[1]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.[doi:10.7685/jnau.201707033]
 XIE Linjie,LI Sha,JIANG Weihua.Cloning and expression profile of nicotinic acetylcholine receptor α5 subunit gene from Leptinotarsa decemlineata[J].Journal of Nanjing Agricultural University,2018,41(2):293-301.[doi:10.7685/jnau.201707033]
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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()
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《南京农业大学学报》[ISSN:1000-2030/CN:32-1148/S]

卷:
41卷
期数:
2018年2期
页码:
293-301
栏目:
出版日期:
2018-03-27

文章信息/Info

Title:
Cloning and expression profile of nicotinic acetylcholine receptor α5 subunit gene from Leptinotarsa decemlineata
作者:
解林杰 李莎 姜卫华
南京农业大学植物保护学院/农作物生物灾害综合治理教育部重点实验室, 江苏 南京 210095
Author(s):
XIE Linjie LI Sha JIANG Weihua
College of Plant Protection/Key Laboratory of Integrated Management of Crop Diseases and Pests, Ministry of Education, Nanjing Agricultural University, Nanjing 210095, China
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PCR
Keywords:
Leptinotarsa decemlineatanicotinic acetylcholine receptorgene cloningsequence analysisquantitative real-time PCR
分类号:
S435.32
DOI:
10.7685/jnau.201707033
摘要:
[目的]烟碱型乙酰胆碱受体(nAChR)在昆虫中枢神经系统兴奋性神经递质突触传导过程中起重要作用,也是新烟碱类杀虫剂的作用靶标。马铃薯甲虫(Leptinotarsa decemlineata)是重要的检疫性马铃薯食叶害虫。根据报道已鉴定得到该虫10个nAChR亚基,本研究的目的是进一步完善该虫nAChR亚基的组成和其表达模式。[方法]依据马铃薯甲虫基因组数据,利用PCR和RACE技术对马铃薯甲虫的nAChR亚基基因进行克隆,通过相似性和系统发育树分析验证该基因的亲缘关系。利用实时定量PCR技术检测该基因在马铃薯甲虫各个发育阶段及成虫头、胸、腹和吡虫啉及噻虫嗪处理的4龄幼虫和成虫的表达水平。[结果]克隆获得了该虫又1个nAChR α亚基基因的全长cDNA,将其命名为Ldα5(GenBank登录号为MF197919)。实时定量PCR结果表明:Ldα5在马铃薯甲虫各个发育阶段及成虫头、胸、腹中都有不同程度的表达量,其中在1龄幼虫和成虫头部的表达量最高;吡虫啉和噻虫嗪处理对Ldα5的表达量有不同影响,在幼虫中,吡虫啉导致Ldα5明显下调,而噻虫嗪无明显影响,在成虫中2种药剂的处理都可明显提高Ldα5的表达量。[结论]本研究结果为明确马铃薯甲虫nAChR亚基的生理功能及该虫对新烟碱类杀虫剂的抗性机制等研究奠定了基础。
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),Leptinotarsa 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.

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备注/Memo

备注/Memo:
收稿日期:2017-07-24。
基金项目:国家公益性行业(农业)科研专项(201103026)
作者简介:解林杰,硕士研究生。
通信作者:姜卫华,副教授,主要从事昆虫分子生物学及生理毒理学研究,E-mail:jwh@njau.edu.cn。
更新日期/Last Update: 1900-01-01