喷雾诱导沉默CYP51和Sdh基因对禾谷丝核菌生长和致病性的影响  

Spray-induced gene silencing of CYP51 and Sdh genes effected the growth and pathogenicity of Rhizoctonia cerealis

作　　者：成永杰 张浩天 孙海燕[2] 曹淑琳 马东方[1] 陈怀谷[2] 李伟[2] CHENG Yongjie;ZHANG Haotian;SUN Haiyan;CAO Shulin;MA Dongfang;CHEN Huaigu;LI Wei(College of Agriculture,Yangtze University,Jingzhou 434025,China;Institute of Plant Protection,Jiangsu Academy of Agricultural Sciences,Nanjing 210014,China)

机构地区：[1]长江大学农学院,荆州434025 [2]江苏省农业科学院植物保护研究所,南京210014

出　　处：《植物病理学报》2023年第1期84-96,共13页Acta Phytopathologica Sinica

基　　金：江苏省重点研发计划项目(BE2019377);国家自然科学基金项目(32072375);国家现代农业产业技术体系(CARS-03)。

摘　　要：由于缺少稳定的遗传转化体系,丝核菌致病相关基因的功能研究受到严重限制。本研究以防治丝核菌病害的常用杀菌剂靶标基因CYP51和Sdh为目标基因,体外合成禾谷丝核菌CYP51的3个同源基因以及Sdh的B、C、D亚基基因的dsRNA,采用喷雾诱导基因沉默(spray-induced gene silencing, SIGS)方法研究这6个基因产生的dsRNA对病菌生长和致病性的影响。结果表明,外源施加靶标基因dsRNA对禾谷丝核菌的生长和致病性均有抑制作用。在被侵染的大麦叶片上,外源dsRNA能够显著抑制病菌中对应基因的表达。100 ng·μL-1浓度的RcCYP51-3-dsRNA在大麦叶片上抑菌效果较为显著,且可以同时抑制3个RcCYP51同源基因的表达;RcSdhD-dsRNA同样可以抑制RcSdh三个亚基的基因表达,在50 ng·μL-1浓度时抑菌效果最好。本研究探索了一种抑制禾谷丝核菌基因表达的方法,为使用SIGS方法研究丝核菌基因功能打下基础。Due to the lack of stable genetic transformation system, researches on the function of pathogenicity-related genes in Rhizoctonia fungi have been severely restricted. CYP51 and Sdh are the target genes of the commonly used fungicides to prevent and treat Rhizoctonia diseases. Wheat sharp eyespot caused by R. cerealis is a stem-base wheat disease that is widely distributed in China. In this study, the dsRNA of three homologous genes of RcCYP51 and B, C, D subunit genes of RcSdh were synthesized in vitro. The influence of these six target genes to growth and pathogenicity of R. cerealis was studied based on the spray-induced gene silencing(SIGS) method. The results indicated that the growth and pathogenicity of R. cerealis were suppressed by the dsRNA of all target genes. On the infected barley leaves, exogenous dsRNA can significantly inhibit the expression of the corresponding gene. Among them, the RcCYP51-3-dsRNA at the concentration of 100 ng·μL-1had a significant inhibitory effect on barley leaves, and can simultaneously inhibit the expression of the three homologous genes of RcCYP51. The RcSdhD-dsRNA can also inhibit the gene expression of the three subunits of RcSdh, and the optimum concentration was 50 ng·μL-1. This study explored a method to inhibit the gene expression in R. cerealis, and provided a useful exploration for using the SIGS method to study the gene function of Rhizoctonia fungi.

关 键 词：小麦纹枯病 基因沉默 甾醇14α-去甲基化酶 琥珀酸脱氢酶 

分 类 号：S432.1[农业科学—植物病理学]