泸县沼水蛙16SrRNA基因克隆与系统发育分析

Sequencing and Phylogenetic Analysis of the 16SrRNA gene of Hylarana Guentheri from Luxian County in Southern China

对采于四川省泸县地区的沼水蛙（Hylaranaguentheri）线粒体16SrRNA基因进行扩增测序。经3对扩增引物分别扩增及测序获得长度为550bp、300bp、1000bp核苷酸序列，拼接后序列长度1601bp。核苷酸相似性分析表明，序列与Genbank公布的福建的沼水蛙线粒体16SrRNA基因（KF185060）核苷酸序列相似性最高（99．8％）；与越南地区沼水蛙的16S序列相似性也较高（99％～99．7％），无明显种间差异，从遗传上证明沼水蛙为广布物种。同时，与亲源关系较近的水蛙类代表物种构建的系统发育树显示。所有沼水蛙聚为一支系，然后与Ranalateralis互为姊妹群。本文同时也对水蛙类的系统发育关系做了初步讨论。

The present paper clones and sequences the mitochondrial 16SrRNA of Hylarana guentheri from Luxian county in Sichuan province. Three nucleotide sequences （550bp, 300bp, 1000bp） have been cloned and sequenced with 3 pairs of primer were assembled into a complete 1601bp contig. The result shows that the sequence was 99.8% identical to that of another H. guentheri sample from Fujian province （KF185060）, and 99% - 99.7% to the ones from Vietnam. This result proves H. guentheri to be a widespread species geneti- cally. Further, the phylogenetic analysis including available 16S sequences of Hylarana species shows that all H. guentheri sequences clustered together as the sister group to that of Rana lateralis. In addition, the phyloge- netical relationship among Hylarana species has been discussed.