性信息素结合蛋白在识别TypeII型茶尺蠖性信息素中的功能研究

性信息素识别是蛾类昆虫求偶定位、交配繁衍过程中的重要环节, 其分子机制一直是昆虫化学生态学领域的前沿热点。性信息素结合蛋白PBPs是昆虫嗅觉感器中第一个与性信息素分子相互作用的靶标蛋白，对性信息素分子的识别至关重要。目前，感知TypeI型性信息素的PBPs研究较为系统，但感知TypeII型性信息素的PBPs研究相对匮乏。本申请拟以重要害虫茶尺蠖为对象，基于已成功鉴定PBPs基因的研究基础，拟先分析PBPs的表达谱；进一步应用原位杂交、免疫组化和单感器记录技术研究PBPs在雄蛾性信息素敏感感器中的表达分布。通过体外配体结合、定点突变和活体水平RNAi技术系统的阐明PBPs在识别茶尺蠖TypeII型性信息素中的生理功能。研究结果有助于揭示蛾类昆虫感受TypeII型性信息素的分子机制，也可为利用PBPs靶向设计合成高效的嗅觉干扰剂调控茶尺蠖求偶行为，进而发展新型害虫防控技术提供科学依据。

Species-specific sex pheromone recognition is crucial to mate finding and a successful reproduce in moth species, and its molecular mechanism is always the research hot topic in chemical ecology, in which pheromone binding proteins (PBPs) as the first target protein to interact with sex pheromones in the olfactory sensillum, which play key roles in binding and discriminating different pheromone components. To date, the physiological functions of PBPs related to Type II sex pheromones remain unclear in comparison to various information from PBPs in the recognition of Type I pheromones. In the current project proposal, we mainly focus on the physiological functions of PBPs from Ectropis obliqua Prout, one of the most serious tea pests secreting Type II sex pheromones. First, based on the previously successful identification of EoblPBPs gene sequences, the expression profiles among different tissues in male and female moths and during different developmental stages will be analyzed. Secondly, the chemosensillum localization of PBPs in pheromone-sensitive sensilla will be investigated applied to use the technical of single sensillum recording, in situ hybridizations and immunocytochemistry. Finally, using ligand-binding assay, site-directed mutagenesis in vitro in combination with RNA interference in vivo to evaluate the physiological functions of PBPs in perception of sex pheromones in E.obliqua. Our study answers the call by addressing further research on olfaction molecular mechanism in the moth species secreting Type II sex pheromones, which should be lay the foundation to explore RNAi with PBPs as target proteins for mating behavior regulation, eventually, contributes towards development of new technical solutions in pest management.