新杀菌化合物氮-（1-萘基）吩嗪-1-甲酰胺抑制水稻纹枯病菌的主靶标及分子机理研究

水稻纹枯病是水稻的重要病害，每年由其造成的产量损失可达45%。生产上常采用井冈霉素防治，但长期使用易导致药效下降，开发新型杀菌剂十分紧迫。本课题组以拮抗细菌SU8活性成分吩嗪-1-甲酰胺为母体，经结构改造获得抑制纹枯病菌活性更高的氮-（1-萘基）吩嗪-1-甲酰胺，其EC50 4.25μg/ml，远优于井冈霉素，但抑菌机理尚不明确，影响了开发和应用。针对这个问题，课题组深入研究发现其作用机理具有多靶标，究竟何种靶标主导抑菌？分子机理怎样？亟待解释，因此，拟从以下3个方面进行研究。（1）经超微结构观察、荧光标记以及免疫荧光等亚细胞定位，明确主导靶标；（2）利用iTraq技术解析病菌差异显著的蛋白质，明确作用靶蛋白，药与靶蛋白应激关系；（3）采用Autodock技术、分子动力学模拟和结合自由能计算NNPCN与靶标的结合机制。该研究可为纹枯病新型杀菌剂的创制提供理论依据。

Rice sheath blight is an important disease of rice and damages related to it result in 45% in the rice field each year. The occurrence of the disease was controlled by Jinggangmycin in production; nevertheless, control effect was decreased because of its long-term usage. Therefore, the creation of novel fungicide is extremely urgent. N-(naphthalen-1-yl)phenazine-1-carboxamide has better effect of inhibition (EC50 was 4.25 μg/ml) after structure modification of phenazine-1-carboxamide in previous researches, is better than Jinggangmycin, which showed that it has higher exploitation potential. However, the inhibition mechanism of N-(naphthalen-1-yl)phenazine-1-carboxamide is still not clear that affects the exploitation and application of new pesticide. In order to solve this problem, our group found that the mechanism of N-(naphthalen-1-yl)phenazine-1-carboxamide is multiple targets, Which target dominates antifungal? How to inhibit pathogen at the molecular level? It needs to be explained. Thus, the research mainly carries on the study from following three aspects. (1) The major targets are clearly define through the subcellular organelles experiments of ultrastructural observation, fluorescence labeling and immunofluorescence verification; (2) The influence of this compound on proteins with significant difference, finding target proteins, relation of compound and proteins, functional proteins are studied by iTraq technology; (3) The bond mechanism of N-(naphthalen-1-yl)phenazine-1-carboxamide and target were explained by autodock technology, molecular dynamics simulation and binding free energy. It would provide theoretical basis for the creation of a novel fungicide for sheath blight.