苹果蠹蛾体内wolbachia wsp基因克隆与序列分析

Cloning and sequence analysis of the Wolbachia wsp gene to detect of Wolbachia in Cydia pomonella

【目的】对苹果蠹蛾Cydia pomonella L.体内共生菌Wolbachia进行分子生物学鉴定,确定该虫体内Wolbachia的进化位置,为进一步探讨Wolbachia对其生殖作用的调控机制提供理论依据。【方法】应用Wolbachia的wsp基因特异引物,通过PCR扩增法检测了苹果蠹蛾10个地理种群(新疆伊犁、吐鲁番、和田、石河子、奎屯、哈密、库尔勒、阿拉尔、喀什、和甘肃张掖)感染Wolbachia的状况,并对阿拉尔种群体内的Wolbachia的wsp基因进行测序和序列分析。【结果】苹果蠹蛾10个地理种群全部感染了tWolbachia,利用wsp基因的特异性引物从阿拉尔种群体内扩增出了617 bp的Wolbachia的wsp基因片段(GenBank登录号为KC832324),系统发育分析结果表明,苹果蠹蛾体内感染的Wolbachia属于A群Dor亚群,与锤角细蜂亲缘关系较近。【结论】苹果蠹蛾体内普遍感染了Wolbachia,属于A群Dor亚群。

[Objectives] The evolution position of Wolbachia in Cydiapornonella （Lepidoptera：Tortricidae） was identified by molecular biology in order to further provide theoretic basis for the regulatory mechanisms of reproductive role of Wolbachia. [Methods] Wolbachia infection in ten populations of Cydia pomonella were detected by using a specific primer for the Wolbaehia wsp gone and PCR. These populations were from Yili, Tulufan, Hetian, Shihezi, Kuitun, Hami, Korla, Alar and Kashi in Xinjiang and Zhangye in Gansu. The Wolbachia wsp gene of from Alar populations of C. pomonella was amplified and sequenced. [Results] All populations were infected with Wolbachia. Using the specific primers, a 617 bp region of the Wolbachia wsp gene （GenBank accesstion number is KC832324） was amplified from Alar populations. Phylogenetic analysis showed that Wolbachia in C. pomonella belongs to the Dot subgroup of the A group, and had the closest genetic relationship to Trichopria drosophilae. [Conclusion] Wolbachia infection was more common in the C. pomonella, and belong to Dot subgroup in A group.