RESUMO
Heat stress poses a major threat to plant productivity and crop yields. The induction of heat shock proteins (HSPs) by heat shock factors is a principal defense response of plants exposed to heat stress. In this study, we identified and analyzed the heat stress-induced Whirly1 (SlWHY1) gene in tomato (Solanum lycopersicum). We generated various SlWHY1-overexpressing (OE) and SlWHY1-RNA interference (RNAi) lines to investigate the role of WHIRLY1 in thermotolerance. Compared with the wild type (WT), the OE lines showed less wilting, as reflected by their increased membrane stability and soluble sugar content and reduced reactive oxygen species (ROS) accumulation under heat stress. By contrast, RNAi lines with inhibited SlWHY1 expression showed the opposite phenotype and corresponding physiological indices under heat stress. The heat-induced gene SlHSP21.5A, encoding an endoplasmic reticulum-localized HSP, was upregulated in the OE lines and downregulated in the RNAi lines compared with the WT. RNAi-mediated inhibition of SlHSP21.5A expression also resulted in reduced membrane stability and soluble sugar content and increased ROS accumulation under heat stress compared with the WT. SlWHY1 binds to the elicitor response element-like element in the promoter of SlHSP21.5A to activate its transcription. These findings suggest that SlWHY1 promotes thermotolerance in tomato by regulating SlHSP21.5A expression.
Assuntos
Proteínas de Ligação a DNA/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Choque Térmico/genética , Proteínas de Plantas/genética , Solanum lycopersicum/genética , Termotolerância/genética , Arabidopsis/genética , Proteínas de Arabidopsis , Clorofila , Proteínas de Ligação a DNA/metabolismo , Proteínas de Choque Térmico/metabolismo , Resposta ao Choque Térmico/genética , Temperatura Alta , Solanum lycopersicum/fisiologia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Espécies Reativas de Oxigênio/metabolismo , Termotolerância/fisiologia , Nicotiana/genéticaRESUMO
In plants, the chilling response involves decreased photosynthetic capacity and increased starch accumulation in chloroplasts. However, the mechanisms that modulate these processes remain unclear. We found that the SlWHY1 gene is significantly induced by chilling stress (4°C) in tomato. Three SlWHY1 overexpression (OE) lines grew better than the wild type (WT) under chilling stress; the OE plants retained intact photosynthetic grana lamellae and showed enhanced hydrolysis of starch. By contrast, RNAi lines that inhibited SlWHY1 were more affected than the corresponding WT cultivar. Their grana lamellae were damaged and starch content increased. The psbA gene encodes the key photosystem II (PSII) protein D1. We show that SlWHY1 binds to the upstream region (A/GTTACCCT/A) of SlpsbA and enhances the de novo synthesis of D1 in chloroplasts. Additionally, SlWHY1 regulates the expression of the starch-degrading enzyme α-amylase (SlAMY3-L) and the starch synthesis-related enzyme isoamylase gene (SlISA2) in the nucleus, thus modulating the starch content in chloroplasts. We demonstrate that SlWHY1 enhances the resistance of tomato to chilling stress by maintaining the function of PSII and degrading starch. Thus, overexpression of WHY1 may be an effective strategy for enhancing resistance to chilling stress of chilling-sensitive crops in agricultural production.
Assuntos
Adaptação Fisiológica , Temperatura Baixa , Complexo de Proteína do Fotossistema II/metabolismo , Proteínas de Plantas/metabolismo , Solanum lycopersicum/fisiologia , Amido/metabolismo , Estresse Fisiológico , Sequência de Bases , Cloroplastos/metabolismo , Regulação da Expressão Gênica de Plantas , Solanum lycopersicum/genética , Modelos Biológicos , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Ligação Proteica , Espécies Reativas de Oxigênio/metabolismoRESUMO
High temperature has become a major abiotic stress that limits crop productivity. Heat shock transcription factors (HSFs) and heat shock proteins (HSPs) play important roles in enhancing thermotolerance of plants. SUMOylation is an important post-translational modification in regulating cellular functions in eukaryotes. SIZ1, a well-characterized SUMO E3 ligase, mediates the process of SUMOylation. In this study, SUMO conjugations were clearly induced by high temperature. Overexpression of SIZ1 SUMO E3 ligase (SlSIZ1) in tomato could enhance the tolerance to heat stress in tomato. The RNA interference (RNAi) plants were more wilted than the wild type with heat treatment. Under heat stress, SlSIZ1 could decrease the accumulation of reactive oxygen species (ROS) and induce some genes of HSF and HSP transcription. Furthermore, overexpression of SlSIZ1 could increase the level of Hsp70 under high temperature. Yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays showed that SlSIZ1 could interact with SlHsfA1 to mediate the SUMOylation of SlHsfA1 and consequently enhance thermotolerance of tomato. In conclusion, overexpression of SlSIZ1 enhanced heat tolerance by regulating the activities of HsfA1 and increasing the content Hsp70.