白背飞虱气味结合蛋白与感染南方水稻黑条矮缩病的水稻互作及选择机理研究

Herbivorous insects use high specificity and sensitivity olfactory system to locate a suitable host via perceiving host volatiles. In recent years,it is the research hotspot that the biochemical and molecular mechanisms of animal olfactory communication, which possesses important scientific significance and application value. Odorant binding protein (OBP) play a crucial role on plant volatiles perception in herbivorous insect. OBP genes have been found in a lot of herbivorous insects in recent years. What the distinct OBPs tissue expression characteristics, physiological function and differentiation? The project intends to take Sogatella furcifera as the object of this research , which is one of important rice pests and vector of southern rice black-streaked dwarf virus.We first adopt bioinformatics and molecular cloning of S. furcifera OBP cDNA full-length, and then to investigate the OBP tissue expression pattern by quantitative real-time PCR technology and in situ hybridization, and use in vitro expression,binding experiments, RNAi technology combined behavioural bioassay and protein three-dimensional structure analysis to study the OBP physiology function and differentiation. The findings may further clarify the mechanism of interaction between OBP and normal rice plant or southern rice black-streaked dwarf virus-infected rice plant volatiles, as well as for provide a scientific basis of the development of a new type of pest behavior blockers or interference agent.

植食性昆虫通过具有极高特异性和灵敏性的嗅觉系统感知寄主的气味挥发物从而定位合适的寄主，其生化和分子机制是近年动物嗅觉通讯研究的前沿热点，具有重要的科学意义和应用价值。气味结合蛋白（OBP）在昆虫气味挥发物感受中起重要作用，近年已在多种植食性昆虫的触角中发现OBP基因。不同 OBP 的组织表达特征如何，它们在功能上又有什么样的分化？本项目拟以重要水稻及南方水稻黑条矮缩病媒介害虫白背飞虱为对象，利用生物信息学和分子克隆的方法获得白背飞虱OBP基因全序列，进而通过实时定量PCR技术和原位杂交探索OBP的组织定位，并通过 OBP的体外表达、结合实验和RNAi结合行为生测、蛋白三维结构解析研究 OBP的生理功能及分化。研究结果可为进一步明确 OBP与正常水稻植株和病毒感染水稻挥发物组分间的互作机制，以及为开发基于嗅觉感受的新型害虫行为阻断剂或干扰剂，提供科学依据。

植食性昆虫通过具有极高特异性和灵敏性的嗅觉系统感知寄主的气味挥发物从而定位合适的寄主，其生化和分子机制是近年动物嗅觉通讯研究的前沿热点，具有重要的科学意义和应用价值。气味结合蛋白（OBP）在昆虫气味挥发物感受中起重要作用，近年已在多种植食性昆虫的触角中发现OBP基因。不同 OBP 的组织表达特征如何，它们在功能上又有什么样的分化？本项目以重要水稻及南方水稻黑条矮缩病媒介害虫白背飞虱为对象，首先建立固相微萃取平台，结合气-质谱联用技术搜集并鉴定了感染SRBSDV水稻植株和正常植株的挥发物共84种。其中有28种有显著差异。 随后，利用转录组测序、生物信息学和分子克隆的方法获得白背飞虱12个OBP基因全序列，进而通过实时定量PCR技术检测出一个成虫触角高表达的OBP基因SfurOBP11。蛋白免疫荧光定位表明SfurOBP11在嗅觉感器淋巴液中高表达；随后通过 OBP的体外表达、结合实验探明水稻中的酮类和萜烯类有可能是白背飞虱用于搜索并定位水稻植株的主要挥发物。最后RNA干扰结合行为生测表明SfurOBP11在白背飞虱选择定位水稻植株行为中起到关键作用。研究结果可为进一步明确 OBP与正常水稻植株和病毒感染水稻挥发物组分间的互作机制，以及为开发基于嗅觉感受的新型害虫行为阻断剂或干扰剂，提供科学依据。