南京农业大学学报2018，Vol.41Issue(2)：293-301,9.DOI:10.7685/jnau.201707033
马铃薯甲虫烟碱型乙酰胆碱受体α5亚基基因的克隆及其表达模式分析
Cloning and expression profile of nicotinic acetylcholine receptor α5 subunit gene from Leptinotarsa decemlineata
摘要
Abstract
[Objectives]Nicotinic acetylcholine receptor(nAChR)plays an important role in excitatory transmission at cholinergic synapse in insect central nerve systems and is also the target of neonicotinoid insecticides. The Colorado potato beetle(CPB),Leptino-tarsa decemlineata,is an important worldwide quarantine defoliator.The ten nAChR subunit genes in L.decemlineata had been obtained from previous reports,namely Ldα1-Ldα4,Ldα6-Ldα10 and Ldβ1. The aim of the study is to further improve the composition and expression profile of the nAChR subunits of L.decemlineata. [Methods]The nAChR α subunit gene was cloned by polymerase chain reaction(PCR)and rapid amplification of cDNA ends(RACE)based on bioinformatic analysis of L.decemlineata genome dataset. The cloned gene was subjected to phylogenetic relationship analysis by constructing phylogenetic tree. Quantitative real-time PCR was applied to the investigation of the expression levels of different expression profiles of the Ldα5 in different development stages or body parts of adults and under imidacloprid and thiacloprid treatments. [Results]In this study,the full-length cDNA of a new nAChR α subunit gene from L.decemlineata was cloned.It was named as Ldα5(GenBank accession:MF197919).The quantitative real-time PCR results showed Ldα5 had the highest expression level in 1st instar larvae and heads of adults respectively. In the 4th instar larvae exposed to the insecticides tested,imidacloprid significantly decreased the expression level of Ldα5 and thiamethoxam had no effects, whereas in the adults,the two insecticides led to the up-regulation of Ldα5 significantly. [Conclusions]The findings may provide basis for further study on the function of nAChR subunits and the molecular mechanisms of target resistance to neonicotinoids of L.decemlineata.关键词
马铃薯甲虫/烟碱型乙酰胆碱受体/基因克隆/序列分析/实时定量PCRKey words
Leptinotarsa decemlineata/nicotinic acetylcholine receptor/gene cloning/sequence analysis/quantitative real-time PCR分类
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解林杰,李莎,姜卫华..马铃薯甲虫烟碱型乙酰胆碱受体α5亚基基因的克隆及其表达模式分析[J].南京农业大学学报,2018,41(2):293-301,9.基金项目
国家公益性行业(农业)科研专项(201103026) （农业）
