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PubMedDate not supplied

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.

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.

Flavonoids↗
PubMed2003

Effects of fludioxonil and pyrimethanil, two fungicides used against Botrytis cinerea, on carbohydrate physiology in Vitis vinifera L.

In Vitis vinifera L, photosynthesis and photosynthate partitioning are affected in the presence of fludioxonil and pyrimethanil, two fungicides commonly used in vineyards against Botrytis cinerea Pers. However, the effects were found to be different according to the model studied: plantlets (cv Chardonnay) grown in vitro, fruiting cuttings (cv Chardonnay) and plants grown in vineyards (cvs Chardonnay, Pinot noir and Pinot Meunier). In the plantlets grown in vitro, both fungicides decreased gas exchanges, photosynthetic pigment and starch concentrations in the leaves, whereas soluble carbohydrates transiently accumulated, suggesting that plantlets mobilised starch in response to photosynthesis inhibition caused by fungicides. In the fruiting cuttings, the fungicides did not affect photosynthesis, although fludioxonil caused starch decrease in parallel with sucrose accumulation, suggesting that the fungicide effects were of lower intensity than in vitro. Conversely, in vineyard, the two fungicides stimulated photosynthesis and increased pigment concentrations in the three vine cultivars tested. In the meantime, glucose, fructose and starch levels of the leaves declined after fungicide exposure, whereas sucrose accumulated, indicating that sucrose synthesis increased in the leaves following the fungicide treatment. Among the three varieties, Chardonnay was the most sensitive to the fungicides as revealed by the intensity of the responses and the longer period for recovery. In vineyard, the results suggested that the two fungicides, in addition to inhibiting B cinerea development, had a beneficial effect on vine physiology through the stimulation of leaf carbon nutrition, which may further enable the plant to rapidly make use of its defence reactions.

Botrytis↗
PubMed2003

AFLP analysis of genetic relationships among aromatic grapevines (Vitis vinifera).

Genotypic diversity has been detected among aromatic grapevines (Vitis vinifera) by molecular markers (AFLPs). The 22 primer-pairs generated a total of 1,331 bands of which 564 (40%) were polymorphic over all the genotypes. The bootstrap analysis pointed out that a large number of polymorphic bands (200-400) has to be used for a better estimation of the genetic distances among genotypes; 383 polymorphic AFLP bands were used for the cluster and the principal coordinate analyses because they did not present missing data across all the genotypes. The cluster analysis (UPGMA), based on polymorphic AFLP markers, revealed no relationship between the Moscato and Malvasia grapevines. The Malvasias, unlike the Moscatos distinguished by their distinct muscat aroma, have to be considered a more complex group because it includes muscat and non-muscat grapevines. The principal coordinate analysis (PCO) confirmed the pattern of the cluster analysis only for those varieties which presented a low coefficient of dissimilarity, while for the other varieties there was no correspondence between the two analyses. The pattern of aggregation among aromatic grapevines in the cluster and principal coordinate analyses does not support any classification that might include an aromatic grapevine group in V. vinifera. Even though some synonyms and homonyms are present among aromatic grapevines (V. vinifera), genetic diversity exists among genotypes in AFLP markers.

Cluster Analysis↗
PubMed2004

Mapping 245 SSR markers on the Vitis vinifera genome: a tool for grape genetics.

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.

Agriculture↗
PubMed2005

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

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.

Base Sequence↗
PubMed2006

Linkage disequilibrium in cultivated grapevine, Vitis vinifera L.

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.

Chromosome Mapping↗
PubMed2005

QTL analysis for fruit yield components in table grapes (Vitis vinifera).

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.

Breeding↗
PubMed2006

Refined mapping of the Pierce's disease resistance locus, PdR1, and Sex on an extended genetic map of Vitis rupestris x V. arizonica.

A framework genetic map based on genomic DNA-derived SSR, EST-derived SSR, EST-STS and EST-RFLP markers was developed using 181 genotypes generated from D8909-15 (female) x F8909-17 (male), the '9621' population. Both parents are half siblings with a common female parent, Vitis rupestris 'A. de Serres', and different male parents (forms of V. arizonica). A total of 542 markers were tested, and 237 of them were polymorphic for the female and male parents. The female map was developed with 159 mapped markers covering 865.0 cM with an average marker distance of 5.4 cM in 18 linkage groups. The male map was constructed with 158 mapped molecular markers covering 1055.0 cM with an average distance of 6.7 cM in 19 linkage groups. The consensus '9621' map covered 1154.0 cM with 210 mapped molecular markers in 19 linkage groups, with average distance of 5.5 cM. Ninety-four of the 210 markers on the consensus map were new. The 'Sex' expression locus segregated as single major gene was mapped to linkage group 2 on the consensus and the male map. PdR1, a major gene for resistance to Pierce's disease, caused by the bacterium Xylella fastidiosa, was mapped to the linkage group 14 between markers VMCNg3h8 and VVIN64, located 4.3 and 2.7 cM away from PdR1, respectively. Differences in segregation distortion of markers were also compared between parents, and three clusters of skewed markers were observed on linkage groups 6, 7 and 14.

Chromosome Mapping↗
PubMed2007

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

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.

Alleles↗
PubMed2005

Heavy metals (Cr, Zn, Ni, V, Pb, Cd) in lingonberries (Vaccinium vitis-idaea L.) and assessment of human exposure in two industrial areas in the Kemi-Tornio region, Northern Finland.

The concentration of Cr, Zn, Ni, V, Pb, and Cd were measured in lingonberries (Vaccinium vitis-idaea L.) sampled at 23 sampling sites around a ferrochrome and stainless steel works and opencast chromium mine in the Kemi-Tornio region, Northern Finland. Two different microwave-assisted digestion procedures were used for sample digestion, i.e., a mixture of HNO3 + H2O2 and a mixture of HNO3 + H2O2 + HCl + HF + H3BO3. According to the results, the digestion procedure with the mixture of HNO3 + H2O2 underestimated especially the Cr concentrations in plant material. The maximum concentrations of Cr (1.3 mg kg(-1), wet weight), Ni (358 microg kg(-1); ww), V (36 microg kg(-1); ww), and Cd (2.4 microg kg(-1); ww) in the immediate vicinity of the point sources were 33, 6, 4, and 8 times higher than the background levels, respectively. The dietary intakes of Cd and Pb were assessed and compared to the health criteria recommendations set by the joint Food and Agriculture Organization and World Health Organization Expert Committee on Food Additives (JECFA). The results showed that, depending on the consumption of lingonberries, human exposure based on the mean concentrations for Pb and Cd varied between 0.04% and 0.07% for Pb and between 0.04% and 0.09% for Cd compared to the tolerable total quantities of 25 microg kg(-1) for Pb and 7 microg kg(-1) for Cd per body weight per week set by JECFA.

Air Pollutants↗
PubMed2003

Impact of flumioxazin herbicide on growth and carbohydrate physiology in Vitis vinifera L.

The impact of flumioxazin herbicide on in vitro-grown grapevine ( Vitis vinifera L. cv. Chardonnay) was investigated. The herbicide treatments (1, 10 or 100 micro M flumioxazin in MM medium) had a negative impact on photosynthesis, as revealed by a reduction in foliar chlorophyll and carotenoid contents, gas exchanges and alteration in plastid structure and, consequently, resulted in a strong inhibition of plantlet growth. Surprisingly, soluble sugars and starch accumulated in all organs, suggesting a stimulation of sugar uptake from the medium. Moreover, photosynthetic activity and starch content partially recovered within 3 weeks of treatment at the weakest herbicide concentration. These results provide new insights into the physiological responses of non-target crops to flumioxazin, showing that flumioxazin is active in photosynthetic tissues of the non-target grapevine via root uptake, which is contrary to what is mentioned in the literature, and that the in vitro-grown plantlet is a good model for investigating the physiological effects of pesticides on crop species.

Benzoxazines↗
PubMed2003

Stress effects of flumioxazin herbicide on grapevine ( Vitis vinifera L.) grown in vitro.

Among the herbicides used in vineyards, the pre-emergence soil-applied flumioxazin (FMX) is a recently synthesized molecule that inhibits chlorophyll biosynthesis in weed species. The aim of this work was to characterize the effects of FMX on non-target grapevine ( Vitis vinifera L. cv. Chardonnay) plantlets grown in vitro. FMX treatment (from 1 to 100 micro M) represented a stress, as revealed by measurement of several parameters. Stem and leaves underwent dehydration and a decrease in both water- and osmotic-potential. Treated plantlets exhibited concomitant accumulation of soluble carbohydrates in all tissues and of free proline in stems and leaves. Moreover, FMX caused lipid peroxidation and electrolyte leakage in leaf tissues. These results indicate that the herbicide FMX is toxic for grapevine grown in vitro. In addition to inhibiting protoporphyrinogen IX oxidase, it causes water stress and membrane alteration in tissues and, as a consequence, generates the accumulation of carbohydrates and free proline.

Benzoxazines↗
PubMed2003

Transformation of apple ( Malus domestica Borkh.) with the stilbene synthase gene from grapevine ( Vitis vinifera L.) and a PGIP gene from kiwi ( Actinidia deliciosa).

The objective of the present research was to introduce genes with antifungal potential into the commercially important apple cvs. Elstar and Holsteiner Cox in order to establish resistance against fungal diseases. The gene encoding the stilbene synthase (Vst1) from Vitis vinifera L., responsible for the synthesis of the phytoalexin resveratrol in grapevine, and the gene for a polygalacturonase-inhibiting protein (PGIP) from kiwi ( Actinidia deliciosa) were transferred into Holsteiner Cox and Elstar via Agrobacterium tumefaciens-mediated transformation. A total of nine transgenic Holsteiner Cox clones and one transgenic E clone carrying the stilbene-synthase gene as well as three transgenic Holsteiner Cox lines harbouring the polygalacturonase-inhibiting protein from Kiwi were identified via polymerase chain reaction and Southern blot analysis. High performance liquid chromatography analysis revealed the accumulation of a resveratrol-derivate, a glycoside, in transgenic Vst1 plants.

Acclimatization↗
PubMed2003

High-efficiency biolistic co-transformation and regeneration of 'Chardonnay' (Vitis vinifera L.) containing npt-II and antimicrobial peptide genes.

A reliable and efficient system for transformation and regeneration of 'Chardonnay' (Vitis vinifera L.) plants via microprojectile bombardment was developed. Improvements over the previous biolistic transformation system included: (1) the use of gold particles for bombardment; (2) step-wise selection at 10 then 15 mg/l kanamycin; and (3) embryo induction at 27 degrees C. Embryogenic cell cultures were either bombarded with pBI426, which contains the reporter gene gus (uidA) coding for beta-glucuronidase (GUS), or were co-bombarded with pSAN237 carrying the npt-II (neomycin phosphotransferase II) selectable marker gene, and a second plasmid with an antimicrobial peptide gene. A large number of transient (7,883 +/- 1,928) and stable (46 +/- 32) blue spots per plate at 2 and 95 days after bombardment, respectively, were obtained according to GUS expression analyses. A total of 447 putative transgenic embryos was harvested from 84 bombarded plates. From these embryos, 242 (54%) were regenerated into plants within the first year of the experiment. Southern blot analyses confirmed integration of the transgenes into the grape genome. Co-transformation was tested with four separate antimicrobial constructs. The co-transformation frequency of unlinked genes was 48% as measured by polymerase chain reaction (PCR), and 56% as estimated by dot blot hybridization. Expression of the gus gene, and PCR and Southern blot analyses of npt-II and antimicrobial genes from regenerated plants document stable transformation of 'Chardonnay' and establish the parameters for highly-efficient biolistic transformation in V. vinifera.

Anti-Bacterial Agents↗
PubMed2003

Chitosan improves development, and protects Vitis vinifera L. against Botrytis cinerea.

We evaluated the potential of chitosan both to stimulate plant development and to induce protection from Botrytis cinerea in Vitis vinifera L. plantlets. The presence of 1.75% (v/v) chitogel in the culture medium was the optimal concentration for in vitro grapevine plantlet growth, as determined by measurements on enhancement of root and shoot biomass. Photosynthesis and related parameters were also stimulated in chitogel-treated plantlets. Chitogel reduced the development of Botrytis cinerea and induced cytological alterations to the pathogen. When challenged with the fungus, a significant decrease in disease incidence was observed in plants growing on medium supplemented with chitogel. Furthermore, exogenous foliar applications of chitogel to plantlets growing on chitogel-free medium sensitized them so as to be protected against Botrytis cinerea attack. Our results indicate that chitogel can be used in the vineyard as a means to attain protection against Botrytis cinerea and that its application may counteract the wide use of chemical pesticides.

Botrytis↗
PubMed2006

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

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.

Acyltransferases↗
PubMed2002

Immunocharacterization of Vitis vinifera L. ferredoxin-dependent glutamate synthase, and its spatial and temporal changes during leaf development.

The grapevine (Vitis vinifera L.) partial fragment of cDNA clone pGOGAT1 [Loulakakis and Roubelakis-Angelakis (1997) Physiol Plant 101:220-228], encoding the ferredoxin-dependent glutamate synthase (Fd-GOGAT; EC 1.4.7.1), was overexpressed in Escherichia coli cells. A hybrid between the Fd-GOGAT fragment and maltose-binding protein was purified and used to raise a polyclonal antibody in a rabbit. The prepared antibody appeared to be specific towards Fd-GOGAT; it recognized a protein band of approximately 160 kDa on nitrocellulose blots after SDS-PAGE of total proteins from leaves, internodes, roots and calluses, and precipitated most of the enzyme activity present in grapevine protein extracts. The quantity of Fd-GOGAT protein was substantially higher in leaves than in other grapevine tissues tested, coincident with a similar distribution of the enzyme specific activity. Intracellular localization studies revealed that both the enzyme activity and the 160-kDa immunoreactive protein were associated with the chloroplastic fraction. Furthermore, the accumulation of Fd-GOGAT, glutamine synthetase (GS) and glutamate dehydrogenase (GDH), at the activity and protein levels, was monitored during leaf development of field-grown plants, from the stage of the newly expanding leaf to the senescing old leaf. Both the specific activity and quantity of the 160-kDa polypeptide of Fd-GOGAT were higher in the mature, full sized leaves and substantially lower in young and senescing leaves. GS specific activity and immunoreactive protein followed the same trend as Fd-GOGAT, while GDH showed opposite developmental patterns of accumulation. The biological significance of the presence of Fd-GOGAT in the various grapevine tissues and its physiological role during early development and natural senescence of the leaves are discussed.

Amino Acid Oxidoreductases↗
PubMed2002

Myb-related genes of the Kyoho grape ( Vitis labruscana) regulate anthocyanin biosynthesis.

Partial cDNAs of myb-related regulatory genes were isolated from the tetraploid Kyoho grape ( Vitis labruscana: V. labrusca x V. vinifera) and the expression patterns of the corresponding genes were studied. Since MybA gene expression is closely related to coloring and/or ripening of the berry (expression increases strongly with the commencement of coloring and berry softening, and is detected only in berry skin and flesh), full-length cDNAs for the gene were isolated from a mature-berry cDNA library. Three different species of MybA were identified from the cDNA sequences. Delivery of these cDNAs to somatic embryos of grape led to the induction of reddish-purple spots and UDP-glucose:flavonoid 3- O-glucosyltransferase (UFGT) gene expression in non-colored embryos. The UFGT transcript was not detected in control embryos, while other structural genes for anthocyanin biosynthesis were expressed in both control and pigmented embryos. In addition, introduction of the UFGT gene induced the same reddish-purple spots in embryos. In contrast, treatment with the leucoanthocyanidin dioxygenase (LDOX) gene failed to induce these spots. Our results strongly suggest that MybA genes are involved in the regulation of anthocyanin biosynthesis in the grape via expression of the UFGT gene.

Amino Acid Sequence↗