Differences in Anthocyanin Accumulation Profiles between Teinturier and Non-Teinturier Cultivars during Ripening.

Anthocyanins are vital components of plant secondary metabolites, and are also the most important coloring substances in wine. Teinturier cultivars are rich in anthocyanins. However, the differences in anthocyanin accumulation and profiles between teinturier and non-teinturier cultivars have not been reported. In this study, Yan 73 and Dunkelfelder were selected as the experimental materials, and three non-teinturier cultivars were used for comparison. LC-MS and qRT-PCR were used to determine the individual anthocyanin contents and the relative gene expression. The results show that the total anthocyanin content of the teinturier cultivars was considerably higher than that in non-teinturier cultivars, and the levels of individual anthocyanins increased gradually during ripening. Lower ratios of modified anthocyanins were found in the teinturier cultivars, which was not only due to the high expression level of VvUFGT and VvGST4, but also due to the relatively low expression of VvOMT in these cultivars. Cluster analysis of gene expression and anthocyanin accumulation showed that VvUFGT is related to anthocyanin accumulation, and that AM1 is related to the synthesis and transport of methylated anthocyanins. Our results will be useful for further clarifying the pathways of anthocyanin synthesis, modification, and transport in teinturier cultivars.

Exogenous 24-Epibrassinolide alleviates oxidative damage from copper stress in grape (Vitis vinifera L.) cuttings.

Copper (Cu) stress is the most common abiotic stress experienced in vineyards owing to the copper-based fungicides application. Plant hormones, including 24-Epibrassinolide (EBR), may alleviate the adverse impacts of heavy metal stress on plants. We investigated the effects of EBR pretreatment on root morphological parameters, active oxygen metabolism, osmolytes contents, antioxidant enzyme activity, endogenous phytohormone contents, and ascorbate-glutathione (AsA-GSH) cycle activity of one-year-old grape (Vitis vinifera L.) cuttings under Cu stress. Pretreatment with EBR significantly enhanced root morphological parameters (total root length, root surface area, root diameter, root volume, and tip number), increased soluble protein and proline contents, and significantly decreased the contents of H2O2, O2⋅-, and malondialdehyde (MDA) in roots and leaves. EBR pretreatment increased the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase oxidase (POD), and the contents of the endogenous phytohormones abscisic acid, jasmonic acid, and salicylic acid in the leaves. In addition, EBR regulated the balance of the AsA-GSH cycle by increasing the activities of monodehydroascorbate reductase (MDHAR), glutathione peroxidase (GR), ascorbate peroxidase (APX), and dehydroascorbate reductase (DHAR), and the contents of the antioxidant ascorbate (AsA) and dehydroascorbic acid (DHA), but the contents of glutathione (GSH) and oxidized glutathione (GSSG) decreased. Among the treatments tested, pretreatment with 0.10 mg/L EBR showed the optimal performance for alleviation of Cu toxicity. The results show that exogenous brassinosteroids reduce oxidative damage and improve the tolerance of Cu stress of grapevine cuttings.

Effect of Three Training Systems on Grapes in a Wet Region of China: Yield, Incidence of Disease and Anthocyanin Compositions of Vitis vinifera cv. Cabernet Sauvignon.

Grapevine training systems determine the suitability for grape varieties in a specific growing region. We evaluated the influence of three training systems, Single Guyot (SG), Spur-pruned Vertical Shoot-Positioned (VSP), and Four-Arm Kniffin (4AK), on the performance of grapes and vines of Vitis vinifera L. cv. Cabernet Sauvignon in the 2012 and 2013 growing seasons in a wet region of central China. 4AK was the most productive system in comparison to SG and VSP. SG and VSP had lower disease infections of leaves and berries, especially in the mid- and final stage of berry ripening. Three training systems had no impact on berry maturity. PLS-DA (Partial Least Squares-Discriminant) analysis showed that the relatively dry vintage could well discriminate three training systems, but the wet vintage was not. A wet vintage of 2013 had more accumulation of 3'5'-substituted and acylated anthocyanins, including malvidin-3-O-(6-O-acetyl)-glucoside, malvidin-3-O-glucoside, and petunidin-3-O-(cis-6-O-coumaryl)-glucoside, etc. With regard to the effect of training systems, 4AK grapes had the lowest concentrations of total anthocyanins and individual anthocyanins, SG and VSP differed according to the different vintages, and showed highest concentration of total individual anthocyanins in 2012 and 2013, respectively. Generally, VSP benefited the most, contributing to significantly highest levels of total individual anthocyanins, and major anthocyanin, including malvidin-3-O-glucoside and malvidin-3-O-(6-O-acetyl)-glucoside, and the grapes obtained from VSP presented significantly highest proportion of 3'5'-substituted anthocyanins. With regard to the ratios of 3'5'/3'-substituted, methoxylated/non-methoxylated and acylated/non-acylated anthocyanins, the significantly higher levels were also shown in VSP system. In summary, VSP was the best training system for Cabernet Sauvignon to accumulate relatively stable individual anthocyanins in this wet region of China and potentially in other rainy regions.

Brassinosteroids are involved in controlling sugar unloading in Vitis vinifera 'Cabernet Sauvignon' berries during véraison.

Sugar unloading in grape berries is a crucial step in the long-distance transport of carbohydrates from grapevine leaves to berries. Brassinosteroids (BRs) mediate many physiological processes in plants including carbohydrate metabolism. Here, 'Cabernet Sauvignon' (Vitis vinifera L.) grape berries cultivated in clay loam fields were treated with an exogenous BR (24-epibrassinolide; EBR), a BR synthesis inhibitor (brassinazole; Brz), Brz + EBR (sprayed with EBR 24 h after a Brz treatment), and deionized water (control) at the onset of véraison. The EBR treatment sharply increased the soluble sugars content in the berries, but decreased it in the skins. The EBR and Brz + EBR treatments significantly promoted the activities of both invertases (acidic and neutral) and sucrose synthase (sucrolytic) at various stages of ripening. The mRNA levels of genes encoding sucrose metabolic invertase (VvcwINV), and monosaccharide (VvHT3, 4, 5 and 6) and disaccharide (VvSUC12 and 27) transporters were increased by the EBR and/or Brz + EBR treatments. Generally, the effects of the Brz treatment on the measured targets contrasted with the effects of the EBR treatments. The EBR and Brz treatments inhibited the biosynthesis of the endogenous BRs 6-deoxocastastarone and castasterone. Both EBR and Brz + EBR treatments increased the brassinolide contents, down-regulated the expression of genes encoding BRs biosynthetic enzymes BRASSINOSTEROID-6-OXIDASE and DWARF1, (VvBR6OX1 and VvDWF1) and induced BR receptor gene BRASSINOSTEROID INSENSITIVE 1 (VvBRI1) expression in deseeded berries. Together, these results show that BRs are involved in controlling sugar unloading in grape berries during véraison.

Melatonin treatment of pre-veraison grape berries to increase size and synchronicity of berries and modify wine aroma components.

A comprehensive investigation was carried out to determine the effect of exogenous melatonin treatment of pre-veraison grapes on grape berries and its wines. Two melatonin treatments of pre-veraison grape berries increased the weight of the berries by approximately 6.6%. Meanwhile, this melatonin treatment could be beneficial in the reduction of underripe and overripe fruits and in enhancing the synchronicity of the berries. In addition, there were significant differences in the volatile compound composition between the wine produced from the melatonin-treated berries and the wines made from untreated berries. The wine from melatonin-treated pre-veraison grape berries had stronger fruity, spicy, and sweet sensory properties, compared to the wines made from untreated berries. Prolonging the treatment through repeated applications can enhance these effects and under different seasonal conditions, more pronounced effects on the grape quality and wine properties can be observed.

Promoting effect of foliage sprayed zinc sulfate on accumulation of sugar and phenolics in berries of Vitis vinifera cv. Merlot growing on zinc deficient soil.

The effect of foliage sprayed zinc sulfate on berry development of Vitis vinifera cv. Merlot growing on arid zone Zn-deficient soils was investigated over two consecutive seasons, 2013 and 2014. Initial zinc concentration in soil and vines, photosynthesis at three berry developmental stages, berry weight, content of total soluble solids, titratable acidity, phenolics and expression of phenolics biosynthetic pathway genes throughout the stages were measured. Foliage sprayed zinc sulfate showed promoting effects on photosynthesis and berry development of vines and the promotion mainly occurred from veraison to maturation. Zn treatments enhanced the accumulation of total soluble solids, total phenols, flavonoids, flavanols, tannins and anthocyanins in berry skin, decreasing the concentration of titratable acidity. Furthermore, foliage sprayed zinc sulfate could significantly influence the expression of phenolics biosynthetic pathway genes throughout berry development, and the results of expression analysis supported the promotion of Zn treatments on phenolics accumulation. This research is the first comprehensive and detailed study about the effect of foliage sprayed Zn fertilizer on grape berry development, phenolics accumulation and gene expression in berry skin, providing a basis for improving the quality of grape and wine in Zn-deficient areas.

The ameliorative effects of exogenous melatonin on grape cuttings under water-deficient stress: antioxidant metabolites, leaf anatomy, and chloroplast morphology.

Grapes are an important economic crop and are widely cultivated around the world. Most grapes are grown in arid or semi-arid regions, and droughts take a heavy toll in grape and wine production areas. Developing effective drought-resistant cultivation measures is a priority for viticulture. Melatonin, an indoleamine, mediates many physiological processes in plants. Herein, we examined whether exogenously applied melatonin could improve the resistance of wine grape seedlings grown from cuttings to polyethylene glycol-induced water-deficient stress. The application of 10% polyethylene glycol (PEG) markedly inhibited the growth of cuttings, caused oxidative stress and damage from H2 O2 and O2∙-, and reduced the potential efficiency of Photosystem II and the amount of chlorophyll. Application of melatonin partially alleviated the oxidative injury to cuttings, slowed the decline in the potential efficiency of Photosystem II, and limited the effects on leaf thickness, spongy tissue, and stoma size after application of PEG. Melatonin treatment also helped preserve the internal lamellar system of chloroplasts and alleviated the ultrastructural damage induced by drought stress. This ameliorating effect may be ascribed to the enhanced activity of antioxidant enzymes, increased levels of nonenzymatic antioxidants, and increased amount of osmoprotectants (free proline). We conclude that the application of melatonin to wine grapes is effective in reducing drought stress.

Phenolic profiles and antioxidant properties of young wines made from Yan73 (Vitis vinifera L.) and Cabernet Sauvignon (Vitis vinifera L.) grapes treated by 24-epibrassinolide.

The grape berries of two varieties, Yan73 (Vitis vinifera L.) and Cabernet Sauvignon (CS) (Vitis vinifera L.) were treated with 0.40 mg/L 24-epibrassinolide (EBR), 1.00 mg/L brassinazole (Brz), and deionized water (control), at the veraison period. The EBR treatment significantly increased total phenolic content (TPC), total tannin content (TTC) and total anthocyanin content (TAC) of Yan73 and CS wines, whereas Brz treatment decreased TPC, total flavonoid content (TFC), TAC in the two wines. Moreover, the content of most of the phenolic compounds identified by HPLC-DAD/ESI-MS in EBR-treated wines was significantly higher than that in control. The antioxidant capacities, which determined using DPPH, ABTS and HRSA methods, of the wines were increased by EBR treatment as well. There was a good correlation between the antioxidant capacity and phenolic content. The results demonstrated that EBR could enhance the phenolic compounds and antioxidant capacity of Yan73 and CS wines, but the effects may vary by different cultivars.

Regulating the secondary metabolism in grape berry using exogenous 24-epibrassinolide for enhanced phenolics content and antioxidant capacity.

The objective of this study is to investigate the influence of application of exogenous 24-epibrassinolide (EBR) on the antioxidant capacity, anthocyanins and phenolics content of Vitis vinifera grape berry (cvs. Yan 73 and Cabernet Sauvignon). The grapevine clusters were sprayed with 0 (control), 0.10, 0.40, or 0.80 mg/l of 24-epibrassinolide during veraison, respectively. The EBR application increased the activities of phenylalanine ammonia-lyase (PAL) and UDP-glucose: flavonoid 3-O-glucosyltransferase (UFGT), the content of total phenolics, tannins, flavonoids and anthocyanins, individual anthocyanins and the antioxidant capacity of matured grape skins in both varieties. The application also increased the content of flavonoids and anthocyanins in Yan73 pulp. Compared to the other treatments, the treatment of EBR at 0.40 mg/l had significantly higher level than the control in all above assays. Our results indicated that the exogenous EBR treatment can significantly promote grape ripening and enhance anthocyanins and other phenolics contents and antioxidant capacity in the grape skin.

Exogenously applied abscisic acid to Yan73 (V. vinifera) grapes enhances phenolic content and antioxidant capacity of its wine.

Yan73 is a 'teinturier' red wine variety cultivated in China and widely used in winemaking to strengthen red wine colour. The objective of this study was to evaluate the effect of exogenous abscisic acid (ABA) applied to the grapevine cluster on the antioxidant capacity and phenolic content of the wine made from Yan73. Two hundred mg/l ABA was applied on Yan73 grapevine cluster during veraison. As they mature, these ABA-treated and untreated grape berries were transformed into wines, respectively, and the phenolic content and antioxidant capacity of these wines were compared. The results showed that phenolic content (total phenolics, tannins, flavonoids and anthocyanins) and antioxidant capacity were higher in the wine produced with ABA-treated Yan73 grapes than those in the wine from untreated grapes. Compared to Cabernet Sauvignon wine, Yan73 wine had higher phenolic content and stronger antioxidant capacity. These strongly suggest that exogenously applied ABA to Yan73 grapes can enhance phenolic content and antioxidant capacity of its wine, and Yan73 wine has the higher utilization value and potential for development.

Impact of cover crops in vineyard on the aroma compounds of Vitis vinifera L. cv Cabernet Sauvignon wine.

This study compared the influence of different cover crops with clean tillage on wine aroma compounds of 5-year-old Cabernet Sauvignon vines. White clover, alfalfa, and tall fescue were used in the vineyard and compared with clean tillage. Aroma compounds of wine were analysed by solid-phase microextraction gas chromatography-mass spectrometry (SPME-GC/MS). Forty-seven volatile compounds were identified and quantified. Wines made from grapes grown with various cover crops had higher levels of aroma compounds. Ethyl acetate, isoamyl acetate, ethyl octanoate, ethyl hexanoate, phenylethyl acetate, isoamyl alcohol, linalool, citronellol, ß-damascenone, α-ionone, and 5-amyl-dihydro-2(3H)-furan were the impact odorants of sample wines. Wines from cover crop also had higher contents of these impact odorants than the control. For different cover crops, alfalfa sward yielded the highest levels, followed by the tall fescue treatment. According to the data analysis of aroma compounds and sensory assess, permanent cover crop may have the potential to improve wine quality.