淡色库蚊越冬蚊抗寒性及转录组和蛋白质组的研究

Culex pipiens pallens Coquillett is an important vector for disease transmission, overwinters as diapause female adult. Overwintering mosquitoes play a key role in the whole life history of mosquitoes and are important for the continuation of populations. Our group has mastered the overwintering sites and overwintering habits of Cx. pipiens pallens in Shandong province. However, the cold resistance and molecular regulation mechanism of Cx. pipiens pallens for wintering are still unclear. Therefore, the overwintering adults are chosen as the research object in this study. Using the principle of thermal resistance, the hardiness of them are analyzed by measuring the supercooling point and freezing point and combining with lethal time of 50% (LT50). Furthermore, PCR-RFLP technology and RNA-Seq high-throughput sequencing are used. Differences in proteomics analysis will be found between the non-overwintering and overwintering adults by iTRAQ technology and LC-MS/MS. According to the bioinformatics analysis method, the differentially expressed genes and proteins are selected, and the expression levels of them are verified and analyzed by PCR. We are going to explore cold resistance of overwintering adults and provide data support for the molecular regulation mechanism of Cx. pipiens pallen overwintering, with a view to providing new ideas for the control of vector mosquitoes and control of mosquito-borne diseases.

淡色库蚊是重要的疾病传播媒介，以雌成蚊滞育越冬，越冬蚊对种群的延续发展有重要意义，本课题组已掌握淡色库蚊在山东地区的越冬场所和越冬习性。目前对淡色库蚊越冬蚊的抗寒性和分子调控机制尚不清楚，因此本项目拟以淡色库蚊越冬蚊为研究对象，利用热电阻的原理，测定越冬蚊的过冷却点、结冰点和致死中时，阐释淡色库蚊越冬蚊的抗寒性；同时利用PCR-RFLP技术、RNA-Seq高通量测序、iTRAQ定量技术、生物信息学、PCR验证和质谱多反应监测技术等研究手段，对淡色库蚊越冬蚊的转录组和蛋白质组进行系统分析，探索与抗寒性相关的蛋白和基因。结果可为淡色库蚊越冬的分子调控机制提供数据支持，为淡色库蚊越冬蚊的预测预报和有效防治方法提供新的思路。

淡色库蚊是我国北方主要的家栖性蚊种，可传播班氏丝虫病及流行性乙型脑炎等蚊媒传染病，是重要的疾病传播媒介。淡色库蚊以雌成蚊越冬，越冬蚊在蚊虫的整个生活史中起关键作用，对种群的延续发展有重要意义。项目通过比较蛋白质组学、转录组学和16SrRNA测序等技术，获得淡色库蚊越冬蚊特异表达和差异表达的蛋白，揭示低温胁迫下淡色库蚊肠道菌群和基因表达水平的变化，筛选出受低温影响较大的10个基因和细菌属，简单阐述淡色库蚊对低温响应的分子调控策略，为研究淡色库蚊的越冬机制提供数据支撑。资助期内发表多篇相关论文。未来拟在前期研究基础上进一步验证特异基因和细菌属，并将研究结果应用到越冬蚊的防治中。