昆虫的烟碱型乙酰胆碱受体（nAChRs）极其多样丰富，在神经系统中发挥着极其重要的作用。多杀菌素的主要作用靶标是nAChRs，但具体的作用位点目前尚不明确，西花蓟马对多杀菌素的抗性具有种群差异性，其抗性机理有待探明。本项目利用高通量RNA测序即RNA-Seq技术，对多杀菌素敏感和抗性西花蓟马品系（抗性倍数>10,000倍）进行差异表达分析，鉴定、筛选与抗性有关的差异表达基因；主要应用RT-PCR及RACE技术克隆所有nAChRs亚基基因全长；采用定向突变、爪蟾卵母细胞体外表达和双电极电压钳技术验证敏感和抗性受体亚基的功能差异，找出与抗性相关的突变位点，研发抗性早期分子检测技术；对于表达差异显著的胆碱能系统相关基因进行荧光定量分析，采用RNAi技术研究差异基因在抗性中作用。研究结果将明确多杀菌素类药剂在nAChRs上的功能位点，阐明西花蓟马对多杀菌素的抗性机理，为西花蓟马的抗性治理提供依据。

Nicotinic acetylcholine receptors (nAChRs) in insects are diversified and play a number of essential physiological roles in the central nervous system. The primary target of spinosad appears to be nAChRs but the target site is unclear. The resistance of Frankliniella occidentalis to spinosad differs in different populations and the resistance mechanisms need to be proved. A next-generation RNA sequencing (RNA-Seq) will be performed in the present study to find and identify differentially expressed genes between spinosad-resistant (>10,000-fold) and susceptible strains of F. occidentalis. The full-length of all the subunits genes of nAChRs will be cloned by using RT-PCR and RACE and the allele sequence will be compared. The functional difference in resistant and susceptible strains is to verify by using site mutation, in vitro expression of subunits including stable mutations in Xenopus oocytes, and two electrode voltage clamp techniques to find the resistance related mutations and then to develop a molecular method for early resistance. As for the cholinergic genes with significant difference in expression quantity will be tested by real-time fluorescent quantitative PCR and their roles in resistance is studied by RNA interference technique. The results will make clear the functional site of spinosad in nAChRs, clarify the resistance mechanism of spinosad in F. occidentalis, and then provide evidence for resistance management. In addition, the results will be important for prolonging and protecting the life of spinosad in field application.