - indexed for MEDLINE]

418: Life Sci. 1999;64(8):627-42.

Diet enriched with procyanidins enhances antioxidant activity and reduces
myocardial post-ischaemic damage in rats.

Facino RM, Carini M, Aldini G, Berti F, Rossoni G, Bombardelli E, Morazzoni P.

Istituto Chimico Farmaceutico Tossicologico, University of Milan, Italy.
[email protected]

Aim of this work was to study the efficacy of procyanidins from Vitis vinifera
seeds, a standardized mixture of polyphenol antioxidants, on cardiac mechanics
following ischemia/reperfusion stunning in the rat, after 3 weeks
supplementation. Young and aged male rats were fed a diet enriched with
procyanidins complexed (1:3 w/w) with soybean lecithin (2.4%); control animals
(CTR-young and CTR-aged) received an equal amount of lecithin and 2 additional
groups of animals the standard diet. At the end of the treatment, the total
plasma antioxidant defense (TRAP), vitamin E, ascorbic acid and uric acid were
determined in plasma and the hearts from all groups of animals subjected to
moderate ischemia (flow reduction to 1 ml/min for 20 min) and reperfusion (15
ml/min for 30 min). In both young and aged rats supplemented with procyanidins
the recovery of left ventricular developed pressure (LVDP) at the end of
reperfusion was 93% (p < 0.01) and 74% (p < 0.01) of the preischemic values and
the values of coronary perfusion pressure (CPP) were maintained close to those
of the preischemic period. Also creatine kinase (CK) outflow was restrained to
baseline levels, while a 2-fold increase in prostacyclin (6-keto-PGF1alpha) in
the perfusate from hearts of young and aged rats was elicited during both
ischemia and reperfusion. In parallel, procyanidins significantly increased the
total antioxidant plasma capacity (by 40% in young and by 30% in aged rats) and
the plasma levels of ascorbic acid, while tend to reduce vitamin E levels; no
significant differences were observed in uric acid levels. The results of this
study demonstrate that procyanidins supplementation in the rat (young and aged)
makes the heart less susceptible to ischemia/reperfusion damage and that this is
positively associated to an increase in plasma antioxidant activity.

PMID: 10069526 [PubMed - indexed for MEDLINE]

419: Plant Physiol. 1999 Feb;119(2):621-6.

Purification and characterization of a NADPH-dependent aldehyde reductase from
mung bean that detoxifies eutypine, a toxin from eutypa lata1

Colrat S, Latche A, Guis M, Pech JC, Bouzayen M, Fallot J, Roustan JP.

Ecole Nationale Superieure Agronomique Unite, Associee, Institut National de la
Recherche Agronomique, BP 107, 31326 Castanet-Tolosan cedex, France.

Eutypine (4-hydroxy-3-[3-methyl-3-butene-1-ynyl] benzaldehyde) is a toxin
produced by Eutypa lata, the causal agent of eutypa dieback in the grapevine
(Vitis vinifera). Eutypine is enzymatically converted by numerous plant tissues
into eutypinol (4-hydroxy-3-[3-methyl-3-butene-1-ynyl] benzyl alcohol), a
metabolite that is nontoxic to grapevine. We report a four-step procedure for
the purification to apparent electrophoretic homogeneity of a eutypine-reducing
enzyme (ERE) from etiolated mung bean (Vigna radiata) hypocotyls. The purified
protein is a monomer of 36 kD, uses NADPH as a cofactor, and exhibits a Km value
of 6.3 &mgr;M for eutypine and a high affinity for 3- and 4-nitro-benzaldehyde.
The enzyme failed to catalyze the reverse reaction using eutypinol as a
substrate. ERE detoxifies eutypine efficiently over a pH range from 6.2 to 7.5.
These data strongly suggest that ERE is an aldehyde reductase that could
probably be classified into the aldo-keto reductase superfamily. We discuss the
possible role of this enzyme in eutypine detoxification.

PMID: 9952458 [PubMed - as supplied by publisher]

420: J Allergy Clin Immunol. 1999 Feb;103(2 Pt 1):262-6.

Rhinoconjunctivitis and asthma caused by vine pollen: a case report.

Feo Brito F, Martinez A, Palacios R, Mur P, Gomez E, Galindo PA, Borja J,
Martinez J.

Section of Allergy, Hospital Complex, Ronda del Carmen, Ciudad Real, Spain.

BACKGROUND: The vine (Vitis vinifera) is a cultivated plant that is found in
some European and American countries. Its pollen gathers in small quantities
during a short pollination period in the months of May and June. Allergy to vine
pollen has not been previously documented. OBJECTIVE: We sought to describe a
case report of allergy to vine pollen documented on the basis of anamnesis,
cutaneous, provocation, and specific IgE determination tests. METHODS: An
allergenic extract was obtained from collected V vinifera pollens by aqueous
standard procedures. Pollen counts and pollination periods of this and other
common pollens in the area where the patient became symptomatic were studied.
Cutaneous tests and the presence of specific IgE to the pollen extracts were
performed by prick, CAP, and RAST techniques. Bronchial and conjunctival tests
with the involved pollen extracts were also carried out to identify the
sensitizing allergens. Five healthy subjects and 5 pollinic patients were used
as control subjects and underwent the same tests. RESULTS: Skin prick test
responses with vine pollen at different concentrations were positive for the
studied patient and negative for the control subjects. Patient serum revealed a
total IgE titer of 334 IU/mL and a specific IgE value of 1.3 PRU/mL (RAST class
2) to vine pollen. Bronchial and conjunctival provocation test responses were
also positive when the patient was challenged with V vinifera extract.
CONCLUSION: Exposure to the pollen of the vineyard plants (V vinifera) can
induce immunologic sensitization and rhinoconjunctivitis/asthma.

Publication Types:
Case Reports

PMID: 9949317 [PubMed - indexed for MEDLINE]

421: Plant J. 1998 Nov;16(3):335-43.

A novel NADPH-dependent aldehyde reductase gene from Vigna radiata confers
resistance to the grapevine fungal toxin eutypine.

Guillen P, Guis M, Martinez-Reina G, Colrat S, Dalmayrac S, Deswarte C, Bouzayen
M, Roustan JP, Fallot J, Pech JC, Latche A.

Ecole Nationale Superieure Agronomique de Toulouse, Tolosan, France.

Eutypine, 4-hydroxy-3-(3-methyl-3-butene-1-ynyl) benzyl aldehyde, is a toxin
produced by Eutypa lata, the causal agent of eutypa dieback of grapevines. It
has previously been demonstrated that tolerance of some cultivars to this
disease was correlated with their capacity to convert eutypine to the
corresponding alcohol, eutypinol, which lacks phytotoxicity. We have thus
purified to homogeneity a protein from Vigna radiata that exhibited
eutypine-reducing activity and have isolated the corresponding cDNA. This
encodes an NADPH-dependent reductase of 36 kDa that we have named Vigna radiata
eutypine-reducing enzyme (VR-ERE), based on the capacity of a recombinant form
of the protein to reduce eutypine into eutypinol. The strongest homologies
(86.8%) of VR-ERE at the amino acid level were found with CPRD14, a
drought-inducible gene of unknown function, isolated from Vigna unguiculata and
with an aromatic alcohol dehydrogenase (71.7%) from Eucalyptus gunnii.
Biochemical characterization of VR-ERE revealed that a variety of compounds
containing an aldehyde group can act as substrates. However, the highest
affinity was observed with 3-substituted benzaldehydes. Expression of a VR-ERE
transgene in Vitis vinifera cells cultured in vitro conferred resistance to the
toxin. This discovery opens up new biotechnological approaches for the
generation of grapevines resistant to eutypa dieback.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 9881154 [PubMed - indexed for MEDLINE]

422: Plant Physiol. 1998 Nov;118(3):783-92.

Changes in cell wall composition during ripening of grape berries

Nunan KJ, Sims IM, Bacic A, Robinson SP, Fincher GB.

Department of Plant Science, University of Adelaide, Waite Campus, Glen Osmond,
SA 5064, Australia (K.J.N., G.B.F.).

Cell walls were isolated from the mesocarp of grape (Vitis vinifera L.) berries
at developmental stages from before veraison through to the final ripe berry.
Fluorescence and light microscopy of intact berries revealed no measurable
change in cell wall thickness as the mesocarp cells expanded in the ripening
fruit. Isolated walls were analyzed for their protein contents and amino acid
compositions, and for changes in the composition and solubility of constituent
polysaccharides during development. Increases in protein content after veraison
were accompanied by an approximate 3-fold increase in hydroxyproline content.
The type I arabinogalactan content of the pectic polysaccharides decreased from
approximately 20 mol % of total wall polysaccharides to about 4 mol % of wall
polysaccharides during berry development. Galacturonan content increased from 26
to 41 mol % of wall polysaccharides, and the galacturonan appeared to become
more soluble as ripening progressed. After an initial decrease in the degree of
esterification of pectic polysaccharides, no further changes were observed nor
were there large variations in cellulose (30-35 mol % of wall polysaccharides)
or xyloglucan (approximately 10 mol % of wall polysaccharides) contents.
Overall, the results indicate that no major changes in cell wall polysaccharide
composition occurred during softening of ripening grape berries, but that
significant modification of specific polysaccharide components were observed,
together with large changes in protein composition.

PMID: 9808722 [PubMed - as supplied by publisher]

423: Cytobios. 1998;93(373):123-8.

Antidotal effect of grape juice (Vitis vinifera) on ochratoxin A caused
hepatorenal carcinogenesis in mice (Mus musculus).

Jeswal P.

Department of Botany, Bhagalpur University, India.

Oral administration of ochratoxin A to young weanling mice (Mus musculus) caused
several haematological changes and induced hepatoma and renal carcinoma.
Concurrent administration of berry and leaf juice of the common grape (Vitis
vinifera) to mice together with ochratoxin A significantly reduced the hepatic
and renal damage caused by ingestion of this mycotoxin. None of the animals
receiving berry/leaf juice of V. vinifera showed the formation of hepatorenal
carcinoma whereas 25% of animals receiving only ochratoxin A developed well
differentiated renal carcinoma and hepatic lesions.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 9734345 [PubMed - indexed for MEDLINE]

424: Virology. 1998 Sep 1;248(2):432-44.

Population diversity in grapevine yellow speckle viroid-1 and the relationship
to disease expression.

Szychowski JA, Credi R, Reanwarakorn K, Semancik JS.

Department of Plant Pathology, University of California Riverside, Riverside,
California, 92521, USA.

Vitis vinifera cultivars Zinfandel-1A and Mission were found to harbor different
grapevine yellow speckle viroid-1 (GYSVd-1) variants and characterized to define
the relationship to yellow speckle (YS) and vein-banding (VB) diseases. Products
from the left terminal (T1), pathogenic (P), and a portion of the central (C)
domains of Zinfandel-1A and Mission displayed distinct single-stranded
conformation polymorphism (SSCP) patterns, presumably reflecting nucleotide
changes in the P domain. The two selections were shown to contain homogeneous
populations of type 1 and type 2 GYSVd-1 variants described in Australia.
Symptoms of YS were induced only in vines containing the type 2 variant by
treatment at a constant temperature of 32 degreesC in continuous light. SSCP of
Pagadebit selections from Italy revealed the nonsymptomatic variant was
essentially identical to Zinfandel-1A, whereas symptomatic selections were
unlike any other previously described. Nucleotide sequence confirmed that
nonsymptomatic selections from Italy contained the GYSVd-1 type 1 variant. A
total of 43 changes were spread throughout the T1, C, V, and T2 domains from
symptomatic selections. This study establishes the Australian type 1 variant as
the non-symptom-inducing form of GYSVd-1 and type 2 as the symptom-inducing
variant. The distinct symptom-inducing variant from Italy is proposed as a new
type 3 variant of GYSVd-1. Copyright 1998 Academic Press.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 9721250 [PubMed - indexed for MEDLINE]

425: J Nat Prod. 1998 Jul;61(7):881-6.

Structure of a hemicellulose B fraction in dietary fiber from the seed of grape
variety Palomino (Vitis vinifera cv. palomino).

Igartuburu JM, Pando E, Rodriguez-Luis F, Gil-Serrano A.

Departamento de Quimica Organica, Facultad de Ciencias, Universidad de Cadiz,
Apdo 40. 11510, Puerto Real, Cadiz, Spain. [email protected]

The structure of one of the hemicellulose B fractions (HB-1) extracted from the
seeds of the grape variety Palomino (Vitis vinifera cv. Palomino) has been
studied by means of methylation analysis, 1H NMR, 13C NMR, and partial acid
hydrolysis. This hemicellulose seems to be a homogeneous polysaccharide with an
apparent molecular weight of 35 000. Its structure is that of an acidic
arabinoxylan, a linear chain of beta-D-xylopyranosyl units, bonded together by
(1-->4) glycosidic links, containing a single L-arabinofuranosyl,
alpha-D-xylopyranosyl and 4-O-methyl-alpha-D-glucopyranuronosyl residues joined
by glycosidic links to position 2 of the xylose units of the main chain, in
proportions of one branch to every seven units of xylose.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 9677268 [PubMed - indexed for MEDLINE]

426: J Nat Prod. 1998 Jul;61(7):876-80.

Structure of a hemicellulose A fraction in dietary fiber from the seed of grape
variety Palomino (Vitis vinifera cv. palomino).

Igartuburu JM, Pando E, Rodriguez-Luis F, Gil-Serrano A.

Departamento de Quimica Organica, Facultad de Ciencias, Universidad de Cadiz,
Apdo 40. 11510, Puerto Real, Cadiz, Spain. [email protected]

The structure of one of the hemicellulose A fractions (HA-1) extracted from the
seeds of the grape variety Palomino (Vitis vinifera cv. Palomino) has been
studied by means of methylation, 1H NMR, 13C NMR, and partial hydrolysis. This
hemicellulose seems to be a homogeneous polysaccharide with an apparent
molecular weight of 68 500. Its structure is that of an acidic xylan, a linear
chain of beta-D-xylopyranosyl units, bonded together by (1-->4) glycosidic
links, containing single alpha-D-xylopyranosyl and
4-O-methyl-alpha-D-glucopyranuronosyl residues joined by glycosidic links to
position 2 of the xylose units of the main chain, in proportions of one residue
to every 25 units of xylose.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 9677267 [PubMed - indexed for MEDLINE]

427: Planta Med. 1998 May;64(4):343-7.

Sparing effect of procyanidins from Vitis vinifera on vitamin E: in vitro
studies.

Maffei Facino R, Carini M, Aldini G, Calloni MT, Bombardelli E, Morazzoni P.

Istituto Chimico Farmaceutico Tossicologico, Milan, Italy.
[email protected]

The sparing/recycling effect of a highly purified, high molecular weight
fraction of catechin oligomers (procyanidins) from Vitis vinifera seeds on
alpha-tocopherol was studied in both homogeneous solution and in heterogeneous
phase (phosphatidylcholine liposomes and red blood cells). By HPLC and electron
spin resonance (ESR) spectroscopy we evidenced that tocopheroxyl radical,
induced by reaction of alpha-tocopherol with the stable radical DPPH
(2,2-diphenyl-1-picrylhydrazyl) is recycled by procyanidins. In addition
procyanidins significantly and dose-dependently spare vitamin E from consumption
(HPLC monitoring) during the autooxidation phase of the HO-induced peroxidation
of phosphatidylcholine, by 23% at the lowest concentration (0.5 microM) and by
65.5% at 3 microM. In this membrane model the combination of 0.5 microM
procyanidins and 2 microM alpha-tocopherol results in a marked delay in the
appearance of conjugated dienes in respect to the single antioxidants
(synergistic interaction), while catechin showed to be active only at 5 microM.
In red blood cells oxidatively stressed by UVB exposure, procyanidins at 0.1-1.0
microM prevent vitamin E loss, markedly decrease membrane lipid peroxidation,
linearly related to the concentration of vitamin E in the membranes, and
significantly delay the onset of hemolysis (catechin protects between 5 and 10
microM).

PMID: 9619118 [PubMed - indexed for MEDLINE]

428: J Nat Prod. 1998 May;61(5):655-7.

Isolation, identification, and antioxidant activity of three stilbene glucosides
newly extracted from vitis vinifera cell cultures

Waffo Teguo P, Fauconneau B, Deffieux G, Huguet F, Vercauteren J, Merillon JM.

Groupe d'Etude des Substances Naturelles a Interet Therapeutique, EA 491,
Faculte des Sciences Pharmaceutiques, Universite de Bordeaux 2, 3 place de la
Victoire, 33000 Bordeaux, France, and Centre d'Etudes et de Recherche sur les
Xenob.

Suspension cultures of Vitis vinifera L. (Vitaceae) produce many hydroxylated
stilbene glucosides found in red wine. From these cells, we isolated and
characterized glycosylated stilbenes, (Z)-piceatannol
(3,5,3',4'-tetrahydroxystilbene) -3-O-beta-d-glucopyranoside (6) and (E)- and
(Z)-resveratrol (3,5, 4'-trihydroxystilbene)-4'-O-beta-d-glucopyranoside (2 and
7, respectively), which have not previously reported to be constituents of Vitis
vinifera or wine. The ability of these compounds to act as radical scavengers
was investigated using 1,1 diphenyl-2-picryl-hydrazyl, a stable free radical.
Antioxidant activities were assessed by their capacity to prevent Cu2+-induced
lipid peroxidation in human low-density lipoprotein.

PMID: 9599270 [PubMed - as supplied by publisher]

429: J Biol Chem. 1998 Apr 10;273(15):9224-33.

Cloning and characterization of Vitis vinifera UDP-glucose:flavonoid
3-O-glucosyltransferase, a homologue of the enzyme encoded by the maize Bronze-1
locus that may primarily serve to glucosylate anthocyanidins in vivo.

Ford CM, Boss PK, Hoj PB.

Department of Horticulture, Viticulture and Oenology, the University of
Adelaide, Glen Osmond SA 5064, South Australia.

We report here the cloning and optimized expression at 16 degrees C and the
characterization of a Vitis vinifera UDP-glucose:flavonoid
3-O-glucosyltransferase, an enzyme responsible for a late step in grapevine
anthocyanin biosynthesis. The properties of this and other UDP-glucose:flavonoid
3-O-glucosyltransferases, homologues of the product encoded by the maize
Bronze-1 locus, are a matter of conjecture. The availability of a purified
recombinant enzyme allowed for the unambiguous determination of the
characteristics of a flavonoid 3-O-glucosyltransferase. Kinetic analyses showed
that kcat for glucosylation of cyanidin, an anthocyanidin substrate, is 48 times
higher than for glucosylation of the flavonol quercetin, whereas Km values are
similar for both substrates. Activity toward other classes of substrates is
absent. Cu2+ ions strongly inhibit the action of this and other
glucosyltransferases; however, we suggest that this phenomenon in large part is
due to Cu2+-mediated substrate degradation rather than inhibition of the enzyme.
Additional lines of complementary biochemical data also indicated that in the
case of V. vinifera, the principal, if not only, role of UDP-glucose:flavonoid
3-O-glucosyltransferases is to glucosylate anthocyanidins in red fruit during
ripening. Other glucosyltransferases with a much higher relative activity toward
quercetin are suggested to glucosylate flavonols in a distinct spatial and
temporal pattern. It should be considered whether gene products homologous to
Bronze-1 in some cases more accurately should be referred to as
UDP-glucose:anthocyanidin 3-O-glucosyltransferases.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 9535914 [PubMed - indexed for MEDLINE]

430: Plant Physiol. 1997 Nov;115(3):1039-48.

Characterization and expression of caffeoyl-coenzyme A 3-O-methyltransferase
proposed for the induced resistance response of Vitis vinifera L.

Busam G, Junghanns KT, Kneusel RE, Kassemeyer HH, Matern U.

Lehrstuhl fur Biochemie der Pflanzen, Institut fur Biologie II, Universitat
Freiburg, Germany.

Cell-suspension cultures of Vitis vinifera L. cv Pinot Noir accumulated
resveratrol upon fungal elicitation, and the activity of
S-adenosyl-L-methionine:trans-caffeoyl-coenzyme A 3-O-methyl-transferase
(CCoAOMT), yielding feruloyl-CoA, increased to a transient maximum at 12 to 15
h. CCoAOMT cDNA was cloned from the elicited cells and was shown to encode a
polypeptide highly homologous to CCoAOMTs from cells of Petroselinum species or
Zinnia species. The expression of the cDNA in Escherichia coli revealed that
grapevine CCoAOMT methylates both caffeoyl- and 5-hydroxyferuloyl-coenzyme A and
is probably involved in phenolic esterification and lignification. Commercial
plant activators induce the disease-resistance response of test plants and are
considered to mimic the action of salicylic acid. Among these chemicals,
2,6-dichloroisonicotinic acid and benzo(1,2,3)-thiadiazole-7-carbothioic acid
S-methyl ester provoke systemic acquired resistance (SAR) and were also shown to
induce the expression of class III chitinase in grapevine. The SAR response is
classified by an unchanged phenotype of tissues, but the mechanistic basis is
unknown. Treatment of the cultured V. vinifera cells with either fungal elicitor
or low concentrations of salicylic acid and 2,6-dichloroisonicotinic acid,
respectively, raised the CCoAOMT or stilbene synthase transcript abundance,
suggesting that grapevine is capable of the SAR response, whereas
benzo(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester was ineffective. The
data imply for the first time (to our knowledge) that the expression of
phenyl-propanoid genes in grapevine is induced by SAR activators without
phenotypic consequences and suggest a role for CCoAOMT and stilbene synthase in
the disease-resistance response leading beyond the level of pathogenesis-related
proteins as markers of the SAR.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 9390437 [PubMed - indexed for MEDLINE]

431: Plant Physiol. 1997 Nov;115(3):1029-38.

Differential expression of chitinases in Vitis vinifera L. responding to
systemic acquired resistance activators or fungal challenge.

Busam G, Kassemeyer HH, Matern U.

Lehrstuhl fur Biochemie der Pflanzen, Institut fur Biologie II, Universitat
Freiburg, Germany.

The concept of systemic acquired resistance (SAR) enables a novel approach to
crop protection, and particular pathogenesis-related proteins, i.e. an acidic
chitinase, have been classified as markers of the SAR response. Basic class I
(VCHIT1b) and a class III (VCH3) chitinase cDNAs were cloned from cultured Vitis
vinifera L. cv Pinot Noir cells and used to probe the induction response of
grapevine cells to salicylic acid or yeast elicitor. Furthermore, the cells were
treated with the commercial SAR activators 2,6-dichloroiso-nicotinic acid or
benzo(1,2,3)-thiadiazole-7-carbothioic acid S-methyl ester. Elicitor or
salicylic acid induced both VCHIT1b and VCH3 transcript abundances, whereas
2,6-dichloroiso-nicotinic acid or benzo(1,2,3)-thiadiazole-7-carbothioic acid
S-methyl ester enhanced exclusively the expression of VCH3. To assess the
systemic sensation of chitinase expression, single leaves of Vitis vinifera L.
cv Pinot Noir or Vitis rupestris plants were inoculated with Plasmopara viticola
spore suspensions, and the VCH3 and VCHIT1b mRNA amounts in the infected versus
the adjacent, healthy leaf were monitored. Two VCH3 mRNA maxima were observed 2
and 6 d postinoculation in the infected, susceptible V. vinifera tissue, whereas
in the healthy leaf the transcript increased from low levels d 2 postinoculation
to prominent levels d 6 to 8 postinoculation. The level of VCH3 mRNA increased
also over 4 d in the inoculated, resistant V. rupestris tissue. However,
necrotic spots rapidly limited the infection, and the VCH3 transcript was
undetectable in the upper-stage, healthy leaf. The expression of VCHIT1b
remained negligible under either experimental condition. Overall, the results
suggest that the selective expression of VCH3 might be a reliable indicator of
the SAR response in V. vinifera L.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 9390436 [PubMed - indexed for MEDLINE]

432: Biochem Mol Biol Int. 1997 Sep;42(6):1249-60.

A comparison of the hydroxyl radical scavenging properties of the shark bile
steroid 5 beta-scymnol and plant pycnogenols.

Macrides TA, Shihata A, Kalafatis N, Wright PF.

Department of Medical Laboratory Science, RMIT-University, Melbourne, Victoria,
Australia.

The hydroxyl radical (OH.) quenching abilities of the following compounds were
compared in the deoxyribose degradation system (initiated by the
ferrous-ascorbate Fenton reaction): (a) 5 beta-scymnol, the hepatoprotective
shark bile sterol, and its mono- and di-sulfate esters; (b) three marketed
pycnogenol preparations (syn: proanthocyanidin--natural plant-derived
polyphenolic bioflavonoids) extracted from pine tree (Pinus maritima) bark and
grape (Vitis vinifera) seeds; and (c) two known hydroxyl radical scavengers,
dimethyl sulfoxide and mannitol, and the peroxyl radical scavenger Trolox (the
alpha-tocopherol analogue). 5 beta-scymnol was a more potent OH. quencher than
dimethyl sulfoxide, mannitol and Trolox, and markedly more potent than the
pycnogenol preparations. Increased sulfation of 5 beta-scymnol progressively
reduced its free radical scavenging activity, thus clearly attributing the
potent OH. quenching properties to its novel tri-alcohol-substituted aliphatic
side chain. The favourable interaction of these bile steroids with reactive
oxygen species in an aqueous environment, makes them attractive candidates for
evaluation as protective agents against disorders in which oxidative stress is
implicated.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 9305543 [PubMed - indexed for MEDLINE]

433: Plant Physiol. 1997 Jul;114(3):771-8.

A class IV chitinase is highly expressed in grape berries during ripening.

Robinson SP, Jacobs AK, Dry IB.

Commonwealth Scientific and Industrial Research Organization, Division of
Horticulture, Adelaide, Australia. [email protected]

Chitinase activity increased markedly at the onset of ripening in grape (Vitis
vinifera L.) berries and continued to increase throughout the sugar accumulation
phase of berry development. In contrast, beta-1,3-glucanase activity was not
detected in grape berries at any stage of development. Two closely related
chitinase cDNAs (VvChi4A and VvChi4B) were cloned from grapes. Sequence and
Southern analysis indicate that these two clones may represent alleles of the
same gene. The predicted proteins are acidic and have a signal peptide followed
by a cysteine-rich, chitin-binding domain and a catalytic region. An analysis of
their sequences indicates that they are class IV chitinase. The deduced protein
sequence of VvChi4A has a high level of identity with the 32- and 28-kD
chitinases present as haze proteins in wine. Expression of VvChi4 was high in
berries and low in flowers but was not detected in leaves, roots, or seeds. No
expression was detected in berries 2 to 8 weeks postflowering, but expression
was high 12 to 16 weeks postflowering, which coincided with sugar accumulation
and an increase in chitinase activity. Constitutive expression of VvChi4 appears
to be fruit-specific and induced at high levels in grapes during ripening.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 9232868 [PubMed - indexed for MEDLINE]

434: Plant Physiol. 1997 Jul;114(3):759-69.

Identification and characterization of a fruit-specific, thaumatin-like protein
that accumulates at very high levels in conjunction with the onset of sugar
accumulation and berry softening in grapes.

Tattersall DB, van Heeswijck R, Hoj PB.

Department of Horticulture, Viticulture, and Oenology, University of Adelaide,
Glen Osmond, SA, Australia.

The protein composition of the grape (Vitis vinifera cv Muscat of Alexandria)
berry was examined from flowering to ripeness by gel electrophoresis. A protein
with an apparent molecular mass of 24 kD, which was one of the most abundant
proteins in extracts of mature berries, was purified and identified by amino
acid sequence to be a thaumatin-like protein. Combined cDNA sequence analysis
and electrospray mass spectrometry revealed that this protein, VVTL1 (for V.
vinifera thaumatin-like protein 1), is synthesized with a transient signal
peptide as seen for apoplastic preproteins. Apart from the removal of the
targeting signal and the formation of eight disulfide bonds, VVTL1 undergoes no
other posttranslational modification. Southern, northern, and western analyses
revealed that VVTL1 is found in the berry only and is encoded by a single gene
that is expressed in conjunction with the onset of sugar accumulation and
softening. The exact role of VVTL1 is unknown, but the timing of its
accumulation correlates with the inability of the fungal pathogen powdery mildew
(Uncinula necator) to initiate new infections of the berry. Western analysis
revealed that the presence of thaumatin-like proteins in ripening fruit might be
a widespread phenomenon.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 9232867 [PubMed - indexed for MEDLINE]

435: Ecotoxicol Environ Saf. 1997 Jun;37(1):24-9.

Uptake and transformation of benzene and toluene by plant leaves.

Ugrekhelidze D, Korte F, Kvesitadze G.

Durmishidze Institute of Plant Biochemistry, Academy of Sciences of Georgia,
Digomi, Tbilisi, Georgia.

The [1-6(14)C]benzene and [1-(14)C]toluene vapors penetrate into hypostomatous
leaves of Acer campestre, Malus domestica, and Vitis vinifera from both sides,
whereas hydrocarbons are more intensively absorbed by the stomatiferous side and
more actively taken up by young leaves. Benzene and toluene conversion in leaves
occurs with the aromatic ring cleavage and their carbon atoms are mainly
incorporated into nonvolatile organic acids, while their incorporation into
amino acids is less intensive. Intact spinach chloroplasts oxidize benzene, and
this process is strongly stimulated in light. Oxidation of benzene by spinach
chloroplasts or by enzyme preparation from spinach leaves is almost completely
inhibited by 8-oxyquinoline or sodium diethyldithiocarbamate, and slightly
affected by alpha, alpha'-dipyridyl. Benzene oxidation by enzyme preparation is
significantly stimulated by NADH and NADPH; in their presence, the benzene
hydroxylation product, phenol, is formed in a determinable amount. It is
supposed that the enzyme performing the first step of oxidative transformation
of benzene in plant leaves contains copper as the prosthetic group.

PMID: 9212332 [PubMed - indexed for MEDLINE]

436: Clin Chem. 1997 Jun;43(6 Pt 1):1092-3.

Antioxidant activity of the stilbene astringin, newly extracted from Vitis
vinifera cell cultures.

Merillon JM, Fauconneau B, Teguo PW, Barrier L, Vercauteren J, Huguet F.

Publication Types:
Letter
Research Support, Non-U.S. Gov't

PMID: 9191572 [PubMed - indexed for MEDLINE]

437: Plant Physiol. 1997 May;114(1):185-191.

CO2 and Water Vapor Exchange across Leaf Cuticle (Epidermis) at Various Water
Potentials.

Boyer JS, Wong SC, Farquhar GD.

Environmental Biology Group, Research School of Biological Sciences, The
Australian National University, Canberra, ACT 2601, Australia.

Cuticular properties affect the gas exchange of leaves, but little is known
about how much CO2 and water vapor cross the cuticular barrier or whether low
water potentials affect the process. Therefore, we measured the cuticular
conductances for CO2 and water vapor in grape (Vitis vinifera L.) leaves having
various water potentials. The lower leaf surface was sealed to force all gas
exchange through the upper surface, which was stoma-free. In this condition both
gases passed through the cuticle, and the CO2 conductance could be directly
determined from the internal mole fraction of CO2 near the compensation point,
the external mole fraction of CO2, and the CO2 flux. The cuticle allowed small
amounts of CO2 and water vapor to pass through, indicating that gas exchange
occurs in grape leaves no matter how tightly the stomata are closed. However,
the CO2 conductance was only 5.7% of that for water vapor. This discrimination
against CO2 markedly affected calculations of the mole fraction of CO2 in leaves
as stomatal apertures decreased. When the leaf dehydrated, the cuticular
conductance to water vapor decreased, and transpiration and assimilation
diminished. This dehydration effect was largest when turgor decreased, which
suggests that cuticular gas exchange may have been influenced by epidermal
stretching.

PMID: 12223698 [PubMed - as supplied by publisher]

438: Nat Genet. 1997 May;16(1):84-7.

Comment in:
Nat Genet. 1997 May;16(1):1.
Nat Genet. 1997 May;16(1):4-5.

The parentage of a classic wine grape, Cabernet Sauvignon.

Bowers JE, Meredith CP.

Department of Viticulture and Enology, University of California, Davis 95616,
USA.

The world's great wines are produced from a relatively small number of classic
European cultivars of Vitis vinifera L Most are thought to be centuries old and
their origins have long been the subject of speculation. Among the most
prominent of these cultivars is Cabernet Sauvignon, described as "the world's
most renowned grape variety for the production of fine red wine". Although now
grown in many countries, Cabernet Sauvignon derives its fame from its long
association with the Bordeaux region of France, where it has been grown at least
since the 17th century. We present microsatellite DNA evidence for the
hypothesis that Cabernet Sauvignon is the progeny of two other Bordeaux
cultivars, Cabernet franc and Sauvignon blanc. Likelihood ratios support this
hypothesis to a very high degree of probability. A close relationship between
Cabernet Sauvignon and Cabernet franc has been suspected but the genetic
contribution of Sauvignon blanc, despite its similar name, is a surprise.

Publication Types:
Historical Article
Research Support, Non-U.S. Gov't

PMID: 9140400 [PubMed - indexed for MEDLINE]

439: DNA Seq. 1997;8(1-2):109-12.

Nucleotide sequence of grapevine (Vitis vinifera) cDNA similar to SNAP proteins.

Matsumoto S, Dry IB, Thomas M.

Department of Biology, Faculty of Education, Gifu University, Japan.
[email protected]

A cDNA clone (VS1) homologous to SNAP proteins was isolated from a grapevine
cDNA library. The cDNA insert was 1167 bp long and contained a single open
reading frame coding for 289 amino acids. The amino acid sequence of VS1 shows
similarity (35%-45%) to SNAP proteins from various sources.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 9522131 [PubMed - indexed for MEDLINE]

440: Life Sci. 1997;61(21):2103-10.

Comparative study of radical scavenger and antioxidant properties of phenolic
compounds from Vitis vinifera cell cultures using in vitro tests.

Fauconneau B, Waffo-Teguo P, Huguet F, Barrier L, Decendit A, Merillon JM.

Center for Study and Research on Xenobiotics, Faculty of Pharmacy, Poitiers,
France.

Vitis vinifera cell suspensions were used to isolate and characterize the
flavonoids (anthocyanins, catechins) and non-flavonoids (stilbenes) found in red
wine. Furthermore, we showed that astringin is produced although this stilbene
has not previously been reported to be a constituent of V. vinifera or wine. The
ability of these compounds to act as radical scavengers was investigated using
1,1-diphenyl-2-picryl-hydrazyl (DPPH), a stable free radical. Antioxidant
activities were assessed by their capacity to prevent Fe2+-induced lipid
peroxidation in microsomes and their action on Cu2+-induced lipid peroxidation
in low-density lipoproteins. The results showed that astringin has an important
antioxidant effect similar to that of trans-resveratrol, and a higher radical
scavenger activity than the latter. Astringinin appeared to be more active.
These data indicate that phenolic compounds (stilbenes, catechins, anthocyanins)
exhibit interesting properties which may account in part for the so-called
"French paradox," i.e. that moderate drinking of red wine over a long period of
time can protect against coronary heart disease.

Publication Types:
Comparative Study

PMID: 9395251 [PubMed - indexed for MEDLINE]

441: Biofactors. 1997;6(4):403-10.

Polyphenols produced during red wine ageing.

Brouillard R, George F, Fougerousse A.

Laboratoire de Chimie des Polyphenols, Universite Louis Pasteur, Faculte de
Chimie, Strasbourg, France.

Over the past few years, it has been accepted that a moderate red wine
consumption is a factor beneficial to human health. Indeed, people of France and
Italy, the two major wine-producing European countries, eat a lot of fatty foods
but suffer less from fatal heart strokes than people in North-America or in the
northern regions of Europe, where wine is not consumed on a regular basis. For a
time, ethanol was thought to be the "good" chemical species hiding behind what
is known as the "French paradox". Researchers now have turned their
investigations towards a family of natural substances called "polyphenols",
which are only found in plants and are abundant in grapes. It is well known that
these molecules behave as radical scavengers and antioxidants, and it has been
demonstrated that they can protect cholesterol in the LDL species from
oxidation, a process thought to be at the origin of many fatal heart attacks.
However, taken one by one, it remains difficult to demonstrate which are the
best polyphenols as far as their antioxidant activities are concerned. The main
obstacle in that kind of research is not the design of the chemical and
biological tests themselves, but surprisingly enough, the limited access to
chemically pure and structurally elucidated polyphenolic compounds. In this
article, particular attention will be paid to polyphenols of red wine made from
Vitis vinifera cultivars. With respect to the "French paradox", we address the
following question: are wine polyphenolic compounds identical to those found in
grapes (skin, pulp and seed), or are there biochemical modifications
specifically taking place on the native flavonoids when a wine ages? Indeed,
structural changes occur during wine conservation, and one of the most studied
of those changes concerns red wine colour evolution, called "wine ageing". As a
wine ages, it has been demonstrated that the initially present grape pigments
slowly turn into new more stable red pigments. That phenomenon goes on for
weeks, months and years. Since grape and wine polyphenols are chemically
distinct, their antioxidant activities cannot be the same. So, eating grapes
might well lead to beneficial effects on human health, due to the variety and
sometimes large amounts of their polyphenolic content. However, epidemiological
surveys have focused on wines, not on grapes....

Publication Types:
Review

PMID: 9388306 [PubMed - indexed for MEDLINE]

442: Planta Med. 1996 Dec;62(6):495-502.

Procyanidines from Vitis vinifera seeds protect rabbit heart from
ischemia/reperfusion injury: antioxidant intervention and/or iron and copper
sequestering ability.

Maffei Facino R, Carini M, Aldini G, Berti F, Rossoni G, Bombardelli E,
Morazzoni P.

Istituto Chimico Farmaceutico Tossicologico, Milan, Italy.

An isolated rabbit heart Langendorff preparation paced electrically was used to
evaluate the effects of a highly purified, high molecular weight fraction of
oligomeric procyanidines isolated from Vitis vinifera seeds on myocardial
reperfusion injury after 40 minutes of low flow (1 ml/min) ischemia. Infusion of
the heart with 100 or 200 micrograms/ml procyanidines dose-dependently reduced
ventricular contracture during ischemia (LVEDP values decreased by 28% and 51%),
decreased coronary perfusion pressure (CPP), improved cardiac mechanical
performance upon reperfusion, increased the release of 6-keto-PGF1 alpha into
the perfusate in both the pre-ischemic and the reperfusion periods (by 68% at
200 micrograms/ml), and suppressed rhythm irregularity. This antiarrhythmogenic
action was confirmed in a more severe model of ischemia (flow rate 0.2 ml/ min).
The cardioprotective agent allopurinol infused at 20 micrograms/ml had effects
on the contractility and on the release of 6-keto-PGF1 alpha comparable to those
of 200 micrograms/ml procyanidines. The results of the second part of this study
show that procyanidines are potent scavengers of several reactive oxygen species
involved in the ischemia/reperfusion damage: the superoxide anion (IC50 = 5.64
microM: rate constant K = 7.55 x 10(5) M-1 s-1, determined by the phenazine
methosulfate/NADH method); the hydroxyl radical (IC50 = 28 microM; rate constant
K = 1.2 x 10(12) M-1 s-1, determined by the electron spin resonance
spectroscopy); peroxyl radicals (IC50 = 0.025 microM and 0.35 microM, determined
using two different lipid substrates, phosphatidylcholine liposomes and methyl
linoleate micelles by UV spectroscopy at 233 nm). Finally, procyanidines
interact with Fe2+ and Cu2+ ions (the catalysts of HO. radicals production)
giving rise to strong complexes, with stability constants (log K) ranging from
9.35 to approximately 9.

PMID: 9000880 [PubMed - indexed for MEDLINE]

443: Tsitol Genet. 1996 Nov-Dec;30(6):33-7.

[The genetic polymorphism of the grape studied by RAPD analysis]

[Article in Russian]

Sivolap IuM, Balashova IA, Troshin LP.

Genetic polymorphism in plant of 3 genera of Vitaceae family, 11 species of
Vitis genera, 10 cultivars of Vitis vinifera and 4 cultivars that obtained from
distant hybridization was studied. According to date of RAPD analysis genetic
distances were determined and phylogenetic dendrogram was constructed. The
possibility of using PCR for evaluation of breeding material by the levels of
genetic relations was shown.

Publication Types:
English Abstract

PMID: 9139436 [PubMed - indexed for MEDLINE]

444: HortScience. 1996 Oct;31(6):944-6.

A whole-plant, open, gas-exchange system for measuring net photosynthesis of
potted woody plants.

Miller DP, Howell GS, Flore JA.

Department of Horticulture, Michigan State University, East Lansing 48824, USA.

Chambers were constructed to measure gas exchange of entire potted grapevines
(Vitis vinifera L.). The plant enclosures were constructed from Mylar film,
which is nearly transparent to photosynthetically active radiation. Maintaining
a slight, positive, internal pressure allowed the Mylar chambers to inflate like
balloons and required no other means of support. The whole-plant, gas-exchange
chamber design and construction were simple and inexpensive. They were assembled
easily, equilibrated quickly, and did not require cooling. They allowed for the
measurement of any plants in a relatively short period. This system would enable
the researcher to make replicated comparisons of treatment influences on
whole-plant CO2 assimilation throughout the growing season. While CO2
measurement was the focus of this project, it would be possible to measure
whole-plant transpiration with this system.

PMID: 11540961 [PubMed - indexed for MEDLINE]

445: Phytochemistry. 1996 Oct;43(3):621-4.

Mono- and diglycosides of (E)-6,9-dihydroxymegastigma-4,7-dien-3-one in Vitis
vinifera wine.

Baltenweck-Guyot R, Trendel JM, Albrecht P, Schaeffer A.

Laboratoire de Chimie Organique des Substances Naturelles, URA 31 associee au
CNRS, Institut de Chimie, Universite Louis Pasteur, Strasbourg, France.

Two beta-D-glucopyranosides and two
6-O-beta-D-apiofuranosyl-beta-D-glucopyranosides of
(E)-6,9-dihydroxymegastigma-4,7-dien-3-one were isolated from Vitis vinifera cv.
Gewurztraminer wine and their structures were established by NMR spectroscopy.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 8987582 [PubMed - indexed for MEDLINE]

446: Plant Physiol. 1996 Aug;111(4):1059-1066.

Analysis of the Expression of Anthocyanin Pathway Genes in Developing Vitis
vinifera L. cv Shiraz Grape Berries and the Implications for Pathway Regulation.

Boss PK, Davies C, Robinson SP.

Cooperative Research Centre for Viticulture, P.O. Box 145, Glen Osmond, South
Australia 5064.

Anthocyanin synthesis in Vitis vinifera L. cv Shiraz grape berries began 10
weeks postflowering and continued throughout berry ripening. Expression of seven
genes of the anthocyanin biosynthetic pathway (phenylalanine ammonia lyase
[PAL], chalcone synthase [CHS], chalcone isomerase [CHI],
flavanone-3-hydroxylase [F3H], dihydroflavonol 4-reductase [DFR],
leucoanthocyanidin dioxygen-ase [LDOX], and UDP glucose-flavonoid 3-o-glucosyl
transferase [UFGT]) was determined. In flowers and grape berry skins, expression
of all of the genes, except UFGT, was detected up to 4 weeks postflowering,
followed by a reduction in this expression 6 to 8 weeks postflowering.
Expression of CHS, CHI, F3H, DFR, LDOX, and UFGT then increased 10 weeks
postflowering, coinciding with the onset of anthocyanin synthesis. In grape
berry flesh, no PAL or UFGT expression was detected at any stage of development,
but CHS, CHI, F3H, DFR, and LDOX were expressed up to 4 weeks postflowering.
These results indicate that the onset of anthocyanin synthesis in ripening grape
berry skins coincides with a coordinated increase in expression of a number of
genes in the anthocyanin biosynthetic pathway, suggesting the involvement of
regulatory genes. UFGT is regulated independently of the other genes, suggesting
that in grapes the major control point in this pathway is later than that
observed in maize, petunia, and snapdragon.

PMID: 12226348 [PubMed - as supplied by publisher]

447: Plant Mol Biol. 1996 Aug;31(5):983-92.

Characterization of Vitis vinifera L. glutamine synthetase and molecular cloning
of cDNAs for the cytosolic enzyme.

Loulakakis KA, Roubelakis-Angelakis KA.

Department of Biology, University of Crete, Greece.

Grapevine (Vitis vinifera L.) glutamine synthetase (GS) was analysed into two
distinct classes of isoforms; one of them was present in both leaf and root
tissues while the other one showed leaf specificity. Western blot analysis
revealed that grapevine GS consists of three types of polypeptides of distinct
size and differential tissue specificity. Two structurally distinct cDNA clones,
pGS1;1 and pGS1;2, encoding grapevine GS were isolated from a cell suspension
library and characterized. Both clones contained open reading frames encoding
for polypeptides of 356 amino acids with a predicted molecular mass of about 39
kDa. Although the coding sequences of pGS1;1 and pGS1;2 were 84% similar, their
5'- and 3'-untranslated sequences showed only 40% similarity. The coding
sequences of the two clones and the derived amino acid sequences showed higher
homology to cytosolic than to chloroplastic GSs of other higher plants
indicating that the cDNAs isolated encode for cytosolic isoforms of grapevine
GS. Southern blot analysis suggested the existence of more than two GS genes in
the grapevine genome. In northern blots both clones were hybridized to mRNAs of
about 1.4 kb that are differentially expressed in the various tissues. Supply of
nitrate or ammonium in the cell suspension culture medium, as a sole nitrogen
source, resulted in differential response of the pGS1;1- and pGS1;2-related
genes.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 8843941 [PubMed - indexed for MEDLINE]

448: Nat Biotechnol. 1996 May;14(5):624-8.

Erratum in:
Nat Biotechnol 1996 Nov;14(11):1521.

Establishment of an Agrobacterium-mediated transformation system for grape
(Vitis vinifera L.): the role of antioxidants during grape-Agrobacterium
interactions.

Perl A, Lotan O, Abu-Abied M, Holland D.

Department of Fruit Tree Breeding and Molecular Genetics, Volcani Center,
Bet-Dagan, Israel. [email protected]

Very short exposures of embryogenic calli of Vitis vinifera cv. Superior
Seedless grape plants to diluted cultures of Agrobacterium resulted in plant
tissue necrosis and subsequent cell death. Antibiotics used for Agrobacterium
elimination or as plant selectable markers were not responsible for this
necrotic response. Rather, cell death seemed to be oxygen-dependent and
correlated with elevated levels of peroxides. Therefore, we studied the effects
on necrosis of various combinations of antioxidants during and after
grape-Agrobacterium cocultivation. The combination of polyvinylpolypyrrolidone
and dithiothreitol was found to improve plant viability. Tissue necrosis was
completely inhibited by these antioxidants while Agrobacterium virulence was not
effected. These treatments enabled the recovery of stable transgenic grape
plants resistant to hygromycin.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 9630955 [PubMed - indexed for MEDLINE]

449: Plant Physiol. 1996 May;111(1):275-83.

Sugar accumulation in grape berries. Cloning of two putative vacuolar invertase
cDNAs and their expression in grapevine tissues.

Davies C, Robinson SP.

Cooperative Research Centre for Viticulture, South Australia, Australia.
[email protected]

During grape berry (Vitis vinifera L.) ripening, sucrose transported from the
leaves is accumulated in the berry vacuoles as glucose and fructose. To study
the involvement of invertase in grape berry ripening, we have cloned two cDNAs
(GIN1 and GIN2) from berries. The cDNAs encode translation products that are 62%
identical to each other and both appear to be vacuolar forms of invertase. Both
genes are expressed in a variety of tissues, including berries, leaves, roots,
seeds, and flowers, but the two genes have distinct patterns of expression. In
grape berries, hexose accumulation began 8 weeks postflowering and continued
until the fruit was ripe at 16 weeks. Invertase activity increased from
flowering, was maximal 8 weeks postflowering, and remained constant on a per
berry basis throughout ripening. Expression of GIN1 and GIN2 in berries, which
was high early in berry development, declined greatly at the commencement of
hexose accumulation. The results suggest that although vacuolar invertases are
involved in hexose accumulation in grape berries, the expression of the genes
and the synthesis of the enzymes precedes the onset of hexose accumulation by
some weeks, so other mechanisms must be involved in regulating this process.

PMID: 8685267 [PubMed - indexed for MEDLINE]

450: Gene. 1996 Feb 2;168(1):87-92.

The amino-acid sequence similarity of plant glutamate dehydrogenase to the
extremophilic archaeal enzyme conforms to its stress-related function.

Syntichaki KM, Loulakakis KA, Roubelakis-Angelakis KA.

Department of Biology, University of Crete, Heraklion, Greece.

A cDNA clone encoding grapevine (Vitis vinifera L. cv Sultanina)
NAD(H)-glutamate dehydrogenase (GDH) was isolated from a cDNA expression library
by immunoscreening with a polyclonal antibody raised against grapevine GDH.
Nucleotide sequence analysis revealed an open reading frame (ORF) encoding a
precursor protein of 411 amino acids (aa) with a calculated molecular mass of
44.517 kDa. The deduced aa sequence showed relatively higher homology to GDH
from archaebacteria species, than to those from eukaryotes and eubacteria. This
resemblance indicated a functional and/or evolutionary relationship in this
class of enzymes which might be relevant to the stress-related function of plant
GDH. We have shown that the bacterially produced plant GDH was thermostable.

PMID: 8626071 [PubMed - indexed for MEDLINE]

451: J Gen Virol. 1996 Jan;77 ( Pt 1):155-61.

Variation of viroid profiles in individual grapevine plants: novel grapevine
yellow speckle viroid 1 mutants show alterations of hairpin I.

Polivka H, Staub U, Gross HJ.

Institut fur Biochemie, Bayerische Julius-Maximilians-Universitat, Biozentrum,
Am Hubland, Wurzburg, Germany.

This is the first report which gives a general survey about viroid variant
composition in a vineyard and within single plants. A German vineyard with
20-year-old grapevines (Vitis vinifera) of the cultivars 'Bacchus' and 'Kerner'
was analysed for viroid infections. Only grapevine yellow speckle viroid 1
(GYSVd1) and the grapevine isolate of hop stunt viroid (HSVdg) were detected.
Both viroids occur in several sequence variations. Eighteen novel GYSVd1
variants and two previously published HSVdg main variants with six new minor
variants were found. They were randomly spread in the vineyard. The distribution
of GYSVd1 and HSVdg main variants and their accompanying subvariants differed
even in neighbouring plants. We conclude that these individual viroid variant
profiles are the result of 20 years of independent evolution, i.e. mutation and
selection, in each single plant. Four of the nine GYSVd1 main variants were
mutated in the inverted repeats bordering the central conserved region. These
base substitutions decreased the thermodynamic stability of a metastable
structure called hairpin I.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 8558124 [PubMed - indexed for MEDLINE]

452: J Biol Chem. 1995 Mar 3;270(9):4368-74.

The plant inorganic pyrophosphatase does not transport K+ in vacuole membrane
vesicles multilabeled with fluorescent probes for H+, K+, and membrane
potential.

Ros R, Romieu C, Gibrat R, Grignon C.

Departament de Biologia Vegetal, Facultat de Ciencies Biologiques, Universitat
de Valencia, Burjassot (Valencia), Spain.

It has been claimed that the inorganic pyrophosphatase (PPase) of the plant
vacuolar membrane transports K+ in addition to H+ in intact vacuoles (Davies, J.
M., Poole, R. J., Rea, P. A., and Sanders, D. (1992) Proc. Natl. Acad. Sci.
U.S.A. 89, 11701-11705). Since this was not confirmed using the purified and
reconstituted PPase consisting of a 75-kDa polypeptide (Sato, M.H., Kasahara,
M., Ishii, N., Homareda, H., Matsui, H., and Yoshida, M. (1994) J. Biol. Chem.
269, 6725-6728), these authors proposed that K+ transport by the PPase is
dependent on its association with other membrane components lost during
purification. We have examined the hypothesis of K+ translocation by the PPase
using native vacuolar membrane vesicles from Vitis vinifera suspension cells,
multilabeled with fluorescent probes for K+, H+, and membrane potential. This
material contained a high proportion of right-side-out, tightly sealed vesicles,
exhibiting high PPase activity which was strongly stimulated by uncouplers and
K+. Proton pumping occurred in response to pyrophosphate addition in the absence
of K+. No K+ incorporation into the vesicles could be observed after PPase
energization in the presence of K+, although H+ transport was highly stimulated.
The hydrolytic activity was stimulated by a protonophore and by a H+/K+
exchanger but not by the K+ ionophore valinomycin. No evidence could be obtained
supporting the operation of an endogenous K+/H+ exchanger capable to dissipate
the putative active K+ flux generated by the PPase. We conclude that PPase in
native vacuolar membrane vesicles does not transport K+.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 7876200 [PubMed - indexed for MEDLINE]

453: Arch Virol. 1995;140(1):157-64.

Coat protein gene sequence of an Austrian isolate of grapevine fanleaf virus.

Brandt S, Ibl M, Himmler G.

Institut fur Angewandte Mikrobiologie, Universitat fur Bodenkultur, Wien,
Austria.

The nucleotide sequence of the coat protein cistron of an Austrian isolate of
grapevine fanleaf virus (GFLV-FC) from Vitis vinifera cv. French Colombard was
determined. It shows small differences at the RNA level as well as at the
protein level compared to the sequences of already published grapevine fanleaf
virus strains. The differences may be a result of the natural variation among
virus populations or a consequence of selection in a special host plant. As the
virus RNA sequence reported here was isolated directly from its natural woody
host by an immunocapture-PCR technique, passage of the virus through a
herbaceous host could be avoided and a possible bias introduced by a different
host environment was excluded. The sequence similarity of known GFLV coat
protein cistrons to the sequence described is as high as among the other
strains.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 7646340 [PubMed - indexed for MEDLINE]

454: Eur J Histochem. 1995;39(1):69-74.

Effect of fosetyl-A1 on peroxidase from grapevine (Vitis vinifera) cells.

Lopez-Serrano M, Ferrer MA, Ros Barcelo A, Pedreno MA.

Department of Plant Biology (Plant Physiology), University of Murcia, Spain.

Grapevine (Vitis vinifera cv. Monastrell) suspension cell cultures were treated
with fosetyl-A1, a widely used systemic fungicide for grapevine diseases caused
by oomycetes, and examined at the electron microscope level for peroxidase
cytochemistry. The results showed that treatment with fosetyl-A1 provokes an
activation of both vacuolar sap and tonoplast-located peroxidase, already
described as due to the basic peroxidase isoenzyme, B5, which was previously
characterized as a constitutive marker of disease resistance against Plasmopara
viticola in axillary bud cultures of Vitis spp. This activation of peroxidase
isoenzyme B5, as seen at the electron microscope level, was confirmed by
cytophotometric methods, but is in contrast with the unchanged enzyme level
determined by biochemical methods. These results suggest a metabolic activation
of peroxidase isoenzyme B5 as a consequence of fosetyl-A1 treatment, probably
due to an acidification of the vacuole. This response was accompanied by the
appearance of myelin-like structures inside the cytoplasm and osmiophylic-bodies
inside the mitochondria. However, the latter structural changes cannot easily be
related to the above described specific peroxidase response.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 7612959 [PubMed - indexed for MEDLINE]

455: Plant Physiol. 1994 Aug;105(4):1375-1383.

Catalase Is Differentially Expressed in Dividing and Nondividing Protoplasts.

Siminis CI, Kanellis AK, Roubelakis-Angelakis KA.

Department of Biology, University of Crete, P.O. Box 1470, 71110 Heraklio,
Crete, Greece (C.I.S., K.A.R.-A).

Based on our previous results that peroxidase is induced in dividing tobacco
protoplasts but it is not expressed in the nondividing grapevine (Vitis vinifera
L.) protoplasts during culture (C.I. Siminis, A.K. Kanellis, K.A.
Roubelakis-Angelakis [1993] Physiol Plant 87: 263-270), we further tested the
hypothesis that oxidative stress may be implicated in the recalcitrance of plant
protoplasts. The expression of catalase, a major defense enzyme against cell
oxidation, was studied during isolation and culture of mesophyll protoplasts
from the recalcitrant grapevine and regenerating tobacco (Nicotiana tabacum L.).
Incubation of tobacco leaf strips with cell wall-degrading enzymes resulted in a
burst of catalase activity and an increase in its immunoreactive protein; in
contrast, no such increases were found in grapevine. The cathodic and anodic
catalase isoforms consisted exclusively of subunits [alpha] and [beta],
respectively, in tobacco, and of subunits [beta] and [alpha], respectively, in
grapevine. The catalase specific activity increased only in grapevine
protoplasts during culture. The ratio of the enzymatic activities to the
catalase immunoreactive protein declined in dividing tobacco protoplasts and
remained fairly constant in nondividing tobacco and grapevine protoplasts during
culture. Also, in dividing tobacco protoplasts the de novo accumulation of the
catalase [beta] subunit gave rise to the acidic isoenzymes, whereas in
nondividing tobacco and grapevine protoplasts, after 8 d in culture, only the
basic isoenzymes remained due to de novo accumulation of the [alpha] subunit.
The pattern of catalase expression in proliferating tobacco leaf cells during
callogenesis was similar to that in dividing protoplasts. The different
responses of catalase expression in dividing and nondividing tobacco and
grapevine mesophyll protoplasts may indicate a specificity of catalase related
to induction of totipotency.

PMID: 12232292 [PubMed - as supplied by publisher]

456: Arzneimittelforschung. 1994 May;44(5):592-601.

Free radicals scavenging action and anti-enzyme activities of procyanidines from
Vitis vinifera. A mechanism for their capillary protective action.

Maffei Facino R, Carini M, Aldini G, Bombardelli E, Morazzoni P, Morelli R.

Istituto Chimico Farmaceutico Tossicologico, Milan, Italy.

The scavenging by procyanidines (polyphenol oligomers from Vitis vinifera seeds,
CAS 85594-37-2) of reactive oxygen species (ROS) involved in the onset (HO
degrees) and the maintenance of microvascular injury (lipid radicals R degrees,
RO degrees, ROO degrees) has been studied in phosphatidylcholine liposomes
(PCL), using two different models of free radical generation: a) iron-promoted
and b) ultrasound-induced lipid peroxidation. In a) lipid peroxidation was
assessed by determination of thiobarbituric acid-reactive substances (TBARS); in
b) by determination of conjugated dienes, formation of breakdown carbonyl
products (as 2,4-dinitrophenylhydrazones) and loss of native
phosphatidylcholine. In the iron-promoted (Fenton-driven) model, procyanidines
had a remarkable, dose-dependent antilipoperoxidant activity (IC50 = 2.5
mumol/l), more than one order of magnitude greater than that of the monomeric
unit catechin (IC50 = 50 mumol/l), activity which is due, at least in part, to
their metal-chelating properties. In the more specific model b), which
discriminates between the initiator (hydroxyl radical from water sonolysis) and
the propagator species of lipid peroxidation (the peroxyl radical, from
autooxidation of C-centered radicals), procyanidines are highly effective in
preventing conjugated diene formation in both the induction (IC50 = 0.1 mumol/l)
and propagation (IC50 = 0.05 mumol/l) phases (the scavenging effect of
alpha-tocopherol was weaker, with IC50 of 1.5 and 1.25 mumol/l). In addition,
procyanidines at 0.5 mumol/l markedly delayed the onset of the breakdown phase
(48 h), totally inhibiting during this time the formation of degradation
products (the lag-time induced by alpha-tocopherol was only of 24 h at 10
mumol/l concentration). The HO degrees entrapping capacity of these compounds
was further confirmed by UV studies and by electron spin resonance (ESR)
spectroscopy, using DMPO as spin trapper: procyanidines markedly reduced, in a
dose-dependent fashion, the signal intensity of the DMPO-OH radical spin adduct
(100% inhibition at 40 mumol/l). The results of the second part of this study
show that procyanidines, in addition to free radical scavenging action, strongly
and non-competitively, inhibit xanthine oxidase activity, the enzyme which
triggers the oxy radical cascade (IC50 = 2.4 mumol/l). In addition procyanidines
non-competitively inhibit the activities of the proteolytic enzymes collagenase
(IC50 = 38 mumol/l) and elastase (IC50 = 4.24 mumol/l) and of the glycosidases
hyaluronidase and beta-glucuronidase (IC50 = 80 mumol/l and 1.1 mumol/l),
involved in the turnover of the main structural components of the extravascular
matrix collagen, elastin and hyaluronic acid.(ABSTRACT TRUNCATED AT 400 WORDS)

PMID: 8024628 [PubMed - indexed for MEDLINE]

457: Plant Mol Biol. 1994 Mar;24(5):743-55.

Cloning and molecular analysis of structural genes involved in flavonoid and
stilbene biosynthesis in grape (Vitis vinifera L.).

Sparvoli F, Martin C, Scienza A, Gavazzi G, Tonelli C.

Dipartimento di Genetica e di Biologia dei Microorganismi, Universita degli
Studi di Milano, Italia.

Genes involved in flavonoid and stilbene biosynthesis were isolated from grape
(Vitis vinifera L.). Clones coding for phenylalanine ammonia-lyase (PAL),
chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase
(F3H), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin dioxygenase (LDOX)
and UDP glucose:flavonoid 3-O-glucosyl transferase (UFGT), were isolated by
screening a cDNA library, obtained from mRNA from seedlings grown in light for
48 h using snapdragon (Antirrhinum majus) and maize heterologous probes. A cDNA
clone coding for stilbene synthase (StSy) was isolated by probing the library
with a specific oligonucleotide. These clones were sequenced and when the
putative products were compared to the published amino acid sequence for
corresponding enzymes, the percentages of similarity ranged from 65% (UFGT) to
90% (CHS and PAL). The analysis of the genomic organization and expression of
these genes in response to light shows that PAL and StSy genes belong to large
multigene families, while the others are present in one to four copies per
haploid genome. The steady-state level of mRNAs encoded by the flavonoid
biosynthetic genes as determined in young seedlings is coordinately induced by
light, except for PAL and StSy, which appear to be constitutively expressed.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 8193299 [PubMed - indexed for MEDLINE]

458: Free Radic Biol Med. 1994 Mar;16(3):355-62.

Oxidative stress in recalcitrant tissue cultures of grapevine.

Benson EE, Roubelakis-Angelakis KA.

Department of Biology, University of Crete, Heraklion, Greece.

Thiobarbituric acid reactive substances (TBARS), and fluorescent compounds with
spectral characteristics typical of products associated with oxidative stress in
senescent and aging plant and animal cells, were detected in tissue cultures of
the recalcitrant grapevine Vitis vinifera L. cultivar, Sultanina. These
compounds increased during the early stages of dedifferentiation (callogenesis)
of nodal stem explants. Catalase activity was not detected in the original
explant, but was induced during callogenic dedifferentiation. Conversely,
superoxide dismutase activity was detectable in the original explant, but
diminished during the first week of callus induction. Transfer to callus
induction medium promoted a large increase in the sulfhydryl content of nodal
tissues. TBARS and fluorescent products accumulated in Sultanina callus during
long-term culture (over 6 months). The possibility that oxidative stress may
contribute to culture recalcitrance in this vine is discussed.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 8063199 [PubMed - indexed for MEDLINE]

459: Anal Biochem. 1993 Nov 1;214(2):484-9.

A narrow-bore HPLC method for the identification and quantitation of free,
conjugated, and bound polyamines.

Kotzabasis K, Christakis-Hampsas MD, Roubelakis-Angelakis KA.

Department of Biology, University of Crete, Heraklion, Greece.

A modified sensitive high-pressure liquid chromatography (HPLC) method, using a
narrow-bore column (2.1 x 200 mm, C-18, 5-microns particle size) with a
methanol:water gradient (55-84%, v/v), is described for direct analysis of
benzoylated free (S), soluble-conjugated (SH), and insoluble-bound (PH)
polyamines in plant tissues. Regression curves for each fraction of polyamines
allow a precise quantitative determination of putrescine, cadaverine,
spermidine, spermine, and agmatine in all fractions (S, SH, PH) of a crude
extract. A comparison of the results to the corresponding values for dansylated
polyamines separated by thin-layer chromatography and quantitated by fluorometry
is also presented. S, SH, and PH polyamines as benzoylated derivatives from
leaves of Vitis vinifera L. cv Dogridge were characterized and quantitatively
determined with this improved sensitive HPLC method.

PMID: 8109737 [PubMed - indexed for MEDLINE]

460: Plant Physiol. 1993 May;102(1):205-211.

Purification and Characterization of Geranyl Diphosphate Synthase from Vitis
vinifera L. cv Muscat de Frontignan Cell Cultures.

Clastre M, Bantignies B, Feron G, Soler E, Ambid C.

Ecole Nationale Superieure Agronomique, 145 Avenue de Muret, F-31076 Toulouse
Cedex, France.

A geranyl diphosphate synthase (EC 2.5.1.1), which catalyzes the formation of
geranyl diphosphate from dimethylallyl diphosphate and isopentenyl diphosphate,
was isolated from Vitis vinifera L. cv Muscat de Frontignan cell cultures.
Purification of the enzyme was achieved successively by ammonium sulfate
precipitation and chromatography on DEAE-Sephacel, hydroxylapatite, Mono Q,
Phenyl Superose, Superose 12, and preparative nondenaturing polyacrylamide gels.
The enzyme formed only geranyl diphosphate as a product. In all cases, neither
neryl diphosphate, the cis isomer, nor farnesyl diphosphate was detected. The
enzyme showed a native molecular mass of 68 [plus or minus] 5 kD as determined
by gel permeation. On sodium dodecyl sulfate polyacrylamide gels, geranyl
diphosphate synthase purified to electrophoretic homogeneity migrated with a
molecular mass of 66 [plus or minus] 2 kD. Michaelis constants for isopentenyl
diphosphate and dimethylallyl diphosphate were 8.5 and 56.8 [mu]M, respectively.
The enzyme required Mn2+ and Mg2+ as cofactors and its activity was enhanced by
Triton X-100. Inorganic pyrophosphate, aminophenylethyl diphosphate, and geranyl
diphosphate had inhibitory effects on the enzyme.

PMID: 12231811 [PubMed - as supplied by publisher]

461: Indian J Exp Biol. 1993 May;31(5):482-4.

Control of citrinin caused nephrotoxicosis through aqueous leaf extract of Vitis
vinifera L., mercurious corrossivus and cortisone.

Bilgrami KS, Jeswal P.

University Department of Botany, Bhagalpur University, India.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 8359859 [PubMed - indexed for MEDLINE]

462: Nature. 1993 Jan 14;361(6408):153-6.

Disease resistance results from foreign phytoalexin expression in a novel plant.

Hain R, Reif HJ, Krause E, Langebartels R, Kindl H, Vornam B, Wiese W, Schmelzer
E, Schreier PH, Stocker RH, et al.

Bayer AG, Institut fur Biotechnologie, Germany.

Although phytoalexins have long been inferred to be important in the defence of
plants against fungal infection, there are few reports showing that they provide
resistance to infection. Several plants, including grapevine, synthesize the
stilbene-type phytoalexin resveratrol when attacked by pathogens. Stilbenes with
fungicidal potential are formed in several unrelated plant species, such as
peanut (Arachis hypogaea), grapevine (Vitis vinifera) and pine (Pinus
sylvestris). Stilbene biosynthesis only specifically requires the presence of
stilbene synthase. Furthermore, the precursor molecules for the formation of
hydroxy-stilbenes are malonyl-CoA and p-coumaroyl-CoA, both present in plants.
To investigate the potential of stilbene biosynthetic genes in a strategy of
engineering pathogen resistance, we isolated stilbene synthase genes from
grapevine, where they are expressed at a high level, and transferred them into
tobacco. We report here that regenerated tobacco plants containing these genes
are more resistant to infection by Botrytis cinerea. This is, to our knowledge,
the first report of increased disease resistance in transgenic plants based on
an additional foreign phytoalexin.

PMID: 8421520 [PubMed - indexed for MEDLINE]

463: Res Microbiol. 1992 Oct;143(8):791-806.

[Determination of an ELISA test on tissues of grapevine affected by yellow
disease]

[Article in French]

Caudwell A, Kuszala C.

INRA, Station de Recherches sur les Mycoplasmes et les Arbovirus des Plantes,
Dijon, France.

We report here results concerning the best part of the grapevine to section, the
best sampling period and an important adaptation of the extraction media for the
diagnosis of a grapevine yellow disease by ELISA. The addition of Triton-X100,
or better of Chaps enabled us to get clear results for the sensitive Vitis
vinifera scion varieties. Until now, results have not been obtained for the
symptomless root stock varieties. However, antigen concentration by molecular
filtration may be a way to solve the problem of assaying root stocks.

Publication Types:
English Abstract
In Vitro

PMID: 1298032 [PubMed - indexed for MEDLINE]

464: Biochem J. 1992 Sep 1;286 ( Pt 2):623-6.

pH-induced kinetic co-operativity of a thylakoid-bound polyphenol oxidase.

Valero E, Garcia-Carmona F.

Departamento de Quimica-Fisica, Escuela Universitaria Politecnica de Albacete,
Universidad de Castilla, La Mancha, Albacete, Spain.

A study of the catecholase activity of a latent plant polyphenol oxidase,
extracted and purified from the chloroplast membranes of grapes (Vitis vinifera
cv. Airen), revealed for the first time a lag phase above pH 5.0, whereas a
steady-state rate was reached immediately when pH values were lower, thus
suggesting the hysteretic nature of the enzyme. During steady state, the enzyme
showed negative co-operativity concomitant with the presence of the lag period,
and followed classical Michaelis-Menten kinetics under more acid pH conditions.
Statistical analysis of these data showed a minimal value for the extreme Hill
coefficient of 0.54 at pH 6.0. This kinetic behaviour of polyphenol oxidase has
been interpreted in terms of the pH-induced 'slow' transition mechanism reported
by Ricard, Noat & Nari [(1984) Eur. J. Biochem. 145, 311-317] in which the
conformational change does not affect the active site of the enzyme.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 1530593 [PubMed - indexed for MEDLINE]

465: Plant Physiol. 1992 Aug;99(4):1619-1625.

Aberrant Processing of Polyphenol Oxidase in a Variegated Grapevine Mutant.

Rathjen AH, Robinson SP.

Commonwealth Scientific and Industrial Research Organization Division of
Horticulture, GPO Box 350, Adelaide, South Australia 5001 Australia.

Bruce's Sport is a mutant grapevine (Vitis vinifera L.) with green and white
variegated fruit derived from the Sultana variety. The white regions of tissue
have decreased polyphenol oxidase (PPO) activity resulting in a reduced capacity
for browning. Active PPO from Sultana grapes was purified and had an apparent
molecular weight of 40,000 on sodium dodecyl sulfate-polyacrylamide gel
electrophoresis. Western blots indicated that mature Sultana grapes contained a
single 40-kilodalton PPO, and young Sultana berries also had small quantities of
a 60-kilodalton protein. Bruce's Sport grapes had much less of the 40-kilodalton
PPO and greater amounts of the 60-kilodalton band. Protease digestion of Bruce's
Sport extracts decreased the proportion of the 60-kilodalton protein and
increased the 40-kilodalton band. A cDNA clone of grape PPO was used to probe a
northern blot of Sultana and Bruce's Sport RNA and hybridized to a 2.2-kilobase
transcript in both grapevines. The level of PPO mRNA was high in the early
stages of berry development but then declined. The results suggest that in
grapevine the active 40-kilodalton form of PPO is synthesized as a precursor
protein of at least 60 kilodaltons, and normal processing is interrupted in
Bruce's Sport resulting in the accumulation of the 60-kilodalton inactive
preform of PPO.

PMID: 16669082 [PubMed - as supplied by publisher]

466: J Chromatogr. 1992 Mar 20;595(1-2):269-81.

Analytical methods for monoterpene glycosides in grape and wine. II. Qualitative
and quantitative determination of monoterpene glycosides in grape.

Voirin SG, Baumes RL, Sapis JC, Bayonove CL.

INRA, Institut des Produits de la Vigne, Laboratoire des Aromes et des
Substances Naturelles, Montpellier, France.

Free and glycosidically bound terpenes of five Vitis vinifera grape cultivars
(muscat of Alexandria, muscat of Frontignan, muscat of Hamburg, muscat Ottonel
and Gewurztraminer) were investigated. The free and bound fractions were
separated by selective retention on Amberlite XAD-2 resin. The glycosidic
fractions were analysed by gas chromatography and gas chromatography-mass
spectrometry using either enzymic hydrolysis and subsequent analysis of the
released aglycones or trimethylsilyl (TMS) and trifluoroacetyl derivatives. The
known monoterpenyl, benzyl and 2-phenylethyl beta-D-glucopyranosides,
beta-rutinosides, 6-O-alpha-L-arabinofuranosyl-beta-D-glucopyranosides and
6-O-beta-D-apiofuranosyl-beta-D-glucopyranosides were determined. A number of
other glycosides were detected and the structures of some of them, mainly
apiosylglucosides and glucosides with aglycones in higher oxidation state than
linalol, were tentatively identified using the mass spectra of their TMS and TFA
derivatives and the results obtained from the analysis of their aglycones.

PMID: 1577909 [PubMed - indexed for MEDLINE]

467: Plant Physiol. 1992 Feb;98(2):774-776.

Hysteresis and Cooperative Behavior of a Latent Plant Polyphenoloxidase.

Valero E, Garcia-Carmona F.

Departamento de Quimica-Fisica, E. U. Politecnica de Albacete, Universidad de
Castilla-La Mancha, E-02006 Albacete, Spain.

Appearance of a lag period dependent on pH in the expression of the catecholase
activity of a polyphenoloxidase extracted in a latent state from Airen grape
(Vitis vinifera L.) berries, is revealed, suggesting the hysteretic nature of
the enzyme. The lag time was independent of enzyme concentration, indicating
that slow pH-induced conformational changes in the protein must occur during
assay. Results obtained by varying substrate concentration show that the system
presents hyperbolic or cooperative kinetics depending on the pH of the assay.

PMID: 16668711 [PubMed - as supplied by publisher]

468: Biochimie. 1992 Feb;74(2):143-8.

Oxidation of hydroquinone by both cellular and extracellular grapevine
peroxidase fractions.

Zapata JM, Calderon AA, Munoz R, Ros Barcelo A.

Department of Plant Biology (Plant Physiology), University of Murcia, Spain.

The oxidation of hydroquinone by two peroxidase (EC 1.11.1.7) fractions obtained
from the cells and spent medium of cell cultures of grapevine (Vitis vinifera cv
Monastrell) has been studied, and their comparative efficacy (kcat/KM ratio)
studied in both the H2O2-consuming and hydroquinone-consuming reactions. While
the efficacy in the H2O2-consuming reaction is practically identical for both
enzyme fractions, the cellular peroxidase has five-fold more efficacy in the
hydroquinone-consuming reaction than the peroxidase located in the spent medium.
Screening of cellular peroxidases capable of oxidizing hydroquinone on
polyacrylamide gels, by means of a staining reaction based on the nucleophilic
attack of 4-aminoantipyrine on p-benzoquinone in acidic media, reveals that all
the cellular peroxidase isoenzymes are capable of oxidizing hydroquinone,
probably yielding a quinone-diimine as a product of the staining reaction. Since
isoperoxidases found in cellular fractions are also present in the spent medium,
the values found for the different efficacies in the hydroquinone-consuming
reaction must be considered as the results of the different proportions in which
each peroxidase isoenzyme was found in the two fractions. The localization of a
benzoquinone-generating system of high efficacy inside the plant cell, and
probably located in vacuoles, is discussed with respect to the harmful role
which the quinone/semiquinone pair might play in cell death, as part of the
hypersensitive response expressed within the mechanism of plant disease
resistance.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 1316172 [PubMed - indexed for MEDLINE]

469: Plant Physiol. 1991 Dec;97(4):1483-1486.

Vacuolar Release of 1-(Malonylamino)cyclopropane-1-Carboxylic Acid, the
Conjugated Form of the Ethylene Precursor.

Pedreno MA, Bouzayen M, Pech JC, Marigo G, Latche A.

Ecole Nationale Superieure Agronomique, 145, avenue de Muret, F-31076 Toulouse
Cedex, France.

The mechanisms underlying the vacuolar retention or release of
1-(malonylamino)cyclopropane-1-carboxylic acid (MACC), the conjugated form of
the ethylene precursor, has been studied in grape (Vitis vinifera) cells grown
in vitro using the technique of compartmental analysis of radioisotope elution.
Following its accumulation in the vacuole, M[2,3-(14)C]ACC could be released
from cells when the vacuolar pH was artificially lowered by external buffers
from its initial value of 6.2 to below the critical pH of 5.5. Successive
release and retention of vacuolar MACC could be achieved by switching the
vacuolar pH from values lower and higher than 5.5. The rate constant of efflux
was highly correlated with the vacuolar pH. In plant tissues having low vacuolar
pH under natural conditions, e.g. apple fruits (pH 4.2) and mung bean hypocotyls
(pH 5.3), an efflux of M[2,3-(14)C]ACC also occurred. Its rate constant closely
corresponded to the theorical values derived from the correlation established
for grape cells. Evidence is presented that the efflux proceeded by passive
lipophilic membrane diffusion only when MACC was in the protonated form. In
contrast to other organic anions like malic acid, the mono and diionic species
could not permeate the tonoplast, thus indicating the strict dependence of MACC
retention upon the ionic status of the molecule and the absence of
carrier-mediated efflux.

PMID: 16668574 [PubMed - as supplied by publisher]

470: J Bacteriol. 1991 Oct;173(20):6547-52.

Polygalacturonase is a virulence factor in Agrobacterium tumefaciens biovar 3.

Rodriguez-Palenzuela P, Burr TJ, Collmer A.

Department of Plant Pathology, Cornell University, Ithaca, New York 14853-5908.

Agrobacterium tumefaciens biovar 3 causes both crown gall and root decay of
grapes. All biovar 3 strains, regardless of their tumorigenicity, produce in
culture a single polygalacturonase with a pI around 4.5. A. tumefaciens biovar 3
strain CG49 was mutagenized with Tn5 by using pSUP2021 as a suicide vector. A
mutant strain, CG50, lacking polygalacturonase activity was isolated. The
mutation was due to a single Tn5 insertion in an 8.5-kb EcoRI fragment that also
contained the polygalacturonase structural gene. The polygalacturonase-encoding
pehA gene was cloned in Escherichia coli by using the plasmid pBluescript as a
vector. Activity-stained isoelectric focusing gel analysis demonstrated that E.
coli cells harboring the pehA+ recombinant plasmid pCPP2067 produced a
polygalacturonase in culture with the same pI as the enzyme produced by CG49.
The pehA gene was localized within a 2.5-kb HindIII-SalI fragment. This fragment
was used as a probe in Southern hybridization analysis and showed that no
closely related genes are present in A. tumefaciens biovars 1 or 2, Rhizobium
leguminosarum, or Bradyrhizobium japonicum. The polygalacturonase mutant was
unable to induce root decay in grapes (Vitis vinifera cv. Chardonnay) and was
substantially less tumorigenic than the wild type in grape stems when low levels
of inoculum were used, although both strains were equally tumorigenic in potato
disc assays. The results indicate that polygalacturonase is a virulence factor
in both the root decay and crown gall incited in grapes by A. tumefaciens biovar
3.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 1655716 [PubMed - indexed for MEDLINE]

471: Plant Physiol. 1991 Sep;97(1):104-111.

Plant NAD(H)-Glutamate Dehydrogenase Consists of Two Subunit Polypeptides and
Their Participation in the Seven Isoenzymes Occurs in an Ordered Ratio.

Loulakakis KA, Roubelakis-Angelakis KA.

Department of Biology, University of Crete, P.O. Box 1470, 711 10 Heraklio,
Crete, Greece.

The structure and function of NAD(H)-glutamate dehydrogenase in plants was
studied by using grapevine (Vitis vinifera L. cv Sultanina) callus grown under
different nitrogen sources. The enzyme consists of two subunit-polypeptides,
alpha and beta, with similar antigenic properties but with different molecular
mass and charge. The two polypeptides have molecular masses of 43.0 and 42.5
kilodaltons, respectively. The holoenzyme is hexameric and is resolved into
seven isoenzymes by native gel electrophoresis. Two-dimensional native/SDS-PAGE
revealed that the 1 and 7 isoenzymes are homohexamers and the isoenzymes 2
through 6 are hybrids of the two polypeptides following an ordered ratio. The
total quantity of alpha- and beta-polypeptides and the isoenzymic pattern was
altered by the exogenous nitrogen source. The sample derived from callus grown
on nitrate or glutamic acid contained a slightly greater amount of
beta-polypeptide and of the more cathodal isoenzymes, whereas alpha-polypeptide
and the more anodal isoenzymes predominated in callus grown in the presence of
either ammonium or glutamine. The anabolic reaction was correlated with the
alpha- and the catabolic reaction with the beta-polypeptide; this could suggest
that each isoenzyme exhibits anabolic and catabolic function of different
magnitude. The isoenzymic patterns did not obey the expected binomial
distribution proportions.

PMID: 16668355 [PubMed - as supplied by publisher]

472: Mol Plant Microbe Interact. 1991 Mar-Apr;4(2):163-72.

Role of T-region borders in Agrobacterium host range.

Paulus F, Huss B, Tinland B, Herrmann A, Canaday J, Otten L.

Department of Molecular Plant Pathology, Institute of Plant Molecular Biology,
Strasbourg, France.

The limited host range AB3 strain of Agrobacterium tumefaciens induces tumors by
transferring two T-regions, TA and TB. TA is a deleted version of the well-known
biotype I octopine TL-region that lacks the iaa and ipt genes, but carries an
intact oncogene, gene 6b, and typical left and right border sequences. TB
carries two iaa genes that together code for the synthesis of indoleacetic acid.
Gene 6b and the iaa gene act synergistically when transferred in a coinoculation
experiment. The TA-region of the limited host range isolate Ag57 is related to
the TA-region of AB3, but differs from it at several positions. The most
significant difference is the absence of the right border region. In spite of
this, Ag57 and the exconjugant strain C58C9(pTiAg57) induce normal tumors on
Nicotiana rustica and Vitis vinifera. Various experiments indicate that gene 6b
of the Ag57 TA-region is active and transferred in spite of the absence of the
right border. On N. tabacum, C58C9(pTiAg57) is nononcogenic but becomes
oncogenic when the pTiAg57 TA-region is restored by the right TA border sequence
of pTiAB3. Thus, the right TA border sequence of the biotype III limited host
range strains is required for tumor induction on some hosts, but not on others.

Publication Types:
Comparative Study
Research Support, Non-U.S. Gov't

PMID: 1932812 [PubMed - indexed for MEDLINE]

473: FEBS Lett. 1990 Oct 1;271(1-2):236-8.

Prenyltransferase compartmentation in cells of Vitis vinifera cultivated in
vitro.

Feron G, Clastre M, Ambid C.

Ecole Nationale Superieure Agronomique, Toulouse, France.

Two prenyltransferases were located in cell cultures of Vitis vinifera. A
geranyl pyrophosphate synthase (EC 2.5.1.1) was associated with plastid-like
membranes whereas a farnesyl pyrophosphate synthase (EC 2.5.1.10) was found to
be soluble.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 2226808 [PubMed - indexed for MEDLINE]

474: Plant Physiol. 1990 Sep;94(1):109-113.

Immunocharacterization of NADH-Glutamate Dehydrogenase from Vitis vinifera L.

Loulakakis CA, Roubelakis-Angelakis KA.

Department of Biology, University of Crete, P. O. Box 1470, 711 10 Heraklion,
Greece.

Rabbit antiserum was raised against NADH-glutamate dehydrogenase (GDH) isoenzyme
1, purified from leaves of Vitis vinifera L. cv Soultanina and its specificity
was tested. This antiserum was used for immunocharacterization of the GDH from
leaf, shoot, and root tissues. The antiserum recognized the seven isoenzymes of
NADH-GDH and precipitated all the enzyme activity from the three tissues tested.
Western blot following SDS-PAGE revealed the same protein band for the three
tissues, with a molecular mass of 42.5 kilodaltons corresponding to NADH-GDH
subunit. Results, based on the immunological studies, revealed that NADH-GDH
from leaf, shoot, and root tissues are closely related proteins. Furthermore,
addition of ammonium ions to the culture medium of in vitro grown explants
resulted in a significant increase in NADH-GDH activity in root, shoot, and leaf
tissues.

PMID: 16667675 [PubMed - as supplied by publisher]

475: Acta Physiol Pharmacol Bulg. 1990;16(3):50-4.

Antiexudative and capillaritonic effects of procyanidines isolated from grape
seeds (V. Vinifera).

Zafirov D, Bredy-Dobreva G, Litchev V, Papasova M.

Institute of Physiology, Bulgarian Academy of Sciences, Sofia.

A purified and enriched fraction, containing procyanidines, was isolated from
grape seeds of Bulgarian sorts of Vitis Vinifera. The effects of procyanidines
on the local oedema, produced by subplantar injection of carrageenin and dextran
in the hind rat paw, were studied. We also tested their effect on the capillary
permeability using intravenous injection of Evans blue and local irritation by
xylene. Procyanidines at a dose of 2 mg/kg applied orally three times daily for
6 days inhibited the carrageenin-induced hind paw oedema. On the same schedule
of administration these compounds inhibited the dextran-induced oedema 4 hours
after the development of the process. Procyanidines stabilized the capillary
wall and prevented the increase of capillary permeability caused by local
cutaneous application of xylene.

PMID: 1714228 [PubMed - indexed for MEDLINE]

476: Plant Physiol. 1989 Dec;91(4):1501-1506.

Effects of Ambient and Acute Partial Pressures of Ozone on Leaf Net CO(2)
Assimilation of Field-Grown Vitis vinifera L.

Roper TR, Williams LE.

Department of Viticulture and Enology, University of California, Davis,
California 93648.

Mature, field-grown Vitis vinifera L. grapevines grown in open-top chambers were
exposed to either charcoal-filtered air or ambient ozone partial pressures
throughout the growing season. Individual leaves also were exposed to ozone
partial pressures of 0.2, 0.4, or 0.6 micropascals per pascal for 5 hours. No
visual ozone damage was found on leaves exposed to any of the treatments.
Chronic exposure to ambient O(3) partial pressures reduced net CO(2)
assimilation rate (A) between 5 and 13% at various times throughout the season
when compared to the filtered treatment. Exposure of leaves to 0.2 micropascals
per pascal O(3) for 5 hours had no significant effect on A; however, A was
reduced 84% for leaves exposed to 0.6 micropascals per pascal O(3) when compared
to the controls after 5 hours. Intercellular CO(2) partial pressure (c(i)) was
lower for leaves exposed to 0.2 micropascals per pascal O(3) when compared to
the controls, while c(i) of the leaves treated with 0.6 micropascals per pascal
of 0(3) increased during the fumigation. The long-term effects of ambient O(3)
and short-term exposure to acute levels of O(3) reduced grape leaf
photosynthesis due to a reduction in both stomatal and mesophyll conductances.

PMID: 16667208 [PubMed - as supplied by publisher]

477: Plant Physiol. 1989 Apr;89(4):1136-1140.

Net CO(2) Assimilation and Carbohydrate Partitioning of Grapevine Leaves in
Response to Trunk Girdling and Gibberellic Acid Application.

Roper TR, Williams LE.

Department of Viticulture and Enology, University of California, Davis,
California 93648.

Leaf net CO(2) assimilation rate (A), stomatal conductance (g(s)), carboxylation
efficiency, and foliar nonstructural carbohydrates were measured on mature,
field-grown Vitis vinifera L. (cv Thompson Seedless) vines that had been trunk
girdled, sprayed with gibberellic acid, or both, shortly after anthesis.
Girdling reduced A, g(s), and carboxylation efficiency when measured 2 weeks
after imposition of the treatments. Diurnal measurements indicated that A of
girdled vines was less than that of control vines between 1000 and 1800 hours.
Gibberellic acid mitigated the depressing effect of girdling on g(s) during the
same diurnal measurements. The concentrations of foliar carbohydrates were
greatest for the girdled vines, followed by the combination treatment and were
lowest for the control vines. Foliar carbohydrates were greater for girdled
vines 4 weeks after the treatments were imposed, however, by this time there was
no significant difference in A between the control and girdled vines. Two and 4
weeks after the experiment was initiated root carbohydrate concentrations were
less for the girdled vines when compared to the control vines. The data indicate
that the reduction in A of girdled grapevines is not associated with the
accumulation of leaf nonstructural carbohydrates following the girdling
treatment.

PMID: 16666676 [PubMed - as supplied by publisher]

478: Ann Pharm Fr. 1989;47(4):229-34.

[Flavonoids of 3 cultivars vine leaves, Vitis vinifera L. var. tinctoria
(Alicante, Carignan, Grand noir). Value in chemical control]

[Article in French]

Diaz Lanza AM, Elias R, Maillard C, Faure R, de Sotto M, Balansard G.

Three flavonoids were isolated and identified from the leaves of 3 cultivars
vine Vitis vinifera L. var. tinctoria (Alicante, Carignan and Grand noir):
hyperin, isoquercitrin and quercetin 3-O-beta-D glucuronic acid. TLC analytic
control and HPLC determination are proposed in this paper.

Publication Types:
English Abstract

PMID: 2637643 [PubMed - indexed for MEDLINE]

479: Plant Physiol. 1988 Nov;88(3):718-724.

Resistance to Water Transport in Shoots of Vitis vinifera L. : Relation to
Growth at Low Water Potential.

Schultz HR, Matthews MA.

Department of Viticulture and Enology, University of California, Davis,
California 95616.

Apparent resistances to water transport in the liquid phase were determined from
measurements of soil, root, basal shoot internode, shoot apex, and leaf water
potentials and water flux in Vitis vinifera (cv White Riesling) during soil
drying. Predawn water potential differences (DeltaPsi) in the shoots accounted
for 20% of the total DeltaPsi between the soil and the shoot apex when plants
were well-watered but increased to about 90% when shoot growth ceased. The
DeltaPsi from soil to root was essentially constant during this period. At low
water potential, the DeltaPsi in the shoot was persistent when transpiration was
low (predawn) or completely prevented (plant bagging). The apparent hydraulic
resistance between the basal shoot internode and most rapidly expanding leaf (or
shoot apex) increased several-fold when water was withheld. Leaf and internode
expansion both exhibited high sensitivity to increasing hydraulic resistance.
Measurements of pneumatic resistance to air flow through frozen internode
segments indicated progressive vapor-filling of vessels as soil drying
progressed. From these observations and others in the literature, it was
suggested that embolization may be a common occurrence and play an important
role in the inhibition of shoot growth at moderate water deficits.

PMID: 16666373 [PubMed - as supplied by publisher]

480: Plant Physiol. 1987 Aug;84(4):1166-1171.

Dynamic Relation between Expansion and Cellular Turgor in Growing Grape (Vitis
vinifera L.) Leaves.

Shackel KA, Matthews MA, Morrison JC.

Department of Viticulture and Enology, University of California, Davis,
California 95616.

Measurements of the growth and water relations of expanding grape (Vitis
vinifera L.) leaves have been used to determine the relationship between leaf
expansion rate and leaf cell turgor. Direct measurement of turgor on the small
(approximately 15 micrometer diameter) epidermal cells over the midvein of
expanding grape leaves was made possible by improvements in the pressure probe
technique. Leaf expansion rate and leaf water status were perturbed by
environmentally induced changes in plant transpiration. After establishing a
steady state growth rate, a step decrease in plant transpiration resulted in a
rapid and large increase in leaf cell turgor (0.25 megapascal in 5 minutes), and
leaf expansion rate. Subsequently, leaf expansion rate returned to the original
steady state rate with no change in cell turgor. These results indicate that the
expansion rate of leaves may not be strongly related to the turgor of the leaf
cells, and that substantial control of leaf expansion rate, despite changes in
turgor, may be part of normal plant function. It is suggested that a strictly
physical interpretation of the parameters most commonly used to describe the
relationship between turgor and growth in plant cells (cell wall extensibility
and yield threshold) may be inappropriate when considering the process of plant
cell expansion.

PMID: 16665579 [PubMed - as supplied by publisher]

481: Plant Physiol. 1987 Jun;84(2):381-385.

Effects of Chilling and ABA on [H]Gibberellin A(4) Metabolism in Somatic Embryos
of Grape (Vitis vinifera L. x V. rupestris Scheele).

Pearce D, Pharis RP, Rajasekaran K, Mullins MG.

Plant Physiology Research Group, Department of Biology, University of Calgary,
Calgary, Alberta, Canada T2N 1N4.

Previous work has indicated that changes in gibberellin (GA) metabolism may be
involved in chilling-induced release from dormancy in somatic embryos of grape
(Vitis vinifera L. x V. rupestris Scheele). We have chilled somatic embryos of
grape for 2, 4, or 8 weeks, then incubated them with [(3)H]GA(4) (of high
specific activity, 4.81 x 10(10) becquerel per millimole) for 48 hours at 26
degrees C. Chilling had little effect on the total amount of free [(3)H]GA-like
metabolites formed during incubation at 26 degrees C, but did change the
relative proportions of individual metabolites. The amount of highly
water-soluble [(3)H] metabolites formed at 26 degrees C decreased in embryos
chilled for 4 or 8 weeks. The concentration of endogenous GA precursors (e.g.,
GA(12) aldehyde-, kaurene-, and kaurenoic acid-like substances) increased in
embryos chilled for 4 or 8 weeks. Treatment with abscisic acid (ABA) (known to
inhibit germination in grape embryos) concurrent with [(3)H]GA(4) treatment at
26 degrees C, reduced the uptake of [(3)H] GA(4) but had little effect on the
qualitative spectrum of metabolites. However, in the embryos chilled for 8 weeks
and then treated with ABA for 48 hours at 26 degrees C, there was a higher
concentration of GA precursors than in untreated control embryos. Chilled
embryos thus have an enhanced potential for an increase in free GAs through
synthesis from increased amounts of GA precursors, or through a reduced ability
to form highly water-soluble GA metabolites (i.e., GA conjugates or
polyhydroxylated free GAs).

PMID: 16665447 [PubMed - as supplied by publisher]

482: Br J Nutr. 1987 May;57(3):331-43.

The effects of fruit juices and fruits on the absorption of iron from a rice
meal.

Ballot D, Baynes RD, Bothwell TH, Gillooly M, MacFarlane BJ, MacPhail AP, Lyons
G, Derman DP, Bezwoda WR, Torrance JD, et al.

The effects of the chemical composition of fruit juices and fruit on the
absorption of iron from a rice (Oryza sativa) meal were measured in 234 parous
Indian women, using the erythrocyte utilization of radioactive Fe method. The
corrected geometric mean Fe absorptions with different juices varied between
0.040 and 0.129, with the variation correlating closely with the ascorbic acid
contents of the juices (rs 0.838, P less than 0.01). Ascorbic acid was not the
only organic acid responsible for the promoting effects of citrus fruit juices
on Fe absorption. Fe absorption from laboratory 'orange juice' (100 ml water, 33
mg ascorbic acid and 750 mg citric acid) was significantly better than that from
100 ml water and 33 mg ascorbic acid alone (0.097 and 0.059 respectively), while
Fe absorption from 100 ml orange juice (28 mg ascorbic acid) was better than
that from 100 ml water containing the same amount of ascorbic acid (0.139 and
0.098 respectively). Finally, Fe absorption from laboratory 'lemon juice' (100
ml orange juice and 4 g citric acid) was significantly better than that from 100
ml orange juice (0.226 and 0.166 respectively). The corrected geometric mean Fe
absorption from the rice meal was 0.025. Several fruits had little or no effect
on Fe absorption from the meal (0.013-0.024). These included grape (Vitis
vinifera), peach (Prunus persica), apple (Malus sylvestris) and avocado pear
(Persea americana). Fruit with a mild to moderate enhancing effect on Fe
absorption (0.031-0.088) included strawberry (Fragaria sp.) (uncorrected
values), plum (Prunus domestica), rhubarb (Rheum rhaponticum), banana (Musa
cavendishii), mango (Mangifera indica), pear (Pyrus communis), cantaloup
(Cucumis melo) and pineapple (Ananas comosus) (uncorrected values). Guava
(Psidium guajava) and pawpaw (Carica papaya) markedly increased Fe absorption
(0.126-0.293). There was a close correlation between Fe absorption and the
ascorbic acid content of the fruits tested (rs 0.738, P less than 0.0001). There
was also a weaker but significant correlation with the citric acid content (rs
0.55, P less than 0.03). Although this may have reflected a direct effect of
citric acid on Fe absorption, it should be noted that fruits containing citric
acid also contained ascorbic acid (rs 0.70, P less than 0.002).(ABSTRACT
TRUNCATED AT 400 WORDS)

PMID: 3593665 [PubMed - indexed for MEDLINE]

483: Plant Foods Hum Nutr. 1987;37(3):275-81.

Nutritive value of different grape musts (Vitis vinifera L.).

Juhasz O, Dworschak E, Kozma P.

Department of Viticulture, University of Horticulture, Budapest, Hungary.

Musts of 11 different grape sorts were analyzed for titratable acids, total
sugars, nitrogen, vitamin B group (B1, B2, B6, pantothenic acid) and macro- and
micro elements (K, Ca, Mg, Fe, Zn, Cu, Mn, Mo). Analyses were done on samples
collected in 1983 and 1984. The nutritive value of each sample was characterised
by indices based on the nutrient concentrations and recommended dietary
allowances (RDA). The nutritive values of the various grape must sorts were
generally similar to those of the following vintage. Favourable climatic
conditions had a positive effect on the nutritive values.

PMID: 3507685 [PubMed - indexed for MEDLINE]

484: Plant Physiol. 1986 Dec;82(4):1154-1157.

Presence and Identification of Polyamines in Xylem and Phloem Exudates of
Plants.

Friedman R, Levin N, Altman A.

Department of Horticulture, The Hebrew University of Jerusalem, P. O. Box 12,
Rehovot 76100, Israel.

Polyamines were identified by high performance liquid chromatography
(benzoylation) and by thin layer chromatography (dansylation) in xylem exudates
from stems of sunflowers (Helianthus annuus [L.]), mung bean (Vigna radiata [L.]
Wilczek), grapevine (Vitis vinifera [L.] cv Grenache), and orange (Citrus
sinensis [L.] Osbeck, cv Valencia), as well as in phloem sap (using elution into
EDTA) of sunflower and mung bean plants. Putrescine was the major polyamine
detected, ranging in concentrations of 150 to 9200 picomoles per milliliter
exudate, whereas only trace amounts of spermine were detected. High amounts of
putrescine and spermidine were found in EDTA eluates (possibly phloem sap) as
compared with elution into water. Concentrations of putrescine and spermidine in
xylem exudates were related to the physiological conditions of the plants prior
to exudate collection. More putrescine was found in exudates of older than in
younger sunflower plants, and salt stress applied to sunflower plants resulted
in a higher concentration of putrescine and spermidine in the exudate. The
presence and abundance of putrescine and spermidine in xylem and phloem exudates
indicate that polyamines may be translocated in plants. This long-distance
translocation further supports the hypothesis that polyamines have a regulatory
role in plant growth and response to stress.

PMID: 16665153 [PubMed - as supplied by publisher]

485: Virology. 1986 Nov;155(1):39-45.

Separation of viroid RNAs by cellulose chromatography indicating conformational
distinctions.

Semancik JS.

A series of viroids and viroid-like RNAs extracted from citron (Citrus medica),
grapevine (Vitis vinifera), and avocado (Persea americanum) displayed
differential binding capacities to cellulose in the presence of ethanol. This
nonionic interaction was influenced by the presence of magnesium ions suggesting
variations in conformation among the viroid RNAs and corresponding differences
in reactivity with cellulose. The specific elution profiles present a basis for
the subclassification of groups of viroid-like molecules.

Publication Types:
Research Support, Non-U.S. Gov't
Research Support, U.S. Gov't, Non-P.H.S.

PMID: 3776103 [PubMed - indexed for MEDLINE]

486: Plant Physiol. 1986 Aug;81(4):1140-1142.

Changes in the Activity of Catalase (EC 1.11.1.6) in Relation to the Dormancy of
Grapevine (Vitis vinifera L.) Buds.

Nir G, Shulman Y, Fanberstein L, Lavee S.

Department of Olei and Viticulture, Agricultural Research Organization, Volcani
Center, Bet Dagan 50-250, Israel.

Catalase activity in grapevine (Vitis vinifera L.) buds cv. ;Perlette.'
increased to a maximum in October and thereafter decreased within 3 months to
less than half its maximal rate. The decrease in catalase activity coincided
with the decline in temperature during winter. The rate of sprouting of buds
forced at 23 degrees C was negatively related to the activity of catalase.
Artificial chilling of grapevine canes at 5 degrees C resulted in a 25% decrease
of catalase activity in the buds after 3 days and 31% after 17 days. The
activity of catalase increased to the control level only 96 hours after removing
canes from 5 degrees C to room temperature. Efficient buddormancy breaking
agents, such as thiourea and cyanamide decreased catalase activity to 64 and 50%
of the controls respectively, while the activity of peroxidase remained the same
under those conditions. A less efficient dormancy breaking agent
dinitro-ortho-cresol, did not decrease catalase activity.

PMID: 16664957 [PubMed - as supplied by publisher]

487: Ecotoxicol Environ Saf. 1984 Jun;8(3):208-14.

The effect of atmospheric pollution on Vitis vinifera L. pollen ultrastructure
under natural conditions.

Stirban M, Craciun C, Bathory D, Cipleu D.

The ultrastructural modification of pollen grains in Vitis vinifera L. variety
and hybrids in areas of SO2 atmospheric pollution (the main polluting SO2
usually reaches 2.72 mg/m3), nitrogen oxide, and other gases derived from
noniron metal processing factories have been studied. Strains 1001 and 1002,
resistant varieties, do not undergo ultrastructural modifications. Neuburger and
Issabelle, medium resistant ones, have a heterogeneity in ultrastructural
organization from normal forms to forms having both wall covers as well as the
main organelles changed.

PMID: 6734498 [PubMed - indexed for MEDLINE]

488: Food Chem Toxicol. 1984 Apr;22(4):309-13.

Mutagen content of table wines made from various grape species and hybrid
cultivars.

Subden RE, Krizus A, Rancourt D.

The mutagen content of wines produced from the European Vitis vinifera grapes
was compared with that of wines produced from hybrids of V. vinifera and species
indigenous to North America. Mutagens were extracted on an XAD-2 Amberlite resin
column and activated with S-9 and/or faecalase in the Salmonella/microsomal
mutagen assay. All white wines had insignificant mutagen levels. The only red
wine to produce statistically significant reversion frequencies was that made
from the Concord grape. The mutagens were shown to be extracted from the grape
skins during fermentation.

Publication Types:
Research Support, Non-U.S. Gov't

PMID: 6373529 [PubMed - indexed for MEDLINE]

489: Plant Physiol. 1983 Nov;73(3):803-808.

Endogenous Gibberellin-Like Substances in Somatic Embryos of Grape (Vitis
vinifera x Vitis rupestris) in Relation to Embryogenesis and the Chilling
Requirement for Subsequent Development of Mature Embryos.

Takeno K, Koshioka M, Pharis RP, Rajasekaran K, Mullins MG.

Department of Biology, University of Calgary, Calgary, Alberta T2N 1N4 Canada.

Endogenous gibberellin (GA)-like substances were examined in suspension cultures
of somatic embryos of a hybrid grape (Vitis vinifera x Vitis rupestris) during
embryogenesis, and in mature embryos chilled at 4 degrees C, and subsequently
incubated at 26 degrees C with and without abscisic acid (ABA). The extract was
separated into a nonpolar fraction (would contain GA-precursors); a fraction
that would contain free GAs; and a highly H(2)O-soluble fraction (would contain
GA glucosyl conjugates and very polar free GAs). Quantitation after SiO(2)
partition chromatography was accomplished by microdrop and immersion dwarf rice
bioassays. As embryogenesis developed, the free and highly H(2)O-soluble GA-like
substances, expressed on a dry weight basis, decreased (however, they increased
on a per embryo basis). Chilling at 4 degrees C for 1 week greatly increased
activity of free GA-like substances (per g dry weight and per embryo), it then
declined over the next three weeks of chilling. Activity (per g dry weight and
per embryo) in the H(2)O-soluble fraction declined throughout chilling. Activity
in the GA-precursor fraction, however, increased steadily with chilling (per g
dry weight and per embryo). Incubation at 26 degrees C after chilling enhanced
activity in the free GA and H(2)O-soluble fractions (per g dry weight and per
embryo), but activity in the GA-precursor fraction dropped dramatically.
Incubation at 26 degrees C with (+/-) ABA after chilling prevented germination
and maintained high activity for GA precursors and less polar free GAs and low
activity in the polar free GA and H(2)O-soluble fractions.Kaurene and kaurenoic
acid were characterized in the GA-precursor fraction of chilled embryos by
gas-liquid chromatography-mass spectrometry (GLC-MS). The existence of GA(4) and
GA(9) in ABA-treated, chilled embryos was also confirmed by GLC-MS.

PMID: 16663304 [PubMed - as supplied by publisher]

490: Plant Physiol. 1983 Nov;73(3):582-585.

Pathway of Photosynthetic Malate Formation in Vitis vinifera, a C(3) Plant.

Ruffner HP, Brem S, Rast DM.

Institute of Plant Biology, University of Zurich, Zollikerstrasse 107, CH-8008
Zurich, Switzerland.

The time course of intramolecular isotope distribution in phosphoglyceric acid
and serine was determined after exposure of grape leaf discs (Vitis vinifera L.)
to (14)CO(2) (1000 microliters per liter) for variable metabolic periods, and
the labeling patterns were compared with the respective isotope distribution in
the C(1-3) fragment of malic acid. The results clearly support the classical
concept of a close precursor-product relationship between photosynthetic
phosphoglycerate and malic acid. Under the assimilatory conditions used in this
study, there was no indication of an immediate carbon transfer from serine to
malate as has been suggested for C(3) plants (Kent et al. 1974 Plant Physiol 53:
491-495) because of a coincident labeling of these compounds in Vicia faba.
According to our data, previous evidence in favor of this hypothetical pathway
is based largely on an unusual (14)C distribution in serine, due to an extreme
suppression of photorespiration, as well as on arbitrary comparisons between
compounds of divergent kinetic characteristics and consequently different
degrees of metabolic label randomization.

PMID: 16663263 [PubMed - as supplied by publisher]

491: Plant Physiol. 1983 Nov;73(3):579-581.

Determination of C Distribution in Photosynthetic Serine and Phosphoglycerate
from Grape Leaves.

Brem S, Ruffner HP, Rast DM.

Institute of Plant Biology, University of Zurich, Zollikerstrasse 107, CH-8008
Zurich, Switzerland.

Serine and phosphoglyceric acid are the classical marker intermediates of
photorespiration and reductive carbon assimilation in C(3) plants. The present
paper introduces a new and fast method for the determination of (14)C
distribution in these compounds by selective elimination of C-3 (NaIO(4)) or C-1
(ninhydrin/ceric sulfate). Reproducibility of the procedure was found to be
better than +/-1% upon degradation of [U-(14)C]serine and [U-(14)C]glycerate
standards.Determination of labeling patterns in (dephosphorylated)
phosphoglycerate and serine, extracted from grape leaf discs (Vitis vinifera L.)
after administration of (14)CO(2) for 1 minute showed that (14)C distribution in
serine changes considerably with changing assimilatory conditions.
Interpretations of data basing on unusual labeling patterns of serine must
therefore be accepted with caution.

PMID: 16663262 [PubMed - as supplied by publisher]

492: J Pharm Belg. 1983 Jan-Feb;38(1):41-6.

[Anthocyanosides extracted from Vitis vinifera, Vaccinium myrtillus and Pinus
maritimus. I. Elastase-inhibiting activities in vitro. II. Compared
angioprotective activities in vivo]

[Article in French]

Jonadet M, Meunier MT, Bastide J, Bastide P.

Publication Types:
Comparative Study
English Abstract
In Vitro

PMID: 6553084 [PubMed - indexed for MEDLINE]

493: Plant Physiol. 1978 Sep;62(3):344-347.

Enzymes of Krebs-Henseleit Cycle in Vitis vinifera L: III. In Vivo and In Vitro
Studies of Arginase.

Roubelakis KA, Kliewer WM.

Department of Viticulture and Enology, University of California, Davis,
California 95616.

The presence of arginase (EC 3.5.3.7) in various tissues from Vitis vinifera L.
cultivars was demonstrated by both in vivo and in vitro enzyme assays. Initial
velocities determined by the two methods were in close agreement. Optimum
conditions for maximum enzyme activity were 25 to 30 millimolar l-arginine,
about 1 millimolar Mn(2+) (pH 9.4 to 9.8), and incubation temperature of 37 to
38 C. l-Arginine hydrolysis was linear with increasing sliced fresh tissue up to
500 milligrams for in vivo assay, and with enzyme extract equivalent up to about
200 milligrams of fresh tissue for in vitro. Similarly, l-arginine hydrolysis
was linear with incubation time for the first 45 minutes for in vivo assay and
for the first 20 minutes for in vitro.

PMID: 16660514 [PubMed - as supplied by publisher]

494: Plant Physiol. 1978 Sep;62(3):340-343.

Enzymes of Krebs-Henseleit Cycle in Vitis vinifera L: II. Arginosuccinate
Synthetase and Lyase.

Roubelakis KA, Kliewer WM.

Department of Viticulture and Enology, University of California, Davis,
California 95616.

Arginosuccinate (ASA) synthetase and lyase activities were detected in extracts
from Vitis vinifera L. cv. Chenin blanc mature leaves and seedlings. Optimum
reaction conditions for ASA synthetase were 10 millimolar l-citrulline, 7.5
millimolar l-aspartate, 3 to 4 millimolar ATP, 12 millimolar Mg(2+) (pH 7.5 to
8.0), enzyme extract up to equivalent of about 200 milligrams of fresh tissue,
and incubation temperature of 38 to 40 C. Optimum reaction conditions for ASA
lyase were 4 millimolar ASA-K salt (pH 7.3 to 7.8), amount of extract up to
equivalent of about 180 milligrams of fresh tissue, and incubation temperature
of 38 to 40 C.

PMID: 16660513 [PubMed - as supplied by publisher]

495: Plant Physiol. 1978 Sep;62(3):337-339.

Enzymes of Krebs-Henseleit Cycle in Vitis vinifera L: I. Ornithine
Carbamoyltransferase: Isolation and Some Properties.

Roubelakis KA, Kliewer WM.

Department of Viticulture and Enology, University of California, Davis,
California 95616.

Ornithine carbamoyltransferase (OCT) activity was detected in extracts from
mature leaves, fruit, germinating seeds, and seedlings of Vitis vinifera L.
Michaelis-Menten constants for OCT were 3.5 millimolar for carbamyl phosphate
and 5.5 millimolar for l-ornithine. Concentrations of l-ornithine greater than
10 millimolar slightly inhibited the enzyme, whereas carbamyl phosphate at
concentrations greater than the optimal (about 10 millimolar) did not affect OCT
activity. l-Citrulline formation was linear with incubation period for the first
25 minutes and with increasing amounts of enzyme up to an equivalent of about
200 milligrams of fresh tissue. The optimum pH for in vitro OCT activity was
between 8.4 and 8.8, and the optimum incubation temperature was 38 C.

PMID: 16660512 [PubMed - as supplied by publisher]

496: Plant Physiol. 1978 Jan;61(1):127-130.

Control of Flowering in the Grapevine (Vitis vinifera L.): Formation of
Inflorescences in Vitro by Isolated Tendrils.

Srinivasan C, Mullins MG.

Department of Agronomy and Horticultural Science, University of Sydney, New
South Wales, 2006, Australia.

Tendrils produced from shoot tips of grapevine (Vitis vinifera L.) cultured in
vitro on Nitsch's medium developed into inflorescences when 5 to 10 mum
benzyladenine (BA) or 6-(benzylamino)-9-(2-tetrahydropyranyl)-9H-purine (PBA)
were applied directly to the tendril tips. Inflorescences did not form on
tendrils if the cytokinins were supplied in the agar. Tendrils cultured in
agitated liquid medium containing BA, PBA, or zeatin riboside showed profuse
branching and tendrils were transformed into inflorescences. Calyx and corolla
(calyptra) stamens and pistils developed normally in the presence of both zeatin
riboside and PBA, but micro- and macrosporogenesis were absent.Inflorescences
were formed by tendrils from five cultivars (Muscat of Alexandria, Shiraz,
Carbernet Sauvignon, Wortley Hall, and Sultana syn. Thomson Seedless) and also
on tendrils from 12- to 15-week-old-seedlings.

PMID: 16660225 [PubMed - as supplied by publisher]

497: Plant Physiol. 1977 Oct;60(4):543-547.

Factors Affecting the Extraction of Intact Ribonucleic Acid from Plant Tissues
Containing Interfering Phenolic Compounds.

Newbury HJ, Possingham JV.

CSIRO Division of Horticultural Research, Box 350, G.P.O., Adelaide, South
Australia, 5001.

Using conventional methods it is impossible to extract RNA as uncomplexed intact
molecules from the leaves of grapevines (Vitis vinifera L.) and from a number of
woody perennial species that contain high levels of reactive phenolic compounds.
A procedure involving the use of high concentrations of the chaotropic agent
sodium perchlorate prevents the binding of phenolic compounds to RNA during
extraction. Analyses of the phenolics present in plant tissues used in these
experiments indicate that there is a poor correlation between the total phenolic
content and the complexing of RNA. However, qualitative analyses suggest that
proanthocyanidins are involved in the tanning of RNA during conventional
extractions.

PMID: 16660134 [PubMed - as supplied by publisher]

498: Plant Physiol. 1976 Oct;58(4):468-472.

Changes in Anthocyanin and Phenolics Content of Grapevine Leaf and Fruit Tissues
Treated with Sucrose, Nitrate, and Abscisic Acid.

Pirie A, Mullins MG.

Department of Agronomy and Horticultural Science, University of Sydney, New
South Wales, 2006 Australia.

Sucrose (0.04 to 0.12 m) induces accumulation of both total phenolics and
anthocyanin in leaf discs of grapevine (Vitis vinifera L.) incubated in
intermittent light. Abscisic acid (20 muM) and 2-chloroethyl phosphonic acid (60
mum) act synergistically with the sucrose to enhance its induction of both total
phenolics and anthocyanin. The magnitude of this interaction depends on leaf
age. Nitrate (30 mm) inhibits sucrose induction of phenolics and anthocyanin.
Levels of total phenolics and anthocyanin changed independently.

PMID: 16659699 [PubMed - as supplied by publisher]

499: Plant Physiol. 1975 Sep;56(3):370-372.

The Influence of Temperature on Malic Acid Metabolism in Grape Berries: I.
Enzyme Responses.

Lakso AN, Kliewer WM.

Department of Viticulture and Enology, University of California, Davis,
California 95616.

Phosphoenolpyruvate (PEP) carboxylase activity in immature ;Carignane' grape
berries (Vitis vinifera L.) had a temperature optimum of about 38 C, whereas
malic enzyme activity rose with increasing temperature between 10 and 46 C. In
vitro temperature inactivation rates for the PEP carboxylase were markedly
greater than for the malic enzyme activity. From the simultaneous action of
malic acid-producing enzymes (PEP carboxylase and malic dehydrogenase) and malic
acid-degradating enzyme (malic enzyme) systems at different temperatures, the
greatest tendency for malic acid accumulation in immature grape berries was at
20 to 25 C. Time-course measurements of enzymic activity from heated, intact
berries revealed greater in vivo temperature stability for the malic enzyme
activity than for the PEP carboxylase activity.

PMID: 16659305 [PubMed - as supplied by publisher]

500: Plant Physiol. 1974 Feb;53(2):258-260.

Rapid Estimates of Relative Water Content.

Smart RE.

Department of Agriculture, Viticultural Research Station, Griffith, New South
Wales, Australia.

Relative water content may be accurately estimated using the ratio of tissue
fresh weight to tissue turgid weight, termed here relative tissue weight. That
relative water content and relative tissue weight are linearly related is
demonstrated algebraically. The mean value of r(2) for grapevine (Vitis vinifera
L. cv. Shiraz) leaf tissue over eight separate sampling occasions was 0.993.
Similarly high values were obtained for maize (Zea mays cv. Cornell M-3) (0.998)
and apple (Malus sylvestris cv. Northern Spy) (0.997) using a range of leaf
ages. The proposal by Downey and Miller (1971. Rapid measurements of relative
turgidity in maize (Zea mays L.). New Phytol. 70: 555-560) that relative water
content in maize may be estimated from water uptake was also investigated for
grapevine leaves; this was found to be a less reliable estimate than that
obtained with relative tissue weight. With either method, there is a need for
calibration, although this could be achieved for relative tissue weight at least
with only a few subsamples.

PMID: 16658686 [PubMed - as supplied by publisher]

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