氟啶胺是一种广谱高效的二硝基苯胺类杀菌剂，应用广泛，前人研究表明该药剂作用于线粒体氧化磷酸化过程，但其分子靶标未知。氟啶胺现已登记防治水稻恶苗病，近期我们以水稻恶苗病菌为研究对象，通过氟啶胺处理菌体的转录组分析和定量PCR验证，筛选到3个潜在靶蛋白（ATP合成酶的三个亚基：ATP5、ATPb和ATPh），为了揭示氟啶胺分子靶标及其与靶蛋白互作的关键氨基酸位点，本项目拟构建以上潜在靶基因表达载体，原核表达和纯化，体外研究其与氟啶胺的亲和性，确定靶蛋白；利用同源模建和分子对接方法预测氟啶胺与靶蛋白互作的氨基酸位点；构建靶基因的氨基酸点突变、敲除（或沉默）、回复和过表达等突变体，测定突变体对氟啶胺的敏感性和突变体中ATP合成酶的活性；构建靶基因不同氨基酸点突变表达载体，原核表达和纯化，体外验证其与氟啶胺的亲和性。研究成果将为科学用药提供指导，为基于作用靶标的新农药分子合理设计提供理论依据。

Fluazinam is a broad-spectrum and highly effective dinitroaniline fungicide. It has been widely used. Previous studies have shown that this fungicide acts on mitochondrial oxidative phosphorylation, but its molecular target is unknown. Fluazinam has been registered to control rice bakanae disease. Recently, we have used Fusarium fujikuroi as the research object. Three potential target proteins (three subunits: ATP5, ATPb and ATPh of ATP synthase) were screened by transcriptome analysis and quantitative PCR verification of fluazinam-treated cells. In order to reveal the target of the fluazinam molecule and its key amino acid interaction with the target protein, this project intends to construct the expression, vector prokaryotic expression and purification of above potential target genes, study its nucleophilicity with fluazinam in vitro to determine the target protein; use homologous modeling and molecular docking methods to predict the amino acid position of the interaction between fluazinam and the target protein; construct the target gene mutants by using amino acid site-directed, knockout or silence, reply and overexpress methods, determining the sensitivity of the mutant to fluazinam and the activity of ATP synthase in the mutant; constructing a target gene with different amino acid point mutation expression vectors, prokaryotic expression and purification, and verifying the nucleophilicity of vectors with fluazinam in vitro. This study will provided scientific guidance for using pesticide safely and reasonably and theoretical basis for rational design of new pesticide based on target.