东亚飞蝗谷胱甘肽S-转移酶RNA干扰效率研究
The RNA interference efficiency of glutathione Stransferases from Locusta migratoria manilensis
刘婷1，秦国华1，2张建珍1，马恩波1**
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作者单位：1 山西大学应用生物学研究所太原030006； 2 山西大学环境与资源学院太原030006
中文关键词：谷胱甘肽硫转移酶， 东亚飞蝗， RNA干扰， 时间效应
英文关键词：glutathione Stransferases (GSTs), Locusta migratoria manilensis, RNAi, time effect
中文摘要： 东亚飞蝗Locusta migratoria manilensis (Meyen)是我国主要的农业害虫之一，已发现东亚飞蝗对某些农药产生了抗性，其抗性机制可能与谷胱甘肽硫转移酶（GST）代谢解毒相关。本研究利用特异性引物合成东亚飞蝗GST 4个不同家族基因的双链RNA（dsRNA），将dsRNA注射到东亚飞蝗幼虫体内，采用Real time RT-PCR技术测定了干扰不同时间后目的基因mRNA的表达水平。结果表明，4个不同家族GST的沉默效应具有时间差异。来自delta家族的LmGSTd1和sigma家族的LmGSTs5基因在注射dsRNA后12 h时mRNA量就已显著下降；而来自theta家族的LmGSTt1和unknown家族的LmGSTu1基因在注射24 h后mRNA水平才呈现显著下降。本研究对后续东亚飞蝗GST功能及抗性机制研究提供了基础资料和依据，同时对其它昆虫RNA干扰研究具有一定的借鉴作用。
英文摘要：T he migratory locust, Locusta migratoria manilensis (Meyen) (Orthoptera: Acridoidea), is one of the most important pests in our country. Resistance of this locust to some insecticides has been detected and the resistance mechanism hypothesized to involve glutathione Stransferases (GST). In this study, doublestranded RNA of four different classes of GST was synthesized using specific gene primers and injected into the 2nd instar nymphs of L. m. manilensis. The effects of silencing time on target GSTs mRNA expression was assayed by real time RT-PCR. The results show that the expression of LmGSTd1 and LmGSTs5 were reduced significantly 12 h after injection, whereas LmGSTt1 and LmGSTu1 were reduced significantly 24 h after injection. These results provide a foundation for further study of GST function and mechanisms in locusts.