PCR was performed using universal primers (27F-1513R) for 16S rRNA gene of endosymbiotic bacteria from Cicadella viridis,followed by agarose gel electrophoresis,cloning,sequencing,sequence alignment and analysis.The phylogenetic trees were generated by the Bayesian and maximum-likelihood methods.The results showed that Sulcia sp.was classified as Flavobacteriaceae,Flavobacteriales,Flavobacteria Bacteroidetes.The 16S rRNA gene sequences of Sulcia sp.got 13 new haplotypes and 14 variable sites,and identity rate and genetic distance of which were 99.40％-99.80％ and 0.001-0.004,respectively.The phylogenetic tree revealed that Sulcia sp.was associated with Sulcia muelleri detected from the leafhoppers of Cicadellinae.Sulcis sp.had a low degree of variation,and all haplotypes belonged to Sulcia muelleri species.Sodalis sp.belonged to Enterobacteriaceae,Enterobacteriales,Gammaproteobacteria,Proteobacteria.The 16S rRNA gene sequences of Sodalis sp.obtained 5 new haplotypes and 20 variable sites in this study,and identity rate and genetic distance of which were 96.10％-99.70％ and 0.002-0.040,respectively.The phylogenetic tree showed that Sodalis sp.was most closely related to Sodalis glossinidius detected from the Tsetse flies.Sodalis sp.has a high degree of variation,and all haplotypes belong one genus.%用细菌通用引物27F-1513R对大青叶蝉内共生菌的16S rRNA基因进行PCR扩增,琼脂糖凝胶电泳,克隆、测序、序列比对与分析,用贝叶斯法和极大似然法构建系统发育树.结果表明:从大青叶蝉体内鉴定出Sulcia muelleri和Sodalis sp.2种内共生菌,Sulcia muelleri隶属于拟杆菌门Bacteroidetes黄杆菌纲Flavobacteria黄杆菌目Flavobacteriales黄杆菌科Flavobacteriaceae,该内共生菌获得13个新单倍型,14个变异位点,其序列间的一致率为和遗传距离分别为99.40％-99.80％和0.001-0.004;系统发育树显示,Sulcia muelleri与大叶蝉亚科中检测到的Sulcia muelleri聚在一起,其变异程度低,所有单倍型为同一个种.Sodalis sp.属于变形杆菌门Proteobacteria伽马变形菌纲Gammaproteobacteria肠杆菌目Enterobacteriale肠杆菌科Enterobacteriaceae,本研究获得5个新单倍型,20个变异位点,其序列间的一致率为96.10％-99.70％,遗传距离范围为0.012-0.040;系统发育树显示,Sodalis sp.与采采蝇检测到的Sodalis glossinidius聚在一起,其变异程度相对较大,属于同一个属.
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