番茄第9号染色体上抗晚疫病基因的发掘及表达模式

Research on the excavation and response patterns of genes resistant to late blight located in the No.9 chromosome of tomato

为提供番茄晚疫病抗病育种候选基因,本研究以高抗晚疫病的CLN2037E自交系和感病5#自交系为实验材料,在前期使用分子标记将CLN2037E中存在的抗晚疫病基因定位在第9号染色体的基础上,利用晚疫病病菌诱导CLN2037E的cDNA文库、番茄基因组CDS数据库、本地Blast程序及实时荧光定量PCR技术,发掘与第9号染色体上EST最佳匹配的CDS以及对晚疫病病菌的响应模式。结果发掘出6条抗晚疫病的候选基因,其中Solyc09g097960、Solyc09g082810和Solyc09g065760在5#自交系和CLN2037E自交系中均响应晚疫病病菌,Solyc09g092030和Solyc09g090430在2个自交系中分别被抑制和不响应晚疫病病菌,Solyc09g008670STBZ的表达在5#自交系中被抑制,而在CLN2037E中被诱导。为验证该基因的功能,成功构建Solyc09g008670基因的过表达和VIGS(Virus Induced Gene Silencing)沉默载体。

To provide candidate genes resistant to late blight fungus for disease resistance breeding,the high resistance inbred line CLN2037 Eand susceptible inbred line 5#of tomato were used as experimental materials.Based on the previous results that the late blight resistance genes were located in the No.9chromosome of CLN2037 Eby using molecular marker technique,we excavated the candidate genes optimally matching the EST in No.9chromosome and revealed their response patterns by employing the differential cDNA library,tomato genome CDS database,bioinformatics technology and real time fluorescence quantitative PCR.The results showed that：A total of six candidate genes were found resistant to late blight.Solyc09g097960,Solyc09g082810 and Solyc09g065760 were all induced in the 5#and CLN2037 Etomato inbred lines;Solyc09g092030 was inhibited in both lines.Solyc09g090430 was not response to late blight in these two inbred lines;The expression of Solyc09g008670 was suppressed in the 5#inbred line,but accelerated in CLN2037 E.Furthermore,in order to verify its function,overexpression and VIGS vectors of Solyc09g008670 were constructed.The results will provide the theory basis for understanding resistance mechanism and resistance breeding.