基于黄瓜CsCER1和CsWAX2基因的烷类蜡质合成调控机制研究

Most land plants have a wax layer which covers their aerial parts to protect them from biotic and abiotic stresses. Alkanes as the main components of cucumber waxes, the mechanism of its biosynthesis and regulation are still unclear. We have previously demonstrated that CsCER1 and CsWAX2 genes are critical in cucumber alkanes biosynthesis. In this project, we will identify the function of CsCER1 and CsWAX2 genes by crispr gene editing technology. Then we will screen the possible regulator upstream to CsCER1 and CsWAX2 gene by using yeast one-hybrid Screening, moreover, to verify the function of this gene through the cucumber genetic transformation experiment. Finally, we will identify novel interacting proteins in CsCER1 and CsWAX2 protein complex by yeast two-hybrid. Our overarching goal is to understand the molecular mechanisms of CsCER1 and CsWAX2 regulation in cucumber alkanes biosynthesis to provide insights and theoretical basis in improving cucumber stress resistance.

蜡质作为黄瓜表皮的覆盖层，在保护植物抵御非生物及生物胁迫方面起到了重要作用。烷类作为蜡质的主要成分，有关其在黄瓜中的合成机制及调控途径尚不明确。申请人前期研究表明，CsCER1和CsWAX2在黄瓜烷类蜡质合成中起着关键作用，本项目拟在前期工作基础上，通过CRISPR基因编辑技术将CsCER1和CsWAX2基因分别敲除，进一步验证CsCER1和CsWAX2基因在烷类合成中的作用；其次通过构建酵母单杂文库筛选调控CsCER1和CsWAX2基因表达的上游转录因子，并利用黄瓜遗传转化验证该转录因子在调控烷类蜡质合成中的功能；再者，通过构建酵母双杂文库，筛选CsCER1和CsWAX2的互作蛋白。本研究旨在解析CsCER1和CsWAX2在黄瓜烷类蜡质中的作用机制和调控途径，为黄瓜抗逆品种改良提供理论依据。

蜡质作为黄瓜表皮的覆盖层，在保护植物抵御非生物及生物胁迫方面起到了重要作用。申请人前期研究表明，CsCER1在黄瓜烷类蜡质合成中起着关键作用，但其调控机理尚不清楚。本研究通过酵母单杂文库筛选探究了黄瓜CsCER1 基因上游的转录因子，共找到34个阳性克隆，后经基因亚细胞定位预测、转录组分析、基因组序列比对筛选出7个候选转录因子。我们通过生信分析从7个候选转录因子中选择CsCOL5作为目标基因，并对其进行黄瓜转基因功能验证，目前已得到阳性植株，其表皮蜡质成分需要进一步测定，该研究为解析黄瓜蜡质形成的分子调控网络奠定了基础。