【作者】 董世楠； 朱峰； 徐莉； 沈中元；

【Author】 1Dong Shinan1,3,Shen Zhongyuan*2,3,Xu Li2,3,Zhu Feng1,3 (1.College of Biotechnology and Environmental Engineering,Jiangsu University of Science and Technology,Zhenjiang 212018,China;2.Sericultural Research Institute,Chinese Academy of Agricultural Sciences,Zhenjiang 212018,China;3.Key Laboratory of Genetic Improvement of Silkworm and Mulberry,Ministry of Agriculture,Zhenjiang 212018,China)

【机构】 江苏科技大学生物与环境工程学院； 农业部蚕桑遗传改良重点开放实验室； 中国农业科学院蚕业研究所；

【摘要】 对家蚕微粒子虫镇江株(Nosema bombycis Zhenjiang)、柞蚕微孢子虫(Nosema antheraeae)、棉铃虫微孢子虫(Nosema heliothidis)、从菜粉蝶(Pieris rapae)分离的微孢子虫、从蓝叶虫(Phyllobrotica armta)分离的微孢子虫、从桑尺蠖(Hemerophila atrilineata)分离的微孢子虫以及从家蚕分离的微孢子虫的SSU rRNA基因进行PCR扩增、克隆和测序,得到该7种微孢子虫的SSU rRNA基因的全序列。采用Phylip软件的邻位相连法(Neighbour-Joining)和MEGA4.0软件的UPGMA法,以微孢子虫的SSU rRNA基因序列构建系统发育树,通过对新微孢子虫的聚类分析,以及基因长度、G+C含量、相似性和分化度等方面的分析,确定了4种新分离的微孢子虫的分类地位。研究结果表明,从菜粉蝶分离的微孢子虫、从蓝叶虫分离的微孢子虫和从桑尺蠖分离的微孢子虫与微粒子属(Nosema)微孢子虫较近,因此,将这3种微孢子虫归为Nosema属,暂分别定名为Nosema sp.PR、Nosema sp.PA、Nosema sp.HA;从家蚕中分离的微孢子虫与内网虫属(Endoreticulatus)微孢子虫关系相近,将其归为Endoreticulatus属,暂定名为Endoreticulatus sp.Zhenjiang。研究结果同时显示,用微孢子虫的SSU rRNA基因分析微孢子虫的进化关系及其分类地位是可行的。更多还原

【Abstract】 The complete SSU rRNA (small subunit rRNA) gene sequences of seven microsporidia including Nosema bombycis Zhenjiang,Nosema antheraeae,Nosema heliothidis,and four new microsporidia isolated from Pieris rapae,Phyllobrotica armta,Hemerophila atrilineata and Bombyx mori respectively,were obtained by PCR amplification,cloning and sequencing.Two phylogenetic tree based on SSU rRNA sequences had been constructed by using Neighbour-Joining of Phylip software and UPGMA of MEGA4.0 software.The taxonomic status of four new microsporidia were determined through analysis of phylogenetic relationship,length,G+C content,identity and divergence for the SSU rRNA sequence.The results showed that the microsporidia isolated from Pieris rapae,Phyllobrotica armta and Hemerophila atrilineata have close phylogenetic relationship with the Nosema,while another microsporidium isolated from Bombyx mori is closely related to the Endoreticulatus.So,we temporarily classify three species of microsporidia to genus Nosema,as Nosema sp.PR,Nosema sp.PA,Nosema sp.HA.Another is temporarily classified into genus Endoreticulatus,as Endoreticulatus sp.Zhenjiang.The result indicated as well that it is feasible and valuable to elucidate phylogenetic relationships and taxonomic status of microsporidian species by analyzing information from SSU rRNA sequence of microsporidia.更多还原