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Induction of de novo volatile terpene biosynthesis via cytosolic and plastidial pathways by methyl jasmonate in foliage of Vitis vinifera L.

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.

Acetates↗

Antioxidant activity in lingonberries (Vaccinium vitis-idaea L.) and its inhibitory effect on activator protein-1, nuclear factor-kappaB, and mitogen-activated protein kinases activation.

Lingonberry has been shown to contain high antioxidant activity. Fruits from different cultivars of lingonberry (Vaccinium vitis-idaea L.) were evaluated for fruit quality, antioxidant activity, and anthocyanin and phenolic contents. The fruit soluble solids, titratable acids, antioxidant capacity, and anthocyanin and phenolic contents varied with cultivars. Lingonberries contain potent free radical scavenging activities for DPPH*, ROO*, *OH, and O2*- radicals. Pretreatment of JB6 P+ mouse epidermal cells with lingonberry extracts produced a dose-dependent inhibition on the activation of activator protein-1 (AP-1) and nuclear factor-kappaB (NF-kappaB) induced by either 12-O-tetradecanoylphorbol-13-acetate (TPA) or ultraviolet-B (UVB). Lingonberry extract blocked UVB-induced phosphorylation of the mitogen-activated protein kinase (MAPK) signaling members ERK1, ERK2, p38, and MEK1/2 but not JNK. Lingonberry extract also prevented TPA-induced phosphorylation of ERK1, ERK2, and MEK1/2. Results of soft agar assays indicated that lingonberry extract suppressed TPA-induced neoplastic transformation of JB6 P(+) cells in a dose-dependent manner. Lingonberry extract also induced the apoptosis of human leukemia HL-60 cells in a dose-independent manner. These results suggest that ERK1, ERK2, and MEK1/2 may be the primary targets of lingonberry that result in suppression of AP-1, NF-kappaB, and neoplastic transformation in JB6 P(+) cells and causes cancer cell death by an apoptotic mechanism in human leukemia HL-60 cells.

Antioxidants↗

Development of activated carbon using vine shoots (Vitis vinifera) and its use for wine treatment.

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.

Carbon↗

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

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.

Chlorophyll↗

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

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.

Alkynes↗

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

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.

Anthocyanins↗

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

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.

Carbohydrates↗

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

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.

Antioxidants↗

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.

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.

Carbohydrate Metabolism↗

Induction of cell death in Caco-2 human colon carcinoma cells by ellagic acid rich fractions from muscadine grapes (Vitis rotundifolia).

Possible anticancer mechanisms exerted by polyphenolic compounds contained in fruits and vegetables include antioxidant activity, the inhibition of proliferation, and the induction of apoptosis in cancer cells. This study examined the effects of four isolated polyphenolic extracts from red muscadine grapes (Vitis rotundifolia) on vital cell parameters and the induction of apoptosis in Caco-2 colon carcinoma cells. The magnitude of effects in cell culture was then correlated to polyphenolic composition and antioxidant capacity. Whereas anticancer effects of individual polyphenolic compounds have been demonstrated multiple times, information relating to anticancer effects of polyphenolic extracts is not available in abundance. All four extracts induced apoptosis, decreased cell number, and caused alterations in cell cycle kinetics in a concentration-dependent manner. The efficacy of the polyphenolics on vital cell parameters correlated well to the presence of ellagic acid glycosides and flavonoids and also to the antioxidant capacity. This study demonstrated the anticancer properties of ellagic acid rich extracts from red muscadine juice.

Antioxidants↗

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

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.

Chromatography, High Pressure Liquid↗

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

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.

Chromatography, High Pressure Liquid↗

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).

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.

Acetates↗

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

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 (véraison) 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 véraison; 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 véraison 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.

Flavonoids↗

Extractable amounts of trans-resveratrol in seed and berry skin in Vitis evaluated at the germplasm level.

Extractable amounts of resveratrol in berry skins and seeds were studied in 120 grape (Vitis) germplasm cultivars during two consecutive years to determine the distribution of resveratrol among the main grape genotypes. Interspecific rootstock cultivars had much higher extractable amounts of resveratrol in skin and seed than all other grape genotypes studied in both years. Extremely high extractable amounts of resveratrol in berry skins [>100 microg g(-1) of skin fresh weight (FW)] and seeds (>20 microg g(-1) of seed FW) were observed on two rootstock cultivars obtained from hybrids of V. monticula x V. riparia. Extractable amounts of resveratrol in berries of rootstock cultivars that are the descendants of V. riparia were also very high. The cultivated European type (V. vinifera) cultivars and their hybrids with V. labrusca had relatively low levels of extractable resveratrol in berry skin and seed, and the extractable amounts of resveratrol in berry skin and seeds were, with a few exceptions, <2 microg g(-1) of skin or seed FW. Extractable amounts of resveratrol in berry skin and seeds were closely related with fruit traits or purpose of uses and climate. Significantly higher extractable amounts of resveratrol in berry skin were found in seeded cultivars than in seedless ones, in both berry skin and seeds in winemaking grapes than in table grapes, and in red grapes than in green ones. Moreover, rainfall during fruit development resulted in higher extractable amounts of resveratrol in berry skin, whereas resveratrol synthesis and accumulation in grape seeds were not related to climate change.

Fruit↗

Characterization of phenolic compounds from lingonberry (Vaccinium vitis-idaea).

Phenolic compounds from the lingonberry (Vaccinium vitis-idaea) were identified using LC-TOFMS, LC-MS/MS, and NMR experiments. The compounds were extracted from the plant material using methanol in an ultrasonicator and further isolated and purified using solid-phase extraction and preparative liquid chromatographic techniques. A total of 28 phenolic compounds were at least tentatively identified, including flavonols, anthocyanidins, catechins and their glycosides, and different caffeoyl and ferulic acid conjugates. This is apparently the first report of coumaroyl-hexose-hydroxyphenol, caffeoyl-hexose-hydroxyphenol, coumaroyl-hexose-hydroxyphenol, quercetin-3-O-alpha-arabinofuranoside, kaempferol-pentoside, and kaempferol-deoxyhexoside in the plant, and the flavonol acylglycosides quercetin-3-O-[4' '-(3-hydroxy-3-methylglutaroyl)]-alpha-rhamnose and kaempferol-3-O-[4' '-(3-hydroxy-3-methylglutaroyl)]-alpha-rhamnose are presented here for the first time ever. In addition, more detailed structure in comparison to earlier reports is described for some compounds previously known to exist in lingonberry.

Anthocyanins↗

A hemicellulose B fraction from grape skin (Vitis vinifera, Palomino variety).

The structure of a hemicellulose B fraction (B-1) isolated from grape skins (Vitis vinifera) of the Palomino variety has been studied by methylation analysis, (1)H NMR and (13)C NMR spectroscopy, and partial acid hydrolysis. Hemicellulose B-1 appeared to be homogeneous by gel filtration with a weight-average molecular weight of 22 600. This polysaccharide is a linear xyloglucan chain composed of xylopyranosyl and glucopyranosyl residues linked by beta-(1-->4) glycosidic bonds. Attached to this backbone, 4-O-methyl-D-glucuronopyranosyl acid, D-glucopyranosyl, and L-fucopyranosyl residues occur at position 2 in a ratio of one residue for every five units of xylose in the main chain, with D-xylopyranosyl residues attached at position 6 of glucose units in a ratio of one residue for every two glucose-derived moieties in the main chain.

Carbohydrate Sequence↗

Resveratrol derivatives from the roots of Vitis thunbergii.

Investigating the constituents of the roots of Vitis thunbergii led to the isolation of four new resveratrol derivatives, vitisinols A-D (1-4), together with (+)-epsilon-viniferin, (-)-viniferal, ampelopsin C, miyabenol A, (+)-vitisin A, and (+)-vitisin C. The structures of these 10 compounds were established by spectroscopic (NMR and MS) analyses. All of the isolated compounds, except 1, were tested for their antiplatelet and antioxidative activities.

Antioxidants↗