棉铃虫5-HT1和5-HT2基因的克隆及表达模式分析  

Cloning and expression analysis of the 5-HT1 and 5-HT2 genes in Helicoverpa armigera

作　　者：王惠鑫 陈淑婷 王凯 郜晓妍 谢桂英[1] 陈文波 赵新成[1] WANG Huixin;CHEN Shuting;WANG Kai;GAO Xiaoyan;XIE Guiying;CHEN Wenbo;ZHAO Xincheng(Henan International Laboratory for Green Pest Control,College of Plant Protection,Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]河南省害虫绿色防控国际联合实验室,河南农业大学植物保护学院,郑州450002

出　　处：《植物保护》2023年第1期147-156,163,共11页Plant Protection

基　　金：国家自然科学基金(32001912)。

摘　　要：5-羟色胺(5-hydroxytryptamine,5-HT)是昆虫体内一种重要的生物胺,其通过结合特异性的G蛋白偶联受体在昆虫体内发挥不同的神经调控作用。本研究基于转录组测序数据结合RT-PCR技术鉴定克隆了棉铃虫5-HT受体基因5-HT1和5-HT2,采用生物信息学方法分析其序列特征,利用实时荧光定量PCR(RT-qPCR)技术检测了其在幼虫和成虫不同组织内的表达特征。结果显示,棉铃虫体内分别表达5-HT的2类受体基因:5-HT1A、5-HT1B、5-HT2A和5-HT2B。系统进化树分析表明,棉铃虫5-HT受体基因分为3个不同分支,而5-HT1和5-HT2可明显分别分成2个不同分支,各分支均与家蚕和烟草天蛾5-HT相应受体基因具有较高的同源性。RT-qPCR结果显示,在幼虫期,4种5-HT受体基因都在头部显著高表达,成虫期除在脑部高表达外,在触角或腹部也有高表达。其中5-HT1A和5-HT1B在雌成虫触角表达量显著高于雄成虫触角,5-HT1A和5-HT2B在雌成虫腹部表达量显著高于雄成虫腹部,而5-HT1B在雄成虫腹部表达量显著高于雌成虫腹部。以上结果表明5-HT1和5-HT2可能参与棉铃虫脑部神经调控,触角嗅觉感受及生殖发育等活动。该结果为进一步明确棉铃虫5-HT受体的功能奠定了基础。5-hydroxytryptamine is an important biogenic amine as neurotransmitter,which regulates various biological functions in insects by binding and activating specific G protein-coupled receptors.To clarify the types and expression profiles of 5-HT1 and 5-HT2 receptor genes in the cotton bollworm,Helicoverpa armigera,5-HT1 and 5-HT2 were cloned by using RT-PCR protocol based on the transcriptome data,and their sequence characteristics were predicted and analyzed by bioinformatics method.Real-time quantitative PCR(RT-qPCR)was used to compare the relative expression levels of 5-HT1 and 5-HT2 in different tissues of larva,male and female adults.The results showed that two 5-HT1(5-HT1A,5-HT1B)and two 5-HT2(5-HT2A,5-HT2B)subtype receptor genes were identified and cloned from H.armigera:Phylogenetic analysis showed that H.armigera 5-HT receptor genes were clustered in three different clades,and 5-HT1 and 5-HT2 were further divided into two distinct branches,respectively.Each H.armigera 5-HT subtype had a high homology with the 5-HT receptor genes of Bombyx mori and Manduca sexta.The RT-qPCR results showed that 5-HT1 and 5-HT2 were expressed at the highest level in H.armigera heads during larval stage.During the adult stage,they were not only highly expressed in brains,but also highly expressed in antennae and abdomens.The expression levels of 5-HT1A and 5-HT1B were significantly higher in female antennae than in male ones;the expression level of 5-HT1A and 5-HT2B were significantly higher in female abdomens than in male,whereas the expression level of 5-HT1B was significantly higher in male abdomens than in female.These results indicated that 5-HT1 and 5-HT2 receptors might play a role in diverse physiological and biochemical processes,such as neural regulation of brain,olfactory perception of antennae and reproductive development in H.armigera,which established the foundation for further study on the functions of the 5-HT receptors in H.armigera.

关 键 词：棉铃虫 5-HT受体 实时荧光定量PCR 表达量 

分 类 号：S435.622.3[农业科学—农业昆虫与害虫防治]