麦长管蚜羧酸酯酶基因cDNA片段克隆及吡虫啉胁迫对其表达的影响  

Cloning of Carboxylesterase Gene cDNA Fragments in Sitobion avenae (Fabricius) and Its Expression Analysis Under Imidacloprid Stress

作　　者：白微微[1] 高海峰[1] 张航[1] 杨安沛[1] 李广阔[1] BAI Wei-wei;GAO Hai-feng;ZHANG Hang;YANG An-pei;LI Guang-kuo(Institute of Plant Protection, Xinjiang Academy of Agricultural Sciences~Key Laboratory of Integrated Pest Management on Crop in Northwestern Oasis, Ministry of Agriculture of P. R. China, Urumqi 830091)

机构地区：[1]新疆农业科学院植物保护研究所农业部西北荒漠绿洲作物有害生物综合治理重点实验室,乌鲁木齐830091

出　　处：《生物技术通报》2018年第6期109-114,共6页Biotechnology Bulletin

基　　金：农业部西北荒漠绿洲作物有害生物综合治理重点实验室开放基金(KFJJ20150106)

摘　　要：羧酸酯酶在昆虫对杀虫剂的解毒代谢过程中发挥重要作用,本研究旨在分析吡虫啉胁迫对麦长管蚜羧酸酯酶基因表达的影响。采用同源克隆方法克隆麦长管蚜羧酸酯酶基因cDNA片段,实时荧光定量PCR技术检测羧酸酯酶基因在不同吡虫啉剂量下的表达量变化。扩增所得麦长管蚜羧酸酯酶基因cDNA片段大小为392 bp,命名为SaEST 3(GenBank登录号KY 441614),该片段编码130个氨基酸残基,分子量14 kD,等电点4.93。序列同源性比对及生物信息学分析表明SaEST3推导的氨基酸序列与豌豆长管蚜、麦双尾蚜、夹竹桃蚜、桃蚜的羧酸酯酶氨基酸序列相似性较高,分别为94%、85%、80%和80%。实时荧光定量PCR结果显示,不同吡虫啉处理剂量下,SaEST3 m RNA的相对表达量均上调。克隆得到的基因片段为麦长管蚜羧酸酯酶基因片段,吡虫啉对麦长管蚜羧酸酯酶基因SaEST3表达有一定的影响。Carboxylesterase plays an important role in insect detoxification and metabolism processes of insecticides. This research aims at analyzing the expression levels of carboxylesterase gene in Sitobion avenae（Fabricius）under imidacloprid stress. Homologous cloning strategy was used to clone carboxylesterase gene cDNA fragment,and its expression variations under different dose of imidacloprid were analyzed with qRT-PCR. A carboxylesterase gene cDNA fragment of 392 bp,named SaEST 3（GenBank accession number is KY 441614）,was amplified from S. avenae（Fabricius）using homologous cloning strategy. The gene sequence coded a polypeptide of 130 amino acid residues,the calculated molecular mass was 14 kD and the theoretical isoelectric point（p I）was 4.93. Homology alignment and bioinformatics analysis suggested that the amino acid sequence of SaEST3 was highly identical to that of carboxylesterase gene from Acyrthosiphon pisum（94%）,Diuraphis noxia（85%）,Aphis nerii（80%）,and Myzus persicae（80%）. The results of qRT-PCR showed that the relative expression of SaEST 3 m RNA was up-regulated under different dose of imidacloprid. In a summary,the cloned sequence was carboxylesterase gene of S. avenae（Fabricius）,and imidacloprid stress may affect the expression of SaEST 3 gene in S. avenae（Fabricius）.

关 键 词：羧酸酯酶 麦长管蚜 吡虫啉 克隆 基因表达 

分 类 号：S433.39[农业科学—农业昆虫与害虫防治]