Best Procedures for Leaf and Stem Water Potential Measurements in Grapevine: Cultivar and Water Status Matter.

The pressure chamber is the most used tool for plant water status monitoring. However, species/cultivar and seasonal effects on protocols for reliable water potential determination have not been properly tested. In four grapevine cultivars and two times of the season (early season, Es; late season, Ls, under moderate drought), we assessed the maximum sample storage time before leaf water potential (Ψleaf) measurements and the minimum equilibration time for stem water potential (Ψstem) determination, taking 24 h leaf cover as control. In 'Pinot gris', Ψleaf already decreased after 1 h leaf storage in both campaigns, dropping by 0.4/0.5 MPa after 3 h, while in 'Refosk', it decreased by 0.1 MPa after 1 and 2 h in Es and Ls, respectively. In 'Merlot' and 'Merlot Kanthus', even 3 h storage did not affect Ψleaf. In Es, the minimum Ψstem equilibration was 1 h for 'Refosk' and 10 min for 'Pinot gris' and 'Merlot'. In Ls, 'Merlot Kanthus' required more than 2 h equilibration, while 1 h to 10 min was sufficient for the other cultivars. The observed cultivar and seasonal differences indicate that the proposed tests should be routinely performed prior to experiments to define ad hoc procedures for water status determination.

Effect of Cocoa Roasting on Chocolate Polyphenols Evolution.

Cocoa and chocolate antioxidants might contribute to human health through, for instance, blood flow improvement or blood pressure and glycemia reduction, as well as cognitive function improvement. Unfortunately, polyphenol content is reduced during cocoa fermentation, drying, roasting and all the other phases involved in the chocolate production. Here, we investigated the evolution of the polyphenol content during all the different steps of chocolate production, with a special emphasis on roasting (3 different roasting cycles with 80, 100, and 130 °C as maximum temperature). Samples were followed throughout all processes by evaluating the total polyphenols content, the antioxidant power, the epicatechin content, and epicatechin mean degree of polymerization (phloroglucinol adducts method). Results showed a similar trend for total polyphenol content and antioxidant power with an unexpected bell-shaped curve: an increase followed by a decrease for the three different roasting temperatures. At the intermediate temperature (100 °C), the higher polyphenol content was found just after roasting. The epicatechin content had a trend similar to that of total polyphenol content but, interestingly, the mean degree of polymerization data had the opposite behavior with some deviation in the case of the highest temperature, probably due to epicatechin degradation. It seems likely that roasting can free epicatechin from oligomers, as a consequence of oligomers remodeling.

Effect of extraction time on content, composition and sensory perception of proanthocyanidins in wine-like medium and during industrial fermentation of Cabernet Sauvignon.

BACKGROUND: The research objectives focused on the extraction of grape tannins during extended maceration. Skins and seeds were extracted separately in a wine-like medium. In parallel, the same grapes were fermented in industrial tanks. The content and structural characteristics of extractable proanthocyanidins (PAs) were determined spectrophotometrically and using UHPLC-DAD-MS/MS, respectively. Skin, seed extracts and fermented wines were characterized in chemical and sensorial terms after different extraction durations. RESULTS: The extraction of high molecular-weight PAs (HMWPs) from seeds increased for up to 20 days, whereas low molecular-weight PAs (LMWPs) reached a plateau earlier. The extraction of HMWPs and LMWPs from skins reached a maximum at the first sampling. Sensory evaluation confirmed greater astringency and bitterness of seed extracts with increasing time. Neither seed nor skin extracts differed statistically in terms of the mean degree of polymerization (mDP) and percentage of galloylation (%G) on different extraction days (except for seeds at the first sampling). During industrial maceration, HMWPs and LMWPs increased up to 12.7% alcohol (9 days of maceration); thereafter, the increase was not significant, whereas the mDP, %G and percentage of prodelphinidins did not significantly change after 11.4% alcohol. There were positive correlations with the wine astringency and PA content. CONCLUSION: Looking at both simulated and industrial maceration, it can be concluded that, with a longer maceration time, the increase in HMWP content was more evident than PA structural changes. The increasing content of tannins from seeds played an important role in the greater astringency and bitterness of Cabernet Sauvignon macerated at length. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Grapevine water relations and rooting depth in karstic soils.

Environmental sustainability of viticulture is negatively affected by prolonged droughts. In limestone dominated regions, there is limited knowledge on grapevine water status and on methods for accurate evaluation of actual water demand, necessary to appropriately manage irrigation. During a dry vintage, we monitored plant and soil water relations in old and young vines of Istrian Malvasia on Karst red soil. The vineyard with young vines was additionally subdivided into two areas, based on their soil type, 1) karst silty-clay loam, and 2) mixture of crushed rocks and karst silty-clay loam (stony soil). Seasonal changes in exploited water resources were estimated via analysis of oxygen isotope composition (δ18O) of rainfall, deep soil water, and xylem sap. We hypothesized that plants are able to thrive during drought thanks to the water stored in deep soil layers, while they rely less on superficial soil horizons. Our results show that vines growing on karstic substrates have deep roots securing the use of stable water sources during summer, with consequent favourable plant water status. In fact, both young and mature vines approached the threshold of severe water stress, but never surpassed it, as midday leaf water potentials were >-1.3MPa in all study sites. Vines roots showed flexible water uptake, i.e. the ability to absorb water from deep or shallow soil horizons during drought and after late-summer thunderstorms, which was particularly evident in vines growing on the stony soil. In fact, precipitations of 20mm were enough for plant water status recovery, due to fast infiltration. On the other hand, at least 50mm of rainfall were necessary to induce water status recovery in more compact soil (karst silty-clay loam). Our findings provide new knowledge on the rooting depth and water needs of vines growing on shallow soils overlying fractured limestone bedrock.

Enhancement of Istrian Malvasia wine aroma and hydroxycinnamate composition by hand and mechanical leaf removal.

BACKGROUND: Leaf removal is a viticultural practice that promotes the biosynthesis of several important grape constituents by improving fruit zone microclimate. The purpose of this study was to assess the effects of hand and mechanical leaf removal, applied at the pea-size stage of berry development, on fruit zone microclimate, volatile aroma compounds, hydroxycinnamates and sensory characteristics of Istrian Malvasia (Vitis vinifera L.) wines. Three different sunlight exposure conditions were applied: hand leaf removal (HLR), mechanical leaf removal (MLR) and untreated control (UC). RESULTS: Both leaf removal treatments, and especially the more intense HLR, significantly increased the concentration of varietal thiol 3-sulfanylhexan-1-ol, monoterpenes, ß-damascenone and esters. The higher concentration of these aromas contributed to the improvement of wine sensory quality, as expressed by more enhanced floral, fruity and tropical sensory attributes in leaf removal treatments. Hydroxycinnamates were increased only by HLR, a treatment with a greater degree of fruit exposure to sunlight than MLR. CONCLUSION: Leaf removal applied at the pea-size stage of berry development in a season characterized by abundant rainfall improves both the chemical composition and sensory quality of Istrian Malvasia wine, even when performed by machine, implying that this technique might be successfully applied in large-scale viticultural production. © 2018 Society of Chemical Industry.

UHPLC-MS/MS determination of varietal thiol precursors in Sauvignon Blanc grapes.

Varietal thiol precursors in grapes are subject to metabolic changes during post-harvest treatments. Metabolic activity should therefore be limited after sampling to understand their biosynthesis in the berry and genetic regulation. In this study, berries were frozen in liquid nitrogen immediately after harvesting, transported in dry ice, stored briefly at -80 °C, cryo-milled and extracted without being thawed in cold methanol in a ratio of 1:4 (w/v). A UHPLC-MS/MS method for quantitative determination of the thiol precursors 3-S-glutathionylhexan-1-ol (G3MH), 3-S-cysteinylhexan-1-ol (Cys3MH), 4-S-glutathionyl-4-methylpentan-2-one (G4MMP) and 4-S-cysteinyl-4-methylpentan-2-one (Cys4MMP), glutathione, oxidized glutathione and L-methionine in grapes was developed. Reference material was provided through synthesis of precursors and their deuterium labelled analogues. The average thiol precursor content in grapes in 2013-15 was in the range 8-16 µg kg-1 for G3MH, 1-6 µg kg-1 for Cys3MH, 1-4 µg kg-1 for Cys4MMP and 0.3 µg kg-1 for G4MMP. In 2013 and 2014, the highest precursor content in mature Sauvignon Blanc grapes from vineyards located in Italy regarded G3MH, followed by Cys3MH, Cys4MMP and G4MMP. In 2015, G3MH was again the most abundant precursor, but followed by Cys4MMP, Cys3MH and G4MMP.

Combined Effects of Early Season Leaf Removal and Climatic Conditions on Aroma Precursors in Sauvignon Blanc Grapes.

Early leaf removal around the cluster zone is a common technique applied in cool climate viticulture, to regulate yield components and improve fruit quality. Despite the increasing amount of information on early leaf removal and its impact on total soluble solids, anthocyanins, and polyphenols, less is known regarding aroma compounds. In order to verify the hypothesis that defoliation, applied before or after flowering, could impact the biosynthesis of thiol precursors, we performed a two year (2013 and 2014) experiment on Sauvignon blanc. We provided evidence that differential accumulation of thiol precursors in berries is affected by the timing of defoliation, and this impact was related to modifications in the biosynthetic pathway. Furthermore, the possible interaction between leaf removal treatment and seasonal weather conditions, and its effect on the biosynthesis of volatile precursors are discussed. Our results suggested that in Sauvignon blanc the relative proportion of 4-S-glutathionyl-4-methylpentan-2-one (G-4MSP) and 3-S-glutathionylhexan-1-ol (G-3SH) precursors can be affected by defoliation, and this could be related to the induction of two specific genes encoding glutathione-S-transferases (VvGST3 and VvGST5), while no significant effects on basic fruit chemical parameters, polyphenols, and methoxypyrazines were ascertained under our experimental conditions.

Plant protection product residues in white grapes and wines of "Malvasia Istriana" produced in Istria.

Monitorting of plant protection product residues was performed in 12 grape and 66 wine samples of "Malvasia Istriana" variety, produced in Istria winegrowing region of Croatia and Slovenia. The samples were analysed for the presence of 169 different active compounds using two multiresidual analytical methods: gas chromatography coupled with mass spectrometry and liquid chromatography coupled with tandem mass spectrometry. Residues were found in 58.3% of all the inspected grape samples and in 28.8% of all the inspected wine samples. Beside that contents of residues in grapes were below 10% of maximum residue level values and they should not represent any risk for "Malvasia Istriana" grape or wine consumers.

Impact of Leaf Removal, Applied Before and After Flowering, on Anthocyanin, Tannin, and Methoxypyrazine Concentrations in 'Merlot' (Vitis vinifera L.) Grapes and Wines.

The development and accumulation of secondary metabolites in grapes determine wine color, taste, and aroma. This study aimed to investigate the effect of leaf removal before flowering, a practice recently introduced to reduce cluster compactness and Botrytis rot, on anthocyanin, tannin, and methoxypyrazine concentrations in 'Merlot' grapes and wines. Leaf removal before flowering was compared with leaf removal after flowering and an untreated control. No effects on tannin and anthocyanin concentrations in grapes were observed. Both treatments reduced levels of 3-isobutyl-2-methoxypyrazine (IBMP) in the grapes and the derived wines, although the after-flowering treatment did so to a greater degree in the fruit specifically. Leaf removal before flowering can be used to reduce cluster compactness, Botrytis rot, and grape and wine IBMP concentration and to improve wine color intensity but at the expense of cluster weight and vine yield. Leaf removal after flowering accomplishes essentially the same results without loss of yield.

Plant protection product residues in red grapes and Teran PTP wine.

Between 2011 and 2013, a total of 73 grape and 82 wine samples from winegrowing district Kras (Karst) were analysed for plant protection product (PPP) residues. The samples were analysed for the presence of 215 different active compounds using two multiresidual analytical methods: gas chromatography coupled with mass spectrometry and liquid chromatography coupled with tandem mass spectrometry. Residues were found in 91.8% of all grape samples and in 67.1% of all wine samples. In all grape samples, residues were below maximum residue levels (MRLs) and 58.9% of all grape samples contained active substances at a level of less then 10% of the MRL. Nearly 33% of all Teran PTP wines did not contain PPP residues at all. Residues diminish during processing of grapes to wine, so their content in wine was low. Therefore, it could be concluded that grapes and wine in the winegrowing district Kras are safe for consumers.

Classification of grape berries according to diameter and total soluble solids to study the effect of light and temperature on methoxypyrazine, glutathione, and hydroxycinnamate evolution during ripening of Sauvignon blanc (Vitis vinifera L.).

Grape berries were classified according to diameter and total soluble solids (TSS) to study the effect of light and temperature on methoxypyrazines (MPs), glutathione (GSH), and hydroxycinnamates (HCAs) during the ripening of Sauvignon blanc. The light exposure of the fruiting zone was modified within leaf and lateral removal at the phenological stage berry of peppercorn size and no removal (control). In comparison to the control, the concentration of 3-isobutyl-2-methoxypyrazine (IBMP) was below the limit of detection in leaf removal 2 weeks before harvest. Leaf removal had no significant influence on GSH and HCAs in the grape juice at harvest. Berry diameter significantly influenced the concentration of IBMP in the grape juice and did not influence the concentration of GSH and HCAs. At harvest, the concentrations of IBMP in grape juices of similar TSS in the control were 12.6 and 5.2 ng/L in 15.5 and 13.5 mm berry diameter classes, respectively. Furthermore, the study showed that berries of the same diameter were not at the same physiological ripening level (not the same TSS).

Determination of glutathione content in grape juice and wine by high-performance liquid chromatography with fluorescence detection.

A modified preparation of sample was developed for the determination of glutathione content in grape juice and wine by high-performance liquid chromatography with fluorescence detection, using on-line pre-column derivatization. Ice-cold deoxygenated methanol was used to deactivate the oxidation enzymes in juices or wines and keep the glutathione stable. The optimum recovery of glutathione content in grape juice and wine was obtained when either the sample of grape juice or wine was mixed in ice-cold deoxygenated methanol in the ratio 10:90 (v:v) and further diluted in sodium acetate buffer in the ratio 1:1 (v:v). The optimized method was validated for linearity, limit of detection, limit of quantification, precision and uncertainty. According to the validation data the method is appropriate for the determination of glutathione content in grape juice and wine. Glutathione contents in grape juices made from White Muscat grapes and Sauvignon Blanc wines were analysed. The average glutathione content in 28 young Sauvignon Blanc wines was 12.5 mg L(-1).

Using LC-MSMS to assess glutathione levels in South African white grape juices and wines made with different levels of oxygen.

The effect of oxygen on the levels of glutathione, an important antioxidant, in must and wine was studied using a novel LC-MSMS method for the analysis of reduced and oxidized glutathione. This study found that the storage of grape juice at high SO2 and ascorbic acid levels at -20 degrees C did not lead to a decrease in reduced glutathione levels. The effect of varying the oxygen levels in South African white grape juices, which included a reductive treatment (less than 0.3 mg/L dissolved O2 added), a control treatment (between 1.0 and 1.5 mg/L dissolved O2 added), and an oxidative treatment (3.5-4 mg/ L dissolved O2 added, without SO2) on reduced glutathione levels in the juice and resulting wine was also investigated. A custom build press was used to press whole bunches of two different Sauvignon Blanc and Colombard grapes. Alcoholic fermentation and oxygen additions to the must led to lower reduced glutathione levels in the wine. Reduced glutathione levels were only significantly higher in the wine made from reductive juice that had the highest initial reduced glutathione levels in the grapes.