WRKY转录因子在一氧化氮诱导的采后番茄果实抗病信号转导中的作用机制

Disease is a main restrict factor for postharvest fruit storage and transportation, research on disease-resistant mechanism and developing safe, effective, simple and economic disease-resistant methods are very important for theoretical and practical approach. Many studies have shown that nitric oxide (NO) as an important message molecule can induce disease resistant ability in postharvest tomato fruit, but it is not well known that how WRKY transcription factors participate in this process and their roles in signal transduction pathway. Based on our well research background on NO and disease resistant pathway, genomics and proteomics are applied to figure out key disease-related WRKY genes/proteins. Antisense LeNPR1 tomato (already got) and antisense LeWRKY tomato (will be developed in this project) are constructed to reveal the interaction mechanism in NO-induced disease resistance. In addition, spectroscopical method and some new biomolecular methods can help to reveal possible mechanism of NO- -WRKY signal transduction pathway.

采后病害是导致果蔬贮运损失的主要原因，探索果蔬采后诱导抗病机制，完善安全高效、简便经济的采后防病技术，具有重要理论和实践意义。一氧化氮（NO）作为重要的信号分子能够诱导采后番茄果实抗病能力提高，而WRKY转录因子参与调控植物抗性反应多个关键基因，与NO调控途径有交叉，但WRKY转录因子在NO诱导的抗病途径中的作用和地位尚不清楚。本项目拟在前期研究基础上，借助转录组学和蛋白组学手段筛选NO诱导番茄抗病过程中关键抗病WRKY基因或蛋白，进而构建关键抗病WRKY基因的转基因材料，结合已获得的转反义LeNPR1番茄材料，利用原核表达、荧光探针标记、活性电泳、分子模建等技术，结合光谱学方法，研究WRKY转录因子参与调控NO诱导的番茄抗病过程中关键基因的转录与表达，逐步揭示WRKY转录因子参与NO调控的采后果实抗病诱导的可能机制，为进一步完善番茄抗病信号网络、构建果蔬采后病害绿色防控技术体系奠定基础。

NO 诱导采后果蔬抗病与WRKY和NPR1 密切相关，解析其中的“NO-WRKY-MAPK”网络关系，对探索绿色、广谱、高效、简便的果蔬采后病害防控策略具有重要意义。本项目利用申请人已获得的基因编辑SlNPR1基因敲除番茄材料、SlWRKY46、SlWRKY80过表达番茄材料，结合SNP处理以及接菌处理，以及生物信息学、RNA-Seq、实时定量PCR、CRISPR/Cas9基因编辑、亚细胞定位、扫描电镜、透射电镜、生理生化等相关学科成熟新技术，层次分明的确定了NO 调控番茄果实抗病中WRKY46、SlWRKY80、NPR1 的作用及互相调控模式，揭示了WRKY46、80和NPR1在NO诱导抗病性中的重要性，以及WRKY46、WRKY80和NPR1之间的相互作用，为实现果蔬采后病害的绿色简易防控奠定理论基础。