薇甘菊颈盲蝽化学通讯及其嗅觉相关基因克隆、功能分析

The applicant has found that Pachypeltis sp. is an important natural enemy of quarantined forestry plant Mikania micrantha. As the preparative work, we investigated its biology and ecological habits, initially identified the existence of sex pheromone and host plant volatile inforchemicals to regulate this insect behavior and analyzed hindgut and host plant volatiles. The objectives of this study were to make isolation and identification of sex pheromone and host volatile inforchemicals; to determine the role in regulating coordinately the insect behavior by the composition of sex pheromone and host volatile inforchemicals; to reveal the chemical communication characteristics of Pachypeltis sp. and identify the molecular mechanisms of chemical communication odors by building antennae cDNA library, identifying olfactory genes related to the chemical communication, using Fluorescence Displacement Method to assay the binding capacity of antennae -specific protein for identification of chemical communication, making a homology modeling to predict three-dimensional structure of the proteins and the key binding sites of the odor molecule; to be a new technique and theoretical support for the control of Mikania micrantha by developing adults attractant and proposing the method of trapping Pachypeltis sp..

薇甘菊颈盲蝽（Pachypeltis sp.）为申请者发现的一种专性取食林业检疫性有害植物薇甘菊（Mikania micrantha）的重要天敌，现已掌握其生物学、生态学习性，初步确定调控颈盲蝽行为的性信息素和薇甘菊挥发性化学信息物质存在,并解析其后肠及薇甘菊挥发性成分。本研究将分离、鉴定调控颈盲蝽行为的性信息素和薇甘菊挥发性化学信息活性物质；复合性信息素和薇甘菊挥发性化学信息物质，确定其协同调控颈盲蝽行为的作用能力；构建薇甘菊颈盲蝽触角cDNA文库，发掘与化学通讯相关的嗅觉基因，荧光竞争结合实验检测触角特异性表达嗅觉基因蛋白识别化学通讯气味物质的结合能力，同源建模预测其蛋白的三维结构和识别气味分子的关键结合位点，揭示薇甘菊颈盲蝽化学通讯特性及其嗅觉识别化学通讯气味物质的分子机制；研制薇甘菊颈盲蝽成虫引诱剂、人工诱集方法，为助迁、聚集颈盲蝽防控薇甘菊提供新技术和理论支撑。

薇甘菊Mikania micrantha H.B.K.属菊科Compositae 假泽兰属Mikania 攀缘草本植物，广泛分布于我国的广东、广西、福建、海南、云南、香港、澳门、台湾地区，2013 年1 月9 日再次被国家林业局公告确定为唯一的全国林业检疫性有害植物，其入侵的危害、危险性已是极为严重。依托前期研究工作基础，进行薇甘菊天敌资源收集，首次发现薇甘菊颈盲蝽Pachypeltis micranthus Mu et Liu (Hemiptera: Miridae) 取食薇甘菊。前期研究既表明了薇甘菊颈盲蝽专性取食薇甘菊，同时也发现薇甘菊颈盲蝽存在局部聚集取食薇甘菊的现象，为了揭示其专一性和局部聚集取食机制，2014年获得本项研究计划，为此，解析确定了调控薇甘菊颈盲蝽行为的寄主挥发性化学信息活性物质香叶烯，鉴定出了薇甘菊颈盲蝽性信息素十六烷酸，初步阐明了性信息素与寄主挥发性化学信息活性物质对颈盲蝽行为调控作用机理，掌握了薇甘菊颈盲蝽化学通讯特性，研制出了薇甘菊颈盲蝽成虫引诱剂2个，并提出了人工诱集颈盲蝽持续控制薇甘菊技术；进一步构建了薇甘菊颈盲蝽触角cDNA 文库，发掘筛选、基因克隆、表达了薇甘菊颈盲蝽触角特异性表达的嗅觉基因13个，荧光竞争结合实验检测触角特异性表达嗅觉基因蛋白识别化学通讯气味物质的结合能力，同源建模预测其嗅觉基因蛋白PmicOBP6和PmicCSP1三级结构和识别化学通讯活性物质分子的关键识别位点，初步阐明薇甘菊颈盲蝽触角特异性表达嗅觉基因蛋白的嗅觉识别功能，为揭示薇甘菊颈盲蝽识别化学通讯气味物质的分子机制提供理论基础。本项目取得研究成果，为进一步开发利用薇甘菊颈盲蝽持续控制薇甘菊提供研究基础，也为相关学科相应学术研究提供了可以参考的借鉴，具有重要的学术价值。