华南地区柑橘木虱与柑橘粉虱内共生菌检测及其Wolbachia共生菌的系统发育关系分析
Endosymbiont detection and phylogeny of Wolbachia in Diaphorina citri and Dialeurodes citri
孙秀新1 ** 师沛琼1 许炜明1 覃振强2 任顺祥1 邱宝利1***
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DOI：10.7679/j.issn.2095-1353.2016.096
作者单位：1. 广东省生物农药创制与应用重点实验室，广东省农业害虫生物防治工程技术研究中心，华南农业大学昆虫学系，广州 510640；2. 广西农业科学院甘蔗研究所，南宁530007
中文关键词：柑橘木虱，柑橘粉虱，内共生菌，Wolbachia，系统进化
英文关键词：citrus psyllid, citrus whitefly, endosymbionts, Wolbachia, phylogeny
中文摘要：
【目的】 探明不同地理种群的柑橘木虱Diaphorina citri Kuwayama和柑橘粉虱Dialeurodes citri Ashmead体内昆虫内共生菌的种类及其感染率，并以Wolbachia共生菌为代表，对其系统发育关系进行分析，为今后自共生菌角度研发柑橘木虱和柑橘粉虱的新型防控技术奠定基础。【方法】 以16S rDNA、23S rDNA以及wsp为目标基因，利用PCR技术检测采自于广州、湛江、南宁、桂林、厦门的柑橘木虱以及采自广州的柑橘粉虱体内共生菌的种类及其感染率；利用多位点序列分型（MLST）技术和MEGA 5.0软件对不同昆虫样本中的Wolbachia进行系统发育关系分析。【结果】 本研究采集的柑橘木虱和柑橘粉虱均含有原生共生菌Portiera和次生共生菌 Wolbachia、Cardinium、Rickettsia，但该3种次生共生菌在不同木虱与粉虱种群的感染率有所不同；Arsenophonus只在广州和湛江种群的柑橘木虱中检出。基于wsp基因及MLST基因序列的Wolbachia系统发育分析表明，华南地区柑橘木虱和柑橘粉虱体内的Wolbachia均属于Wolbachia的 B 大组Con亚组。【结论】 不同地理种群的柑橘木虱与柑橘粉虱体内感染的共生菌种类及其感染率不同；Wolabchia共生菌与柑橘木虱寄主不存在协同进化关系，在同一采集点存在Wolbachia通过柑橘寄主在柑橘木虱之间、柑橘木虱与柑橘粉虱之间水平传播的可能性。
英文摘要：
[Objectives] To survey the infection rates of different symbiotic bacteria in different geographic populations of the citrus psyllid and citrus whitefly, and to analyze the phylogenetic relationships between different Wolbachia populations in order to provide endosymbiont-based scientific support for the continuous management of the citrus psyllid and citrus whitefly. [Methods] Endosymbionts in citrus psyllids from Guangzhou, Zhanjiang, Xiamen, Guilin and Nanning, and citrus whiteflies from Guangzhou, were detected using PCR. The phylogenetic relationships of Wolbachia were analyzed using a phylogenetic tree constructed with MEGA 5.0 based on the Wolbachia wsp and MLST genes detected in citrus psyllids and citrus whiteflies. [Results] All citrus psyllid and whitefly populations in the current study were infected with the primary endosymbiont Portiera and the secondary endosymbionts Wolbachia, Cardinium, Rickettsia, but infection rates of these three endosymbionts varied between different geographical populations. Arsenophonus was only detectable in Guangzhou and Zhanjiang psyllid populations. Phylogenetic analysis revealed that Wolbachia endosymbionts in all citrus psyllid and whitefly populations were very similar and belonged to the Con group of the Wolbachia B supergroup. [Conclusion] The species and infection rates of endosymbionts varied among different geographical populations of the citrus psyllid and whitefly. There is no apparent coevolutionary relationship between Wolbachia and its psyllid hosts. Indeed, the high homology of Wolbachia in different hosts suggests plant-mediated horizontal transmission of Wolbachia between different psyllid hosts.