我国西部6省9地牛带绦虫rDNA-ITS1序列测定及分析

目的利用分子生物学技术鉴定我国6省9地是否存在牛带绦虫亚洲亚种。方法取成虫标本孕节2~3片,酚氯仿法提取DNA,PCR法扩增rDNA-ITS1片段,并纯化、克隆此片段后作序列测定。利用BioEdit,clustalx,PHYLIP,Treeview软件处理后构建系统发育树。结果广西融水(RS)、宾阳(BY)牛带绦虫标本与贵州都匀(DY),云南大理(DL)牛带绦虫标本序列基本一致,同源性为98%~99%;新疆乌什(WS)、西藏拉萨(LS)、内蒙古(NM)、云南西双版纳(BN)牛带绦虫标本与贵州从江(CJ)牛带绦虫标本序列基本一致,同源性为98%~99%。系统发育树显示:RS、BY、DL和DY标本遗传距离较近;WS、LS、CJ、NM和BN标本遗传距离较近。而两组间遗传距离较远。结论RS、BY、DL和DY四地存在牛带绦虫亚洲亚种;WS、LS、CJ、NM和BN五地存在传统牛带绦虫。

Detection and analysis on the sequence of internal transcribed space-1(ITS1) in ribosomal DNA of Taenia saginata from nine regions of six provinces in western China

Using molecular biology techniques,it was attempted to determine whether the subspecies of Taenia saginata exists in western part of China,gravid proglottids were obtained from adult worms of Taenia saginata collected from 9 regions of 6 provinces in western China.,DNAs of worms were extracted with phenol-chloroform,and the rRNA-ITSI was amplified by PCR. Meanwhile,the sequence assay was made after cloning of rRNA-ITSI and analysis with BioEdit,Clustalx,PHYLIP and Treeview softwares to establish the phylogenetic tree. It was found that the sequences of rRNA-ITSI in samples of T saginata collected from Rongshui and Bingyang of Guangxi were consistent genetically with those collected from Duyun of Guizhou and Dali of Yunnan with the homologies of 98%-99%;while those collected from Wushi of Xingjiang,Lasa of Tiber,Inner Mongolia and Xishuangbanna of Yunnan were consistent with that of Congjiang of Guizhou genetically also with the homologies of 98-99%.From these observations,it is concluded that T saginata in Lasa,inner Mongolia and Xishuangbanna were identical genetically to Wushi and Congjiang. It is probable that the subspecies of T saginata asiatica exist in Rongshui,Bingyang,Dali and Duyun 4 districts,while T saginata collected in other 5 districts are of the classical T saginata.