山东不同地区韭菜迟眼蕈蚊共生菌Wolbachia 的检测及鉴定

【目的】为了揭示山东省韭菜迟眼蕈蚊Bradysia odoriphaga Yang et Zhang种群共生菌 Wolbachia 的感染率及其分类地位，探讨该共生菌对韭菜迟眼蕈蚊的潜在影响。【方法】利用线粒体细胞色素氧化酶I（mtCOI）基因引物（LCO1490/HCO2198），通过扩增测序和序列比对对采自山东省12个地区的根蛆种群进行了分类鉴定。在上述基础上，利用 Wolbachia 的16S rDNA和 wsp 基因特异引物（分别为16S-F/16S-R和81F/691R）对鉴别出的11个韭菜迟眼蕈蚊种群体内Wolbachia 感染情况进行了PCR检测；对感染个体体内 Wolbachia 依据16S rDNA基因片段序列进行分类鉴定。【结果】山东省12个根蛆种群中，11个种群为韭菜迟眼蕈蚊种群。基于 Wolbachia 的16S rDNA基因特异引物检测结果发现，这些韭菜迟眼蕈蚊种群广泛感染 Wolbachia （感染率为6.67%~93.33%），而利用wsp 基因特异引物检测的感染率（0.00%~40.00%）相对较低些。基于 Wolbachia 的16S rDNA基因构建系统发育树表明，这些韭菜迟眼蕈蚊种群感染的Wolbachia 全部属于A组。【结论】确定了 Wolbachia 在山东省韭菜迟眼蕈蚊体内的感染率及其分类地位，为研究 Wolbachia 对韭菜迟眼蕈蚊生物学及生态学的影响奠定了基础。

Detection and identification of Wolbachia in Bradysia odoriphaga (Diptera: Sciaridae) populations from Shandong Province,China

【Aim】 This study aims to determine the infection rate and taxonomic identity of the endosymbiont Wolbachia in the populations of Bradysia odoriphaga Yang et Zhang in Shandong Province, so as to reveal the potential effects of this endosymbiont on B. odoriphaga. 【Methods】 The species identity of 12 chive maggot populations collected in Shandong was first identified based on the mitochondrial cytochrome oxidase I (mtCOI) gene. Two pairs of primers (16S-F/16S-R and 81F/691R) were used to detect the infection of Wolbachia in the 11 populations identified as B. odoriphaga. The Wolbachia infecting these B. odoriphaga populations were further identified based on the 16S rDNA sequence. 【Results】 Compared with the infection rates of Wolbachia (ranging from 6.67% to 93.33%) based on 16S-F/16S-R primers, the infection rate of Wolbachia based on 81F/691R primers was relatively low (0.00%-40.00%). Phylogenetic tree based on the 16S rDNA sequence showed that all of the Wolbachia detected belonged to group A. 【Conclusion】 This study reveals the infection rate of Wolbachia in B. odoriphaga in Shandong and their taxonomic position, and these results provide the basis for future study on the effects of Wolbachia on the biology and ecology of B. odoriphaga.