绿盲蝽性信息素受体的鉴定和功能研究

The green mirid bug (GMB), Apolygus lucorum is an important agricultural pest in China. The mechanism of olfactory recognition in the GMB is a hot issue in neurology and chemical ecology, and the functions of pheromone receptors attract more attentions because of their special importance. Previous studies have demonstrated the presence of intraspecific sex pheromones in Mirada. And also the component and biological activity of the pheromones has also been clearly studied. However, few studies have been focused on the molecular and neural mechanisms of pheromone perceiving in the GMB so far. So we analyzed the odorant receptor genes of GMB in our previous study and first proved the GMB also have sensitive and specific pheromone receptors. Therefore, we speculate GMB encode its sex pheromone components through a series of specific receptors. The receptors determine the function of corresponding receptor nerves; achieve mutual recognition between males and females. In this project, we will perform a systematic study on the functions of sex pheromone receptors and their corresponding olfactory receptor neurons in the GMB. The candidate pheromone receptors will be predicted and identified by transcriptome sequencing and bioinformatics analysis. Functional study of the pheromone receptors will be carried out both in vitro and in vivo. After conducting single-sensillum recording, we will clarify the function of peripheral nerve participating in sex pheromone detection. Eventually we will clarify the molecular and neural mechanism by which the GMB can specifically recognize its mate. Moreover, this study will provide a theoretical basis on which we can screen new behavioral disturbance agents that are more effective than sex pheromones at molecular level.

绿盲蝽作为我国重要的农业害虫，其嗅觉识别的机制是神经学和化学生态学研究的热点，性信息素受体功能更是重要且前沿的科学问题。前期研究已证明盲蝽科昆虫存在种内异性联系的性信息素，其组分和生物活性也已经明确，但关于其性信息素感受的机制尤其是性信息素受体的研究还未见报道。申请人在前期研究中对绿盲蝽气味受体基因进行了分析；首次发现了绿盲蝽也具有灵敏特异的性信息受体。据此我们推测绿盲蝽也是通过特异的气味受体对性信息素成分进行编码，并决定对应受体神经的功能，从而实现雌雄个体间的相互识别。因此本研究将对绿盲蝽的性信息素受体展开系统研究：采用转录组测序和生物信息学分析鉴定候选的性信息素受体；通过体外表达和体内RNAi技术，确定绿盲蝽性信息素受体功能；通过单细胞记录，明确参与性信息素识别的外周神经功能。最终阐明绿盲蝽雌雄识别高度特异性的分子和神经机制，为下一步从分子水平筛选比性信息素更有效的行为干扰剂奠定基础。

绿盲蝽作为我国重要的农业害虫，其嗅觉识别的机制是神经学和化学生态学研究的热点，性信息素受体功能更是重要且前沿的科学问题。前期研究已证明盲蝽科昆虫存在种内异性联系的性信息素，其组分和生物活性也已经明确，但关于其性信息素感受的机制尤其是性信息素受体的研究还未见报道。本项目中我们对绿盲蝽的性信息素受体进行了鉴定和系统的功能研究。测定了绿盲蝽、苜蓿盲蝽和中黑盲蝽3种盲蝽的触角转录组，通过比较转录组学结合表达模式分析鉴定了4个候选的绿盲蝽性信息素受体基因；克隆了绿盲蝽4个候选性信息素受体基因的全长序列，在爪蟾卵母细胞表达系统中表达全部候选性信息素受体基因，用双电极电压钳记录受体对性信息素组分的反应，证明了这4个候选受体均为性信息素受体，可被绿盲蝽的不同性信息素组分激活；利用触角电位记录和单细胞记录技术研究了绿盲蝽触角和感器对性信息素组分的反应；对绿盲蝽性信息素受体进行RNA干扰，结合行为学记录对性信息素受体的体内功能进行了验证。通过以上研究，我们在分子、神经和行为三个水平全面解析绿盲蝽识别性信息素的机制，阐明绿盲蝽雌雄个体相互识别高度特异性的分子和神经机制，为下一步从分子水平筛选比性信息素更有效的行为干扰剂奠定基础。受本项目资助共发表SCI论文8篇，发表中文核心期刊论文3篇。获得国家发明专利授权5项，另外还申请国家发明专利授权5项。圆满完成了本项目的研究目标。