VvERF117 positively regulates grape cold tolerance through direct regulation of the antioxidative gene BAS1.

Cold stress significantly threatens grape quality, yield, and geographical distribution. Although ethylene-responsive factors (ERFs) are recognized for their pivotal roles in cold stress, the regulatory mechanisms of many ERFs contributing to tolerance remain unclear. In this study, we identified the cold-responsive gene VvERF117 and elucidated its positive regulatory function in cold tolerance. VvERF117 exhibits transcriptional activity and localizes to the nucleus. VvERF117 overexpression improved cold tolerance in transgenic Arabidopsis, grape calli, and grape leaves, whereas VvERF117 silencing increased cold sensitivity in grape calli and leaves. Furthermore, VvERF117 overexpression remarkably upregulated the expression of several stress-related genes. Importantly, BAS1, encoding a 2-Cys peroxidase (POD), was confirmed as a direct target gene of VvERF117. Meanwhile, compared to the wild-type, POD activity and H2O2 content were remarkably increased and decreased in VvERF117-overexpressing grape calli and leaves, respectively. Conversely, VvERF117 silencing displayed the opposite trend in grape calli and leaves under cold stress. These findings indicate that VvERF117 plays a positive role in cold resistance by, at least in part, enhancing antioxidant capacity through regulating the POD-encoding gene VvBAS1, leading to effective mitigation of reactive oxygen species.

Telomere-to-telomere and gap-free genome assembly of a susceptible grapevine species (Thompson Seedless) to facilitate grape functional genomics.

Grapes are globally recognized as economically significant fruit trees. Among grape varieties, Thompson Seedless holds paramount influence for fresh consumption and for extensive applications in winemaking, drying, and juicing. This variety is one of the most efficient genotypes for grape genetic modification. However, the lack of a high-quality genome has impeded effective breeding efforts. Here, we present the high-quality reference genome of Thompson Seedless with all 19 chromosomes represented as 19 contiguous sequences (N50 = 27.1 Mb) with zero gaps and prediction of all telomeres and centromeres. Compared with the previous assembly (TSv1 version), the new assembly incorporates an additional 31.5 Mb of high-quality sequenced data with annotation of a total of 30 397 protein-coding genes. We also performed a meticulous analysis to identify nucleotide-binding leucine-rich repeat genes (NLRs) in Thompson Seedless and two wild grape varieties renowned for their disease resistance. Our analysis revealed a significant reduction in the number of two types of NLRs, TIR-NB-LRR (TNL) and CC-NB-LRR (CNL), in Thompson Seedless, which may have led to its sensitivity to many fungal diseases, such as powdery mildew, and an increase in the number of a third type, RPW8 (resistance to powdery mildew 8)-NB-LRR (RNL). Subsequently, transcriptome analysis showed significant enrichment of NLRs during powdery mildew infection, emphasizing the pivotal role of these elements in grapevine's defense against powdery mildew. The successful assembly of a high-quality Thompson Seedless reference genome significantly contributes to grape genomics research, providing insight into the importance of seedlessness, disease resistance, and color traits, and these data can be used to facilitate grape molecular breeding efforts.

Effects of ultraviolet and infrared radiation absence or presence on the aroma volatile compounds in winegrape during veraison.

Light environmental factors have been identified to influence grape aromas greatly. Among them, the effect of non-visible light on grape aroma compounds has scarcely been investigated during grape growth and development. In the present study, ultraviolet (UV) or infrared (IR) radiation was eliminated in the grape bunch zone, and the grape bunches were irradiated with UV or IR light in vitro. The effect of UV and IR radiation on the grape aroma profile of the Cabernet Sauvignon variety was assessed by headspace solid phase microextraction gas chromatography mass spectrometer (HS-SPME GC-MS). A total of 16 aroma compounds were identified in the grape berries under UV radiation absence (UV-) or IR radiation absence (IR-). They were classified into aliphatic alcohols, aliphatic acids, benzenolds, aldehydes, and monoterpenes. A total of 23 aroma compounds were identified in the grape berries under UV radiation presence (UV+) or IR radiation presence (IR+), which were classified into aliphatic alcohols, aliphatic ketones, aliphatic esters, aliphatic acids, monoterpenes, aldehydes, volatile phenols, and other volatiles. Linalool and hexanal aroma compounds were the most responsive to UV- and UV+, according to OPLS-DA analysis. Hexanal was increased by UV- and decreased by UV+, thus was negatively correlated with UV radiation. Benzaldehyde and 2-decanone were also found as the main differing aroma compounds according to VIP scores in the IR- and IR+, respectively. The significant differences of aroma compounds in three UV and IR intensities were also observed by headspace gas chromatography-ion mobility spectrometry (HS-GC-IMS). The content of acetic acid, 2-methylbutanal, and pentanal were reduced with the radiation intensity increase, and the content of 2-3-butanedione, butyl acetate, and 1-hexanol was enhanced, especially with UV radiation. This study improves our understanding of the non-visible light role in volatile aroma compound accumulation and further expands on the useful wavelength for plant growth and development. Our study provides a theoretical basis for non-visible light field management and indoor plant growth applications.

Emerging roles of brassinosteroids and light in anthocyanin biosynthesis and ripeness of climacteric and non-climacteric fruits.

Anthocyanins are important pigments that contribute to fruit quality. The regulation of anthocyanin biosynthesis by several transcription factors via sophisticated regulatory networks has been studied in various plants. Brassinosteroids (BRs), a new class of plant hormone, are involved in regulating anthocyanin biosynthesis in fruits. Furthermore, light directly affects the synthesis and distribution of anthocyanins. Here, we summarize the recent progress toward understanding the impact of BR and light on anthocyanin biosynthesis in climacteric and non-climacteric fruits. We review the BR and light signaling pathways and highlight the important transcription factors that are associated with the synthesis of anthocyanins, such as BZR1 (brassinazole-resistant 1, BR signaling pathway), HY5 (elongated hypocotyl 5) and COP1 (constitutively photomorphogenic 1, light signal transduction pathway), which bind with the target genes involved in anthocyanin synthesis. In addition, we review the mechanism by which light signals interact with hormonal signals to regulate anthocyanin biosynthesis.

Effects of UVA disappearance and presence on the acylated anthocyanins formation in grape berries.

Ultraviolet A (UVA), the major component of the UV, plays a crucial role in formatting the characteristics of color in wine grapes by influencing its anthocyanin composition and contents. Results showed that anthocyanin biosynthesis was suppressed by UVA screening and enhanced by irradiation. The acetylation and p-coumaroylation of anthocyanins were more pronounced and showed positive correlation with a* and negative correlation with L*, b*, C*, and h, thereby leading to changes in color. Weighted gene co-expression network analysis showed that two modules (red and turquoise) were significantly related to the acetylation and p-coumaroylation of peonidin. In addition, relative gene expression assays and correlation analysis also indicated that VvMYBA1 might influence anthocyanin accumulation by directly regulating VvOMT expression and increasing the flux to the vacuole through VvGST4. In conclusion, the results helped in improving our understanding of the role of UVA in skin color formation.

Involvement of Anthocyanin Biosynthesis of Cabernet Sauvignon Grape Skins in Response to Field Screening and In Vitro Culture Irradiating Infrared Radiation.

To study the role of infrared (IR) radiation in the color change of the grape berry, field screening (IR-) and in vitro culture irradiation (IR+) were used. Acylated anthocyanin biosyntheses, including the biosynthesis of malvidin 3-O-glucoside, peonidin 3-O-glucoside, and their derivatives (acetylation and p-coumaroylation), were inhibited by IR-. IR+ promoted the biosynthesis of malvidin 3-O-glucoside and its derivatives, and IR+ inhibited the biosynthesis of peonidin 3-O-glucoside and its derivatives. WGCNA analysis revealed that the red module positively correlated with the flavonoid pathway. The hub genes were related to the anthocyanin pathway, including VvF3'5'H, VvANS, VvOMT1, VIT_18s0001g09400, and VvGST4. Further, the results revealed that transcription factors like RLK-Pelle, MYB, and C2H2 families were involved in response to IR radiation. Therefore, these results provide a complete understanding of IR radiation in grape skin color formation and the prospect of using supplemental light to improve the overall color of berries.

The Expression of Aroma Components and Related Genes in Merlot and Marselan Scion-Rootstock Grape and Wine.

Rootstocks were bred and selected from several species in order to enhance the resistance against biotic or abiotic stresses. There are few studies on the effect of rootstocks on aroma and related gene expression. This study focused on the effects of three rootstocks, Kober 5BB (5BB), 1103 Paulsen (1103P), and Selection Oppenheim (SO4), on the aroma and volatile-related gene expression levels of Merlot and Marselan berries and wines. These three rootstocks reduced the total aroma content of Merlot wine. 5BB upregulated VvLoXA and showed increased C6 alcohols. 1103P enhanced the linalool from Merlot berry, with marked upregulation of VvLinNer1. Conversely, rootstocks increased the total aroma content of Marselan berry, verified by the related expression levels of volatile-related genes. For Marselan berry, 5BB and 1103P upregulated five VvGTs and nine genes from the LOX and MEP pathway. 1103P increased the contents of C6 alcohols, C6 aldehydes, and citronellol from Marselan berry. Compared to 5BB and SO4, rootstock 1103P provided berries of better quality and richer aroma volatiles to Merlot and Marselan, while all three of the rootstocks had a significant effect on scion-rootstocks.

Characterization of 24-epibrassinolide-mediated modulation of the drought stress responses: Morphophysiology, antioxidant metabolism and hormones in grapevine (Vitis vinifera L.).

Drought stress is one of the major abiotic stresses that limit grape growth and yield. Brassinosteroids (BRs) are a class of phytohormones essential for plant growth, development, and adaptation to environmental stress. This study aimed to reveal the physiological and biochemical mechanisms of exogenous BRs in alleviating the drought stress in grapevines. Two-year-old grape seedlings (Vitis vinifera L.) were sprayed with 24-epibrassinolide (EBR), a synthetic analog of BRs, and then subjected to drought treatment. The results showed that exogenous EBR significantly mitigated the reduction of photosynthetic pigment contents and photosystem II efficiency and decreased the damage to chloroplasts when grape seedlings were subjected to drought stress. Drought stress resulted in the accumulation of reactive oxidative species (ROS) and an increase in lipid peroxidation. A reduction in oxidative damage was observed in EBR-pretreated plants, which was probably due to the elevated antioxidant system. Exogenous EBR improved the activities of superoxide dismutase (14%), catalase (18%), peroxidase (17%), and ascorbate peroxidase (9%), and promoted the accumulation of ascorbic acid (10%) and glutathione (7%) under drought stress. EBR pretreatment also promoted autophagic activity, which contributed to the degradation of damaged chloroplasts. Moreover, EBR pretreatment increased the concentrations of abscisic acid, jasmonic acid, auxin, and gibberellic acid. Taken together, exogenous EBR could ameliorate the deleterious effects of drought stress by up-regulating photosynthetic capacity, antioxidant system, autophagic activity, and hormone concentrations.

Exogenous Putrescine Alleviates Drought Stress by Altering Reactive Oxygen Species Scavenging and Biosynthesis of Polyamines in the Seedlings of Cabernet Sauvignon.

Climate change imposes intensive dry conditions in most grape-growing regions. Drought stress is one of the most devastating abiotic factors threatening grape growth, yield, and fruit quality. In this study, the alleviation effect of exogenous putrescine (Put) was evaluated using the seedlings of Cabernet Sauvignon (Vitis vinifera L.) subjected to drought stress. The phenotype, photosynthesis index, membrane injury index (MII), and antioxidant system, as well as the dynamic changes of endogenous polyamines (PAs) of grape seedlings, were monitored. Results showed that drought stress increased the MII, lipid peroxidation, and the contents of reactive oxygen species (ROS) (H2O2 and O2 -), while it decreased the antioxidant enzyme activity and the net photosynthesis rate (Pn). However, the application of Put alleviated the effects of drought stress by altering ROS scavenging, enhancing the antioxidant system, and increasing the net Pn. Put distinctly increased the activity of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT), as well as the contents of ascorbic acid (AsA) and glutathione (GSH). Meanwhile, exogenous Put also promoted the metabolism of endogenous PAs by upregulating their synthetic genes. Our results confirmed that the exogenous application of Put can enhance the antioxidant capacity as well as alter the PA pool, which provides better drought tolerance for Cabernet Sauvignon seedlings.

Effects of leaf removal on hexose accumulation and the expression of sugar unloading-related genes in syrah grapes.

Leaf removal (LR) around the cluster zone is a common practice for regulating grape quality. The purpose of this study was to assess the effects of cluster-zone leaf removal, applied at the pea-size stage of berry development, on berry soluble sugar, organic acid and phenolic compound, sugar unloading-related gene expression of Vitis. vinifera L. Syrah. Four different severities of leaf removal were applied: no leaf removal (Control), removing 2 leaves above the cluster (LR1), removing 4 leaves above the cluster (LR2), and removing 6 leaves above the cluster (LR3). The three leaf removal treatments (LR), especially removing 4 leaves (LR2), resulted in significantly higher reducing sugar, soluble sugar (glucose, fructose and sucrose), total anthocyanin and citric acid contents as compared to the control group during ripening for both vintages. At harvest, the LR treatments increased the transcript abundance of most sugar unloading-related genes. In addition, VvHT3, VvHT5, VvSUC11, VvSUC12, VvSS and VvcwINV were positively correlated with both reducing sugar contents and soluble sugar contents. Our results suggest that removing 4 leaves above the cluster is useful for improving the quality of Syrah (Vitis vinifera L.) grapes in cool climate regions with excessive leaves. These findings provide insights into the molecular basis of the relationship between leaf removing and hexose (glucose and fructose) accumulation in the grape berries.

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.

Whole-genome sequencing reveals rare off-target mutations in CRISPR/Cas9-edited grapevine.

The CRISPR (clustered regularly interspaced short palindromic repeats)-associated protein 9 (Cas9) system is a powerful tool for targeted genome editing, with applications that include plant biotechnology and functional genomics research. However, the specificity of Cas9 targeting is poorly investigated in many plant species, including fruit trees. To assess the off-target mutation rate in grapevine (Vitis vinifera), we performed whole-genome sequencing (WGS) of seven Cas9-edited grapevine plants in which one of two genes was targeted by CRISPR/Cas9 and three wild-type (WT) plants. In total, we identified between 202,008 and 272,397 single nucleotide polymorphisms (SNPs) and between 26,391 and 55,414 insertions/deletions (indels) in the seven Cas9-edited grapevine plants compared with the three WT plants. Subsequently, 3272 potential off-target sites were selected for further analysis. Only one off-target indel mutation was identified from the WGS data and validated by Sanger sequencing. In addition, we found 243 newly generated off-target sites caused by genetic variants between the Thompson Seedless cultivar and the grape reference genome (PN40024) but no true off-target mutations. In conclusion, we observed high specificity of CRISPR/Cas9 for genome editing of grapevine.

Effect of ammonium and nitrate supplies on nitrogen and sucrose metabolism of Cabernet Sauvignon (Vitis vinifera cv.).

BACKGROUND: The quality of wine is highly dependent on the quality of berries. Development of berries is influenced by the type and ratio of different nitrogen supplies in the soil. To understand the impact of varying sources and levels of nitrate and ammonium on sucrose and nitrogen metabolism of Vitis vinifera cv. Cabernet Sauvignon, we tested nutrient solutions with four NO3 - -N:NH4 + -N ratios (100:0, 75:25, 50:50, 0:100) through the root system. RESULTS: The form and quantity of nitrogen affected berries and leaves with source-sink relationships. Soluble sugar levels were significantly higher in plants treated with mixed nitrogen sources (75:25 and 50:50) compared to single nitrogen sources (100:0 and 0:100). In particular, treating plants with mixed nitrogen source at a 75:25 ratio resulted in 22% higher fructose levels in berries compared to the 50:50 treatment. In addition, mixed nitrogen treatments resulted in significantly higher amino acid levels and protein content. Mixed nitrogen substrates also increased the expression of enzymes involved in both nitrogen and sucrose metabolism. CONCLUSION: Plants did not maximize the nitrogen supply when single form nitrogen was provided, and the mixed nitrogen substrates consistently increased the amount of available carbon and nitrogen in the berries and leaves. We found that NO3 - -N:NH4 + -N ratio of 75:25 was the optimum formula for improving nitrogen and sucrose metabolism, and reducing the competition between nitrogen and sucrose. By examining the nutrient utilization of plants cultivated with different nitrogen forms, the present study provides insights into improving cultivation and production practices. © 2020 Society of Chemical Industry.

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.

Exogenous 24-Epibrassinolide Interacts with Light to Regulate Anthocyanin and Proanthocyanidin Biosynthesis in Cabernet Sauvignon (Vitis vinifera L.).

Anthocyanins and proanthocyanidins (PAs) are crucial factors that affect the quality of grapes and the making of wine, which were stimulated by various stimuli and environment factors (sugar, hormones, light, and temperature). The aim of the study was to investigate the influence of exogenous 24-Epibrassinolide (EBR) and light on the mechanism of anthocyanins and PAs accumulation in grape berries. Grape clusters were sprayed with EBR (0.4 mg/L) under light and darkness conditions (EBR + L, EBR + D), or sprayed with deionized water under light and darkness conditions as controls (L, D), at the onset of veraison. A large amount of anthocyanins accumulated in the grape skins and was measured under EBR + L and L treatments, whereas EBR + D and D treatments severely suppressed anthocyanin accumulation. This indicated that EBR treatment could produce overlay effects under light, in comparison to that in dark. Real-time quantitative PCR analysis indicated that EBR application up-regulated the expression of genes (VvCHI1, VvCHS2, VvCHS3, VvDFR, VvLDOX, VvMYBA1) under light conditions. Under darkness conditions, only early biosynthetic genes of anthocyanin biosynthesis responded to EBR. Furthermore, we also analyzed the expression levels of the BR-regulated transcription factor VvBZR1 (Brassinazole-resistant 1) and light-regulated transcription factor VvHY5 (Elongated hypocotyl 5). Our results suggested that EBR and light had synergistic effects on the expression of genes in the anthocyanin biosynthesis pathway.

Iron Supply Affects Anthocyanin Content and Related Gene Expression in Berries of Vitis vinifera cv. Cabernet Sauvignon.

Anthocyanins are important compounds for red grape and red wine quality, and can be influenced by supply of nutrients such as nitrogen, phosphorus, potassium, zinc, and iron. The present work aims to gain a better understanding of the effect of iron supply on anthocyanins concentration in grape berries. To this end, own-rooted four-year-old Cabernet Sauvignon grapevines (Vitis vinifera) were fertigated every three days with 0, 23, 46, 92, and 184 µM iron (Fe) from ferric ethylenediamine di (o-hydroxyphenylacetic) acid (Fe-EDDHA) in a complete nutrient solution. Fe deficiency or excess generally led to higher concentrations of titratable acidity and skin/berry ratio, and to lower reducing sugar content, sugar/acid ratio, pH, berry weight, and concentration of anthocyanins. Most of the individual anthocyanins detected in this study, except cyanidin-3-O-glucoside, delphinidin-3-O-glucoside, and cyanidin-3-O-(6-O-coumaryl)-glucoside, in moderate Fe treatment (46 µM) grapes were significantly higher than those of other treatments. Genes encoding chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), leucoanthocyanidin dioxygenase (LDOX), and anthocyanin O-methyltransferase (AOMT) exhibited higher transcript levels in berries from plants cultivated with 46 µM Fe compared to the ones cultivated with other Fe concentrations. We suggest that grape sugar content, anthocyanins content, and transcriptions of genes involved in anthocyanin biosynthesis were correlated with Fe supply concentrations.

Tissue-Specific Expression Analysis of Anthocyanin Biosynthetic Genes in White- and Red-Fleshed Grape Cultivars.

Yan73, a teinturier (dyer) grape variety in China, is one of the few Vitis vinifera cultivars with red-coloured berry flesh. To examine the tissue-specific expression of genes associated with berry colour in Yan73, we analysed the differential accumulation of anthocyanins in the skin and flesh tissues of two red-skinned grape varieties with either red (Yan73) or white flesh (Muscat Hamburg) based on HPLC-MS analysis, as well as the differential expression of 18 anthocyanin biosynthesis genes in both varieties by quantitative RT-PCR. The results revealed that the transcripts of GST, OMT, AM3, CHS3, UFGT, MYBA1, F3'5'H, F3H1 and LDOX were barely detectable in the white flesh of Muscat Hamburg. In particular, GST, OMT, AM3, CHS3 and F3H1 showed approximately 50-fold downregulation in the white flesh of Muscat Hamburg compared to the red flesh of Yan73. A correlation analysis between the accumulation of different types of anthocyanins and gene expression indicated that the cumulative expression of GST, F3'5'H, LDOX and MYBA1 was more closely associated with the acylated anthocyanins and the 3'5'-OH anthocyanins, while OMT and AM3 were more closely associated with the total anthocyanins and methoxylated anthocyanins. Therefore, the transcripts of OMT, AM3, GST, F3'5'H, LDOX and MYBA1 explained most of the variation in the amount and composition of anthocyanins in skin and flesh of Yan73. The data suggest that the specific localization of anthocyanins in the flesh tissue of Yan73 is most likely due to the tissue-specific expression of OMT, AM3, GST, F3'5'H, LDOX and MYBA1 in the flesh.

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.