棉铃虫普通气味受体基因HarmOR9和HarmOR29的克隆和组织表达分析  

Cloning and tissue expression analysis of general odorant receptor genes HarmOR9 and HarmOR29 in the cotton bollworm,Helicoverpa armigera( Lepidoptera: Noctuidae)

作　　者：刘宁灿 张进[2] 王桂荣[2] 刘杨[2] 严善春[1] 

机构地区：[1]东北林业大学林学院,哈尔滨150040 [2]中国农业科学院植物保护研究所植物病虫害生物学国家重点实验室,北京100193

出　　处：《昆虫学报》2014年第5期522-529,共8页Acta Entomologica Sinica

基　　金：国家自然科学基金重点项目(31230062);国家自然科学基金青年基金项目(31201578);北京市自然科学基金项目(6132028)

摘　　要：【目的】对棉铃虫Helicoverpa armigera 2个普通气味受体基因的cDNA全长进行分析,明确这两个普通气味受体基因在不同组织中的表达分布,为进一步的功能研究奠定基础。【方法】利用PCR结合RACE技术克隆棉铃虫两条普通气味受体基因的cDNA全长;利用不同的生物信息学软件对序列进行结构预测、序列比对和进化树分析;利用半定量RT-PCR检测其在棉铃虫成虫不同组织中的表达。【结果】获得两条棉铃虫气味受体基因的全长序列,并命名为HarmOR9和HarmOR29(GenBank登录号分别为KJ188252和KJ188253)。序列分析显示,HarmOR9全长1 206 bp,编码401个氨基酸;HarmOR29全长1 188 bp,编码395个氨基酸。选择已报道的鳞翅目昆虫烟青虫Heliothis assulta、家蚕Bombyx mori、烟芽夜蛾Heliothis virescens和棉铃虫的气味受体与本实验克隆得到的两个气味受体基因的编码产物进行序列比对和进化树分析,结果显示这两个气味受体与性信息素受体区别明显,并与其他普通气味受体聚类在一起。半定量RT-PCR的结果显示HarmOR9与HarmOR29都主要在触角中高表达且无雌雄间差异,HarmOR29在其他组织中均不表达;而HarmOR9在雄虫下唇须中有微量表达,在其他组织中均不表达。【结论】本研究从棉铃虫中克隆得到2个气味受体基因HarmOR9和HarmOR29的cDNA全长,其编码产物具有气味受体的典型特征并且属于普通气味受体。明确了这两个气味受体基因都在棉铃虫成虫的触角中高表达,且无雌雄差异,推测其可能参与了棉铃虫普通气味的识别过程。[ Aim] This study aims to clone two general odorant receptor genes, analyze their sequences and investigate their expression profiles in different tissues of adults of the cotton bollworm, Helicoverpaarmigera. The findings will provide basic knowledge for further functional study of these two odorant receptor genes. [ Method] We cloned the full-length cDNA sequences of two odorant receptor genes byPCR and RACE techniques. The structure prediction, sequence alignment and phylogenetic analysis of the coding products of these two odorant receptor genes were performed using different bioinformaticssoftwares. The expression profiles of these two genes in different tissues were investigated by using semi- quantitative RT-PCR. [ Results ] Two full-length cDNA sequences encoding odorant receptors wereobtained and named as HarmOR9 and HarmOR29 （GenBank accession number KJ188252 and KJ188253, respectively）. Sequence analysis showed that the full-length of HarmOR9 gene is 1 206 bp,which encodes a polypeptide of 401 amino acids, and the full-length of HarmOR29 gene is 1 188 bp, which encodes a polypeptide of 395 amino acids. Sequence alignment and phylogenetic analysis of thecoding products of these two odorant receptor genes with reported odorant receptors from lepidopteraninsects including Helicoverpa assulta, Bombyx mori, Heliothis virescens and H. armigera showed that the coding products of these two odorant receptor genes are obviously different from sex pheromone receptors,but cluster together with other general odorant receptors. The results of semi-quantitative PCR showed that these two odorant receptor genes were highly expressed in antennae with no significant differencebetween male and female, HarmOR29 was not expressed in other tissues, and HarmOR9 was slightly expressed in male labial palps but not in other tissues. [ Conclusion ] In this study, two odorant receptorgenes were cloned in H. armigera. Sequence analysis showed that the coding products of these two genes own typical characteristics of odorant rec

关 键 词：棉铃虫 嗅觉受体 基因克隆 组织表达谱 半定量RT—PCR 

分 类 号：Q966[生物学—昆虫学]