P450s和UGTs协同介导棉蚜对新烟碱类杀虫剂抗性的分子机制研究

棉蚜是一种危害严重的农业害虫，棉蚜的防治主要依赖于杀虫剂的应用，目前已对多种杀虫剂产生了高水平抗性。高水平的抗性往往是多个机制导致的，揭示不同抗性机制如何协同作用来实现种群抗药性进化具有重要的意义。申请人前期研究发现P450s和UGTs在抗药性害虫中表达水平均显著升高是一种普遍现象，但是P450s和UGTs在杀虫剂的代谢过程中是否协同发挥作用尚不明确。本项目在鉴定棉蚜候选P450s和UGTs对新烟碱类杀虫剂的代谢能力基础上，拟运用Bac-Bac杆状病毒表达、质谱分析、免疫沉淀等多种方法，在体外表达系统和棉蚜体内解析P450s和UGTs两个代谢途径在杀虫剂代谢中的协同作用，从蛋白-蛋白互作角度揭示P450s和UGTs协同作用的分子机制。本项目的开展对揭示棉蚜多代谢抗性因子协同作用机制具有重要的意义，同时也为棉蚜的精准抗药性治理提供依据。

The cotton aphid, Aphis gossypii Glover, is a destructive global crop pest. Control of A. gossypii has relied on the application of chemical insecticides. Consequently, A. gossypii has developed resistance to numerous insecticides. The high level of insecticide resistance is always determined by multiple resistance mechanisms, it is still unknown the interactions between different mechanisms. In our previous study, we found that the co-overexpression of P450s and UGTs is very common in insecticide-resistant insects. In this project, we will address the general question of whether UGTs and CYPs interact with each other and cooperate to modulate the partner enzyme(s) during the detoxification. We will identify which of the co-overexpressed P450s and UGTs actually metabolize neonicotinoids and their dominant metabolites in A. gossypii. Furthermore, we will use Bac-Bac baculovirus expression, liquid chromatography-mass spectrometry (LC-MS/MS) and immunoblotting analysis to investigate their interactions between P450s and UGTs during neonicotinoids metabolism and reveal the mechanism of their interactions. These studies should be useful for understanding the mechanism of the interactions between multi-metabolic resistance and managing insecticide resistance to control the cotton aphids.