【摘要】 【目的】拟轮枝镰孢(Fusarium verticillioides)是一种危害严重的植物病原真菌，极大降低了粮食产量，还产生2B类致癌物伏马毒素，威胁人畜健康。探究RNA甲基化修饰与伏马毒素之间的联系，解析RNA修饰甲基化转移酶在伏马毒素合成中的作用。【方法】利用HPLC检测了不同地区的拟轮枝镰孢菌株的伏马毒素合成能力，采用QuEChERS前处理结合超高效液相色谱-串联质谱(UPLC-MS/MS)技术建立了m~6A、m~lA、m~5C、Gm、m~7G和Um等6种RNA甲基化修饰检测方法，继而对不同产毒菌株的RNA甲基化修饰进行了检测，并采用生物信息学和RT-qPCR方法确定了与伏马毒素合成相关的RNA甲基化修饰基因。【结果】不同地区的拟轮枝镰孢菌株伏马毒素合成能力具有显著差异，成功建立了RNA甲基化修饰检测方法，并确定m~lA和m~5C RNA甲基化修饰与伏马毒素合成负相关，RT-qPCR发现Fvalyref基因负调控伏马毒素生物合成。【结论】RNA m~5C甲基化修饰与伏马毒素生物合成呈负相关且其Reader基因Fvalyref负调控伏马毒素生物合成。更多还原

【Abstract】 【Objective】Fusarium verticillioides is a serious plant pathogen fungus,which greatly reduces grain yield.It also produces class 2B carcinogen fumonisin,which threatens human and animal health.To explore the relationship between RNA methylation modification and fumonisin,the role of RNA methylated transferase in fumonisin synthesis was analyzed.【Method】 HPLC was applied to determine the fumonisin synthesis ability of F.verticillium strains from different regions.Six kinds of RNA methylation modification detection methods,including m~6A,m~lA,m~5C,Gm,m~7G,and Um,were established by QuEChERS pretreatment combined with ultra-high performance liquid chromatograph-tandem mass spectrometry(UPLC-MS/MS).Then the RNA methylation modifications of different toxin-producing fungi were detected,and the RNA methylation modification genes related to fumonisin synthesis were identified by bioinformatics and RT-qPCR.【Result】It was found that there were significant differences in fumonisin synthesis ability of F.verticillium strains in different regions,the detection method for RNA methylation modification was successfully constructed,the methylation of RNA m~lA and m~5C was negatively correlated with fumonisin synthesis,and RT-qPCR showed that Fvalyref gene negatively regulated fumonisin biosynthesis.【Conclusion】The methylation modification of RNA m~5C and its Reader gene Fvalyref negatively regulate fumonisin biosynthesis.更多还原