小菜蛾腺苷受体基因PxAdoR克隆及杀虫靶标作用

doi:10.16409/j.cnki.2095-039x.2016.06.006

中国生物防治学报 ›› 2016, Vol. 32 ›› Issue (6): 712-720.DOI: 10.16409/j.cnki.2095-039x.2016.06.006

小菜蛾腺苷受体基因PxAdoR克隆及杀虫靶标作用

方鸣^1,2, 柴一秋^1,2, 陈官菊², 章金明³, 吕要斌³, 黄勃¹

1. 安徽农业大学微生物防治省重点实验室, 合肥 230036;
2. 浙江省亚热带作物研究所/温州市虫生真菌资源研究开发重点实验室, 温州 325005;
3. 浙江省农业科学院植物保护与微生物研究所, 杭州 310021

收稿日期:2016-02-04 出版日期:2016-12-08 发布日期:2016-12-07
通讯作者: 柴一秋,女,研究员,E-mail:chai_yiqiu@yeah.net
作者简介:方鸣,男,硕士研究生,E-mail:110266502@qq.com
基金资助:
浙江省重大优先项目（2008C12004-3）；浙江省自然科学基金（LY12C14004）；温州市四新项目（F-DY201509170025）

Cloning and Functional Analysis the PxAdoR Gene of Plutella xylostella Adenosine Receptor

FANG Ming^1,2, CHAI Yiqiu^1,2, CHEN Guanju², ZHANG Jinming³, LÜ Yaobin³, HUANG Bo¹

1. Provincial Key Laboratory of Microbial Control, Anhui Agricultural University, Hefei 230036, China;
2. Zhejiang Institute of Subtropical Crops/Key Laboratory of Entomogenous Fungi Resources Research and Development of Wenzhou, Wenzhou 325005, China;
3. Institute of Plant Protection and Microbiology, Zhejiang Academy of Agricultural Sciences, Hangzhou 310021, China

Received:2016-02-04 Online:2016-12-08 Published:2016-12-07

摘要/Abstract

摘要： 利用蝉花虫草Ophiocordyceps sobolifera代谢产物N6-（2-羟乙基）腺苷（HEA）作用于小菜蛾Plutella xylostella L.获得差异表达的片段，采用RT-PCR和RACE技术从小菜蛾中克隆全长1828 bp的小菜蛾腺苷受体（Plutella xylostella adenosine receptor，PxAdoR）基因，通过生物信息学软件预测和分析蛋白质结构，PxAdoR为分子量49.2 kD，没有信号肽，疏水性不稳定蛋白，具有7次跨膜螺旋结构的一类G蛋白偶联受体（G protein coupled receptor，GPCRs）。利用HEA及哺乳动物腺苷受体A2aR抑制剂SCH58261处理小菜蛾2龄幼虫，结果显示联合使用HEA和SCH58261后36~60 h，可使HEA引起的小菜蛾2龄幼虫的校正死亡率显著降低（42.7%~53.9%，P<0.05）；而联合用药后36、48、60 h小菜蛾2龄幼虫体重增长抑制率由43.3%、38.3%、46.2%显著降低（P<0.05）到13.2%、10.3%、8.6%。本研究表明SCH58261能显著降低HEA引起的小菜蛾2龄幼虫的死亡率和体重增长抑制率，HEA可能是通过PxAdoR而起作用，PxAdoR基因可以作为新的杀虫靶点用于害虫控制研究。

关键词: 腺苷受体, 基因克隆, 腺苷受体抑制剂, 蛋白结构, N6-(2-羟乙基)腺苷

Abstract: Differentially expressed fragments of Plutella xylostella treated with the metabolite N6-(2-hydroxyethyl)-adenosine (HEA) of Ophiocordyceps sobolifera were cloned using RT-PCR and RACE and a whole sequence (1828 bp) of adenosine receptor (PxAdoR) gene was obtained. The protein structure predicted and analyzed using the bioinformatics software is of seven transmembrane helices that belong to the G protein coupled receptors. The protein is hydrophobic and instable, without signal peptide, and of molecular weight of 49.2 kD. To make sure the PxAdoR protein functions, P. xylostella 2nd instar larvae were treated with HEA and the antagonist SCH58261 of mammal adenosine A2a receptor. Corrected mortality of treated P. xylostella larvae was significantly reduced by about 50% at 36 to 60 h. And the rate of weight growth inhibition was reduced significantly from 43.3%, 38.3% and 46.2% to 13.2%, 10.3% and 8.6% at 36, 48 and 60 h, respectively. It is prospective that HEA may act by PxAdoR and PxAdoR may be a new insecticidal target for pest control.

Key words: adenosine receptors, gene clone, adenosine receptor antagonist, protein structure, N6-(2-hydroxyethyl)-adenosine

中图分类号:

S436

方鸣, 柴一秋, 陈官菊, 章金明, 吕要斌, 黄勃. 小菜蛾腺苷受体基因PxAdoR克隆及杀虫靶标作用[J]. 中国生物防治学报, 2016, 32(6): 712-720.

FANG Ming, CHAI Yiqiu, CHEN Guanju, ZHANG Jinming, LÜ Yaobin, HUANG Bo. Cloning and Functional Analysis the PxAdoR Gene of Plutella xylostella Adenosine Receptor[J]. journal1, 2016, 32(6): 712-720.

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小菜蛾腺苷受体基因PxAdoR克隆及杀虫靶标作用

Cloning and Functional Analysis the PxAdoR Gene of Plutella xylostella Adenosine Receptor

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