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Drought is the most severe form of stress experienced by plants worldwide. Cucumber is a vegetable crop that requires a large amount of water throughout the growth period. In our previous study, we identified that overexpression of CsHSFA1d could improve cold tolerance and the content of endogenous jasmonic acid in cucumber seedlings. To explore the functional diversities of CsHSFA1d, we treat the transgenic plants under drought conditions. In this study, we found that the heat shock transcription factor HSFA1d (CsHSFA1d) could improve drought stress tolerance in cucumber. CsHSFA1d overexpression increased the expression levels of galactinol synthase (CsGolS3) and raffinose synthase (CsRS) genes, encoding the key enzymes for raffinose family oligosaccharide (RFO) biosynthesis. Furthermore, the lines overexpressing CsHSFA1d showed higher enzymatic activity of GolS and raffinose synthase to increase the content of RFO. Moreover, the CsHSFA1d -overexpression lines showed lower reactive oxygen species (ROS) accumulation and higher ROS-scavenging enzyme activity after drought treatment. The expressions of antioxidant genes CsPOD2, CsAPX1 and CsSOD1 were also upregulated in CsHSFA1d -overexpression lines. The expression levels of stress-responsive genes such as CsRD29A, CsLEA3 and CsP5CS1 were increased in CsHSFA1d -overexpression lines after drought treatment. We conclude that CsHSFA1d directly targets and regulates the expression of CsGolS3 and CsRS to promote the enzymatic activity and accumulation of RFO to increase the tolerance to drought stress. CsHSFA1d also improves ROS-scavenging enzyme activity and gene expression indirectly to reduce drought-induced ROS overaccumulation. This study therefore offers a new gene target to improve drought stress tolerance in cucumber and revealed the underlying mechanism by which CsHSFA1d functions in the drought stress by increasing the content of RFOs and scavenging the excessive accumulation of ROS. [ABSTRACT FROM AUTHOR]

Initiation and progression of leaf senescence are triggered by various environmental stressors and phytohormones. Jasmonic acid (JA) and darkness accelerate leaf senescence in plants. However, the mechanisms that integrate these two factors to initiate and regulate leaf senescence have not been identified. Here, we report a transcriptional regulatory module centred on a novel tomato WRKY transcription factor, SlWRKY37, responsible for both JA- and dark-induced leaf senescence. The expression of SlWRKY37 , together with SlMYC2 , encoding a master transcription factor in JA signalling, was significantly induced by both methyl jasmonate (MeJA) and dark treatments. SlMYC2 binds directly to the promoter of SlWRKY37 to activate its expression. Knock out of SlWRKY37 inhibited JA- and dark-induced leaf senescence. Transcriptome analysis and biochemical experiments revealed SlWRKY53 and SlSGR1 (S. lycopersicum senescence-inducible chloroplast stay-green protein 1) as direct transcriptional targets of SlWRKY37 to control leaf senescence. Moreover, SlWRKY37 interacted with a VQ motif-containing protein SlVQ7, and the interaction improved the stability of SlWRKY37 and the transcriptional activation of downstream target genes. Our results reveal the physiological and molecular functions of SlWRKY37 in leaf senescence, and offer a target gene to retard leaf yellowing by reducing sensitivity to external senescence signals, such as JA and darkness. [ABSTRACT FROM AUTHOR]

SUMMARY: Cucumber (Cucumis sativus) originated in tropical areas and is very sensitive to low temperatures. Cold acclimation is a universal strategy that improves plant resistance to cold stress. In this study, we report that heat shock induces cold acclimation in cucumber seedlings, via a process involving the heat‐shock transcription factor HSFA1d. CsHSFA1d expression was improved by both heat shock and cold treatment. Moreover, CsHSFA1d transcripts accumulated more under cold treatment after a heat‐shock pre‐treatment than with either heat shock or cold treatment alone. After exposure to cold, cucumber lines overexpressing CsHSFA1d displayed stronger tolerance for cold stress than the wild type, whereas CsHSFA1d knockdown lines obtained by RNA interference were more sensitive to cold stress. Furthermore, both the overexpression of CsHSFA1d and heat‐shock pre‐treatment increased the endogenous jasmonic acid (JA) content in cucumber seedlings after cold treatment. Exogenous application of JA rescued the cold‐sensitive phenotype of CsHSFA1d knockdown lines, underscoring that JA biosynthesis is key for CsHSFA1d‐mediated cold tolerance. Higher JA content is likely to lead to the degradation of CsJAZ5, a repressor protein of the JA pathway. We also established that CsJAZ5 interacts with CsICE1. JA‐induced degradation of CsJAZ5 would be expected to release CsICE1, which would then activate the ICE–CBF–COR pathway. After cold treatment, the relative expression levels of ICE–CBF–COR signaling pathway genes, such as CsICE1, CsCBF1, CsCBF2 and CsCOR1, in CsHSFA1d overexpression lines were significantly higher than in the wild type and knockdown lines. Taken together, our results help to reveal the mechanism underlying heat shock‐induced cold acclimation in cucumber. Significance Statement: Cold acclimation is a universal strategy that improves plant resistance to cold stress. In this study, we discovered that a heat‐shock pre‐treatment improves the tolerance of cucumber seedlings for low‐temperature stress, and that CsHSFA1d and JA play important roles in this cold acclimation process. Our results will help to reveal the mechanism underlying heat shock‐induced cold acclimation in cucumber. [ABSTRACT FROM AUTHOR]

Improving plant water‐use efficiency (WUE) is important to plant survival and crop yield in the context of water limitation. In this study, SlTLFP8 (Tubby‐like F‐box protein 8) was identified as an osmotic‐induced gene in tomato. Transgenic tomato with up‐regulated expression of SlTLFP8 showed enhanced water‐deficient resistance, whereas knockout mutants generated by CRISPR/Cas9 were more sensitive to water deficit. SlTLFP8 overexpression significantly enhanced WUE by suppressing transpiration under both water‐sufficient and water‐deficient conditions. Further study showed that overexpressing SlTLFP8 significantly increased leaf epidermal cell size and thereby decreased stomatal density 10–20%, conversely SlTLFP8 knockout resulted in decreased cell size and thereby increased stomatal density 20–50%. SlTLFP8 overexpression and knockout modulated ploidy levels in leaf cells. Changes in expression of cell cycle related genes also indicated that SlTLFP8 affected cell size and stomatal density through endocycle transition. Despite changes in stomata density and transpiration, altering the expression of SlTLFP8 did not change photosynthesis. Additionally, biomass was not altered and there was little difference in fruit yield for transgenic and wild type lines under water‐sufficient and water‐deficient conditions. Our results demonstrate the effect of SlTLFP8 on endoreduplication and the potential of SlTLFP8 for improvement of WUE. Brief Summery: This work found a new mechanism of TLP (Tubby like protein) response to water‐deficient stress. SlTLFP8, a member of TLP family, regulates water‐deficient resistance by modulating water loss via affecting stomatal density. Expression of SlTLFP8 was induced by osmotic stress. Transgenic tomato lines with SlTLFP8 overexpression or SlTLFP8 knockout showed significantly differences in water‐use efficiency (WUE) and water‐deficient resistance. The difference of leaf water loss caused by transpiration is the main explanation of the difference in WUE and water‐deficient resistance. Additionally, overexpressing SlTLFP8 significantly decreased stomatal density, while SlTLFP8 knockout resulted in increased stomatal density, and SlTLFP8 affected stomatal density through endoreduplication and altered epidermal cell size. Despite changes in stomata density, altering the expression of SlTLFP8 did not result in distinct changes in photosynthesis, biomass and yield of tomato. This work found a new mechanism of TLP (Tubby like protein) response to water‐deficient stress. SlTLFP8, a member of TLP family, regulates water‐deficient resistance by modulating water loss via affecting stomatal density. Expression of SlTLFP8 was induced by osmotic stress. Transgenic tomato lines with SlTLFP8 overexpression or SlTLFP8 knockout showed significantly differences in water‐use efficiency (WUE) and water‐deficient resistance. The difference of leaf water loss caused by transpiration is the main explanation of the difference in WUE and water‐deficient resistance. Additionally, overexpressing SlTLFP8 significantly decreased stomatal density, while SlTLFP8 knockout resulted in increased stomatal density, and SlTLFP8 affected stomatal density through endoreduplication and altered epidermal cell size. Despite changes in stomata density, altering the expression of SlTLFP8 did not result in distinct changes in photosynthesis, biomass and yield of tomato. [ABSTRACT FROM AUTHOR]