基因组学法研究马来酰亚胺类化合物抗Sclerotinia sclerotiorum机制及作用靶标酶

菌核病是典型且严重的世界性真菌病害。目前常用的防治农药大多因使用年限长，已有较强的抗药性。项目合成制备马来酰亚胺类化合物，丰富本课题组已构建的马来酰亚胺类化合物库；研究各化合物对S. sclerotiorum和S. cerevisiae的抗菌活性；利用基因过量表达的S. cerevisiae菌株来考察加药与不加药的抗菌效果及基因表达量不同，并与数据库中常用抗菌剂基因表达量进行比对，初步分析这些化合物的抗菌机制；在此基础上，提取S.sclerotiorum相关酶，验证S.sclerotiorum抗菌机制的正确性，确定抗菌作用靶标酶；通过酶的分离纯化、单晶培养等，初步解析靶标酶及其与马来酰亚胺类化合物结合物的三维结构。项目的完成可为抗真菌农药的作用靶标酶的发现提供新方法和理论基础，同时为设计高活性和抗菌核病菌专一性强的化合物提供科学依据，为马来酰亚胺类抗真菌新型农药的创制提供科学理论基础。

Sclerotinia sclerotiorum is one of the typical fungal plant pathogens, also is a serious worldwide pathogen. Most of the present pesticides for protection and cure against it are old and easy to cause resistance. Some are toxic. The program plans to synthesize more maleimides and their derivatives to establish the chemical library of maleimide. With the compounds, the evaluation of antifungal activities against S. sclerotiorum and Saccharomyces cerevisiae. Using overexpression strains of S. cerevisiae, the antifungal mechnism will be investigated by the comparison the results of antifungal activities with the known compounds. Then the enzymes related with the mechnism in S. sclerotiorum will be separated, purificated and analyzed with the compounds to testify the antifungal mechanism of maleimides in S. sclerotiorum. Finally, the target enzyme will be purified, crystalized and its three-dimensional structure will be elucidated preliminary. The completion of the program will provide new method for the discovery of target enzyme for the antifungal agents and give new base and new idea in finding new pesticides against fungal pathogens.