金龟子绿僵菌致病相关小RNA对寄主蝗虫靶基因调控的研究

项目名称： 金龟子绿僵菌致病相关小RNA对寄主蝗虫靶基因调控的研究

项目编号： No.31471821

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 黄勃

作者单位： 安徽农业大学

项目金额： 87万元

中文摘要： 绿僵菌是一类广泛应用于生物防治的昆虫病原真菌，而影响其防治效果不稳定的一个重要因素是寄主昆虫的免疫抵抗作用。小RNA在寄主与病原菌相互作用中发挥重要调控作用。特别是近期Science杂志报道植物病原真菌小RNA可调控植物的免疫相关靶基因以获得成功侵染。然而，迄今小RNA在昆虫病原真菌对寄主致病过程中的作用未见报道。前期工作中，我们确认金龟子绿僵菌及其寄主蝗虫均存小RNA和小RNA沉默机制。本项目在此基础上，围绕该菌致病相关的小RNA跨物种调控寄主蝗虫靶基因进行研究。采用高通量测序分析和筛选，获得绿僵菌致病相关的小RNA及表达特征；再通过生物信息学方法分析其寄主蝗虫的靶基因；并进一步结合定量PCR、免疫共沉淀、基因敲除与过表和致病力测定等技术手段深入分析绿僵菌小RNA对寄主蝗虫靶基因的调控作用。研究结果将有助于阐明小RNA在昆虫病原真菌致病过程中的作用机理，为菌株改良提供可靠的理论依据。

中文关键词： 金龟子绿僵菌；蝗虫；小RNA；靶基因调控

英文摘要： Metarhizium is an insect pathogenic fungi widely used as a biocontrol agent, the anti-immunity of host insects is one of an important factor for the instability of control efficacy.Small RNAs,as regulators,play an important role in host-microbial interaction. Especially，it have recently been reported that small RNAs of plant pathogenic fungi can regulate immunity-related target genes of plants for a successful infection.However, little is known about the roles of small RNAs in the regulation of pathogenesis from entomopathogenic fungi. In our preliminary studies, the existence of small RNAs and small RNAs machinery in Metarhizium anisopliae and its host locust was confirmed. Based on this work, in-depth study of the critical issue for the cross-kingdom regulation of host target genes by pathogenicity-related small RNAs in Metarhizium will be carried out. Firstly, pathogenicity-related small RNAs in Metarhizium and its expression profiles will be analyzed and identified by high-throughput sequencing approach.Secondly,host target genes of these Metarhizium small RNAs will be analyzed by bioinformatics analysis. Finally, experimental methods,such as Real-time PCR, immunoprecipitation,gene knock-out and small RNA overexpression,will be employed to confirm the regulation of host target genes by Metarhizium small RNAs.The findings should help to understand the regulatory mechanism of pathogenicity of small RNAs in entomopathogenic fungi,also provide a reliable basis for the genetic alteration of the production strain of Metarhizium.

英文关键词： Metarhizium anisopliae;locust;small RNA;regulationt of arget gene