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PubMed2005

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

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.

Antioxidants↗

PubMed2001

Purification and characterization of a O-methyltransferase capable of methylating 2-hydroxy-3-alkylpyrazine from Vitis vinifera L. (cv. Cabernet Sauvignon).

An S-adenosyl-L-methionine-dependent O-methyltransferase capable of methylating 2-hydroxy-3-alkylpyrazine (HP) was purified 7,300-fold to apparent homogeneity with an 8.2% overall recovery from Vitis vinifera L. (cv. Cabernet Sauvignon) through a purification procedure including column chromatography on DEAE-Sepharose FF, Ether-5PW, hydroxyapatite, G2000SW(XL), and DEAE-5PW. The relative molecular mass of the native enzyme estimated on gel permeation chromatography was 85 kDa, and the subunit molecular mass was estimated to be 41 kDa on SDS-polyacrylamide gel electrophoresis. The enzyme also methylates caffeic acid. The Vmax for IBHP and caffeic acid were 0.73 and 175 pkatals/mg, respectively, and the respective Km for IBHP and caffeic acid were 0.30 and 0.032 mm. The optimum pH for IBHP (8.5) was different from that for caffeic acid (7.5).

Amino Acid Sequence↗

PubMedDate not supplied

[Photoprotective properties of melanins from grape (Vitis vinifera) and black tea (Thea sinensis)].

Dark pigments of melanin type were extracted from the rind of ripe grapes Vitis vinifera (ring "Alfa") and pack tea (Thea sinensis). The study of photoprotection activity of the extracted pigments have shown that they intensively absorb radiation in the UV and visible. The increase of the photoprotection effect correlates with the concentration of paramagnetic centers in the melanins. The pigments reduce the number of damaged molecules of plasmid DNA pBR-322 and inhibition the IUV-induced lipid peroxidation.

Camellia↗

PubMed2001

[Studies on chemical constituents of oligostilbenes from Vitis davidii Foex].

OBJECTIVE: To study the chemical constituents of oligostilbenes from Vitis davidii. METHOD: Compounds were isolated with polystyrene resin RA, silica gel and C18 column chromatography. The structures were elucidated by means of spectroscopic evidence. RESULT: Seven compounds were obtained and identified as resveratrol, heyneanol A, ampelopsin E, amurensin B, (+)-epsilon-viniferin, vitisin A and amurensin G. CONCLUSION: All the compounds were isolated from this plant for the first time.

Benzofurans↗

PubMed2005

Effects of plant phenolics and grape extracts from Greek varieties of Vitis vinifera on Mitomycin C and topoisomerase I-induced nicking of DNA.

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.

DNA Damage↗

PubMed2005

Isolation and characterization of a chitinase gene VCH3 promoter from grapevine (Vitis amurensis).

The 1216-bp 5' upstream region of the gene encoding a class III chitinase VCH3 was isolated from grapevine (Vitis amurensis Rupr.) by adaptor-PCR (GenBank accession number AF441123), and the transcriptional start site of the VCH3 gene was identified by primer extension, which corresponds to the second A in the DNA sequence 5'-ATCAAGCAC-3'. Sequence analysis revealed that the VCH3 promoter sequence contains CAAT and TATA motifs that are located at the -122 and -29 nucleotide upstream of the transcriptional start site, respectively. Both motifs are characteristic of the eukaryotic gene promoter. In addition, within the VCH3 promoter, we found two inverse salicylic acid (SA)-responsive cis-acting motifs (TGACG) at -1181 bp and -293 bp upstream of the transcriptional start site, respectively. To characterize the VCH3 promoter, the VCH3 promoter-GUS chimera was constructed and transferred to Nicotiana tobacum cv. NC89 by Agrobacterium tumefaciens-mediated leaf disc transformation. Fluorometric and histochemical analysis of GUS activity in the transgenic tobacco root and leaf treated with SA showed that the VCH3 promoter was induced by SA, which indicates that it may have potential use in genetic engineering.

Agrobacterium tumefaciens↗

PubMed2006

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

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.

Argentina↗

PubMed1992

Monitoring by two-dimensional electrophoresis somatic embryogenesis in leaf and petiole explants from Vitis.

A two-dimensional electrophoretic analysis of the total proteins was carried out in Vitis rupestris as model system in order to characterize the different developmental stages--from callus to plantlets--of somatic embryogenesis events in the grapevine. The patterns of callus, embryogenetic callus, somatic embryos and plantlets derived from leaf and petiole explants were compared. Each differentiation step was characterized by specific peptide spots.

Cells, Cultured↗

PubMed1993

The Ti plasmid from the wide host range Agrobacterium vitis strain Tm4: map and homology with other Ti plasmids.

The octopine/cucumopine (o/c) strains of Agrobacterium vitis are divided into limited (LHR) and wide host range (WHR) strains. We present here the first map of a Ti plasmid from a WHR o/c strain, Tm4. pTiTm4 (252 kb) has homology with the octopine Ti plasmid pTiAch5, the nopaline plasmid pTiC58, and the Ri plasmid pRiHRI. Approximately 155 kb of pTiTm4 is homologous with the LHR o/c Ti plasmids pTiAB3 and pTiAg57, the remaining part is pTiTm4-specific. Our data suggest that the evolution of the o/c Ti plasmids was accompanied by large-scale changes in plasmid structure. The origin of replication of pTiTm4 was localized by a functional assay.

Biological Evolution↗

PubMed1994

Cloning and molecular analysis of structural genes involved in flavonoid and stilbene biosynthesis in grape (Vitis vinifera L.).

Genes involved in flavonoid and stilbene biosynthesis were isolated from grape (Vitis vinifera L.). Clones coding for phenylalanine ammonia-lyase (PAL), chalcone synthase (CHS), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin dioxygenase (LDOX) and UDP glucose:flavonoid 3-O-glucosyl transferase (UFGT), were isolated by screening a cDNA library, obtained from mRNA from seedlings grown in light for 48 h using snapdragon (Antirrhinum majus) and maize heterologous probes. A cDNA clone coding for stilbene synthase (StSy) was isolated by probing the library with a specific oligonucleotide. These clones were sequenced and when the putative products were compared to the published amino acid sequence for corresponding enzymes, the percentages of similarity ranged from 65% (UFGT) to 90% (CHS and PAL). The analysis of the genomic organization and expression of these genes in response to light shows that PAL and StSy genes belong to large multigene families, while the others are present in one to four copies per haploid genome. The steady-state level of mRNAs encoded by the flavonoid biosynthetic genes as determined in young seedlings is coordinately induced by light, except for PAL and StSy, which appear to be constitutively expressed.

Anthocyanins↗

PubMed1996

Characterization of Vitis vinifera L. glutamine synthetase and molecular cloning of cDNAs for the cytosolic enzyme.

Grapevine (Vitis vinifera L.) glutamine synthetase (GS) was analysed into two distinct classes of isoforms; one of them was present in both leaf and root tissues while the other one showed leaf specificity. Western blot analysis revealed that grapevine GS consists of three types of polypeptides of distinct size and differential tissue specificity. Two structurally distinct cDNA clones, pGS1;1 and pGS1;2, encoding grapevine GS were isolated from a cell suspension library and characterized. Both clones contained open reading frames encoding for polypeptides of 356 amino acids with a predicted molecular mass of about 39 kDa. Although the coding sequences of pGS1;1 and pGS1;2 were 84% similar, their 5'- and 3'-untranslated sequences showed only 40% similarity. The coding sequences of the two clones and the derived amino acid sequences showed higher homology to cytosolic than to chloroplastic GSs of other higher plants indicating that the cDNAs isolated encode for cytosolic isoforms of grapevine GS. Southern blot analysis suggested the existence of more than two GS genes in the grapevine genome. In northern blots both clones were hybridized to mRNAs of about 1.4 kb that are differentially expressed in the various tissues. Supply of nitrate or ammonium in the cell suspension culture medium, as a sole nitrogen source, resulted in differential response of the pGS1;1- and pGS1;2-related genes.

Amino Acid Sequence↗

PubMed1993

Rapid divergence of Agrobacterium vitis octopine-cucumopine Ti plasmids from a recent common ancestor.

The octopine/cucumopine (o/c) Ti plasmids of the grapevine-associated Agrobacterium vitis strains constitute a family of related DNA molecules. Restriction maps were established of two limited-host-range o/c Ti plasmids, pTiAg57 and pTiAB3, and of the wide-host-range o/c Ti plasmid pTiHm1. Together with the previously obtained map of the wide-host-range o/c Ti plasmid pTiTm4, about 1000 kb were mapped with a resolution of 0.2 kb, allowing a detailed comparison of the various structures. One region of the o/c Ti plasmids is highly conserved and differs mainly by the presence or absence of relatively small DNA fragments (0.9-2.7 kb); the other region has been modified more extensively and carries large sequences specific for each Ti plasmid type. The sequence similarity within large conserved regions shows that these plasmids have diverged recently and that their evolution was driven by large-scale genetic events rather than single nucleotide changes. These results have important implications for studies on bacterial evolution.

Arginine↗

PubMed1995

Powerful hepatoprotective and hepatotoxic plant oligostilbenes, isolated from the Oriental medicinal plant Vitis coignetiae (Vitaceae).

The methanol extract of the Oriental medicinal plant Vitis coignetiae (Vitaceae) showed hepatoprotective activity in the in vitro assay method using primary cultured rat hepatocytes. Activity-guided fractionation of the extract afforded epsilon-viniferin as an active principle. The protective effect of epsilon-viniferin against mice carbon tetrachloride-induced hepatic injury in mice was shown by serum enzyme assay as well as by pathological examination. In addition to epsilon-viniferin, plant oligostilbenes, ampelopsins A, C, F and the mixture of vitisin A and cis-vitisin A were also present in the extract. Among them, ampelopsin C and the mixture of vitisin A and cis-vitisin A were found to be powerful hepatotoxins.

Animals↗

PubMed2002

Genetic mapping of grapevine ( Vitis vinifera L.) applied to the detection of QTLs for seedlessness and berry weight.

Parental and consensus genetic maps of Vitis vinifera L. (2n = 38) were constructed using a F(1) progeny of 139 individuals from a cross between two partially seedless genotypes. The consensus map contained 301 markers [250 amplification fragment length polymorphisms (AFLPs), 44 simple sequence repeats (SSRs), three isozymes, two random amplified polymorphic DNAs (RAPDs), one sequence-characterized amplified region (SCAR), and one phenotypic marker, berry color] mapped onto 20 linkage groups, and covered 1,002 cM. The maternal map consisted of 157 markers covering 767 cM (22 groups). The paternal map consisted of 144 markers covering 816 cM (23 groups). Differences in recombination rates between these maps and another unpublished map are discussed. The major gene for berry color was mapped on both the paternal and consensus maps. Quantitative trait loci (QTLs) for several quantitative subtraits of seedlessness in 3 successive years were searched for, based on parental maps: berry weight, seed number, seed total fresh and dry weights, seed percent dry matter, and seed mean fresh and dry weights. QTLs with large effects (R(2) up to 51%) were detected for all traits and years at the same location on one linkage group, with some evidence for the existence of a second linked major QTL for some of them. For these major QTLs, differences in relative parental effects were observed between traits. Three QTLs with small effects (R(2) from 6% to 11%) were also found on three other linkage groups, for berry weight and seed number in a single year, and for seed dry matter in 2 different years.

Journal Article↗

PubMed2000

In-vitro selection of Vitis vinifera 'Chardonnay' with Elsinoe ampelina culture filtrate is accompanied by fungal resistance and enhanced secretion of chitinase.

Proembryogenic masses of grapevine (Vitis vinifera L.) 'Chardonnay' (clone 02Ch) were exposed to the culture filtrate of Elsinoe ampelina (deBary) Shear, the causal agent of anthracnose disease. After four or five cycles of recurrent in-vitro selection with medium containing 40% fungal culture filtrate, putative resistant lines RC1 and RC2 respectively, were established. The selected lines inhibited the growth of E. ampelina and Fusarium oxysporium (Schlecht.) (isolated from watermelon) in a dual-culture assay and reduced the growth of mycelium on a conditioned-medium test, thus suggesting the involvement of extracellular compounds in resistance. Sodium dodecyl sulfate-polyacrylamide (SDS-PAGE) gel electrophoresis of extracellular proteins from spent suspension-culture medium showed enhanced secretion of new proteins by selected lines. A 36-kDa protein was immunodetected by a chitinase antiserum. This chitinase continued to express constitutively in differentiated somatic embryos and also in the intercellular fluids of plants regenerated from the selected lines. Somatic embryos from selected lines grew uninhibitedly in a medium containing 40% fungal culture filtrate, whereas non-selected (control) somatic embryos became necrotic and died within a few days. Plants regenerated from selected lines exhibited resistance to infection by E. ampelina in both greenhouse tests and detached leaf bioassays. Results suggest that embryogenic cells can be selected for resistance following in-vitro selection, resulting in resistant plants. Whether or not resistant cells pre-existed in the original embryogenic culture or were induced by the selection pressure could not be determined.

Ascomycota↗

PubMed1999

Cloning and expression of an arginine decarboxylase cDNA from Vitis vinifera L. cell-suspension cultures.

Arginine decarboxylase (ADC; EC 4.1.1.19) is a key enzyme in one of the two pathways to putrescine. We present the first ADC cDNA from a woody perennial plant species, the grapevine (Vitis vinifera L.), which exhibits 70-80% homology with other dicot ADCs. The effects of ammonium, nitrate, and putrescine on ADC specific activity, soluble polyamine levels, ADC-mRNA, endogeneous arginine and ornithine, and arginase specific activity were investigated in suspension cultures of grapevine cells. The addition of NH4+ to cells cultured in NH4(+)-free medium, resulted in a 4-fold increase in ADC activity and concomitantly in a 4-fold increase in putrescine and a 3-fold decrease in arginine. During this period ornithine increased and arginase activity followed a reverse pattern of changes compared with ADC. In contrast, the addition of NO3- did not markedly affect ADC activity, putrescine, arginine and ornithine, but transiently increased arginase activity. The addition of putrescine caused a 4-fold decrease in ADC activity and increased arginine, ornithine and arginase activity. The changes in ADC specific activity were not accompanied by analogous changes in the ADC transcript levels. These results further support the view that ADC regulation is not exhibited, at least for the factors considered in this work, at the transcriptional level.

Blotting, Northern↗

PubMed2004

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

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.

Journal Article↗

PubMed1991

Coordinate- and elicitor-dependent expression of stilbene synthase and phenylalanine ammonia-lyase genes in Vitis cv. Optima.

The mechanisms controlling the induction of stilbene synthase and phenylalanine ammonia-lyase (PAL), two putative key regulatory enzymes of the biosynthetic pathway to stilbene phytoalexins, have been investigated. The induction was studied in cell suspension cultures of grape (Vitis cv. Optima) by treatment with fungal cell wall. Several independent cDNA clones for PAL and stilbene synthase were isolated from a cDNA library of fungal cell wall-induced grape cells and identified by sequence analysis. The stilbene synthase cDNA sequence of pSV21 predicted a protein of 392 amino acids and Mr 42,791, similar in size to that observed experimentally for immunodetected stilbene synthase. The cDNA sequences of pSV21 and pSV25 differed in 76 bp in the coding region. The sequences of grape stilbene synthase cDNAs exhibited significant homology to the sequence reported for the peanut stilbene synthase cDNA. Both PAL and stilbene synthase mRNA, measured by RNA blot hybridizations, were induced within 1 h of addition of fungal cell wall preparations to the cell cultures, rose to a maximum by the sixth hour, then declined slowly over the next 20 h. The activities of PAL and stilbene synthase were also induced in parallel, but reached their maximum at different times after fungal cell wall addition to the cell cultures. The induction patterns of stilbene synthase and PAL in grape and peanut are discussed.

Acyltransferases↗