番茄MADS-box类转录因子新基因ShMADSTF的抗病分子机制

Transcription factors can regulate many genes in plant growth and development and adverse stresses. In previous study, a MADS-box transcription factor named as ShMADSTF was cloned from tomato, which was participated in tomato resistance response to biotrophic fungi powdery mildew and nectrophic fungi Verticillium wilt. But there is few report that MADS-box transcription factor participated in plant disease resistance at present. Based on the hypothesis of MADS-box transcription factor involving in new important biological function, further function analysis on ShGSTU1 will be conducted including: the over-expression ShMADSTF tomato is gained by transgenic method, and diease-resistance function of which are identified; plant hormone signaling pathways of whihch were studied by VIGS; the tomato powdery mildew effectors and endogenous signal molecules interaction with ShMADSTF are identified by applying yeast two-hybrid method to analyze the PTI (PAMP triggered innate immunity) and ETI (effector triggered immunity) signaling pathways which involved in plant immune response; the whole genomic combining spectrum of ShMADSTF was gained by ChIP-Seq method, the molecular regulation mechanism of which was analyzed by bioinformatics. All the results will well elucidate the function and resistance mechanism of tomato transcription factor ShMADSTF and provide theoretical basis for plant breeding and improvement of new resistance varieties.

转录因子在植物生长发育和胁迫应答反应中起着重要的调控作用。课题组研究发现从番茄中克隆的一个MADS-box转录因子ShMADSTF同时参与番茄对活体寄生真菌（白粉菌）和腐生真菌（黄萎菌）的抗性反应。但目前MADS-box转录因子参与植物抗病性报道较少。基于MADS-box转录因子新的重要生物学功能的推测，本项目拟进一步对番茄转录因子ShMADSTF进行研究：采用转基因的方法鉴定该转录因子在植物抗病反应中的功能；VIGS方法分析其参与激素信号途径；借助酵母双杂交系统鉴定与转录因子ShMADSTF直接互作的蛋白分子，分析其在植物抗性反应中参与的PTI和ETI信号通路；采用ChIP-Seq方法获得转录因子ShMADSTF在番茄全基因组的结合谱，生物信息学分析其调控分子机理。最终阐释番茄转录因子ShMADSTF的功能及抗病分子机制，为进一步选育和改良植物抗病新品种奠定理论基础。

转录因子在植物生长发育和胁迫应答反应中起着重要的调控作用。从番茄中克隆的一个MADS-box转录因子ShMADSTF同时参与番茄对活体寄生真菌（白粉菌）和腐生真菌（黄萎菌）的抗性反应。本项目对ShMADSTF基因编码蛋白进行了亚细胞定位，定位在细胞质内；转ShMADSTF基因番茄接种白粉菌后，抗性提高，出现快速HR反应，H2O2累积显著增加，并伴随非正常的吸器；SA激素信号途径参与了ShMADSTF介导的番茄对白粉菌的抗性反应；荧光定量分析表明病程相关基因SlPR1、SlPR2和SlPR5参与ShMADSTF抗性反应途径；构建了酵母双杂交文库，筛选到与ShMADSTF互作的5个蛋白及抗性相关基因RIN4；分离和鉴定了11种番茄及其他植物致病病原菌，包括重要病原菌变红镰刀菌、胶孢炭疽菌、疫霉菌、链格孢菌，黑胫病菌，斑污拟盘多毛孢菌等；对番茄全基因组MADS转录因子基因家族进行了生物信息学分析，得到135个MADS-box转录因子，分为I型(Mα,Mβ,Mγ) 和II型(MIKCC,MIKC*)；抗病番茄LA2172和感病番茄Monnymaker分别接种黄萎菌后，诱导了18个SlMADS基因参与番茄抗病及感病反应，克隆了13个番茄SlMADS候选抗病基因，正在构建超表达载体和转基因番茄植株；完成了番茄黄萎菌和番茄其他不同处理下亲和和非亲和互作的转录组测序，获得参与番茄抗病反应的重要候选基因19个，为番茄抗病育种提供了丰富的基因资源。本项目确定了转录因子ShMADSTF在番茄抗病作用中的功能，分析了转录因子ShMADSTF参与的信号转导途径，并初步阐释了ShMADSTF的抗病分子机制，为进一步选育和改良植物抗病新品种奠定理论基础。基于以上研究成果，本项目获批授权专利8项，软件著作权2项，发表SCI论文5篇，中文核心论文19篇。