Background Grapevine is an economically important crop for which yield and berry quality is strongly affected by climate change. Large variations in drought tolerance exist across Vitis species. Some of these species are used as rootstock to enhance abiotic and biotic stress tolerance. In this study, we investigated the physiological and transcriptomic responses to water deficit of four different genotypes that differ in drought tolerance: Ramsey (Vitis champinii), Riparia Gloire (Vitis riparia), Cabernet Sauvignon (Vitis vinifera), and SC2 (Vitis vinifera x Vitis girdiana). Results Ramsey was particularly more drought tolerant than the other three genotypes. Ramsey maintained a higher stomatal conductance and photosynthesis at equivalent levels of moderate water deficit. We identified specific and common transcriptomic responses shared among the four different Vitis species using RNA sequencing analysis. A weighted gene co-expression analysis identified a water deficit core gene set with the ABA biosynthesis and signaling genes, NCED3, RD29B and ABI1 as potential hub genes. The transcript abundance of many abscisic acid metabolism and signaling genes was strongly increased by water deficit along with genes associated with lipid metabolism, galactinol synthases and MIP family proteins. This response occurred at smaller water deficits in Ramsey and with higher transcript abundance than the other genotypes. A number of aquaporin genes displayed differential and unique responses to water deficit in Ramsey leaves. Genes involved in cysteine biosynthesis and metabolism were constitutively higher in the roots of Ramsey; thus, linking the gene expression of a known factor that influences ABA biosynthesis to this genotype’s increased NCED3 transcript abundance. Conclusion The drought tolerant Ramsey maintained higher photosynthesis at equivalent water deficit than the three other grapevine genotypes. Ramsey was more responsive to water deficit; its transcriptome responded at smaller water deficits, whereas the other genotypes did not respond until more severe water deficits were reached. There was a common core gene network responding to water deficit for all genotypes that included ABA metabolism and signaling. The gene clusters and sub-networks identified in this work represent interesting gene lists to explore and to better understand drought tolerance molecular mechanisms.

Climate change threatens food security, and plant science researchers have investigated methods of sustaining crop yield under drought. One approach has been to overproduce abscisic acid (ABA) to enhance water use efficiency. However, the concomitant effects of ABA overproduction on plant vascular system functioning are critical as it influences vulnerability to xylem hydraulic failure. We investigated these effects by comparing physiological and hydraulic responses to water deficit between a tomato (Solanum lycopersicum) wild type control (WT) and a transgenic line overproducing ABA (sp12). Under well‐watered conditions, the sp12 line displayed similar growth rate and greater water use efficiency by operating at lower maximum stomatal conductance. X‐ray microtomography revealed that sp12 was significantly more vulnerable to xylem embolism, resulting in a reduced hydraulic safety margin. We also observed a significant ontogenic effect on vulnerability to xylem embolism for both WT and sp12. This study demonstrates that the greater water use efficiency in the tomato ABA overproducing line is associated with higher vulnerability of the vascular system to embolism and a higher risk of hydraulic failure. Integrating hydraulic traits into breeding programmes represents a critical step for effectively managing a crop's ability to maintain hydraulic conductivity and productivity under water deficit.

Moderate levels of Cl- have been associated with grapevine salt tolerance. The hypothesis to be tested in this work is: photosynthesis in grapevine is negatively correlated with foliar Cl- concentration. To further test this hypothesis, multiple mild salinity experiments on four different Vitis genotypes (Cabernet-Sauvignon, Riparia Gloire, Ramsey and SC2) were conducted and photosynthesis, ion concentrations and gene expression responses were quantified. The salt-tolerant rootstock Ramsey had greater Cl- exclusion capabilities than V. vinifera cultivars both during rooted cutting greenhouse experiments and three years of field-grafted experiments; SC2 also excluded Cl-. Differential gene expression indicated that salinity affected transcript abundance more in salt-sensitive genotypes (97.7 % of DEGs in the dataset), especially chloroplast-related transcripts. The transcript abundances of known anion transporters were determined and a family of putative B transporters was associated with the Cl- exclusion phenotype. Photosynthesis and growth were maintained in Ramsey and SC2 under mild salinity. However, photosynthesis declined in Cabernet-Sauvignon with isosmotic 20 mM salt concentrations of NaCl, KCl or NaNO3, independent of the salt type. While foliar Cl- concentrations did correlate with salt tolerance during control and NaCl conditions, it was not found to be the cause of photosynthetic decline in Vitis during mild salinity.

Background: VviERF6Ls are an uncharacterized gene clade in Vitis with only distant Arabidopsis orthologs. Preliminary data indicated these transcription factors may play a role in berry development and extreme abiotic stress responses. To better understand this highly duplicated, conserved clade, additional members of the clade were identified in four Vitis genotypes. A meta-data analysis was performed on publicly available microarray and RNA-Seq data (confirmed and expanded with RT-qPCR), and Vitis VviERF6L1 overexpression lines were established and characterized with phenotyping and RNA-Seq. Results: A total of 18 PN40024 VviERF6Ls were identified; additional VviERF6Ls were identified in Cabernet Sauvignon, Chardonnay, and Carménère. The amino acid sequences of VviERF6Ls were found to be highly conserved. VviERF6L transcripts were detected in numerous plant organs and were differentially expressed in response to numerous abiotic stresses including water deficit, salinity, and cold as well as biotic stresses such as red blotch virus, N. parvum, and E. necator. VviERF6Ls were differentially expressed across stages of berry development, peaking in the pre-veraison/veraison stage and retaining conserved expression patterns across different vineyards, years, and Vitis cultivars. Co-expression network analysis identified a scarecrow-like transcription factor and a calmodulin-like gene with highly similar expression profiles to the VviERF6L clade. Overexpression of VviERF6L1 in a Seyval Blanc background did not result in detectable morphological phenotypes. Genes differentially expressed in response to VviERF6L1 overexpression were associated with abiotic and biotic stress responses. Conclusions: VviERF6Ls represent a large and distinct clade of ERF transcription factors in grapevine. The high conservation of protein sequence between these 18 transcription factors may indicate these genes originate from a duplication event in Vitis. Despite high sequence similarity and similar expression patterns, VviERF6Ls demonstrate unique levels of expression supported by similar but heterogeneous promoter sequences. VviERF6L gene expression differed between Vitis species, cultivars and organs including roots, leaves and berries. These genes respond to berry development and abiotic and biotic stresses. VviERF6L1 overexpression in Vitis vinifera results in differential expression of genes related to phytohormone and immune system signaling. Further investigation of this interesting gene family is warranted.

Background: Abscisic acid is a phytohormone involved in water deficit response. Abscisic acid metabolism is regulated by biosynthesis, conjugation, and catabolism. NCED3 is the rate limiting step of abscisic acid biosynthesis and is a key contributor to plant water deficit responses. In this study NCED3 transcript accumulation and abscisic acid metabolism were further characterized as key water deficit responses in four Vitis species (Vitis vinifera (Cabernet Sauvignon), Vitis champinii (Ramsey), Vitis riparia (Riparia Gloire), and Vitis vinifera x Vitis girdiana (SC2)) under three levels of water deficit in leaves and roots. Results: The concentrations of abscisic acid and derivative metabolites increased with water deficit and was dependent upon the species. RNA-Seq and RT-qPCR data were consistent with the changes in abscisic acid metabolite concentrations; the corresponding transcript abundances substantiate NCED3 as a key gene in the water deficit response; however, NCED3 protein concentrations assayed in Western Blots were not affected. Major differences in abscisic acid metabolism at the gene, protein, and metabolite levels were detected between leaves and roots in these four species. NCED3 transcript abundance and abscisic acid concentration in drought-tolerant Ramsey increased earlier and more significantly than the other species during long-term, moderate to severe water deficits but were not stimulated as much by short-term, rapid dehydration. In drought-sensitive Riparia, NCED3 transcript abundance and abscisic acid metabolite concentrations increased to a lower extent than in Ramsey during moderate to severe water deficits, but short-term rapid dehydration induced a significantly higher abscisic acid concentration in Riparia than Ramsey. Conclusions: Grapevine species have distinct abscisic acid metabolism that depends highly on the severity and duration of stress and organ (leaves or roots). This study confirms that abscisic acid metabolism and NCED3 are part of a core water deficit response in Vitis species. Relative quantities of transcripts, proteins, abscisic acid and derivative metabolites were determined, but many aspects of abscisic acid metabolism and water deficit responses warrant additional investigation. This study provides a better understanding of how Vitis is adapted to dry environments, which may be exploited for future breeding programs.

Background VviERF6Ls are an uncharacterized gene clade in Vitis with only distant Arabidopsis orthologs. Preliminary data indicated these transcription factors may play a role in berry development and extreme abiotic stress responses. To better understand this highly duplicated, conserved clade, additional members of the clade were identified in four Vitis genotypes. A meta-data analysis was performed on publicly available microarray and RNA-Seq data (confirmed and expanded with RT-qPCR), and a Vitis VviERF6L1 overexpression line was established and characterized with phenotyping and RNA-Seq. Results A total of 18 PN40024 VviERF6Ls were identified; additional VviERF6Ls were identified in Cabernet Sauvignon, Chardonnay, and Carménère. The amino acid sequences of VviERF6Ls were found to be highly conserved. VviERF6L transcripts were detected in numerous plant organs and were differentially expressed in response to numerous abiotic stresses including water deficit, salinity, and cold as well as biotic stresses such as red blotch virus, N. parvum , and E. necator . VviERF6Ls were differentially expressed across stages of berry development, peaking in the pre-veraison/veraison stage, and retaining conserved expression patterns across different vineyards, years, and Vitis cultivars. Co-expression network analysis identified a scarecrow-like transcription factor and a calmodulin-like gene with highly similar expression profiles to the VviERF6L clade. Overexpression of VviERF6L1 in a Seyval Blanc background did not result in detectable morphological phenotypes. Genes differentially expressed in response to VviERF6L1 overexpression were associated with abiotic and biotic stress responses. Conclusions VviERF6Ls represent a large and distinct clade of ERF transcription factors in grapevine. The high conservation of protein sequence between these 18 transcription factors may indicate these genes originate from a duplication event in Vitis . Despite high sequence similarity and similar expression patterns, VviERF6Ls demonstrate unique levels of expression supported by similar but heterogeneous promoter sequences. VviERF6L gene expression differed between Vitis species, cultivars and organs including roots, leaves and berries. These genes respond to berry development and abiotic and biotic stresses. VviERF6L1 overexpression in Vitis vinifera results in differential expression of genes related to phytohormone and immune system signaling. Further investigation of this interesting gene family is warranted.

Background: VviERF6Ls are an uncharacterized gene clade in Vitis with only distant Arabidopsis orthologs. Preliminary data indicated these transcription factors may play a role in berry development and extreme abiotic stress responses. To better understand this highly duplicated, conserved clade, additional members of the clade were identified in four Vitis genotypes. A meta-data analysis was performed on publicly available microarray and RNA-Seq data (confirmed and expanded with RT-qPCR), and Vitis VviERF6L1 overexpression lines were established and characterized with phenotyping and RNA-Seq. Results: A total of 18 PN40024 VviERF6Ls were identified; additional VviERF6Ls were identified in Cabernet Sauvignon, Chardonnay, and Carménère. The amino acid sequences of VviERF6Ls were found to be highly conserved. VviERF6L transcripts were detected in numerous plant organs and were differentially expressed in response to numerous abiotic stresses including water deficit, salinity, and cold as well as biotic stresses such as red blotch virus, N. parvum , and E. necator . VviERF6Ls were differentially expressed across stages of berry development, peaking in the pre-veraison/veraison stage and retaining conserved expression patterns across different vineyards, years, and Vitis cultivars. Co-expression network analysis identified a scarecrow-like transcription factor and a calmodulin-like gene with highly similar expression profiles to the VviERF6L clade. Overexpression of VviERF6L1 in a Seyval Blanc background did not result in detectable morphological phenotypes. Genes differentially expressed in response to VviERF6L1 overexpression were associated with abiotic and biotic stress responses. Conclusions: VviERF6Ls represent a large and distinct clade of ERF transcription factors in grapevine. The high conservation of protein sequence between these 18 transcription factors may indicate these genes originate from a duplication event in Vitis . Despite high sequence similarity and similar expression patterns, VviERF6Ls demonstrate unique levels of expression supported by similar but heterogeneous promoter sequences. VviERF6L gene expression differed between Vitis species, cultivars and organs including roots, leaves and berries. These genes respond to berry development and abiotic and biotic stresses. VviERF6L1 overexpression in Vitis vinifera results in differential expression of genes related to phytohormone and immune system signaling. Further investigation of this interesting gene family is warranted.

Abscisic acid (ABA) metabolism is complex involving biosynthesis, conjugation, and catabolism. Differences in ABA metabolism (ABA, ABA-related metabolites, and transcripts) in response to water deficit (WD) were detected in leaves and roots of four Vitis species differing in drought tolerance: Vitis vinifera (Cabernet Sauvignon), Vitis champinii (Ramsey), Vitis riparia (Riparia Gloire), and Vitis vinifera x girdiana (SC2). Concentrations of ABA and ABA-related metabolites increased after moderate or severe WD depending on species, organ, and time. Differences in ABA, glycosylated-ABA, and ABA catabolite concentrations, as well as previously investigated related transcript abundances, revealed differences in ABA metabolism pathways among the species and organs. NCED3 was a key gene in the WD response of leaves and roots of all species. NCED3 transcript abundance and ABA concentration in drought-tolerant Ramsey increased earlier and to a greater extent than other species. These species provide informative genetic resources to study ABA metabolism and drought tolerance further.