Biochemical Response and Gene Expression to Water Deficit of Croatian Grapevine Cultivars (Vitis vinifera L.) and a Specimen of Vitis sylvestris.

The biochemical response and gene expression in different grapevine cultivars to water deficit are still not well understood. In this study, we investigated the performance of four traditional Croatian Vitis vinifera L. cultivars ('Plavac mali crni', 'Istrian Malvasia', 'Grasevina', and 'Tribidrag'), and one wild (Vitis vinifera subsp. sylvestris) genotype exposed to water deficit (WD) for nine days under semi-controlled conditions in the greenhouse. Sampling for biochemical and gene expression analyses was performed at days six and nine from the beginning of WD treatment. The WD affected the accumulation of metabolites with a significant increase in abscisic acid (ABA), salicylic acid (SA), and proline in the leaves of the stressed genotypes when the WD continued for nine days. Lipid peroxidation (MDA) was not significantly different from that of the control plants after six days of WD, whereas it was significantly lower (297.40 nmol/g dw) in the stressed plants after nine days. The cultivar 'Istrian Malvasia' responded rapidly to the WD and showed the highest and earliest increase in ABA levels (1.16 ng mg-1 dw, i.e., 3.4-fold increase compared to control). 'Grasevina' differed significantly from the other genotypes in SA content at both time points analyzed (six and nine days, 47.26 and 49.63 ng mg-1 dw, respectively). Proline level increased significantly under WD (up to 5-fold at day nine), and proline variation was not genotype driven. The expression of aquaporin genes (TIP2;1 and PIP2;1) was down-regulated in all genotypes, coinciding with the accumulation of ABA. The gene NCED1 (9-cis-epoxycarotenoid dioxygenase) related to ABA was up-regulated in all genotypes under stress conditions and served as a reliable marker of drought stress. This work suggests that the stress response in metabolite synthesis and accumulation is complex, treatment- and genotype-dependent.

Applying Different Vinification Techniques in Teran Red Wine Production: Impact on Bioactive Compounds and Sensory Attributes.

Six different vinification treatments, including a control treatment (7-day standard maceration) (K7), were performed to study the effects of non-standard techniques on bioactive compounds and sensory attributes of Teran red wine. Pre-fermentative mash cooling (8 °C; 48 h) and heating (50 °C; 48 h) followed by prolonged post-fermentative maceration of 13 days (C15;H15) or 28 days (C30;H30) were applied. In another treatment, after cooling, saignée was performed followed by 13-day prolonged maceration (CS15). Wine phenols and vitamins were analyzed by HPLC-DAD-FLD, minerals by ICP-OES, and sensory analysis was performed using the QDA and 100-point O.I.V./U.I.O.E. methods. Obtained results showed total phenolic concentration was the highest in the H30 treatment. The concentration of anthocyanins, flavan-3-ols and phenolic acids was significantly higher in wines of all vinification techniques compared to the control. Stilbene content was highly affected by pre-fermentative heating. Treatments CS15, H15, C30 and H30 resulted in the highest scores by both the QDA and 100-point sensory methods. The obtained results suggest that advanced non-standard vinification techniques have a significant impact on Teran wine by enhancing its composition of bioactive compounds and improving its sensory profile, which gives it an additional market value. Furthermore, a comprehensive comparison of such techniques applied simultaneously in one study is of substantial importance for additional research in wine production.

Severe Shoot Trimming and Crop Size as Tools to Modulate Cv. Merlot Berry Composition.

Viticulture production is challenged by climate change and the consequent higher accumulation of carbohydrates in grapevine berries, resulting in high-alcoholic wines. This study investigates the application of severe shoot trimming performed at three different stages and crop size management as tools for the modulation of cv. Merlot berry composition, aimed at reducing the sugar content in the berry. In the first study, the effects of severe shoot trimming carried out at three different phenological stages were studied. In the second study, late severe shoot trimming was combined with two crop sizes and regulated by shoot thinning. The obtained results demonstrated that severe shoot trimming in earlier stages of berry development limited the accumulation of both sugars and anthocyanins as compared to the control treatment. However, when severe shoot trimming was performed at late veraison (at approximately 14 Brix), it decreased only the accumulation of sugars, without affecting the accumulation of anthocyanins. The results of the second study showed that the modification of crop size by shoot thinning significantly affected the measured yield parameters, whereas the effect on Brix and anthocyanins was seasonally dependent. It was concluded that among the studied techniques, severe shoot trimming at late veraison is the most effective way to reduce sugar content in the berry without affecting the accumulation of anthocyanins.

Genetic Diversity, Population Structure, and Parentage Analysis of Croatian Grapevine Germplasm.

Croatian viticulture was most extensive at the beginning of the 20th century, when about 400 varieties were in use. Autochthonous varieties are the result of spontaneous hybridization from the pre-phylloxera era and are still cultivated today on about 35 % of vineyard area, while some exist only in repositories. We present what is the most comprehensive genetic analysis of all major Croatian national repositories, with a large number of microsatellite, or simple sequence repeat (SSR) markers, and it is also the first study to apply single nucleotide polymorphism (SNP) markers. After 212 accessions were fingerprinted, 95 were classified as unique to Croatian germplasm. Genetic diversity of Croatian germplasm is rather high considering its size. SNP markers proved useful for fingerprinting but less informative and practical than SSRs. Analysis of the genetic structure showed that Croatian germplasm is predominantly part of the Balkan grape gene pool. A high number of admixed varieties and synonyms is a consequence of complex pedigrees and migrations. Parentage analysis confirmed 24 full parentages, as well as 113 half-kinships. Unexpectedly, several key genitors could not be detected within the present Croatian germplasm. The low number of reconstructed parentages (19%) points to severe genetic erosion and stresses the importance of germplasm repositories.

Crop level vs. leaf removal: Effects on Istrian Malvasia wine aroma and phenolic acids composition.

The impact of crop level and leaf removal on Istrian Malvasia (Vitis vinifera L.) white wine aroma and phenolic acids composition was studied over two vine-growing seasons. Two crop levels were combined with leaf removal or untreated control in two-factorial design. Crop level showed limited impact on aroma compounds in wine and the concentration of some esters was increased by higher crop level. In contrast, leaf removal increased the concentration of several aroma compounds and especially monoterpenes and esters. The concentration of hydroxycinnamic acids in wine was enhanced only by leaf removal, while no consistent impact of the investigated factors on hydroxybenzoic acids was observed. The obtained results suggest that in cases where environmental conditions are not limiting, increasing the crop level under adequate microclimate in fruit zone has no detrimental effects on white wine aroma and hydroxycinnamic acids composition, potentially leading to economically more sustainable grape production.

Targeted UPLC-QqQ-MS/MS profiling of phenolic compounds for differentiation of monovarietal wines and corroboration of particular varietal typicity concepts.

Targeted ultra-performance liquid chromatography with triple quadrupole mass spectrometric (UPLC-QqQ-MS/MS) profiling of phenolic compounds was utilised for varietal differentiation of 173 wines made from four red and six white grape varieties. Among 58 identified phenols many were found relevant as exclusive or partial discriminators between wines. Successful differentiation models were built by linear discriminant analysis with the percentage of correct classification higher than 95% in all cases, with peonidin 3-(6″-acetyl)-glucoside and taxifolin as the most potent differentiators between red, and cis-piceid between white monovarietal wines. Diverse typical colour attributes among the monovarietal wines were tentatively ascribed to the variations in the composition of monomeric anthocyanins. Plavac mali red wine exhibited the most specific composition, and its most typical samples were distinguished by the abundance in trans-fertaric acid, isorhapontin, phlorizin, quercetin 3-rhamnoside, and myricitrin. Despite positive correlations with particular astringent flavonols, the typical astringency of Plavac mali wine remained unresolved.

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.

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.

Pre-fermentative cold maceration, saignée, and various thermal treatments as options for modulating volatile aroma and phenol profiles of red wine.

The effects of six maceration treatments on volatile aroma and phenol composition of Teran red wine were studied: standard maceration (control C), cold pre-fermentation maceration (CPM), saignée (S), pre-fermentation heating with extended maceration (PHT) or juice fermentation (PHP), and post-fermentation heating (POH). PHP wine contained the highest amounts of esters, fatty acids and anthocyanins, and the lowest content of other phenols. Alternative treatments decreased higher alcohols in relation to control C. CPM treatment lowered the extraction of seed tannins, exhibited the highest acetaldehyde, ethyl acetate and C6-compounds levels, and had increased ester levels in relation to control C. POH wine contained the highest concentration of total phenols, flavonoids, monomeric, oligomeric and polymeric flavanols, and color intensity and hue. S and PHT wines contained lower amount of total phenols, but higher than in C and CPM wines. The calculated Odor Activity Values were used to establish significant differences between the treatments.