昆虫聚集过程中会释放和感知聚集信息化合物，气味受体ORs参与识别这类信息物并在调控昆虫聚集行为中发挥重要功能。本申请以新疆棉田优势天敌多异瓢虫为研究对象，收集聚集状态的瓢虫成虫体表、虫卵、排泄物及后肠挥发性气味，通过GC-MS、SSR和昆虫嗅觉仪等鉴定具有电生理活性及聚集功能的信息化合物。依据多异瓢虫基因组及触角转录组数据，利用RT-PCR和qPCR方法筛选触角特异或高表达ORs，通过原位杂交和免疫组化技术对候选ORs进行组织定位。采用爪蟾卵母细胞重组表达和双电极电压钳系统研究候选ORs与潜在聚集信息物的识别关系。通过CRISPR-Cas9体系敲除靶标ORs，利用电生理和行为测试评价缺失ORs对多异瓢虫识别信息物及聚集行为的影响，基于聚集类化合物设计多异瓢虫聚集行为调控剂应用于田间天敌昆虫保育。阐明气味受体调控多异瓢虫聚集行为的分子机制，设计昆虫行为控制策略，助力实现“保益控害”目标。

Aggregation pheromones are released and perceived by insects during the process of aggregation. Odorant receptors (ORs) can recognize such chemical signals and play an important role in regulating insect aggregation behavior. In the current work, volatiles from body surface, hindgut, eggs and excrement of aggregated adults of Hippodamia variegate (Goeze) are collected. Semiochemicals with electrophysiological and aggregation behavioral activities will be identified by using GC-MS GC-EAD, SSR and insect olfactometer. Based on the genome and antennal transcriptome data of H. variegate, antennal-biased or antennal enriched ORs will be identified by RT-PCR and real-time quantitative PCR techniques. In situ hybridization and immunohistochemistry are performed to investigate the tissues location of candidate ORs in antennae. Two-electrode voltage clamp recordings will be conducted to explore the binding feature of the recombinant ORs to the active ligands. The roles of the target ORs will be further investigated using the CRISPR-Cas9 system. After the silence of target ORs, electrophysiological and behavioral changes of CRISPR-Cas9 treated ladybirds will be evaluated. According to aggregation compounds, we will design regulators to control the aggregation behavior of H. variegate. Our findings will clarify the molecular mechanism of odorant receptor regulating the aggregation behavior of ladybirds and provide the theoretical support for the goal of “benefit natural enemies and control pests”.