Molecular identification of a Microhyla heymonsi population distributed in Majiang, Guizhou province in China. Molecular identification of a Microhyla heymonsi population distributed in Majiang, Guizhou province in China

Microhylidae is one of the most speciose groups of anurans. Their tadpoles are difficult to distinguish morphologically because of their small size and unclear differences among species. In this study, DNA was extracted from the Microhylidae tadpoles from Majiang County, Qiandongnan Prefecture, in Guizhou Province. Then the mitochondrial 16S rRNA genes were amplified by PCR technology, and preliminary molecular identification was carried out by referring to the phylogenetic relationship of homologous sequences from Genbank. The results showed that the 16S rRNA gene fragments in this study and 27 Microhyla heymonsi homologous sequences were clustered into a monophyletic lineage. The genetic distances among the sequences of the monophyletic group was smaller than the interspecific ones based on the all sequences used in this study. Therefore, they were preliminarily identified as the tadpoles of Microhyla heymonsi. At the same time, the Microhyla heymonsi lineage was further divided into 6 distinct haplotypes nets. The genetic distances among the sequences of the monophyletic group was higher than the intraspecific ones based on the all sequences used in this study, implying that there was large lineage splitting and divergence within this species. In addition, a "suspicious sequence" in the reference sequences used in this study was preliminarily discussed.