An insight into molecular taxonomy of bufonids, microhylids, and dicroglossid frogs: First genetic records from Pakistan.

The current study was focused on documentation of amphibian assemblage in North Punjab and Islamabad Capital Territory, Pakistan, by using mitochondrial gene sequences of 16S rRNA. Our study entailed 37% of the known amphibian species of the country. We provided a phylogenetic analysis based on 74 newly generated mitochondrial 16S rRNAs from nine species of genus and . We employed the maximum-likelihood inference and Bayesian analysis to assess the taxonomic status of the samples obtained from Pakistan, with respect to other congeneric species from surrounding regions. Our findings confirmed the taxonomic status of South Asian anuran species , , (synonym and in Pakistan. We have reported new country records of genus . ). was previously misidentified as due to dearth of genetic information. We provided the first genetic records of our endemic species . . The results revealed the taxonomic placement of . with respect to its congeners and validated the taxonomic status of . from its type locality (Murree) for the first time. The findings of the present study also indicated the paraphyletic relationship of with species. So, based on our phylogenetic inferences, morphological characters, and habitat preferences, validity of generic status of . is undecided. As our data were not enough to resolve this issue, we suggest sequencing of additional mitochondrial and nuclear genes in the future studies to get a better resolution. We recommend carrying out extensive surveys throughout the country for proper scientific documentation of amphibians of Pakistan. Many new species, some of them might be endemic to Pakistan, are expected to be discovered, and taxonomic status of other species would be resolved.