Search

Ethylene is one crucial phytohormone modulating plants' organ development and ripening process, especially in fruits, but its action modes and discrepancies in non-climacteric grape and climacteric peach in these processes remain elusive. This work is focused on the action mode divergences of ethylene during the modulation of the organ development and ripening process in climacteric/non-climacteric plants. We characterized the key enzyme genes in the ethylene synthesis pathway, VvACO1 and PpACO1, and uncovered that their sequence structures are highly conserved, although their promoters exhibit important divergences in the numbers and types of the cis-elements responsive to hormones, implying various responses to hormone signals. Subsequently, we found the two have similar expression modes in vegetative organ development but inverse patterns in reproductive ones, especially in fruits. Then, VvACO1 and PpACO1 were further validated in promoting fruit ripening functions through their transient over-expression/RNAi-expression in tomatoes, of which the former possesses a weaker role than the latter in the fruit ripening process. Our findings illuminated the divergence in the action patterns and function traits of the key VvACO1/PpACO1 genes in the tissue development of climacteric/non-climacteric plants, and they have implications for further gaining insight into the interaction mechanism of ethylene signaling during the modulation of the organ development and ripening process in climacteric/non-climacteric plants. [ABSTRACT FROM AUTHOR]

The aim of this study was to find out how different postharvest temperatures and MeJA treatments affected the quality of table grapes, their antioxidant properties, and the amount of hydrogen peroxide and malondialdehyde they contained. For the investigation, postharvest Shine Muscat table grapes were treated with low and high temperatures and MeJA at concentrations of 10 and 100 μmol/L. The results indicated that treating grape berries with MeJA at concentrations of 10 and 100 μmol/L effectively reduced weight loss and mitigated the increase in soluble solid content while also mitigating the decrease in berry firmness and titratable acidity. Consequently, this treatment preserved the sensory and nutritional qualities of the berries and extended their shelf life. Meanwhile, the application of MeJA at a concentration of 10 μmol/L demonstrated superior effectiveness compared to the 100 μmol/L concentration and resulted in a significant enhancement of antioxidant activities by increasing levels of superoxide dismutase, catalase, ascorbate peroxidase, and polyphenol oxidase. Furthermore, the levels of hydrogen peroxide and malondialdehyde in the samples increased for all treatments throughout the storage period. Nevertheless, the levels of hydrogen peroxide and malondialdehyde generation following MeJA treatment remained much lower compared to samples treated at room temperature and low temperature. Therefore, the postharvest application of MeJA at a concentration of 10 μmol/L could play a critical role as a stimulator of fruit quality as well as enhance physicochemical parameters and antioxidant activities for extending the shelf life of grapes during storage. [ABSTRACT FROM AUTHOR]

Exogenous GA is widely used to efficiently induce grape seedless berry development for significantly improving berry quality. Recently, we found that VvmiR166s are important regulators of response to GA in grapes, but its roles in GA-induced seedless grape berry development remain elusive. Here, the precise sequences of VvmiR166s and their targets VvREV, VvHB15 and VvHOX32 were determined in grape cv. 'Rosario Bianco', and the cleavage interactions of VvmiR166s-VvHB15/VvHOX32/VvREV modules and the variations in their cleavage roles were confirmed in grape berries. Exogenous GA treatment significantly induced a change in their expression correlations from positive to negative between VvmiR166s and their target genes at the seeds during the stone-hardening stages (32 DAF–46 DAF) in grape berries, indicating exogenous GA change action modes of VvmiR166s on their targets in this process, in which exogenous GA mainly enhanced the negative regulatory roles of VvmiR166s on VvHB15 among all three VvmiR166s-target pairs. The transient OE-VvmiR166a-h/OE-VvHB15 in tobacco confirmed that out of the VvmiR166 family, VvmiR166h/a/b might be the main factors in modulating lignin synthesis through inhibiting VvHB15, of which VvmiR166h-VvHB15-NtPAL4/NtCCR1/NtCCR2/NtCCoAMT5/NtCOMT1 and VvmiR166a/b-VvHB15-NtCAD1 are the potential key regulatory modules in lignin synthesis. Together with the GA-induced expression modes of VvmiR166s-VvHB15 and genes related to lignin synthesis in grape berries, we revealed that GA might repress lignin synthesis mainly by repressing VvCAD1/VvCCR2/VvPAL2/VvPAL3/Vv4CL/VvLac7 levels via mediating VvmiR166s-VvHB15 modules in GA-induced grape seedless berries. Our findings present a novel insight into the roles of VvmiR66s that are responsive to GA in repressing the lignin synthesis of grape seedless berries, with different lignin-synthesis-enzyme-dependent action pathways in diverse plants, which have important implications for the molecular breeding of high-quality seedless grape berries. [ABSTRACT FROM AUTHOR]

Drought stress impedes viticultural plant growth and development by modifying various metabolic pathways. However, the regulatory network response underlying drought stress is not yet clear. In this study, the leaves and roots of "Shine Muscat" ("SM," Vitis labruscana × Vitis vinifera) and "Thompson Seedless" ("TS," V. vinifera L. cv.) were subjected to drought stress to study the regulatory network used by drought stress. Morphophysiological results showed that the malondialdehyde content after 28 days of drought stress increased more significantly in "TS" than "SM." Furthermore, the multiomics analysis studies showed that a total of 3036–6714 differentially expressed genes and 379–385 differentially abundant metabolites were identified in "SM" and "TS" grapevine cultivars under drought stress. Furthermore, the retained intron was the major form of differential alternative splicing event under drought stress. The photosynthesis pathway, antioxidant system, plant hormone signal transduction, and osmotic adjustment were the primary response systems in the two grapevine cultivars under drought stress. We have identified GRIK1, RFS2, and LKR/SDH as the hub genes in the coexpression network of drought stress. In addition, the difference in the accumulation of pheophorbide‐a reveals different drought resistance mechanisms in the two grapevine cultivars. Our study explained the difference in drought response between cultivars and tissues and identified drought stress‐responsive genes, which provides reference data for further understanding the regulatory network of drought tolerance in grapevine. [ABSTRACT FROM AUTHOR]

Grapes are widely cultivated around the world and their quality has distinct regional characteristics. In this study, the qualitative characteristics of the 'Cabernet Sauvignon' grape variety in seven regions, from half-véraison to maturity, were analyzed comprehensively at physiological and transcriptional levels. The results indicated that the quality traits of 'Cabernet Sauvignon' grapes in different regions were significantly different with obvious regionality. Total phenols, anthocyanins, and titratable acids were the main factors of the regionality of berry quality, which were very sensitive to changes in the environment. It should be noted that the changes in titrating acids and total anthocyanin of berries vary greatly from half-véraison to maturity between regions. Moreover, the transcriptional analysis showed that the co-expressed genes between regions characterized the core transcriptome of berry development, while the unique genes of each region reflected the regionality of berries. The differentially expressed genes (DEGs) between half-véraison and maturity can be used to demonstrate that the environment of the regions could promote or inhibit gene expression. The functional enrichment suggested that these DEGs help to understand the interpretation of the plasticity of the quality composition of grapes according to the environment. Taken together, the information generated by this study could contribute to the development of viticultural practices aimed at making better use of native varieties for the development of wines with regional characteristics. [ABSTRACT FROM AUTHOR]

SSR (simple sequence repeat) DNA markers are widely used for genotype DNA identification, QTL mapping, and analyzing genetic biodiversity. However, SSRs in grapes are still in their early stages, with a few primer pairs accessible. With the whole-genome sequencing (WGS) of several grape varieties, characterization of grape SSR changed to be necessary not only to genomics but to also help SSR development and utility. Based on this, we identified the whole-genome SSR of nine grape cultivars ('PN40024', 'Cabernet Sauvignon', 'Carménère', 'Chardonnay', 'Merlot', 'Riesling', 'Zinfandel', 'Shine Muscat', and 'Muscat Hamburg') with whole-genome sequences released publicly and found that there are great differences in the distribution of SSR loci in different varieties. According to the difference in genome size, the number of SSRs ranged from 267,385 (Cabernet Sauvignon) to 627,429 (Carménère), the density of the SSR locus in the genome of nine cultivars was generally 1 per Kb. SSR motif distribution characteristic analysis of these grape cultivars showed that the distribution patterns among grape cultivars were conservative, mainly enriched in A/T. However, there are some differences in motif types (especially tetranucleotides, pentanucleotides, and hexanucleotides), quantity, total length, and average length in different varieties, which might be related to the size of the assembled genome or the specificity of variety domestication. The distribution characteristics of SSRs were revealed by whole-genome analysis of simple repeats of grape varieties. In this study, 32 pairs of primers with lower polymorphism have been screened, which provided an important research foundation for the development of molecular markers of grape variety identification and the construction of linkage maps of important agronomic traits for crop improvement. [ABSTRACT FROM AUTHOR]

Grapes (Vitis sp.) are susceptible to infection by grasshopper and fungus that lead to damaging productivity and economic losses. In this study, we infected the young and old leaf of grapevine by Acrida chinensis and Botrytis cinerea and combined the metabolomic and transcriptomic analysis to comprehend the infection-mediated molecular and metabolic responses. Results indicated that the metabolic reprogramming of lipid, proline, and carbohydrate metabolism is necessary for the biosynthesis of specialized metabolites, which are involved in grapevine defense response. Moreover, the resistance mechanisms of grapevine induced by grasshopper and Botrytis required the hormones crosstalk of abscisic acid (ABA), salicylic acid (SA), ethylene (ETH), brassinosteroid (BR), and jasmonic acid (JA). We also identified the different genes involved in resistance to pathogen and insect attack in transcriptional levels, which encoded pathogenesis-related proteins (PRs), disease resistance proteins (RPS), calcium-binding proteins (CMLs), WRKY transcription factor, abscisic acid-insensitive (ABI), brassinosteroid insensitive 1 (BAK1), ethylene-responsive transcription factor (ERF), leucine-rich repeat receptor-like protein kinases (LRRs), zinc finger protein (ZAT10), chitinase 5, and E3 ubiquitin-protein ligase PUB23 in infected leaves. Furthermore, we nominated various infection-related metabolic biomarkers, including glutathione, proline, aspartyl-L-proline, adenine, histidine, and arginine, that has major effects on the tolerance of leaf-damaging insects and pathogens. Taken together, our research results clarify that there are differences in the defense mechanisms of old leaves and young leaves that lead to different biological stress tolerance and provide deeper insights into grapevine regulatory network mediating defense response against infection caused by grasshopper and B. cinerea, and uncover the resistance mechanisms for functional genomic studies. [ABSTRACT FROM AUTHOR]

Seedlessness is one of the important quality and economic traits favored by grapevine consumers, which are mainly affected by phytohormones, especially gibberellin (GA). GA is widely utilized in seedless berry production and could effectively induce grape seed embryo abortion. However, the molecular mechanism underlying this process, like the role of RNA silencing in the biosynthesis pathway of GA remains elusive. Here, Gibberellin 3-β dioxygenase2 (GA3ox2) as the last key enzyme in GA biosynthesis was predicated as a potential target gene for miR3633a, and two of them were identified as a GA response in grape berries. We also analyzed the promoter regions of genes encoding GA biosynthesis and found the hormone-responsive elements to regulate grape growth and development. The cleavage interaction between VvmiR3633a and VvGA3ox2 was validated by RLM-RACE and the transient co-transformation technique in tobacco in vivo. Interestingly, during GA-induced grape seed embryo abortion, exogenous GA promoted the expression of VvmiR3633a, thereby mainly repressing the level of VvGA3ox2 in seed embryos. We also observed a negative correlation between down-regulated VvGA20ox2/VvGA3ox2 and up-regulated VvGA2ox3/VvGA2ox1, of which GA inactivation was greater than GA synthesis, inhibited active GA content, accompanied by the reduction of VvSOD and VvCAT expression levels and enzymatic activities. These series of changes might be the main causes of grape seed embryo abortion. In conclusion, we have preliminarily drawn a schematic mode of GA-mediated VvmiR3633a and related genes regulatory network during grape seed abortion induced by exogenous GA. Our findings provide novel insights into the GA-responsive roles of the VvmiR3633a-VvGA3ox2 module in the modulation of grape seed-embryo abortion, which has implications for the molecular breeding of high-quality seedless grape berries. [ABSTRACT FROM AUTHOR]

Grapes are subject to a wide range of climatic conditions during their life cycle, but the use of rootstocks can effectively ameliorate the effects of abiotic stress. However, the tolerance mechanism of different grape rootstock varieties varies under various stresses, and systematic research on this aspect is limited. On the basis of previous research, transcriptome sequencing was performed on three tolerant grape rootstock varieties (3309C, 520A, 1103P) and three intolerant grape rootstock varieties (5BB, 101–14, Beta). In total, 56,478,468 clean reads were obtained. One hundred and ten genes only existed in all combinations during P1 with a downregulated trend, and 178 genes existed only in P1 of tolerant grape rootstock varieties. Salt treatment firstly affected the photosynthesis of leaves, and tolerant varieties weakened or even eliminated this effect through their own mechanisms in the later stage. Tolerant varieties mobilized a large number of MFs during the P2 stage, such as hydrolase activity, carboxypeptidase activity, and dioxygenase activity. Carbon metabolism was significantly enriched in P1, while circadian rhythm and flavonoid biosynthesis were only enriched in tolerant varieties. In the intolerant varieties, photosynthesis-related pathways were always the most significantly enriched. There were large differences in the gene expression of the main signal pathways related to salt stress in different varieties. Salt stress affected the expression of genes related to plant abiotic stress, biotic stress, transcription factors, hormones, and secondary metabolism. Tolerant varieties mobilized more bHLH, WRKY, and MYB transcription factors to respond to salt stress than intolerant varieties. In the tolerant rootstocks, SOS was co-expressed. Among these, SOS1 and SOS2 were upregulated, and the SOS3 and SOS5 components were downregulated. The genes of heat shock proteins and the phenylalanine pathway were upregulated in the tolerant varieties. These findings outline a tolerance mechanism model for rootstocks for coping with osmotic stress, providing important information for improving the resistance of grapes under global climate change. [ABSTRACT FROM AUTHOR]

Grapevine downy mildew is the most serious disease of grapevine cultivars that affects the rate of resistance/susceptibility to Plasmopara viticola. In this study, we used the susceptible cultivar "Zitian Seedless" and the resistant cultivar "Kober 5BB" as materials to determine the transcriptome differences and phenotypes of the leaves after inoculation with downy mildew. The differences in microstructures and molecular levels were compared and analyzed. Fluorescence staining and microscopic observations confirmed that hypersensitive cell death occurred around the stomata in "Kober 5BB" infected by downy mildew zoospores. Meanwhile, transcriptomic profiling indicated that there were 11,713 and 6,997 gene expression differences between the resistant and susceptible cultivars at 72 h after inoculation when compared to control (0 h), respectively. The differentially expressed genes of the two cultivars are significantly enriched in different pathways, including response to plant-pathogen interaction, mitogen-activated protein kinase (MAPK) signaling pathway, plant hormone signal transduction, phenylpropanoid, and flavonoid biosynthesis. Furthermore, the results of functional enrichment analysis showed that H2O2 metabolism, cell death, reactive oxygen response, and carbohydrate metabolism are also involved in the defense response of "Kober 5BB," wherein a total of 322 key genes have been identified. The protein interaction network showed that metacaspases (MCAs), vacuolar processing enzymes (VPEs), and Papain-like cysteine proteases (PLCPs) play an important role in the execution of hypersensitive responses (HR). In conclusion, we demonstrated that HR cell death is the key strategy in the process of grape defense against downy mildew, which may be mediated or activated by Caspase-like proteases. [ABSTRACT FROM AUTHOR]

Key message: Hormone treatment enhanced the content of osmotic substances under high-temperature conditions. The effect of ABA and BR treated separately is better than treated together. To determine the effect of abscisic acid (ABA), brassinolide (BR) and ABA + BR on grape quality under high-temperature stress, various metabolites were analyzed. Compared with the control (CK), DL-tryptophan, d-raffinose, geniposidic acid, dodecanedioic acid and polyphenols were found to be higher after ABA treatment. After BR treatment, amino acids and poricoic acid B were higher than in CK. And carbohydrates and amino acids were up-regulated after ABA + BR treatment. BR and ABA + BR treatment also induced higher endogenous ABA and epibrassinolide contents. In addition, treated grape had higher soluble solid concentrations and soluble sugar content, and delayed the degradation of middle lamella and microfibrils. Antioxidant and heat shock-related genes were examined, which significantly increased in treated grape. The finding of this study suggested that ABA, BR and ABA + BR are very useful for alleviating high-temperature damage by increasing the accumulation of osmotic adjustment substances, and endogenous hormones content. [ABSTRACT FROM AUTHOR]

Background: In the breeding of new horticultural crops, fruit shape is an important selection characteristic. A variety of fruit shapes appeared during the gradual process of selection and domestication. However, few studies have been conducted on grape berry shape, especially studies related to mining candidate genes. To discover candidate genes related to grape berry shape, the present study first took the berry shape parameters analyzed by Tomato Analyzer as the target traits and used a genome-wide association analysis to analyze candidate genes. Results: In total, 122 single-nucleotide polymorphism (SNP) loci had significant correlations with multiple berry shape traits in both years, and some candidate genes were further mined. These genes were mainly related to LRR receptor-like serine/threonine-protein kinase (At1g05700 and At1g07650), transcription factors (GATA transcription factor 23-like, transcription factor VIP1, transcription initiation factor TFIID, and MADS-box transcription factor 6), ubiquitin ligases (F-box protein SKIP19 and RING finger protein 44), and plant hormones (indole-3-acetic acid-amido synthetase GH3.6 and ethylene-responsive transcription factor ERF061). In addition, some important SNP loci were associated with multiple berry-shape traits. The study further revealed some genes that control multiple traits simultaneously, indicating that these berry shape traits are subject to the coordinated regulation of some genes in controlling berry shape. Conclusions: In the present work, we identified interesting genetic determinants of grape berry shape-related traits. The identification of molecular markers that are closely related to these berry-shape traits is of great significance for breeding specific berry-shaped grape varieties. [ABSTRACT FROM AUTHOR]

Background: Papain-like cysteine proteases (PLCPs), a large group of cysteine proteases, are structurally related to papain. The members belonging to PLCPs family contribute to plant immunity, senescence, and defense responses in plants. The PLCP gene family has been identified in Arabidopsis, rice, soybean, and cotton. However, no systematic analysis of PLCP genes has been undertaken in grapevine. Since Plasmopara viticola as a destructive pathogen could affect immunity of grapes in the field, we considered that the members belonged to PLCPs family could play a crucial role in defensive mechanisms or programmed cell death. We aimed to evaluate the role of PLCPs in 2 different varieties of grapevines and compared the changes of their expressions with the transcriptional data in response to P. viticola. Results: In this study, 23 grapevine PLCP (VvPLCP) genes were identified by comprehensive bioinformatics analysis. Subsequently, the chromosomal localizations, gene structure, conserved domains, phylogenetic relationship, gene duplication, and cis-acting elements were analyzed. Numerous cis-acting elements related to plant development, hormone, and stress responses were identified in the promoter of the VvPLCP genes. Phylogenetic analysis grouped the VvPLCP genes into nine subgroups. The transcription of VvPLCP in different inoculation time points and varieties indicated that VvPLCP may have vital functions in grapevine defense against Plasmopara viticola. According to transcriptome data and qPCR analysis, we observed the increasing expression levels of VvRD21–1 at 72 h after inoculation in resistant variety, inferring that it was related to grape downy mildew resistance. Meanwhile, 3 genes including VvXBCP1, VvSAG12–1, and VvALP1 showed higher expression at 24 h after pathogen inoculation in the susceptible variety and might be related to the downy mildew phenotype. We nominated these four genes to function during hypersensitive response (HR) process, inferring that these genes could be associated with downy mildew resistance in grapes. Conclusions: Our results provide the reference for functional studies of PLCP gene family, and highlight its functions in grapevine defense against P. viticola. The results help us to better understand the complexity of the PLCP gene family in plant immunity and provide valuable information for future functional characterization of specific genes in grapevine. [ABSTRACT FROM AUTHOR]

Background: Stone-hardening stage is crucial to the development of grape seed and berry quality. A significant body of evidence supports the important roles of MicroRNAs in grape-berry development, but their specific molecular functions during grape stone-hardening stage remain unclear. Results: Here, a total of 161 conserved and 85 species-specific miRNAs/miRNAs* (precursor) were identified in grape berries at stone-hardening stage using Solexa sequencing. Amongst them, 30 VvmiRNAs were stone-hardening stage-specific, whereas 52 exhibited differential expression profiles during berry development, potentially participating in the modulation of berry development as verified by their expression patterns. GO and KEGG pathway analysis showed that 13 VvmiRNAs might be involved in the regulation of embryo development, another 11 in lignin and cellulose biosynthesis, and also 28 in the modulation of hormone signaling, sugar, and proline metabolism. Furthermore, the target genes for 4 novel VvmiRNAs related to berry development were validated using RNA Ligase-Mediated (RLM)-RACE and Poly(A) Polymerase-Mediated (PPM)-RACE methods, and their cleavage mainly occurred at the 9th–11th sites from the 5′ ends of miRNAs at their binding regions. In view of the regulatory roles of GA in seed embryo development and stone-hardening in grape, we investigated the expression modes of VvmiRNAs and their target genes during GA-induced grape seedless-berry development, and we validated that GA induced the expression of VvmiR31-3p and VvmiR8-5p to negatively regulate the expression levels of CAFFEOYL COENZYME A-3-O-METHYLTRANSFERASE (VvCCoAOMT), and DDB1-CUL4 ASSOCIATED FACTOR1 (VvDCAF1). The series of changes might repress grape stone hardening and embryo development, which might be a potential key molecular mechanism in GA-induced grape seedless-berry development. Finally, a schematic model of miRNA-mediated grape seed and stone-hardening development was proposed. Conclusion: This work identified 30 stone-hardening stage-specific VvmiRNAs and 52 significant differential expression ones, and preliminary interpreted the potential molecular mechanism of GA-induced grape parthenocarpy. GA negatively manipulate the expression of VvCCoAOMT and VvDCAF1 by up-regulation the expression of VvmiR31-3p and VvmiR8-5p, thereby repressing seed stone and embryo development to produce grape seedless berries. [ABSTRACT FROM AUTHOR]

Background: 3-Hydroxy-3-methylglutaryl-CoA reductase (HMGR) is a key enzyme in the mevalonate (MVA) pathway, which regulates the metabolism of terpenoids in the cytoplasm and determines the type and content of downstream terpenoid metabolites. Results: Results showed that grapevine HMGR family has three members, such as VvHMGR1, VvHMGR2, and VvHMGR3. The expression of VvHMGRs in 'Kyoho' has tissue specificity, for example, VvHMGR1 keeps a higher expression, VvHMGR2 is the lowest, and VvHMGR3 gradually decreases as the fruit development. VvHMGR3 is closely related to CsHMGR1 and GmHMGR9 and has collinearity with CsHMGR2 and GmHMGR4. By the prediction of interaction protein, it can interact with HMG-CoA synthase, MVA kinase, FPP/GGPP synthase, diphosphate mevalonate decarboxylase, and participates in the synthesis and metabolism of terpenoids. VvHMGR3 have similar trends in expression with some of the genes of carotenoid biosynthesis and MEP pathways. VvHMGR3 responds to various environmental and phytohormone stimuli, especially salt stress and ultraviolet (UV) treatment. The expression level of VvHMGRs is diverse in grapes of different colors and aroma. VvHMGRs are significantly higher in yellow varieties than that in red varieties, whereas rose-scented varieties showed significantly higher expression than that of strawberry aroma. The expression level is highest in yellow rose-scented varieties, and the lowest in red strawberry scent varieties, especially 'Summer Black' and 'Fujiminori'. Conclusion: This study confirms the important role of VvHMGR3 in the process of grape fruit coloring and aroma formation, and provided a new idea to explain the loss of grape aroma and poor coloring during production. There may be an additive effect between color and aroma in the HMGR expression aspect. [ABSTRACT FROM AUTHOR]

Summary: In grape, MYBA1 and MYBA2 at the colour locus are the major genetic determinants of grape skin colour, and the mutation of two functional genes (VvMYBA1 and VvMYBA2) from these loci leads to white skin colour. This study aimed to elucidate the regulation of grape berry coloration by isolating and characterizing VvMYBA2w and VvMYBA2r alleles. The overexpression of VvMYBA2r up‐regulated the expression of anthocyanin biosynthetic genes and resulted in higher anthocyanin accumulation in transgenic tobacco than wild‐type (WT) plants, especially in flowers. However, the ectopic expression of VvMYBA2w inactivated the expression of anthocyanin biosynthetic genes and could not cause obvious phenotypic modulation in transgenic tobacco. Unlike in VvMYBA2r, CA dinucleotide deletion shortened the C‐terminal transactivation region and disrupted the transcriptional activation activity of VvMYBA2w. The results indicated that VvMYBA2r positively regulated anthocyanin biosynthesis by forming the VvMYBA2r‐VvMYCA1‐VvWDR1 complex, and VvWDR1 enhanced anthocyanin accumulation by interacting with the VvMYBA2r‐VvMYCA1 complex; however, R44L substitution abolished the interaction of VvMYBA2w with VvMYCA1. Meanwhile, both R44L substitution and CA dinucleotide deletion seriously affected the efficacy of VvMYBA2w to regulate anthocyanin biosynthesis, and the two non‐synonymous mutations were additive in their effects. Investigation of the colour density and MYB haplotypes of 213 grape germplasms revealed that dark‐skinned varieties tended to contain HapC‐N and HapE2, whereas red‐skinned varieties contained high frequencies of HapB and HapC‐Rs. Regarding ploidy, the higher the number of functional alleles present in a variety, the darker was the skin colour. In summary, this study provides insight into the roles of VvMYBA2r and VvMYBA2w alleles and lays the foundation for the molecular breeding of grape varieties with different skin colour. [ABSTRACT FROM AUTHOR]

Background: Grape buds and leaves are directly associated with the physiology and metabolic activities of the plant, which is monitored by epigenetic modifications induced by environment and endogenous factors. Methylation is one of the epigenetic regulators that could be involved in DNA levels and affect gene expression in response to stimuli. Therefore, changes of gene expression profile in leaves and bud through inhibitors of DNA methylation provide a deep understanding of epigenetic effects in regulatory networks. Results: In this study, we carried out a transcriptome analysis of 'Kyoho' buds and leaves under 5-azacytidine (5-azaC) exposure and screened a large number of differentially expressed genes (DEGs). GO and KEGG annotations showed that they are mainly involved in photosynthesis, flavonoid synthesis, glutathione metabolism, and other metabolic processes. Functional enrichment analysis also provided a holistic perspective on the transcriptome profile when 5-azaC bound to methyltransferase and induced demethylation. Enrichment analysis of transcription factors (TFs) also showed that the MYB, C2H2, and bHLH families are involved in the regulation of responsive genes under epigenetic changes. Furthermore, hormone-related genes have also undergone significant changes, especially gibberellin (GA) and abscisic acid (ABA)-related genes that responded to bud germination. We also used protein-protein interaction network to determine hub proteins in response to demethylation. Conclusions: These findings provide new insights into the establishment of molecular regulatory networks according to how methylation as an epigenetic modification alters transcriptome patterns in bud and leaves of grape. [ABSTRACT FROM AUTHOR]

Background: The B-BOX (BBX) proteins are the class of zinc-finger transcription factors and can regulate plant growth, development, and endure stress response. In plants, the BBX gene family has been identified in Arabidopsis, rice, and tomato. However, no systematic analysis of BBX genes has been undertaken in grapevine. Results: In this study, 24 grapevine BBX (VvBBX) genes were identified by comprehensive bioinformatics analysis. Subsequently, the chromosomal localizations, gene structure, conserved domains, phylogenetic relationship, gene duplication, and cis-acting elements were analyzed. Phylogenetic analysis divided VvBBX genes into five subgroups. Numerous cis-acting elements related to plant development, hormone and/or stress responses were identified in the promoter of the VvBBX genes. The tissue-specific expressional dynamics of VvBBX genes demonstrated that VvBBXs might play important role in plant growth and development. The transcript analysis from transcriptome data and qRT-PCR inferred that 11 VvBBX genes were down-regulated in different fruit developmental stages, while three VvBBX genes were up-regulated. It is also speculated that VvBBX genes might be involved in multiple hormone signaling (ABA, ethylene, GA3, and CPPU) as transcriptional regulators to modulate berry development and ripening. VvBBX22 seems to be responsive to multiple hormone signaling, including ABA, ethylene GA3, and CPPU. Some VvBBX genes were strongly induced by Cu, salt, waterlogging, and drought stress treatment. Furthermore, the expression of VvBBX22 proposed its involvement in multiple functions, including leaf senescence, abiotic stress responses, fruit development, and hormone response. Conclusions: Our results will provide the reference for functional studies of BBX gene family, and highlight its functions in grapevine berry development and ripening. The results will help us to better understand the complexity of the BBX gene family in abiotic stress tolerance and provide valuable information for future functional characterization of specific genes in grapevine. [ABSTRACT FROM AUTHOR]

The sequence and structure of grape SBP-box-like18 (VvSPL18) were identified and characterized to explore its regulatory roles during grape berry development and ripening. Homologous conservation across diverse plant species was observed, and its potential function and modulated roles in grapes were investigated. The results showed that VvSPL18 has an ORF sequence of 1,137 bp, encodes 378 amino acids, and is located on chromosome 14 of grapevine. VvSPL18 has the closest relationship with its homolog in soybeans. The promoter of VvSPL18 contains cis-elements responsive to gibberellins (GA) and salicylic acid (SA), indicating that this gene might respond to these hormones involved in the modulation of grape berry. VvSPL18 is mainly distributed in the nucleus. Expression profiles showed that VvSPL18 is highly expressed only at the veraison stage of the grape berry and is slightly expressed in other phases. RNA-seq data also revealed that VvSPL18 might participate in the modulation of grape berry development and ripening. Treatment with diverse hormones demonstrated that abscisic acid (ABA) had almost no effect on its expression, whereas naphthalene acetic acid (NAA) significantly upregulated its expression at the veraison stage. We also found that VvSPL18 has a GA-responsive cis-element but no NAA-responsive cis-element. GA could promote the expression of VvSPL18 with a peak at an earlier stage than NAA, suggesting that VvSPL18 responds faster to GA than to NAA. This result indicates that VvSPL18 might modulate berry development at this phase through an ABA-independent pathway, and it might directly respond to GA, but indirectly to NAA. Our findings provide insights into the functions of VvSPL18 in mediating grape berry development and ripening. [ABSTRACT FROM AUTHOR]

SPL is a plant-specific transcription factor family. Many researchers reported that SPL members targeted by miR156s could play crucial roles in the modulation of plant growth and development. Although there are similar reports on grapes, till now little is known about grape berry development and ripening. To gain more insight into how grape miR156s (Vv-miR156s) modulated the above given processes of grape berries by mediating their target gene Vv-SPLs, here we identified the precise sequences of Vv-miR156s in ‘Giant Rose’ grape berries, predicted their potential targets, and revealed that the matching degree of various Vv-miR156: Vv-SPL pairs exhibited some discrepancy, implying the divergence of their interaction. Subsequently, we also discovered similar motifs such as ABRE, CGTCA and ERE, which are more specific to berry development and ripening, within the promoters of both Vv-MIR156s and Vv-SPLs. With berry development and ripening, meanwhile, Vv-miR156a, b/c/d, e and f/g/i exhibited an overall increasing expression trend, while their targets showed opposite trends at the corresponding stages. Additionally, exogenous ABA and NAA application promoted or curbed the expression of Vv-miR156s to some extent, before grape berry ripening stage. The cleavage products, sites and frequencies of Vv-miR156a, b/c/d, e, f/g/i and their respective targets (Vv-SPL2, 9, 10, 16) during grape berry development and ripening process were validated by our developed PPM-RACE and modified RLM-RACE together with qRT-PCR, which demonstrated that Vv-miR156s can be involved in the modulation of grape berry development and ripening process by mediating the expression of Vv-SPL2, 9, 10, 16. Our findings lay an important foundation for further recognizing their functions in grape berries, and enrich the knowledge of the regulatory mechanism of miRNA-mediated grape berry development and ripening. [ABSTRACT FROM AUTHOR]

Main conclusion: Grapevine autophagy-related genes (ARGs) include 35 members that have unique evolutionary backgrounds and expression patterns, with some of them responding to abiotic stresses, including copper stress.Autophagy is one of the most crucial self-regulating phenomena in livings organisms, including animals, plants, yeasts, etc. In the genomes of plants, like Arabidopsis, rice, tobacco, and barley, more than 30 autophagy-related genes (ARGs) have been found. These ARGs are involved in plant development, programed cell death, and the stress response process. In plants, and particularly in grapevine, high copper stress results from the application of the Bordeaux mixture, a widely used fungicide. However, the function of autophagy in plant tolerance to copper stress is unknown. Accordingly, in this study, a genome-wide analysis was performed to identify Vitis vinifera ARGs (VvARGs), and 35 VvARGs were detected. A gene family analysis revealed that the tandem and segmental duplication events played significant roles in the VvARG gene family expansion. Moreover, there was more intense signature of purifying selection for the comparison between grape and rice than between grape and Arabidopsis. In response to copper treatment, both the autophagosome number and malondialdehyde concentration increased during the initial 4 h post-treatment, and reached maximal values at 24 h. An expression analysis indicated that most VvARGs responded to copper stress at 4 h post-treatment, and some VvARGs (e.g., VvATG6, VvATG8i, and VvATG18h) exhibited responses to most abiotic stresses. These results provide a detailed overview of the ARGs in grapevine and indicate multiple functions of autophagy in fruit development and abiotic stresses in grapevine. The key ARG (e.g., ATG8i) should be investigated in more detail in grapevine and other plant species. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are endogenous small RNAs of −21 nucleotides that play an important role in diverse plant physiological processes at the post-transcriptional level by directing mRNA cleavage or translational inhibition. Previous studies have indicated that down-regulation of miR398 in response to oxidative stress allows up-regulation of the two target genes, cytosolic CSD1 and chloroplastic CSD2 (copper/zinc superoxide dismutase), resulting in protecting the plants to tolerate oxidative stress. In this study, we provide evidence that grapevine miR398 (Vv-miR398), by regulating the expression of its target genes, VvCSD1 and VvCSD2, mediates responses of grapevine to copper (Cu) stress which have been magnified due to increase in Cu-containing pesticide application. The expression of Vv-miR398 was inhibited by different concentrations of Cu stress; on the other hand, there was a steady increase in the activity of VvCSD1 and VvCSD2 genes. The function of VvCSD1 and VvCSD2 under Cu stress was thoroughly examined by overexpressing the use of the VvCSD1 and VvCSD2 in transgenic tobacco ( Nicotiana tabacum). We found that both the overexpressed transgenic lines had lower Cu sensitivity and higher Cu tolerance compared with the wild type. In addition, lower levels of ROS and higher levels of SOD activities were accumulated in the transgenic lines in comparison with the wild type under the higher Cu conditions. Furthermore, these transgenic tobacco lines also recorded a higher UV and salt tolerance than the WT plants. These results suggested that overexpressing the VvCSDs will enhance the ROS-scavenging systems and protect the plant against more oxidative damage. Also, more investigations in this line are needed that would provide significant improvements in our understanding the resistance of fruit crops to environmental stress. [ABSTRACT FROM AUTHOR]

Chloroplast simple sequence repeat (cpSSR) markers previously reported were used to analyze chloroplast diversity of Chinese grapes. Nine cpSSR primer pairs from the chloroplast genomes of citrus and tobacco (Nicotiana tabacum L.) were found useful for analyzing the grapes chloroplast genome (cpDNA). Selected cpSSR primer pairs showed polymorphism among 64 varieties, with polymorphism information content (PIC) values ranging from 0.522 to 0.659, with an average of 0.590. Results depicted that a total number of 43 alleles were detected on nine loci with an average of 4.77 alleles per locus with highest number of six alleles for marker ARCP2 and CCMP10. Gene diversity explained by these cpSSR makers ranged from 4 to 68% with an average of 63.66% per marker. The genetic relationship suggested by UPGMA based dendrogram divided the genotypes into three main clusters and four subgroups, results showed that 2B is the biggest group. Overall Hong Ma Ge and Da LI Mei Gui Xiung 98% similar, while 64-1-19 was the most diverse as compared to rest of genotypes. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) are critical regulators of various biological and metabolic processes of plants. Numerous miRNAs and their functions have been identified and analyzed in many plants. However, till now, the involvement of miRNAs in the response of grapevine berries to ethylene has not been reported yet. Here, Solexa technology was employed to deeply sequence small RNA libraries constructed from grapevine berries treated with and without ethylene. A total of 124 known and 78 novel miRNAs were identified. Among these miRNAs, 162 miRNAs were clearly responsive to ethylene, with 55 downregulated, 59 upregulated, and 14 unchanged miRNAs detected only in the control. The other 35 miRNAs responsive to ethylene were induced by ethylene and detected only in the ethylene-treated grapevine materials. Expression analysis of 27 conserved and 26 novel miRNAs revealed that 13 conserved and 18 novel ones were regulated by ethylene during the whole development of grapevine berries. High-throughput sequencing and qRT-PCR assays revealed consistent results on the expression results of ethylene-responsive miRNAs. Moreover, 90 target genes for 34 novel miRNAs were predicted, most of which were involved in responses to various stresses, especially like exogenous ethylene treatment. The identified miRNAs may be mainly involved in grapevine berry development and response to various environmental conditions. [ABSTRACT FROM AUTHOR]

Grapevine berry development is a complex and genetically controlled process, with many morphological, biochemical and physiological changes occurring during the maturation process. Research carried out on grapevine berry development has been mainly concerned with wine grape, while barely focusing on table grape. ‘Fujiminori’ is an important table grapevine cultivar, which is cultivated in most provinces of China. In order to uncover the dynamic networks involved in anthocyanin biosynthesis, cell wall development, lipid metabolism and starch-sugar metabolism in ‘Fujiminori’ fruit, we employed RNA-sequencing (RNA-seq) and analyzed the whole transcriptome of grape berry during development at the expanding period (40 days after full bloom, 40DAF), véraison period (65DAF), and mature period (90DAF). The sequencing depth in each sample was greater than 12×, and the expression level of nearly half of the expressed genes were greater than 1. Moreover, greater than 64% of the clean reads were aligned to the Vitis vinifera reference genome, and 5,620, 3,381, and 5,196 differentially expressed genes (DEGs) were identified between different fruit stages, respectively. Results of the analysis of DEGs showed that the most significant changes in various processes occurred from the expanding stage to the véraison stage. The expression patterns of F3’H and F3’5’H were crucial in determining red or blue color of the fruit skin. The dynamic networks of cell wall development, lipid metabolism and starch-sugar metabolism were also constructed. A total of 4,934 SSR loci were also identified from 4,337 grapevine genes, which may be helpful for the development of phylogenetic analysis in grapevine and other fruit trees. Our work provides the foundation for developmental research of grapevine fruit as well as other non-climacteric fruits. [ABSTRACT FROM AUTHOR]

β-glucosidase (BG) was believed to take part in abscisic acid (ABA) synthesis via hydrolysis of ABA glucose ester to release active ABA during plant growth and development. However, there is no genetic evidence available to indicate the role of genes during fruit ripening. Here, the expression patterns of three genes ( VvBG1, VvBG2, and VvBG3) encoding β-glucosidase were analyzed during grape fruit development, and it was found that β-glucosidase activity increased in grape fruit in response to various stresses. Furthermore, to verify the function of β-glucosidase during fruit ripening, heterogeneous expression of the VvBG1 gene in strawberry fruit was validated, and the results showed that the VvBG1 over-expression increased β-glucosidase and promoted the fruit ripening process in strawberry. In addition, we found that ABA contents increased in the VvBG1 over-expression of strawberry fruit, which induced fruit anthocyanin, soluble solid accumulation, and fruit softening. Moreover, genes related to coloring ( CHS, CHI, F3H, and UFGT), softening ( PG1, PL1, and EXP1), and aroma ( SAAT, and QR) were up-regulated. This work will elucidate the specific roles of VvBGs in the synthesis of ABA and provide some new insights into the ABA-controlled grape ripening mechanism. [ABSTRACT FROM AUTHOR]

A 2000-bp 5′-flanking region of VvPAL-like was isolated from 'Summer Black' grapevine by PCR amplification, named pVvPAL-like. To gain a better understanding of the expression and regulatory mechanism of VvPAL-like, a chimeric expression unit consisting of the β-glucuronidase (GUS) reporter gene under the control of a 2000-bp fragment of the VvPAL-like promoter was transformed into tobacco via Agrobacterium tumefaciens. Histochemical staining showed that the full-length promoter directs efficient expression of the reporter gene in cotyledons and hypocotyls, stigma, style, anthers, pollen, ovary, trichomes, and vascular bundles of transgenic plants. A series of 5′ progressive deletions of the promoter revealed the presence of a negative regulatory region (−424 to −292) in the VvPAL-like promoter. Exposure of the transgenic tobacco plants to various abiotic stresses demonstrated that the full-length construct could be induced by light, copper (Cu), abscisic acid (ABA), indole-3-acetic (IAA), methyl jasmonate (MeJA) ( N-1-naphthylphthalamic acid), ethylene, and drought. Furthermore, the ethylene-responsive region was found to be located in the −1461/−930 fragment, while the element(s) for the MeJA-responsive expression may be present in the −424/−292 region in the VvPAL-like promoter. These findings will help us to better understand the molecular mechanisms by which VvPAL-like participates in biosynthesis of flavonoids and stress responses. [ABSTRACT FROM AUTHOR]

GRAS domain genes are a group of important plant-specific transcription factors that have been reported to be involved in plant development. In order to know the roles of GRAS genes in grapevine, a widely cultivated fruit crop, the study on grapevine GRAS ( VvGRAS) was carried out, and from which, 43 were identified from 12× assemble grapevine genomic sequences. Further, the genomic structures, synteny, phylogeny, expression profiles in different tissues of these genes, and their roles in response to stress were investigated. Among the genes, two potential target genes ( VvSCL15 and VvSCL22) for VvmiR171 were experimentally verified by PPM-RACE and RLM-RACE, in that not only the cleavage sites of miR171 on the target mRNA were mapped but also the cleaved fragments and their expressing patterns were detected. Transgenic Arabidopsis plants over expression VvSCL15 showed lower tolerance to drought and salt treatments. [ABSTRACT FROM AUTHOR]

With more copper and copper-containing compounds used as bactericides and fungicides in viticulture, copper homeostasis in grapevine ( Vitis) has become one of the serious environmental crises with great risk. To better understand the regulation of Cu homeostasis in grapevine, grapevine seedlings cultured in vitro with different levels of Cu were utilized to investigate the tolerance mechanisms of grapevine responding to copper availability at physiological and molecular levels. The results indicated that Cu contents in roots and leaves arose with increasing levels of Cu application. With copper concentration increasing, malondialdehyde (MDA) content increased in roots and leaves and the activities of superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) increased to protect the plant itself from damage. The expression patterns of 19 genes, encoding transporters, chaperones, and P-type ATPases involved in copper homeostasis in root and leaf of grapevine seedling under various levels of Cu were further analyzed. The expression patterns indicated that CTr1, CTr2, and CTr8 transporters were significantly upregulated in response both to Cu excess and deficiency. ZIP2 was downregulated in response to Cu excess and upregulated under Cu-deficient conditions, while ZIP4 had an opposite expression pattern under similar conditions. The expression of chaperones and P-type ATPases in response to Cu availability in grapevine were also briefly studied. [ABSTRACT FROM AUTHOR]

Improper application of copper-based fungicides has made copper stress critical in viticulture, necessitating the need to identify substances that can mitigate it. In this study, leaves of 'Shine Muscat' ('SM') grapevine seedlings were treated with CuSO 4 solution (10 mM/L), CuSO 4 + 5-aminolevulinic acid (ALA) (50 mg/L), and distilled water to explore the mitigation effect of ALA. Physiological assays demonstrated that ALA effectively reduced malondialdehyde accumulation and increased peroxidase and superoxide dismutase activities in grapevine leaves under copper stress. Copper ion absorption, transport pathways, chlorophyll metabolism pathways, photosynthetic system, and antioxidant pathways play key roles in ALA alleviated-copper stress. Moreover, expression changes in genes, such as CHLH , ALAD , RCA, and DHAR, play vital roles in these processes. Furthermore, abscisic acid reduction caused by NCED down-regulation and decreased naringenin, leucopelargonidin, and betaine contents confirmed the alleviating effect of ALA. Taken together, these results reveal how grapevine responds to copper stress and the alleviating effects of ALA, thus providing a novel means of alleviating copper stress in viticulture. [Display omitted] • Exogenous application of 5-aminolevulinic acid alleviates copper stress in grapevine. • ALA restores suppressed photosynthesis under copper stress. • Antioxidant systems and phytohormone metabolism were induced to alleviate copper stress. [ABSTRACT FROM AUTHOR]

Motivation: Trans-acting small interfering RNAs (ta-siRNAs) play an essential role in the regulation of plant gene expression, but relevant reports are still limited. Bioinformatic analyses indicate that many ta-siRNA-producing loci (TASs) are present in plants, implying the existence of as yet undiscovered ta-siRNAs and related regulatory pathways. To expand our knowledge of these plant gene regulators, we applied high-throughput computational and experimental methods to grapevine (Vitis vinifera L.).Results: Based on bioinformatic predictions, we identified 49 TASs from 49 055 small RNA clusters. Using RNA degradome analysis, we experimentally validated 5 TASs, 22 ta-siRNAs and 37 ta-siRNA targets. The cis-activities of ta-siRNAs were also confirmed, which suggested an inactive mechanism of TAS transcription, and a produced mechanism of multiple forms of small RNA from same TAS. We examined the conservation of newly identified ta-siRNA regulatory cascades and found that while the cascade related to vviTAS3 was conserved, cascades related to vviTAS7, vviTAS8, vviTAS9 and vviTAS10 were grape-specific. These results broaden the known scope of ta-siRNA regulation.Contact:zhang_chq2002@sohu.comSupplementary information: Supplementary data are available at Bioinformatics online. [ABSTRACT FROM AUTHOR]

Background: Alignment analysis of the Vv-miRNAs identified from various grapevine cultivars indicates that over 30% orthologous Vv-miRNAs exhibit a 1-3 nucleotide discrepancy only at their ends, suggesting that this sequence discrepancy is not a random event, but might mainly derive from divergence of cultivars. With advantages of miR-RACE technology in determining precise sequences of potential miRNAs from bioinformatics prediction, the precise sequences of vv-miRNAs predicted computationally can be verified with miR-RACE in a different grapevine cultivar. This presents itself as a new approach for large scale discovery of precise miRNAs in different grapevine varieties. Methodology/Principal Findings: Among 88 unique sequences of Vv-miRNAs from bioinformatics prediction, 83 (96.3%) were successfully validated with MiR-RACE in grapevine cv. 'Summer Black'. All the validated sequences were identical to their corresponding ones obtained from deep sequencing of the small RNA library of 'Summer Black'. Quantitative RT-PCR analysis of the expressions levels of 10 Vv-miRNA/target gene pairs in grapevine tissues showed some negative correlation trends. Finally, comparison of Vv-miRNA sequences with their orthologs in Arabidopsis and study on the influence of divergent bases of the orthologous miRNAs on their targeting patterns in grapevine were also done. Conclusion: The validation of precise sequences of potential Vv-miRNAs from computational prediction in a different grapevine cultivar can be a new way to identify the orthologous Vv-miRNAs. Nucleotide discrepancy of orthologous VvmiRNAs from different grapevine cultivars normally does not change their target genes. However, sequence variations of some orthologous miRNAs in grapevine and Arabidopsis can change their targeting patterns. These precise Vv-miRNAs sequences validated in our study could benefit some further study on grapevine functional genomics. [ABSTRACT FROM AUTHOR]

B Correction to: BMC Plant Biol 21, 499 (2021) b B https://doi.org/10.1186/s12870-021-03279-w b Following publication of the original article [[1]], Peijie Gong is the co-1st author and one of the corresponding authors of this article. Jun Kang and Peijie Gong contributed equally to this work. [Extracted from the article]

Copper stress is one of the most severe heavy metal stresses in plants. Grapevine has a relatively higher copper tolerance than other fruit crops. However, there are no reports regarding the tolerance mechanisms of the 'Shine Muscat' ('SM') grape to a low degree of excess copper stress (Low-ECS). Based on the physiological indicators and multi-omics (transcriptome, proteome, metabolome, and microRNAome) data, 8 h (h) after copper treatment was the most severe stress time point. Nonetheless, copper stress was alleviated 64 h after treatment. Cu ion transportation, photosynthesis pathway, antioxidant system, hormone metabolism, and autophagy were the primary response systems in 'SM' grapevine under Low-ECS. Numerous genes and proteins, such as HMA5 , ABC transporters, PMM , GME , DHAR , MDHAR , ARGs , and ARPs, played essential roles in the 'SM' grapevine's response to Low-ECS. This work was carried out to gain insights into the multi-omics responses of 'SM' grapevine to Low-ECS. This study provides genetic and agronomic information that will guide better vinery management and breeding copper-resistant grape cultivars. [Display omitted] • Response network of grapevine leaves to Low-ECS were constructed via multi-omics. • Chelation, segregation and efflux of copper were enhanced in leaves under Low-ECS. • Antioxidant systems and hormone metabolism were induced to alleviate copper stress. • Low-ECS enhanced ARGs and ARPs expressions and autophagosome formation. The joint analysis of multi omics and physiological indexes indicated that grapevines mainly responded to Low-ECS by regulating copper transport, antioxidant pathway, hormone metabolism, and autophagy. [ABSTRACT FROM AUTHOR]

Fertilization, a fundamental aspect of a plant's life, has been of great concern for agricultural specialists to minimize the yield gap between actual and potential yield. Around the globe, fertilizers with different NPK ratios are being used to attain a better yield of grape. To find the suitable commercially available fertilizer for quality grape production, a 2 years (2017–2018) study was conducted for the evaluation of 10 fertilizers with different NPK ratios. Commercial fertilizers included were Zhanlan (16:16:16), Garsoni (15:15:15), Acron (16:16:16), Norway (21:7:12), Peters 1 (30:10:10), Nutrivant (14:14:30), Peters 2 (20:20:20), UMAX (15:15:15), G2 (20:20:20), and Yara (15:15:15). The fertilizer application rate was 20 g plant−1, and each was applied at L-29, L-33, and L-36 phenological stages. Chlorophylls, carotenoids, macro/micronutrients in leaf, and anthocyanin derivatives in grape peel were evaluated. Expression levels of 24 genes, including nitrogen, phosphorous, potassium, and anthocyanin pathways in leaf, peel, and pulp were validated by qPCR at L-29, L-33, and L-36 stages. Results indicated that Norway (21:7:12) and Peters 1 (30:10:10) increased carotenoids, chlorophylls, and anthocyanins in leaves, while Zhanlan (16:16:16) improved fruit biochemical attributes, and anthocyanin (cyanidin, delphinidin, petunidin, malvidin, peonidin, and pelargonidin contents). However, a better grape yield was obtained by the application of Peters 1 (30:10:10). Potassium pathway genes were upregulated by Nutrivant (14:14:30), phosphorous pathway genes by Peters 2 (20:20:20), and nitrogen pathway genes by Peters 1 (30:10:10), while Nutrivant (14:14:30) upregulated anthocyanin pathway genes and simultaneously enhanced anthocyanin biosynthesis in berry peels. Results of two years' study concluded that Peters 1 (30:10:10) was proved better to increase yield, while Zhanlan (14:14:30) was superior in improving anthocyanin biosynthesis. [ABSTRACT FROM AUTHOR]

In recent years, more and more reports have shown that the miR156-SPL module can participate in the regulation of anthocyanin synthesis in plants. However, little is known about how this module responds to hormonal signals manipulating this process in grapes. In this study, exogenous GA, ABA, MeJA, and NAA were used to treat the 'Wink' grape berries before color conversion, anthocyanin and other related quality physiological indexes (such as sugar, aroma) were determined, and spatio-temporal expression patterns of related genes were analyzed. The results showed that the expression levels of VvmiR156b/c/d showed a gradually rising trend with the ripening and color formation of grape berries, and the highest expression levels were detected at day 28 after treatment, while the expression level of VvSPL9 exhibited an opposite trend as a whole, which further verifies that VvmiR156b/c/d can negatively regulate VvSPL9. Besides, VvmiR156b/c/d was positively correlated with anthocyanin content and related genes levels, while the expression pattern of VvSPL9 showed a negative correlation. Analysis of promoter cis-elements and GUS staining showed that VvmiR156b/c/d contained a large number of hormone response cis-elements (ABA, GA, SA, MeJA, and NAA) and were involved in hormone regulation. Exogenous ABA and MeJA treatments significantly upregulated the expression levels of VvmiR156b/c/d and anthocyanin structural genes in the early stage of color conversion and made grape berries quickly colored. Interestingly, GA treatment downregulated the expression levels of VvmiR156b/c/d and anthocyanin structural genes in the early color-change period, but significantly upregulated in the middle color-change and ripening stages, therefore GA mainly modulated grape berry coloring in the middle- and late-ripening stages. Furthermore, NAA treatment downregulated the expression levels of VvmiR156b/c/d and anthocyanin structural genes and delayed the peak expression of genes. Meanwhile, to further recognize the potential functions of VvmiR156b/c/d, the mature tomato transient trangenetic system was utilized in this work. Results showed that transient overexpression of VvmiR156b/c/d in tomato promoted fruit coloring and overexpression of VvSPL9 inhibited fruit coloration. Finally, a regulatory network of the VvmiR156b/c/d-VvSPL9 module responsive to hormones modulating anthocyanin synthesis was developed. In conclusion, VvmiR156b/c/d-mediated VvSPL9 participated in the formation of grape color in response to multi-hormone signals. [ABSTRACT FROM AUTHOR]

Background: Grapevine (Vitis vinifera L.), which has important nutritional values and health benefits, is one of the most economically important fruit crops cultivated worldwide. Several studies showed a large number of microRNAs (VvmiRNAs) involved in the modulation of grape growth and development, and many VvmiRNA families have multiple members. However, the way by which various members from the same miRNA family work is unclear, particularly in grapes. Results: In this study, an important conserved VvmiR172 family (VvmiR172s) and their targets were set as a good example for elucidating the interaction degree, mechanism, and spatio-temporal traits of diverse members from the same miRNA family. miR-RACE and Stem-loop RT-PCR were used to identify the spatio-temporal expressions of various members of VvmiR172s; together with RLM-RACE, PPM-RACE, Western blot, transgenic technologies, their interaction degree, and regulation mechanism were further validated. The expression of VvmiR172c was significantly higher than that of VvmiR172a, b, and d and showed a positive correlation with the abundance of VvAP2 cleavage products. These findings indicated that VvmiR172c might be one of the main action factors of the VvmiR172 family in flower development. The ability of VvmiR172c to cleave target genes differed due to divergence in complementary degree with VvAP2 and expression levels of various members. In VvmiR172 transgenic lines, we observed that 35S::VvmiR172c resulted in the earliest and abundant flowering, indicating the strong function of VvmiR172c. In contrast, the non-significant phenotypic changes were detected in the VvAP2 transgenic lines. The qRT-PCR and Western bolt results demonstrated that VvmiR172c plays a major role in targeting VvAP2. Conclusions: VvmiR172 up-regulated the expression of NtFT and decreased the expression of NtFLC. The up/down regulation of VvmiR172c was the most pronounced. The functions of four VvmiR172 members in grape differed, and miR172c had the strongest regulation on AP2. [ABSTRACT FROM AUTHOR]

Background: Bud dormancy is a strategic mechanism plants developed as an adaptation to unfavorable environments. The grapevine (Vitis vinifera) is one of the most ancient fruit vine species and vines are planted all over the world due to their great economic benefits. To better understand the molecular mechanisms underlying bud dormancy between adjacent months, the transcriptomes of 'Rosario Bianco' grape buds of 6 months and three nodes were analyzed using RNA-sequencing technology and pair-wise comparison. From November to April of the following year, pairwise comparisons were conducted between adjacent months. Results: A total of 11,647 differentially expressed genes (DEGs) were obtained from five comparisons. According to the results of cluster analysis of the DEG profiles and the climatic status of the sampling period, the 6 months were divided into three key processes (November to January, January to March, and March to April). Pair-wise comparisons of DEG profiles of adjacent months and three main dormancy processes showed that the whole grapevine bud dormancy period was mainly regulated by the antioxidant system, secondary metabolism, cell cycle and division, cell wall metabolism, and carbohydrates metabolism. Additionally, several DEGs, such as VvGA2OX6 and VvSS3, showed temporally and spatially differential expression patterns, which normalized to a similar trend during or before April. Conclusion: Considering these results, the molecular mechanisms underlying bud dormancy in the grapevine can be hypothesized, which lays the foundation for further research. [ABSTRACT FROM AUTHOR]

Background: Grape is highly sensitive to gibberellin (GA), which is crucial during seed and berry development (SBD) either by itself or by interacting with other hormones, such as auxin, Abscisic acid (ABA), and Cytokinin (CK). However, no systematic analysis of GA metabolic and signal transduction (MST) pathway has been undertaken in grapevine. Results: In this study, total endogenous GA3 content significantly decreased during SBD, and a total of 48 known genes in GA metabolic (GAM; 31) and signal transduction (ST; 17) pathways were identified in this process. In the GAM pathway, out of 31 genes, VvGA20ox1–1, VvGA3ox4–1, and VvGA2ox1–1 may be the major factors interacting at the green-berry stage (GBS) accompanied with higher accumulation rate. GA biosynthesis was greater than GA inactivation at GBS, confirming the importance of seeds in GA synthesis. The visible correlation between endogenous GA3 content and gene expression profiles suggested that the transcriptional regulation of GA biosynthesis pathway genes was a key mechanism of GA accumulation at the stone-hardening stage (SHS). Interestingly, we observed a negative feedback regulation between VvGA3oxs-VvGAI1–4, VvGA2oxs-VvGAI1–4, and VvGID1B-VvGAI1–4 in maintaining the balance of GA3 content in berries. Moreover, 11 miRNAs may be involved in the modulation of GA MST pathway by mediating their target genes, such as VvGA3ox, VvGID1B, and VvGAMYB. Many genes in auxin, ABA, and CK MST pathways were further identified and found to have a special pattern in the berry, and the crosstalk between GA and these hormones may modulate the complex process during SBD through the interaction gene network of the multihormone pathway. Lastly, based on the expression characterization of multihormone MST pathway genes, a proposed model of the GA-mediated multihormone regulatory network during SBD was proposed. Conclusions: Our results provided novel insights into GA-mediated regulatory networks during SBD in grape. The complexity of GA-mediated multihormone ST in SBD was also elucidated, thereby providing valuable information for future functional characterizations of specific genes in grape. [ABSTRACT FROM AUTHOR]

The formation of grape quality and the mechanism of resistance against foreign pathogens affect the storage stability of fruits during post-harvest handling. Ethylene plays a crucial role in regulating the ripeness of fruits and can be used as an exogenous regulator to resist exogenous pathogens. In this study, we used different concentrations of ethephon for treatment of grape fruits before veraison, analyzed the anthocyanin content, soluble solids, titratable acid, and determined fruit firmness and cell wall metabolism-related enzymes during fruit development. Results showed that exogenous ethephon promoted the early coloration of grape fruits and increased the coloring-related genes myeloblastosis A1(MYBA1), myeloblastosis A2(MYBA2), chalcone isomerase (CHI), flavanone 3-hydroxylase (F3H), flavonoid 3'-hydroxylase gene (F3'H), flavonoid 3', 5'hydroxylase (F3'5'H), 3-O-flavonoid glucosyltransferase (UFGT), and glutathione S-transferase (GST), softening related genes Polygalacturonase(PG), pectinate lyases(PL) and Pectin methylesterase(PME, as well as ethylene metabolism pathway-related genes 1-aminocyclopropane-1-carboxylic acid synthase 1(ACS1), 1-aminocyclopropane-1-carboxylic acid oxidase 2 (ACO2), ethylene receptor gene(ETR2), and ethylene-insensitive 3 (EIN3). Ethephon treatment also increased soluble solids and decreased titratable acid in grape fruit. Fruits pretreated with ethephon were inoculated with Botrytis cinerea, which led to resistance in grape fruit through activation of the antioxidant system. The expression levels of disease resistance-related genes including VvPAD4, VvPIP1, VvNAC26, VvDREB, VvAPX, Vvpgip, VvWRKY70, VvMYC2, VvNPR1 also increased in inoculated fruit with pathogen following ethephon pretreatment. Furthermore, we monitored ethylene response factor 1(ERF1) transcription factor, which could interact with protein EIN3 during ethylene signal transduction and mediate fruit resistance against B. cinerea infection. Meanwhile, overexpression of VvERF1 vectorin strawberry fruits reduced the susceptibility to B. cinerea infection. We suggest that ethylene can induce resistance in ripened fruits after B. cinerea infection and provide adequate postharvest care. [ABSTRACT FROM AUTHOR]

The plant-specific IQ67-domain (IQD) protein family members are downstream targets of calcium sensors, known to regulate plant growth and lateral organ polarity, and basal defense response against environmental cues. No systematic study of IQD gene family has been performed on grapevine. The public availability of grapevine genome enables us to perform identification, phylogeny, chromosomal orientation, and gene structure analysis of the IQD genes in grapevine. We identified 49 VvIQD genes (VvIQD1–VvIQD49) and further classified them into eight subgroups based on phylogenetic relationships. The 49 VvIQD genes were assigned to 19 different chromosomal positions. The collinear relationship between grapevine and Arabidopsis IQDs (VvIQD and AtIQD), and within grapevine VvIQDs, was highly conserved. In addition, most of duplicated gene pairs showed Ka/Ks ratio less than 1.00, indicating purifying selection within these gene pairs, implying functional discrepancy after duplication. Transcription profiling of VvIQD genes shed light on their specific role in grapevine tissue and organ development. The qRT-PCR validation of the 49 VvIQD genes in grape berry tissue from cultivars with distinct berry shape during developmental phases suggested candidate genes involved in the shape of grape berries. The subcellular prediction of VvIQD22, VvIQD23, VvIQD38, and VvIQD49 genes validated their localization in the nucleus and plasma membrane. The VvIQD49 protein interaction with VvCaM2 was also verified by bimolecular fluorescence complementation (BiFC) analysis in the plasma membrane. Our findings will be valuable for the functional genomic studies for desirable shape development of grape berries. [ABSTRACT FROM AUTHOR]

Vitality and survival rate of grape seedlings are crucial factors affecting quality of vineyards. There is no comprehensive study describing accurate evaluation of dormant grapevine seedlings' vitality and survival rate. The purpose of this study was to explore the possibility of using molecular information to evaluate viability and survival rate of dormant seedlings before transplanting. After bare roots treatment, 1–5 day expression levels of six HKGs in four buds of tetraploid Kyoho grape (Vitis labruscana: V. labrusca × V. vinifera) seedlings were detected by (Sq.) RT-PCR and qRT-PCR for calibration of the molecular method. The results revealed that HKGs expression indicates vitality and survival of plant, higher expression was strongly linked to higher vitality and survival rate, lower expression was associated with lower vitality, and lowest expression was significantly associated with lowest vitality and survival rate. Moreover, DNA and RNA quality can superficially determine seedling qualities. Finally, the survival rate of the seedlings produced in Juxian-Shandong, Laixi-Shandong, Huailai-Hebei, Suizhong-Liaoning, Changli-Hebei, Guanxian-Shandong, and Zhangjiagang-Jiangsu was 100.00%, 100.00%, 100.00%, 100.00%, 100.00%, 87.77%, and 93.33%, respectively. In conclusion, molecular technique is potential approach for promoting gene information to estimate vitality and survival rate of dormant grape seedlings and might contribute to viticulturists' efforts. [ABSTRACT FROM AUTHOR]

Salinity is ubiquitous abiotic stress factor limiting viticulture productivity worldwide. However, the grapevine is vulnerable to salt stress, which severely affects growth and development of the vine. Hence, it is crucial to delve into the salt resistance mechanism and screen out salt-resistance prediction marker genes; we implicated RNA-sequence (RNA-seq) technology to compare the grapevine transcriptome profile to salt stress. Results showed 2472 differentially-expressed genes (DEGs) in total in salt-responsive grapevine leaves, including 1067 up-regulated and 1405 down-regulated DEGs. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotations suggested that many DEGs were involved in various defense-related biological pathways, including ROS scavenging, ion transportation, heat shock proteins (HSPs), pathogenesis-related proteins (PRs) and hormone signaling. Furthermore, many DEGs were encoded transcription factors (TFs) and essential regulatory proteins involved in signal transduction by regulating the salt resistance-related genes in grapevine. The antioxidant enzyme analysis showed that salt stress significantly affected the superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and glutathione S-transferase (GST) activities in grapevine leaves. Moreover, the uptake and distribution of sodium (Na+), potassium (K+) and chlorine (Cl−) in source and sink tissues of grapevine was significantly affected by salt stress. Finally, the qRT-PCR analysis of DE validated the data and findings were significantly consistent with RNA-seq data, which further assisted in the selection of salt stress-responsive candidate genes in grapevine. This study contributes in new perspicacity into the underlying molecular mechanism of grapevine salt stress-tolerance at the transcriptome level and explore new approaches to applying the gene information in genetic engineering and breeding purposes. [ABSTRACT FROM AUTHOR]

Phytochrome-interacting factors (PIFs), as the basic helix–loop–helix (bHLH) transcription factors, are the primary signaling partners for phytochromes (PHY) that play a key role in PHY-mediated light signal transduction. At present, there are few studies on PIFs in fruit trees. In order to clarify the status of PIFs in grapevines, we identified members of the grape PIFs family and conducted phylogenetic and expression analysis. We identified PIF1, PIF3, PIF4, and PIF7 in PIFs families of the grapevine (Vitis vinifera L.), which were distributed on four different chromosomes with similar gene structures. Except for the closer relationship with PIF1 of citrus, PIFs of grape were distant from the other fruit species such as apple, pear, peach, and strawberry. The VvPIFs (except VvPIF4) were located in the syntenic block with those from Arabidopsisthaliana, Solanum lycopersicum, or Citrus sinensis. In addition to PIF1, all PIFs in grapevines have conserved active PHYB binding (APB) sequences. VvPIF1 has a conserved PIF1-specific active PHYA binding (APA) sequence, while amino acid mutations occurred in the specific APA sequence in VvPIF3. Interestingly, two specific motifs were found in the PIF4 amino acid sequence. The photoreceptor-related elements in the VvPIFs promoter region were the most abundant. PIF1, LONG HYPOCOTYL 5 (HY5) and PIF3, PIF4, GIBBERELLIC ACID INSENSITIVE 1 (GAI1) may interact with each other and participate together in light signal transduction. The relative expression levels of the VvPIFs showed diverse patterns in the various organs at different developmental stages, of which PIF4 was most highly expressed. Prior to maturation, the expression of PIF4 and PIF7 in the skin of the different cultivars increased, while the expression of all PIFs in the flesh decreased. The transcription level of PIFs in grape leaves was sensitive to changes in lighting and shading. Shading treatment was beneficial for enhancing the transcription level of VvPIFs, but the effect on VvPIF3 and VvPIF4 was time-controlled. We concluded that PIFs in grapevines are both conservative and species-specific. The identification and analysis of grape PIFs could provide a theoretical foundation for the further construction of grape light regulation networks. [ABSTRACT FROM AUTHOR]

• Chlorophyll metabolism and photosynthesis in grapevine leaves were hindered during copper deficiency. • Antioxidant systems and flavonoid metabolism were activated. • Altered copper ion transport prioritizes supply to mitochondria to meet energy needs. • Differences in Cu transportation, chlorophyll degradation, flavonoid metabolism between copper deficiency and excess stress. Copper is an essential trace element in grapevine growth, and lack of it will cause damage to grapevines. However, there are few studies on the physiological changes and regulatory mechanisms of chronic copper deficiency stress in grapevines. The 'Shine Muscat' ('SM') seedlings were treated with Hoagland nutrient solution without copper ions to explore the adaptive response and regulatory mechanism for 35 days. Firstly, it was found that 'SM' grapevines were indeed affected by copper deficiency stress through the leaf symptoms and physiological data. Then, 657 up-regulated and 527 down-regulated differentially expressed genes (DEGs) and 72 differentially accumulated metabolites (DAMs, 31 up-regulated and 41 down-regulated) were identified between control and copper deficiency treatment. Copper ion transportation, photosynthetic pathway, antioxidant system, flavonoid metabolism, and phytohormone metabolism were identified as the primary response systems in 'SM' grapevines. Several genes, including FRO4, ABCs, CHLP, APX, GPX , and PK played a crucial role in the response of 'SM' grapevine to copper deficiency stress. Furthermore, there were notable differences in copper uptake and transportation, chlorophyll degradation, flavonoid synthesis and metabolites, and ABA content between copper deficiency and excess stress. These results will be beneficial to gain insights with copper deficiency regulation and stress-responsive strategies in grapevines. [ABSTRACT FROM AUTHOR]

O -methyltransferases (OMTs) are an important group of enzymes involved in the methylation of various secondary metabolites, including flavonoids. However, the features and functions of OMTs have not been comprehensively studied in grape (Vitis vinifera), a rich source of methylated flavonoids. Here, 47 OMT members were identified in grape genome. They were unevenly distributed on grape chromosomes and some genes were tandem duplicated, indicating the role of duplication processes in the expansion of this gene family. Based on the phylogenetic relationship, these OMTs were clustered into CCoAOMT and COMT subclades, which were further supported by the results of conserved motif and gene structure analysis. Correlation analysis revealed that three members (VvCCoAOMT1 , VvCCoAOMT4 , and VvCOMT1) were potentially involved in the synthesis of most methylated flavonoids in the berry skins. Expression profiling based on RNA-seq data and qRT-PCR experiments indicated that VvCCoAOMT1 and VvCCoAOMT4 had specific and high expression in berry skins, and responded to abscisic acid and high temperature treatments; and that VvCOMT1 expression was significantly induced during berry development and UVC treatment. Cis -regulatory element analysis suggested important roles of OMTs in growth, development, and defense against stresses. We further demonstrated the transcriptional regulation of VvCCoAOMT4 by VvMYBA1, a master regulator of grape berry anthocyanin, and verified the protein localization of VvCCoAOMT4 in membrane and nucleus. These findings facilitate a better understanding of the characteristics of OMT gene family, especially of the potential members involved in the formation of O -methylated flavonoids in grape. • Identification and characterization of the OMT family in grape (Vitis vinifera). • Identification of tissue-specific and stress-related OMT s. • Three members (VvCCoAOMT1 , VvCCoAOMT4 , and VvCOMT1) were potentially involved in methylation of flavonoids. • VvCCoAOMT4 was transcriptionally regulated by VvMYBA1, a master regulator of grape berry anthocyanin. • VvCCoAOMT4 protein was localized in membrane and nucleus. [ABSTRACT FROM AUTHOR]

• Bio-organic fertilizer (BOF) had the better effect than organic fertilizer (OF) in grapevine. • BOF increased the soil fertility and improved the diversity and richness of soil microbial community. • BOF promoted the growth of root and seedling of grapevine. Grapevine seedlings need to absorb a lot of nutrients from the soil during the growth process, and appropriate fertilizer can greatly promote the growth of seedlings. The purpose of this study was to investigate the effects of different bio-organic fertilizers (BOF) on shoot and root growth, soil biochemical characteristics, and bacterial communities of 'Shine Muscat' and 'Kyoho' seedlings. Four treatments were established: no fertilizer (CK), chemical fertilizer + organic fertilizer (T1), chemical fertilizer + bio-organic fertilizer 1 (T2, containing Bacillus amyloliquefaciens), chemical fertilizer + bio-organic fertilizer 2 (T3, containing Trichoderma). The results showed that BOF had better effects on growth and rhizosphere environment of grapevine seedling than organic fertilizer (OF) treated group. The shoot growth, chlorophyll content, and root growth with BOF of grapevine seedlings were generally better than those treated with OF. Compared with T1, T2 and T3 made grapevine seedlings meet the fixed drying requirement (main shoot 0.8 m) 1–4 weeks earlier. The expansion of roots and the improvement of root activity under BOF treatment were also beneficial to plant growth. Moreover, transcriptome analysis showed that BOF enhanced the phytohormones and cell wall processes in grapevine root, and up-regulated the expression of cell wall-related genes such as XTH23, EXPs, PME17 , and auxin related genes such as ABCBs and GH3. In addition, BOF increased soil organic matter content, available nitrogen, available potassium, available phosphorus, and enzyme activity (soil catalase, urease, sucrase, and nitrate reductase). The Shannon and Chao values of T2 were higher than those of T1 and CK, which improved the diversity and richness of soil microbial community. As compared to CK, T2 treatment increased the symbiotic nitrogen fixing Proteobacteria by 4.10 %, and reduced Acidobacteria by 9.79 %. Actinobacteria, which is related to the degradation of stubborn polymers, was the dominant group in T2 treatment, accounting for 10.17 %. As compared to CK and T1, Actinobacteria was increased by 6.89 % and 5.66 %. In summary, BOF could promote the growth of grapevine seedlings by improving phenotypic traits, soil fertility, bacterial diversity, richness abundance, and provide theoretical and technical support for grapevine garden construction and tree management. [Display omitted] [ABSTRACT FROM AUTHOR]

• The accumulation of osmoprotectants (e.g., glycine betaine and trehalose) enriched in grapevine in response to salt stress. • As the stress duration was alleviated the concentration of ions significantly increased in source and sink tissues. • The activity of ROS detoxifying enzymes drastically enhanced to counter the lipid peroxidation caused by H 2 O 2 and O 2 –. Salinity is an omnipresent stressor, depleting osmotic potential and affecting the vineyard development and productivity. It is indispensable to screen the grapevine genetic resources for possibilities to mitigate and adapt to the stress conditions. This paper describes the distinct response of the grapevine under progressive salt stress duration (0, 24, 48, and 72 h). Intriguingly, besides the adverse effect of salt stress on grapevine growth, the findings exhibit significant levels of stress responses to diminish the impact of salinity. Discerned are the accumulation of glycine betaine (GB) and trehalose, which can be supposed as distinct grapevine responses associated with the maintenance of high K+ level and, consequently, lower K+/Na+ ratios, with an excellent performance of cell water status and photosynthetic activity as compared to the controls. Moreover, an alleviated level of the oxidative stress marker (MDA) gives rise to superoxide radicals (O 2 −) and hydrogen peroxide (H 2 O 2), while the antioxidative scavenging system is activated to counter the synthesis of reactive oxygen species (ROS). Principal component analysis (PCA) distinguished several salt prediction candidate markers based on physiological and transcriptional indexes in four different clusters. Current findings have elucidated new and fascinating responses of the grapevine to lengthy salt-stress period and provide a list of intended enzymatic targets for candidate gene selection to uplift the abiotic stress tolerance in horticultural crops. [ABSTRACT FROM AUTHOR]

Grape (Vitis vinifera L.) is an important fruit for which light quality plays a significant role in plant growth and development. However, the regulatory mechanisms between light quality and plant growth at the physiological and molecular levels are poorly understood. Therefore, in the present study, we performed multivariate sequencing analysis to investigate the effects of four light qualities: blue (B), red (R), green (G), and white (W, used as a control group) to investigate grapevine morphology at the molecular level, which revealed the possible networking pathway controlling grapevine growth. Our results showed that morphological and physiological parameters, such as plant height, stem diameter, leaf area, and the contents of total chlorophyll, chlorophyll a and b, and carotenoid, were significantly improved under blue and red light compared to those under white light. Photosynthetic measurements showed that the total conductance to water vapor, total conductance to water CO 2 , maximum quantum yield of photosystem (PS)II, apparent electron transfer chain, and net photosynthetic rate were elevated by blue and red light compared to white light. However, there were notable reductions in the intercellular CO 2 concentration, transpiration rate, and non-photochemical quenching coefficient under blue light, which showed an irreversible trend compared to other light qualities. The contents of minerals, such as nitrogen, phosphorous, potassium, magnesium, iron, and zinc, were also improved by blue and red light compared to white light. Multivariate sequencing analysis revealed a total of 1065 metabolites, of which 747 were positive and 318 were negative. Gene ontology and Kyoto Encyclopedia of Genes and Genomes analyses showed that many differentially expressed genes (DEGs) were associated with the biosynthesis of secondary metabolites, including flavones, flavonols, and alkaloids, and metabolic and phenylpropanoid pathways. Further, numerous transcription factors, including MYB (37 DEGs), bHLH (32 DEGs), NAC (31 DEGs), and WRKY (29 DEGs), were reported. Photosynthesis-related genes (PsaD, PsaO, PsbB, PetC, PetE, PetF, PetH, PetJ, and Lhca) played pivotal roles in the photosynthetic pathways. In addition, using weighed gene correlation network analysis, 7 module relationships (MEblack, MEgreenyellow, MEred, MEturquoise, MEgreen, MEyellow, and MEmagneta), 4 metabolites, 14 structural genes, and 36 transcription factor-related genes were found. Conclusively, the present research provides new insights for viticulture in response to light quality and offers new strategies for agriculture. [Display omitted] • Blue light treatment increased photosynthesis and material content of grape seedlings. • After 30 days of continuous light quality treatment, photosynthesis was reduced compared to 15 days. • Blue light, red light and green light affect sugar metabolism and amino acid metabolism to some extent. [ABSTRACT FROM AUTHOR]