No abstract

The NAC transcription factors play vital roles in responding to drought stress in plants; however, the molecular mechanisms remain largely unknown in cucumber. Suppression of CsATAF1 via RNA interference (RNAi) weakened drought stress tolerance in cucumber due to a higher water loss rate in leaves, a higher level of hydrogen peroxide (H2O2) and superoxide radicals (O2·-), increased malondialdehyde (MDA) content, lower Fv/Fm ratios and lower antioxidant enzyme activity. The analysis of root length and stomatal apertures showed that CsATAF1-RNAi cucumber plants were less responsive to ABA. In contrast, CsATAF1-overexpression (OE) plants showed increased drought stress tolerance and sensitivity to ABA. Quantitative PCR (qPCR) analysis showed that expression of several stress-responsive genes was significantly up-regulated in CsATAF1-OE transformants and down-regulated in CsATAF1-RNAi transformants. CsABI5, CsCu-ZnSOD and CsDREB2C were verified as direct target genes of CsATAF1. Yeast one-hybrid analysis and electrophoretic mobility shift assay (EMSA) further substantiated that CsATAF1 bound to the promoters of CsABI5, CsCu-ZnSOD and CsDREB2C. Transient expression in tobacco leaves and cucumber protoplasts showed that CsATAF1 directly up-regulated the expression of CsABI5, CsCu-ZnSOD and CsDREB2C. Our results demonstrated that CsATAF1 functioned as a positive regulator in response to drought stress by an ABA-dependent pathway and decreasing reactive oxygen species (ROS) accumulation in cucumber.

The function of aquaporin (AQP) protein in transporting water is crucial for plants to survive in drought stress. With 47 homologues in tomato (Solanum lycopersicum) were reported, but the individual and integrated functions of aquaporins involved in drought response remains unclear. Here, three plasma membrane intrinsic protein genes, SlPIP2;1, SlPIP2;7 and SlPIP2;5, were identified as candidate aquaporins genes because of highly expressed in tomato roots. Assay on expression in Xenopus oocytes demonstrated that SlPIP2s protein displayed water channel activity and facilitated water transport into the cells. With real-time PCR and in situ hybridization analysis, SlPIP2s were considered to be involved in response to drought treatment. To test its function, transgenic Arabidopsis and tomato lines overexpressing SlPIP2;1, SlPIP2;7 or SlPIP2;5 were generated. Compared with wild type, the over-expression of SlPIP2;1, SlPIP2;7 or SlPIP2;5 transgenic Arabidopsis and tomato plants all showed significantly higher hydraulic conductivity levels and survival rates under both normal and drought conditions. Taken together, this study concludes that aquaporins (SlPIP2;1, SlPIP2;7 and SlPIP2;5) contribute substantially to root water uptake in tomato plants through improving plant water content and maintaining osmotic balance.

NACs are one of the largest transcription factor families in plants and are involved in the response to abiotic stress. BoNAC019, a homologue of AtNAC019, was isolated from cabbage (Brassica oleracea). BoNAC019 was localized in the nucleus and functioned as a transcriptional activator. The expression of BoNAC019 was induced by dehydration, salt, abscisic acid (ABA), and H2O2 treatments. BoNAC019 overexpressing plants were generated to explore the function of BoNAC019 in response to drought stress. Overexpression (OE) of BoNAC019 reduced drought tolerance with lower survival rate, higher water loss rate, lower proline content and ABA content. The seed germination and root length assays of BoNAC019-OE plants showed decreased sensitivity to ABA. Under drought condition, antioxidant enzymes and anthocyanin content decreased in BoNAC019 -OE plants, resulting in the accumulation of more reactive oxygen species (ROS), which cause damage to plants. Several stress-responsive genes, antioxidant enzymatic genes, anthocyanin biosynthetic genes and ABA signaling genes were down-regulated under drought condition while the ABA catabolism genes were induced in BoNAC019-OE plants under both normal and drought conditions. Our results demonstrated that BoNAC019 might participated in regulating drought tolerance by inducing ABA catabolism genes and decreasing ABA content.

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.