To preliminarily define the role of WRKY transcription factors in the incompatible interaction between Cf-12 tomato and Cladosporium fulvum,we analyzed differential expression of WRKY transcription factors in the transcriptome of 4 d and 8 d time points of incompatible interaction,and then conducted phylogenetic analyses and gene silencing by virus inoculation.The results showed that 22 WRKY genes were enriched in the process of incompatible interaction,and the expression of Solyc07g047960.2 at 8 days post inoculation increased by nearly 20 times.According to phylogenetic analysis and expression analysis of WRKY CDS,the expression of some WRKY genes was not significantly different at two time points,however,in Cluster Ⅲl,7 WRKY genes showed significant up-regulated expression.After identification of WRKY gene silenced tomato plants(inoculated by TRV-WRKY),the results of Cladosporium fulvum inoculation showed that the expression of WRKY gene was increased compared with that before inoculation,and the number of hypersensitive necrotic spots on leaf was obviously lower than the control(inoculated by TRV).The conclusion was that the WRKY (Solyc07g047960.2) gene showed a significantly upregulated expression during the incompatible interaction between Cf-12 tomato and Cladosporium fulvum,and was actively involved in the resistance response to the tomato leaf mold disease.%为了明确含Cf-12基因的番茄与叶霉菌非亲和互作过程中WRKY转录因子所扮演的角色,首先对非亲和互作4d和8d转录组中WRKY基因的表达量进行比较分析,然后对相关WRKY基因进行系统进化分析和病毒接种的基因沉默验证.结果表明,非亲和互作过程中共富集到22个WRKY基因,其中Solyc07g047960.2在接菌后8d时的表达量达到近20倍.依据WRKY基因CDS序列的系统进化树以及表达量分析结果,部分WRKY基因的表达量在两时间点差异不大,但ClusterⅢ中有7个WRKY基因呈现明显的上调表达趋势.对WRKY(Solyc07g047960.2)基因沉默(接种TRV-WRKY)后的番茄植株接种叶霉菌进行鉴定,结果表明,接菌后该基因的表达量相对于接菌前所有提升,且叶片上的过敏性坏死斑点数量明显低于对照组(接种TRV).以上结果表明,WRKY(Solyc07 g04 7960.2)基因在含Cf-12基因的番茄与叶霉菌的非亲和互作过程中呈现明显的上调表达趋势,积极地参与到对叶霉菌的抗性反应过程中.
展开▼
