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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。
In order to provide candidate genes for disease resistance in late blight of tomato, CLN2037E inbred lines and susceptible inbred lines of 5 # lines were used as experimental materials in this study. The molecular marker was used to locate the late blight resistance genes in CLN2037E On the basis of chromosome 9, the cDNA library of CLN2037E was induced by Phytophthora infestans, the CDS database of tomato genome, the local Blast program and real-time fluorescence quantitative PCR technique were used to explore the CDS which best matches the EST on chromosome 9, Phytophthora infestans response patterns. Results Six candidate genes for resistance to late blight were identified. Among them, Solyc09g097960, Solyc09g082810 and Solyc09g065760 all responded to P. infestans in 5 # and CLN2037E inbred lines. Solyc09g092030 and Solyc09g090430 were respectively inhibited in two inbred lines Not responsive to Phytophthora infestans, the expression of Solyc09g008670STBZ was inhibited in 5 # inbred lines and was induced in CLN2037E. To verify the function of this gene, Solyc09g008670 gene overexpression and VIGS (Silencing Gene Silencing) vector were successfully constructed.