重庆酉阳分布寒露林蛙的分子鉴定 Molecular Identification of a Rana hanluica Population Distributed in Youyang County, Chongqing City in China

于2014年9月17日在重庆市酉阳县中采集到一蛙科标本，经初步形态鉴定为寒露林蛙。然而，之前文献记载寒露林蛙主要分布在湖南、江西、贵州、广西等地，尚无在重庆分布的记载，因此有必要使用分子生物学研究方法做进一步的物种鉴定。对该标本的肌肉样本进行DNA提取及PCR实验，扩增线粒体16S rRNA基因并进行系统发育分析及分子鉴定。结果表明，酉阳蛙类样本16S rRNA基因序列与3条寒露林蛙16S基因序列聚为一个单系，且与寒露林蛙模式产地湖南双牌一标本的16S rRNA基因序列共享同一个单倍型。因此，初步判断该标本属寒露林蛙(Rana hanluica)，为重庆市两栖动物一新纪录。本文还对部分林蛙类物种的有效性进行了讨论。 In September 17th, 2014, a frog specimen was collected from Youyang County, Chongqing City in China and identified as Rana hanluica morphologically. However, the Rana hanluica frogs are usually recorded in the Hunan province, Jiangxi province, Guizhou province and Guangxi prov-ince in most scientific documentation, and there is no distribution record in Chongqing City. So, it needs further species delimitation based on molecular biological methods. Then DNA extrac-tion and PCR experiments were conducted with the tissue of the specimen to amplify the mito-chondrial 16S rRNA gene for further phylogenetic analysis and molecular identification. The results showed that the 16S rRNA gene sequence from the frog specimen in Youyang was clus-tered into a monophyletic group with 3 homologous sequences of Rana hanluica. And the target sequence shared the same haplotype with a sequence from the type locality of Rana hanluica. Therefore, the Youyang frog specimen was preliminary identified as Rana hanluica. This is a new record of amphibians in Chongqing city. We also discussed the validation of some Rana spe-cies in this study.