非核糖体多肽合成酶(NRPS)是真菌毒素合成的关键酶，但NRPS在炭疽菌致病过程中的作用机制尚未见报道。申请人经研究发现了一株蟋蟀草炭疽菌菌株，该菌株对寄主牛筋草具有生防除草活性，其除草机理与生防菌产生毒素有关。为探究NRPS参与蟋蟀草炭疽菌致病过程中的作用机制，本项目拟通过生物信息学的方法分析蟋蟀草炭疽菌的NRPS基因家族，并获得基因全长序列。采用高通量基因敲除筛选致病缺陷突变体，结合功能互补、定位和超表达技术鉴定NRPS基因的功能；通过突变体致病性和生物学性状的测定，明确NRPS基因与病菌致病力间的关系及对病菌生物学特性的影响；通过QRT-PCR方法进一步分析NRPS基因的表达调控关系；最后通过HPLC分离次级代谢产物并进行生测。预期研究结果将对揭示蟋蟀草炭疽菌致病机制具有重要的意义，为后期生物源除草剂的研制提供新的线索和理论依据。

The non-ribosomal peptide synthase (NRPS) genes are the most important secondary metabolite gene clusters in fungi. But how NRPS genes get involved in pathogenicity of Colletotrichum species is still unknown. In our previous work, a biological control strain Colletotrichum eleusines of Eleusine indica was found, and the mechanism of biocontrol strategy may due to the phytotoxin. In order to elucidate the function of NRPS gene family in C. eleusines， bioinformatics will be conducted to analysis the NRPS gene family. The pathogenicity and toxin production will be defined further by the high-throughput gene knockout strategy. And also, the function mechanisms of NRPS genes in C. eleusines pathogenicity will be further characterized by the gene complementation, over expression and protein location assay. QRT-PCR is performed to monitor the expression patterns of NRPS genes. The high performance liquid chromatography will be used to isolate the toxin. These results will contribute to the findings in elucidating C. eleusines pathogenesis and also provide the new theoretical basis for the exploration of natural products for new bioherbicide.