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,Px Ado R）基因,通过生物信息学软件预测和分析蛋白质结构,Px Ado R为分子量49.2 k D,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A2a R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Px Ado R而起作用,Px Ado R基因可以作为新的杀虫靶点用于害虫控制研究。
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（Px Ado R） 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 k D. To make sure the Px Ado R protein functions, P. xylostella 2nd instar larvae were treated with HEA and the antagonist SCH58261 of mammal adenosine A2 a 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 Px Ado R and Px Ado R may be a new insecticidal target for pest control.
出处
《中国生物防治学报》
CSCD
北大核心
2016年第6期712-720,共9页
Chinese Journal of Biological Control
基金
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)
