用基因表达数量性状基因座定位(eQTL)研究蚊杀虫剂抗性的调控机制

Mosquito-borne diseases cause serious mortality and morbidity in humans, and pose significant threats to public health. Vector control is a major method for the control and prevention of mosquito-borne diseases. The wide use of pyrethroid has resulted in the emergence and spread of resistance in mosquito populations. Our previous researches pointed out that multiple genes are involved in insecticide resistance, including the resistance-upregulated genes and susceptible-upregulated genes. The long-term goal of this research is to elucidate the inter-relationship of resistance-related genes in Culex pipiens pallens mosquitoes, and to identify molecules with critical function to resistance. The expression quantitative trait loci (eQTL) mapping using messenger RNA transcript abundance as quantitative traits to mapping gene expression QTL. It has been used in a variety of species to help elucidate the molecular basis of complex disease. This research will use eQTL mapping approach to identify the critical genes involved in mosquito resistance, to clarify the regulatory function of these genes in insecticide-resistance network and to determine the regulatory network. The specific aims are generate independent segregating populations derived from crosses between deltamethrin-sensitive/resistance Culex pipiens pallens and combining the genome-wide expression profiling from RNA-seq experiments with genetic linkage analysis to identify critical genes and to determine the regulatory network involved in insecticide-resistance. This research will provide critical information on monitoring and management of insecticide resistance in field mosquitoes.

蚊媒病（如：疟疾、登革热等）一直是危害人类健康的重要公共卫生问题。化学防制一直是虫媒病防制的主要方法，然而连续大量杀虫剂的使用导致抗性的发生和发展。媒介抗性是多分子的复杂生物现象，存在多基因的协同作用。本课题组前期鉴定了多个抗药性相关基因，功能分别涉及细胞代谢，信号转导，分子转运，蛋白合成与泛素化降解等通路。当前亟待解决的问题是，探索众多抗性基因间相互作用，为抗性检测和防治提供新靶点。新问世的基因表达的数量性状定位（eQTL mapping）法可用于分析基因之间的调控关系，寻找控制基因表达的上游调控位点，发掘协同作用基因，进而建立基因调控网络，以阐明基因调控的机制。本研究拟首次应用eQTL方法对蚊拟除虫菊酯抗性基因的调控网络进行研究，以期阐明抗药性主效基因及其调控作用，探索抗药性检测的新靶点，为抗药性防治提供新思路。

蚊媒病一直是危害人类健康的重要公共卫生问题。化学防制一直是媒介防制规划中的主要方法，然而连续大量杀虫剂的使用导致媒介抗药性的发生和发展。蚊媒抗药性以及随之产生的环境改变加剧了蚊媒病传播的恶化趋势。蚊媒抗药性机制研究对于蚊媒治理至关重要。.本研究将敏感雌蚊与抗性雄蚊杂交子代自交，构建F2分离种群，随后应用WHO敏感性检测方法并将其抗性表型分为高度敏感组，中度敏感组，低度敏感组和抗性组。测定各组蚊基因表达谱及位点多态性，利用eQTL分析构建蚊媒代谢抗性基因调控网络。研究发现差异表达基因多集中于代谢通路，因此后续主要集中研究杀虫剂代谢相关的3类代谢酶，即细胞色素P450（CYPs）、羧酸酯酶(CCEs)和谷胱甘肽-S-转移酶（GSTs）之间的调控关系，构建了代谢抗性调控网络，筛选获得主要调控基因CYP9AL1，并初步鉴定其在抗性中的作用。本研究结果深化了对媒介抗药性机理的认识，为现场抗药性监测和治理提供了新的思路。本研究结果具有重要的科学意义和潜在的应用价值。