微卫星锚定PCR研究湖北钉螺的遗传变异

STUDIES ON THE GENETIC VARIATION OF ONCOMELANIA HUPENSIS WITH SSR-PCR

目的 对中国不同自然隔离群的湖北钉螺进行遗传变异研究 ,并为其种及种下分类提供依据。 方法 采用微卫星锚定 PCR(SSR- PCR)技术对中国 7省 15地湖北钉螺基因组 DNA进行 PCR扩增 ,根据扩增产物 ,计算遗传距离(D) ,并构建系统进化树。 结果 各地域标本 PCR扩增产物均呈多态性 ,其中中国大陆与中国台湾湖北钉螺之间的遗传距离为 0 .35 1± 0 .0 4 8(0 .2 6 3～ 0 .4 4 5 ) ;中国大陆湖北钉螺山区型之间的遗传距离为 0 .0 4 3;湖区型之间的遗传距离为0 .0 0 0～ 0 .2 17;山区型与湖区型之间的遗传距离为 0 .182～ 0 .30 8;来自湖北省境内 9个地域的湖北钉螺 ,其遗传距离为0 .0 0 0～ 0 .16 7。 结论 根据所采集的湖北钉螺标本和 SSR- PCR的实验结果 ,认为中国大陆湖北钉螺在基因分类水平上至少可分为不同层次的 4类 :1云南洱源、四川天全为一类 ;2安徽贵池、湖南岳阳、湖北荆门、江西新建为一类 ;3湖北武汉、湖北钟祥、湖北阳新、湖北松滋、湖北沙市、湖北石首、湖北应城为一类 ;4湖北赤壁单独为一类。其中 1与 2 34在总体上可分为两大类 ,2 34又可再分为 2 3与 4两类 ,2 3还可再分为单独的两类。此外 ,台湾钉螺为单独的一类。

Objective To detect genetic variation of different isolates of Oncomelania hupensis in China. Methods The simple sequence repeat anchored PCR (SSR PCR) technique was performed to amplify genomic DNA. The genetic distance was measured and dendrogram was constructed. Results The SSR PCR products were polymorphic. The genetic distance between the Chinese Taiwan isolate and the Chinese mainland isolates of O. hupensis was 0.351±0.048(0.263-0.445); The genetic distance between the Chinese mainland isolates of O. hupensis from hilly regions, Yunnan Eryuan and Sichuan Tianquan, was 0.043; The genetic distance among the Chinese mainland isolates of O. hupensis from lake regions was 0.000-0.217; The genetic distance between the Chinese mainland isolates of O. hupensis from hilly regions and that from lake regions was 0.182-0.308; The genetic distance among the Chinese mainland isolates of O. hupensis from the same province, Hubei Province, was 0.000-0.167. Conclusion The Chinese mainland strain of O. hupensis can be divided into four groups in different level: ①Yunnan Eryuan and Sichuan Tianquan;②Anhui Guichi, Hunan Yueyang, Hubei Jingmen and Jiangxi Xinjian;③Hubei Wuhan, Hubei Zhongxiang, Hubei Yangxin, Hubei Songzi, Hubei Shashi, Hubei Shishou and Hubei Yingcheng;④Hubei Chibi. In addition, the Chinese Taiwan isolate can be divided a different group.