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, gunata@univ/​montp2.fr.

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,
mariajose.carmona@upm.es.

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, cramer@unr.edu.

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. cadot@angers.inra.fr

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. gyorgy.horvath@ua.ac.be

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. kereid@interchange.ubc.ca

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, perlx@int.gov.il.

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. claudio.lovisolo@unito.it

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, mav@sun.ac.za.

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.
fulvio.mattivi@iasma.it

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. kbgrape@yahoo.com

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. ea.barka@univ/​reims.fr

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. pioust@iihr.ernet.in

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.
takatoshi.tominaga@oenologie.u/​bordeaux2.fr

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. inoronha@ibmc.up.pt

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. Christopher.Davies@csiro.au

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. mandy.walker@csiro.au

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. plantb7@sun.ac.za

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. soulie@ccr.jussieu.fr

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. mkeller@wsu.edu

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.
renata@botanic.hr

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. bcavagnaro@fca.uncu.edu.ar

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.
gianni.fila@entecra.it

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. nsendogdu@gzi.edu.tr

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.
makdessi/​sweidan@uni/​tuebingen.de

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.
skoundou@agro.auth.gr

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. doligez@ensam.inra.fr

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.
dschatelet@ucdavis.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, parce@bio.puc.cl.

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. g_liak@aua.gr

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. lebon@ensam.inra.fr

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.
fruben@murdoch.edu.au

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.
peter.raspor@bf.uni/​lj.si

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. kunz@ccr.jussieu.fr

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.
scarrico@dq.ua.pt

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.
tricia.franks@adelaide.edu.au

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.
jansen@mail.utexas.edu

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. fujita@nrib.go.jp

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.
kortekam@uni/​hohenheim.de

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. fforallo@usc.es

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. castellarin.simone@spes.uniud.it

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. Roque.Bru@ua.es

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. takatoshi.tominaga@oenologie.u/​bordeaux2.fr

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. sandra.apers@ua.ac.be

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. islasosu@cascabel.ciad.mx

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, pfuendel@botanik.uni/​wuerzburg.de.

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. honda@nubio.nagoya/​u.ac.jp

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. christophe.bertsch@uha.fr

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.
liliya.serazetdinova@sainsburg/​laboratory.ac.uk

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. a.genovese@unifg.it

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. lrobert5@wanadoo.fr

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.
sofiapereira@isa.utl.pt

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. fanizza@agr.uniba.it

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.
Sami.Fattouch@insat.rnu.tn

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. claire.barker@csiro.au

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.
claudio.moser@iasma.it

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. arimoto@cc.okayama/​u.ac.jp

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. adam@evry.inra.fr

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. frperez@uchile.cl

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. b.bornhoff@bafz.de

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. jncui@chem.dlut.edu.cn.

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. thiery@bordeaux.inra.fr

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. m.bernard@latrobe.edu.au

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.
blake.bextine@ucr.edu

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.
fcarrau@fq.edu.uy

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.
satyanarayana_sreemantula@rediffmail.com

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. tricia.franks@adelaide.edu.au

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. chbrenes@itesm.mx

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.
dww5@cornell.edu

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. rita.musetti@uniud.it

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. flotrouillas@ucdavis.edu

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. dwaters@scu.edu.au

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. jzhu@harc/​hspa.com

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. luyingreen@163.net

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. Rosario.Salinas@uclm.es

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. stamgreg@yahoo.gr

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. amberry@ucdavis.edu

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. lesniew@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. adam@evry.inra.fr

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. qaenol@urv.es

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. mnkrasnow@ucdavis.edu

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. ferruccio.berti@unimi.it

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. i.leinonen@scri.sari.ac.uk

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. cvandenboom@milieukeur.nl

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. alisonk@tepapa.govt.nz

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 pa