嗅觉受体在苜蓿盲蝽趋花行为中的功能研究

项目名称： 嗅觉受体在苜蓿盲蝽趋花行为中的功能研究

项目编号： No.31471778

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 张永军

作者单位： 中国农业科学院植物保护研究所

项目金额： 85万元

中文摘要： 寄主植物花香气味物质在昆虫趋花行为中具有重要作用，昆虫嗅觉受体(ORs)在识别该类信息化合物中发挥重要功能。深入研究活性花香气味物质及ORs在识别该类物质中的功能，不仅有助于阐明植食性昆虫趋花行为的嗅觉机制，还可为通过调控害虫嗅觉识别进行害虫治理提供新策略。本申请以苜蓿盲蝽为研究对象，利用顶空法收集棉花花香气味物质，结合GC-EAD和昆虫嗅觉仪鉴定具有电生理及行为活性花香气味物质。在前期触角转录组测序的基础上，通过生物信息学和RACE技术鉴定OR基因，利用原位杂交技术和免疫组化方法研究OR基因的表达定位。通过爪蟾卵母细胞表达系统结合双电极电压钳研究OR蛋白和花香气味分子的对应识别关系。利用RNAi技术沉默OR基因表达，结合GC-EAD和昆虫嗅觉仪，检测缺失表达OR基因对苜蓿盲蝽识别花香气味及趋花行为的影响。阐明苜蓿盲蝽趋花行为的嗅觉机制，为昆虫嗅觉行为调控提供理论依据。

中文关键词： 苜蓿盲蝽；嗅觉受体；气味识别；趋花行为；化学通讯

英文摘要： Floral odors play critical roles in preference behavior of insects to flowering plants, and olfactory receptors (ORs) play important roles in chemoreception of these odor compounds. Exploration of the OR's function in chemical communication could help us clear insect olfactory identification mechanism, and provide a new strategy for insect pests management by regulating the behavior of odor perception. In the present work, the functional floral odors of cotton which were involved in preference behavior of alfalfa plant bug Adelphocoris lineolatus to flowering plants were identified from by using the GC-EAD and insect olfactometer. Based on the constructed antennal transcriptome of A. lineolatus early, the OR genes of alfalfa plant bug will be identified by bioinformatics methods and RACE technique. Then,the recombinant OR proteins will be expressed in Xenopus oocytes system, an immunocytochemical localization method will be carried out to locate the proteins position in antennal sensilla. By two-electrode voltage-clamp assays, the binding relationships between different recombinant ORs and selected floral odors will be investigated. In addition, a RNAi experiment will be conducted to silence the expression of key OR genes, and by qRT-PCR method to detect silence effects. Using GC-EAD and H tube trials detect the silence of candidate OR genes impacts on electrophysiological and behavioral responses of A. lineolatus to cotton floral odors. Thus, we could clear the functional differentiation of ORs in preference behavior of alfalfa plant bug to flowering plants. In generally, clarifying the function and molecular mechanism of ORs in A. lineolatus olfactory perception and exploring the RNAi regulation of insect chemical senses could provide a theoretical basis for design of insect olfactory behavior control agents by using ORs as a target.

英文关键词： Adelphocoris lineolatus;olfactory receptor;odor perception;preference behavior to flowering plants;chemical communication