孙 菲 杨春红 李 波 郑金柱 周成刚 谢丽霞 尹淑艳

(1. 山东农业大学植物保护学院 泰安 271018； 2. 泰安市林业局 泰安 271000； 3.泰安市徂徕山林场 泰安 271000)

针叶小爪螨不同种群内共生菌Wolbachia和Cardinium的检测及系统发育分析*

孙 菲1杨春红1李 波2郑金柱3周成刚1谢丽霞1尹淑艳1

(1. 山东农业大学植物保护学院 泰安 271018； 2. 泰安市林业局 泰安 271000； 3.泰安市徂徕山林场 泰安 271000)

【目的】 针叶小爪螨的山东板栗种群(TSBL)和浙江杉木种群(ZJSM)间存在生殖不亲和，而内共生菌Wolbachia和Cardinium能引起宿主的胞质不亲和等生殖异常现象，明确Wolbachia和Cardinium在针叶小爪螨中的分布情况，为下一步研究其在该螨种群分化中的可能作用及其对该螨的生殖调控作用奠定基础。【方法】 提取单头雌成螨的总DNA，使用特异性引物通过PCR扩增Wolbachia的wsp基因和Cardinium的16S rRNA基因的部分片段，将电泳检测为阳性的PCR产物纯化、测序，以确定所得产物是否为目的基因产物，并进行系统发育分析。【结果】 针叶小爪螨的山东板栗种群(TSBL)和浙江杉木种群(ZJSM)均无Wolbachia和Cardinium感染。所检测的12个针叶小爪螨种群中，Wolbachia和Cardinium的感染率分别是0和8.3%。Cardinium仅在河北麻栎种群中检测到，但种群内感染率较高，为66.7%。感染针叶小爪螨的Cardinium与感染二斑叶螨的Cardinium的亲缘关系较近。【结论】 我国针叶小爪螨中Wolbachia的感染率可能极低，该螨山东板栗种群(TSBL)和浙江杉木种群(ZJSM)间的生殖不亲合与Wolbachia和Cardinium无关。

针叶小爪螨； 种群分化；Wolbachia；Cardinium； 系统发育

Abstract： 【Objective】 There exists reproductive incompatibility between theCastaneamollissimapopulation from Shandong Province (TSBL) and theCunninghamialanceolatapopulation from Zhejiang Province (ZJSM) ofOligonychusununguis. However,WolbachiaorCardinium, as the insect’s endosymbiont, has the ability to alter the reproductive capabilities of its host. This study aims to analyze the distribution ofWolbachiaandCardiniumin differentO.ununguispopulations, and to reveal the effects of endosymbionts onO.ununguisreproductive incompatibility.【Method】 Primers were designed according to the sequences of the gene encoding a surface protein ofWolbachia(wsp) and theCardinium’s 16S rRNA gene, and PCR was performed using the genomic DNA extracted from a single adult femaleO.ununguis. Purified PCR fragments were sequenced and blasted against the databases. Phylogenetic tree was constructed based on these sequences.【Result】 Neither theC.mollissimapopulation from Shandong Province (TSBL) nor theC.lanceolatapopulation from Zhejiang Province (ZJSM) was infected withWolbachia. In all the detected 12 populations ofO.ununguis, the infection rates ofWolbachiaandCardiniumwere 0 and 8.3%, respectively.Cardiniumwas detected only in theQuercusacutissimapopulation from Hebei Province, but the intrapopulation infection rate was high with 66.7%.TheCardiniuminfectingO.ununguiswas closer to that fromTeranychusurticaein phylogenetic relationship.【Conclusion】 The infection rate ofWolbachiamight be very low inO.ununguisin China. The reproductive incompatibility between theC.mollissimapopulation from Shandong Province (TSBL) and theC.lanceolatapopulation from Zhejiang Province (ZJSM) ofO.ununguiswas independent ofWolbachiaandCardinium.

Keywords：Oligonychusununguis； population differentiation；Wolbachia；Cardinium； phylogeny

针叶小爪螨(Oligonychusununguis)是一种分布广、食性广的害螨，广泛分布于亚洲、欧洲、澳洲、美洲的众多国家，寄主植物包括松科(Pinaceae)、柏科(Cupressaceae)、杉……