白蜡虫体内杀雄菌属(Arsenophonus)共生菌的分子检测  

Molecular detection of symbiotic bacteria Arsenophonus from Ericerus pela Chavannes

作　　者：刘魏魏[1] 杨璞[1] 阮永明[1] 陈晓鸣[1] 徐冬丽[1] 李燕飞[1] 

机构地区：[1]中国林业科学研究院资源昆虫研究所、国家林业局资源昆虫培育与利用重点实验室,昆明650224

出　　处：《微生物学报》2012年第8期1002-1010,共9页Acta Microbiologica Sinica

基　　金：国家自然科学基金青年基金(31000983);林业公益性行业科研专项经费项目(201204602);云南省应用基础研究基金(2010ZC235);中国林业科学研究院资源昆虫研究所基本科研业务费专项资金(riricaf200904M-3,riricaf2011006M)~~

摘　　要：【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。[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.

关 键 词：白蜡虫 杀雄菌属共生菌 16S RDNA 性比 绝对定量 

分 类 号：S899.1[农业科学—特种经济动物饲养] Q93[农业科学—畜牧兽医]