SlWRKY33参与外源Spd调控番茄抗盐碱性的功能解析

土壤盐碱化是番茄高产优产的主要限制因素，外源Spd可有效提高番茄抗盐碱性，WRKY转录因子与植物胁迫应答密切相关。项目组前期研究发现SlWRKY33受盐碱胁迫强烈诱导，外源Spd可降低此趋势，但SlWRKY33在Spd增强番茄盐碱抗性中的功能及作用机理尚不清楚，因此，本项目以“金鹏朝冠”番茄为试材，基于转基因和Real-time PCR技术分析SlWRKY33序列特性，通过探究SlWRKY33基因沉默和超表达番茄植株对盐碱胁迫及Spd的响应，明确Spd诱导番茄抗盐碱性中SlWRKY33的功能；拟结合ChIP-Seq、RNA-Seq技术确定SlWRKY33的靶基因；通过亲和纯化-质谱、双分子荧光互补技术确定SlWRKY33的互作蛋白，并研究其互作机制。本研究有望阐明SlWRKY33参与外源Spd调控番茄盐碱胁迫抗性的功能及作用机制，为番茄耐盐碱育种提供新的基因来源和抗逆栽培提供途径。

Soil salinization is one of the main factors limiting the stable and high tomato production in northwest district. Application of exogenous spermidine (Spd) can effectively improve the saline-alkali stress resistance of tomato, and WRKY transcription factors are closely related to the stress responses in plants. In the preliminary study, SlWRKY33 was strongly induced by saline-alkali stress, while exogenous Spd could decrease this trend. However the function and mechanism of SlWRKY33 in enhancing tomato salt-alkaline tolerance by Spd has not been fully elucidated. In this study, tomato cultivar ‘Jinpengchaoguan’ will be used as experimental material in this project. By means of transgenic and Real-tmie RCR technolgy, SlWRKY33 genes will be performed the sequence characteristics. We will characterize the molecular mechanisms of SlWRKY33 by loss-of-function and gain-of-function analysis, identifying its down-stream targets by ChIP-Seq and interacting partners by IP-MASS, BiFC, and Co-IP analysis, and examining the biological functions of its targets and interaction partners under saline-alkali stress plus Spd condition. The present study will clarify the response mechanisms of SlWRKY33 transcription factors in the Spd-mediated resistance of tomato to saline-alkali stress. All those will help to provide new gene for the breeding of saline-alkali stress resistant tomato cultivars, and explorea new way based on Spd in the anti-salinity-alkalinity and high-efficiency tomato cultivation.