白蜡虫体内杀雄菌属(Arsenophonus)共生菌的分子检测

摘要

[Objective] The purpose of this study is to examine the relation between the sex ratio of Ericerus pela and its symbiotic bacterial Arsenophonus.[Methods] The symbiotic bacterial diversity in male Ericerus pela was determined through sequencing 16S rDNA gene library.PCR amplification for Arsenophonus was performed by using two 16S rDNA specific primers and 23S rDNA specific primer.The molecular detection of Arsenophonus in six geographic populations of E.pela, namely Zhaotong, Kunming, Jinkouhe, Hangzhou, Changchun, and Jianghua, were performed by semi-quantitative PCR.The absolute concentrations of Arsenophonus in E.pela of Zhaotong, Kunming, Jinkouhe geographic populations were determined using absolute quantitative real-time PCR.[Results] Two different 16S rDNA sequences were obtained; the sizes were 445bp and 1462bp respectively.A 23S rDNA sequence was obtained, the size was 582bp.Some E.pela individuals of Hangzhou and Jianghua were not infected with Arsenophonus.The contents of Arsenophonus in E.pela of Zhaotong were significantly higher than that of Kunming and Jinkouhe, while the contents of Arsenophonus from the latter two geographic populations were not significantly different.[ Conclusion] Arsenophonus is not responsible for the sex ratio of E.pela.%[目的]研究白蜡虫体内杀雄菌属(Arsenophonus)共生菌的含量与白蜡虫性比之间的关系.[方法]采用16S rDNA文库的方法对白蜡虫雄虫体内的共生菌进行分析,利用杀雄菌属特异的2条16S rDNA引物以及23S rDNA引物进行PCR检测.对昭通、昆明、金口河、杭州、长春、江华6个不同地理种群白蜡虫体内的杀雄菌属共生菌进行半定量分析,并采用荧光定量PCR对昭通、昆明、金口河白蜡虫体内的杀雄菌属共生菌进行绝对定量分析.[结果]在白蜡虫体内首次发现杀雄菌属共生菌.在白蜡虫雌雄虫体内均扩增出杀雄菌属的两条不同长度的16S rDNA序列,分别为445bp和1462bp,并扩增得到长度为582bp的23S rDNA序列.杭州和江华地理种群白蜡虫的一些个体不含杀雄菌属共生菌.昭通地理群的杀雄菌属共生菌含量显著高于金口河和昆明,而昆明和金口河白蜡虫的杀雄菌属含量无显著差异.[结论]白蜡虫体内杀雄菌属共生菌的含量与白蜡虫性比无关.