Changes in the phytoalexin content of various Vitis spp. in response to ultraviolet C elicitation.

The phytoalexin production potential of three American Vitis species and that of three cultivars of Vitis vinifera were evaluated in response to UV-C irradiation. Time course changes in resveratrol, piceid, epsilon-viniferin, and pterostilbene contents were studied within 3 days after a short UV-C irradiation. Results show that the two major stilbenes accumulated as a response to UV-C elicitation are resveratrol and epsilon-viniferin, a resveratrol dehydrodimer, the concentration of both compounds usually reaching quantities >100 microgram/g of fresh weight. In contrast, piceid and pterostilbene were constantly produced in low quantities. Owing to the results obtained, the role of stilbene phytoalexins in the resistance of grapevines to diseases is discussed.     

Isolation of isolectins from Vitis vinifera L. Cv. Chardonnay grape berries

A lectin fraction from Chardonnay grape juice has been isolated by affinity chromatography on a column of p-aminophenyl beta-D-glucoside-derivatized agarose. The lectin fractions agglutinate rabbit and human erythrocytes without serological specificity. None of the usual monosaccharides, glycosides, or glycoproteins inhibit the hemagglutinating activity. Erythroagglutination is only inhibited by nitrophenyl glycosides, p-nitrophenyl beta-D-glucoside being the strongest inhibitor. In SDS-PAGE in the presence of 2-mercaptoethanol and gel filtration HPLC, the lectin fraction gave a single band or peak corresponding to M(r) 13.2-11.9 kDa, thus indicating it to be a monomer. Three bands were observed by isoelectric focusing with pI values of 4.1, 4. 4, and 4.9. The isolectins seem to be glycoproteins since they are bound on a concanavalin A-Sepharose column.     

Assessment of anthocyanins in grape (Vitis vinifera L.) berries using a noninvasive chlorophyll fluorescence method

Anthocyanins (Anths) in grape (Vitis vinifera L.) berries harvested at véraison 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.     

Accumulation and distribution pattern of macro- and microelements and trace elements in Vitis vinifera L. cv. Chardonnay berries

This paper describes the accumulation pattern of 42 mineral elements in Vitis vinifera L. berries during development and ripening and their distribution in berry skin, seeds, and flesh around harvest time. Grape berries were sampled in two different vineyards with alkaline soil and analyzed using a ICP-MS. Although elemental amounts were significantly different in the grapes from the two vineyards, the accumulation pattern and percentage distribution in different parts of the berries were generally quite similar. Ba, Eu, Sr, Ca, Mg, Mn, and Zn accumulate prior to veraison. Al, Ce, Dy, Er, Ga, Gd, Ho, La, Nd, Pr, Sm, Sn, Zr, Th, Tm, U, Y, and Yb accumulate mainly prior to veraison but also during ripening. Ag, As, B, Cd, Cs, Cu, Fe, Ge, Hg, K, Li, Na, P, Rb, Sb, Se, and Tl accumulate progressively during growth and ripening. With regard to distribution, Ba, Ca, Eu, Fe, Mn, P, Sr, and Zn accumulate mainly in the seeds, Al, B, Ga, Sn, and the rare earths analyzed, except for Eu, accumulate mainly in the skin, and Ag, As, Cd, Cs, Cu, Ge, Hg, K, Li, Mg, Na, Rb, Sb, Se, Th, Tl, U, and Zr accumulate mainly in the flesh. A joint representation of the accumulation and distribution patterns for the elements in the berry is also given.     

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.     

Study of soil respiration and fruit quality of table grape (Vitis vinifera L.) in response to different soil water content in a greenhouse

ABSTRACT

To understand the response of grape (Hutai No.8) quality and soil respiration (R_s) to different soil relative water contents (SRWCs), this study was designed with three soil moisture levels (A: 80–95%, B: 60–75%, and C: 40–55% of SRWC) for grape cultivation. Meanwhile, environmental factors, including air temperature (T_a), air relative humidity, and light intensity, were also recorded. The results showed the following: (1) Through the comprehensive analysis of fruit quality by the method of subordinate function, we concluded that the optimum soil moisture treatment was 60–75% SRWC, and the soluble sugars, proanthocyanidin, and resveratrol were most abundant. In addition, vitamin C (Vc) content was the largest under C treatment. (2) Photosynthetic characteristic under high soil moisture was better than those under low soil moisture condition during grape coloring periods, and it was largest under A treatment in 2015. R_s rate was in accordance with the trend of grape photosynthesis. High soil moisture could accelerate the photosynthetic rate of grape leaves and increase R_s. (3) Correlation analysis showed that higher soil moisture and air humidity and lower soil temperature (T_s) and T_a could promote the accumulation of more nutrients in grape berries; it also could increase photosynthetic rate and R_s during grape coloring periods. In conclusion, 60–75% SRWC was the optimum soil moisture condition, which could improve the nutrient contents and accumulate more bioactive substances. Of course, keeping a lower T_s and T_a, as well as higher air humidity, was also necessary.

Abbreviations: SRWC: soil relative water content; A, 90-95% SRWC; B, 70-75% SRWC; C, 40-55% SRWC; Rs: soil respiration; Ta: air temperature; Ts: soil temperature; OPC: proanthocyanidin; TSS: total soluble solids.     

Volatile compounds released by enzymatic hydrolysis of glycoconjugates of leaves and grape berries from Vitis vinifera Muscat of Alexandria and Shiraz cultivars.

Glycoconjugates from Muscat of Alexandria and Shiraz leaves and grape berries were isolated by adsorption on Amberlite XAD-2 resin, and enzymatically released aglycons were analyzed by GC-FID and GC-MS. About 120 aglycons were fully or tentatively identified. Compositional differences were observed between leaves and berries of the two varieties in five aglycon chemical groups: C6 alcohols, aliphatic alcohols, monoterpenes, shikimates, and C(13)-norisoprenoids, which were much more abundant in the leaves than in the berries. The differences observed for C(13)-norisoprenoids were in agreement with their hypothetical independent biosynthesis in leaves and berries. Thus, 3-hydroxy-beta-damascone, an important norisoprenoid aglycon of grape berries, was not detected in leaves, whereas its oxidized derivative, 3-oxo-alpha-damascone, was absent in berries. Compositional differences were also observed between Muscat and Shiraz leaves. 3-Oxo-alpha-ionol was not detected in Shiraz leaves, and its retro derivatives were less abundant than in Muscat of Alexandria leaves. Conversely, in Shiraz leaves the levels of 7,8-dihydroionone derivatives, such as megastigman-3,9-diol and 3-oxo-7,8-dihydro-alpha-ionol, were higher than in Muscat of Alexandria leaves.     

Alterations of flavonoid biosynthesis in young grapevine (Vitis vinifera L.) leaves, flowers, and berries induced by the dioxygenase inhibitor prohexadione-Ca

Prohexadione-Ca is a structural mimic of 2-oxoglutarate, and according to this property, it is able to inhibit dioxygenase enzymes, which require 2-oxoglutarate as a cosubstrate. Such enzymes are involved in flavonoid biosynthesis; therefore, prohexadione-Ca treatment leads to alterations in the flavonoid metabolism in grapevine tissues. Because of the fact that phenolic compounds often are responsible for enhanced plant resistance, modification of phenylpropanoid metabolism using elicitation can be considered as a new potential strategy in plant protection. The phenolic compounds were analyzed by high-performance liquid chromatography combined with chemical reaction detection. Tissue treatment induced the accumulation of unusual flavonoids, which were identified as derivatives of pentahydroxyflavanone, eriodictyol, and luteoliflavan. Concentrations of constitutive flavonoids were also affected by the bioregulator treatment. The alterations of the flavonoid profiles are discussed with respect to substrate preferences of relevant enzymes.     

Influence of plant water status on the production of C13-norisoprenoid precursors in Vitis vinifera L. Cv. cabernet sauvignon grape berries

The influence of irrigation strategy on grape berry carotenoids and C13-norisoprenoid precursors was investigated for Vitis vinifera L. cv. Cabernet Sauvignon. Two irrigation treatments were compared, one in which vines received reduced irrigation applied alternately to either side of the vine (partial rootzone drying, PRD) and a second control treatment in which water was applied to both sides of the vine. Over the two years of the experiments, PRD vines received on average 66% of the water applied to the controls. Initially, the PRD treatment did not alter midday leaf (psiL) and stem (psiS) water potential relative to the control, but decreased stomatal conductance (gs). Continued exposure to the PRD treatment resulted in treated grapevines experiencing hydraulic water deficit relative to the control treatment and induced lowered midday psiL and psiS, which was also reflected in decreased berry weight at harvest. In both irrigation treatments, the most abundant grape berry carotenoids, beta-carotene and lutein, followed the developmental pattern typical of other grape varieties, decreasing post-veraison. At certain points in time, as the fruit approached maturity, the concentration of these carotenoids was increased in fruit of PRD-treated vines relative to the controls. This effect was greater for lutein than for beta-carotene. PRD consistently caused increases in the concentration of hydrolytically released C13-norisoprenoids beta-damascenone, beta-ionone, and 1,1,6-trimethyl-1,2-dihydronaphthalene in fruit at harvest (24 degrees Brix) over two seasons. The effect of the PRD treatment on the concentration of hydrolytically released C13-norisoprenoids was greater in the second of the two seasons of the experiment and was also reflected in an increase in total C13-norisoprenoid content per berry. This suggests that the increases in the concentration of the C13-norisoprenoids in response to PRD were independent of water deficit induced changes in berry size and were not the result of an altered berry surface area to volume ratio.     

Effects of copper exposure in tissue cultured Vitis vinifera.

The present study determined the effects of copper treatment on some biochemical parameters in a closed system. Sauvignon grapevines were cultured in agar and exposed to copper levels ranging from 0.07 to 10 microg Cu/g medium. Chlorophylls, carotenoids, lipids, sucrose, soluble sugars, starch, cellulose, and minerals in root, leaves, and sap were determined. Copper levels over 5 microg Cu/g inhibit root and plantlet development. Copper-exposed plants have higher levels of chlorophylls and carotenoids as well as total lipids. Soluble sugars decrease without changes in starch or sucrose. Copper concentration increases dramatically in roots and leaves. Iron concentrations are lower in leaves, although they increase in roots, with respect to control plants. Sap flow and translocation of essential elements are reduced. Reduction of K translocation can be related to limited use of water by the plant and, thus, reduction in growth and physiological activity.     

Vitisin A content in Chilean wines from Vitis vinifera Cv. Cabernet Sauvignon and contribution to the color of aged red wines

Vitisin A was prepared from malvidin 3-glucoside and pyruvic acid in model wine medium, isolated by countercurrent chromatography, and purified by preparative high-performance liquid chromatography (HPLC). The synthesized compound was used as a reference standard to quantify vitisin A in Chilean wines from Vitis vinifera cv. Cabernet Sauvignon, including a vertical row of wines from the same vineyard over 16 years. Maximum vitisin A content was reached within the first year of storage. Importantly, up to half of the initial amount of vitisin A in young wines was still present in 15 year old wines. Although vitisin A was found to be much more stable as compared to other monomeric C-4 underivatized anthocyanins, it also slowly degrades after reaching its peak concentration. The "color activity concept" was applied to vitisin A, malvidin 3-glucoside, malvidin 3-(6' '-acetylglucoside), and polymeric pigments isolated by countercurrent chromatography in order to estimate their contribution toward the overall color expression of wines. It was found that vitisin A is only a minor contributor to the visually perceived color of aged red wines (color contribution approximately 5%). The major contributor is the polymeric fraction (color contribution approximately 70-90%).     

Survey of Nutrient Status in Vitis vinifera Grapes in North Carolina

Grape (Vitis vinifera L.) production is a rapidly growing industry in North Carolina; however, no local-research-based information is available to support nutrient-management decisions. Field studies were initiated to survey soil and plant nutrient status over a wide range in geography and management conditions in the dominant V. vinifera regions in North Carolina. While the survey data are still being analyzed, significant information was initially obtained to identify aluminum toxicity and phosphorus and potassium deficiency as potential limiting factors to vine health and productivity. Field studies were initiated to quantify soil test and plant nutrient responses to lime, phosphorus, and potassium rates. Responses to phosphorus and potassium application were observed at most sites predicted by soil tests. However, lime responses were not observed in either petiole or blade samples collected at full bloom or veraison. These studies will be continued until sufficient response data are collected to establish soil test and plant nutrient diagnostic criteria for efficient nutrient management of V. vinifera in North Carolina.     

Contribution of benzenemethanethiol to smoky aroma of certain Vitis vinifera L. wines

Benzenemethanethiol, a volatile thiol with a strong empyreumatic aroma reminiscent of smoke, has been identified in boxwood (Buxus sempervirens L.) as well as in red and white Vitis vinifera L. wines. The perception threshold in a model hydroalcoholic solution is approximately 0.3 ng/L. All of the wines analyzed for this study contained this compound in concentrations of several dozen nanograms per liter. The Chardonnay wines had 30-40 ng/L. Sensory discrimination between a wine containing 7 ng/L benzenemethanethiol and the same wine with an additional 4 ng/L is very significant; the difference in smell is described as "empyreumatic". This compound can therefore significantly contribute to the aroma of certain wines (Sauvignon Blanc, Semillon, Chardonnay, etc.) containing concentrations as high as 30-100 times higher than their perception threshold.     

Effect of lime-induced leaf chlorosis on ochratoxin A, trans-resveratrol, and epsilon-viniferin production in grapevine (Vitis vinifera L.) berries infected by Aspergillus carbonarius

Berries of Vitis vinifera L. cv. Merlot, grown on a neutral or calcareous soil, were infected, at phenological phases of veraison and ripening, by a conidial suspension of Aspergillus carbonarius to control ochratoxin A production and trans-resveratrol- and epsilon-viniferin-induced synthesis as affected by the soil lime content. Chlorosis occurrence was evaluated by a visual rating scale at veraison, and the leaves from vines growing on the calcareous soil showed the typical yellowing, whereas those grown on the neutral soil were dark green. Berry mineral element yield was recorded at veraison and ripening. Infection symptoms on berries were more severe at ripening in bunches collected from vines grown in calcareous soil. Ochratoxin A concentration increased at phenological phase of veraison in berries harvested from vines cultivated in calcareous soil. A. carbonarius enhanced trans-resveratrol and epsilon-viniferin production in infected berries more than in the control samples. Moreover, at veraison their concentration in the berries collected from vines grown in calcareous soil was greater than that recorded from berries collected from vines grown in the neutral soil. The lowest symptom severity was observed on berries containing the highest copper concentration.     

Stir bar sorptive extraction applied to volatile constituents evolution during Vitis vinifera ripening

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.     

Antioxidant activities of grape (Vitis vinifera) pomace extracts

Antioxidant-rich fractions were extracted from grape (Vitis vinifera) pomace using ethyl acetate, methanol, and water. The extracts were screened for their potential as antioxidants in different models. The ethyl acetate, methanol, and water extracts showed 76, 87.1, and 21.7% antioxidant activities at 100 ppm, respectively, using the 1,1-diphenyl-2-picrylhydrazyl model system. As the methanol extract of grape pomace showed maximum antioxidant activity among all of the extracts, it was selected to determine its effect on lipid peroxidation, hydroxyl radical scavenging activity, and human low-density lipoprotein (LDL) oxidation. The methanol extract showed 71.7, 73.6, and 91.2% inhibition using the thiobarbituric acid method, hydroxyl radical scavenging activity, and LDL oxidation, respectively, at 200 ppm. Treatment of albino rats of the Wistar strain with a single dose of CCl(4) at 1.25 mL/kg of body weight decreases the activities of catalase, superoxide dismutase (SOD), and peroxidase by 81, 49, and 89%, respectively, whereas the lipid peroxidation value increased nearly 3-fold. Pretreatment of the rats with the methanolic extract of grape pomace at 50 mg/kg (in terms of catechin equivalents) followed by CCl(4) treatment causes restoration of catalase, SOD, and peroxidase by 43.6, 73.2, and 54%, respectively, as compared with control, whereas lipid peroxidation was restored to values comparable with the control. Histopathological studies of the liver of different groups also support the protective effects exhibited by the methanol extract of grape pomace by restoring the normal hepatic architecture. Owing to this property, the studies on grape pomace can be further extended to exploit its possible application for the preservation of food products as well as a health supplement and neutraceutical.     

Effects of flumioxazin herbicide on carbon nutrition of Vitis vinifera L

To evaluate the impact of the herbicide flumioxazin (fmx) on nontarget grapevines, its effects were assessed on fruiting cuttings and field-grown plants. The stress caused by the herbicide differed according to the grapevine model. In cuttings, leaf gas exchange and photosynthetic pigment levels as well as hexose contents decreased, whereas sucrose and starch accumulated, suggesting an inhibition of photosynthesis and an increase of carbohydrate reserves as a response to the fmx-induced stress. Paradoxically, in the field-grown grapevine leaves, fmx caused a stimulation of photosynthesis, an accumulation of photosynthetic pigments and monosaccharides, in parallel with a mobilization of sucrose and starch. These results suggest that fmx reaches grapevine leaves via root uptake and has prolonged effects. In cuttings, fmx generated a toxic effect related to its target, whereas in field-grown plants, fmx had rather positive physiological effects and acts as a signal further stimulating photosynthesis and related parameters.     

Polyphenol glucosylating activity in cell suspensions of grape (Vitis vinifera)

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.     

Procyanidins from Vitis vinifera seeds: in vivo effects on oxidative stress

The purpose of this study was to evaluate the effect of supplementation with procyanidins from Vitis vinifera on markers of oxidative stress. Ten healthy volunteers received a daily dose of 110 mg of procyanidins for 30 days. Fasting venous blood samples were taken before and at the end of the supplementation period and after 7 days of wash-out. The total antioxidant activity and the plasma concentrations of alpha-tocopherol were not modified. Conversely, the levels of alpha-tocopherol in red blood cell membranes increased significantly from 1.8 +/- 0.1 to 2.8 +/- 0.2 mg/g. Similarly, the lymphocyte oxidized DNA [8-oxo-7,8-dihydro-2'-deoxyguanosine/2'-deoxyguanosine ratio] was reduced from 7.23 +/- 2.47 to 2.34 +/- 0.51, and the red blood cell membrane fatty acid composition shifted to a higher level of polyunsaturated fatty acids. On the basis of these results, it may be suggested that dietary procyanidins exert their antioxidant protection in vivo by sparing liposoluble vitamin E and reducing DNA oxidative damage.     

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

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 véraison, 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.