Molecular identification of Herpetofauna from Punjab, Pakistan, using mtDNA genes
DOI:
https://doi.org/10.5281/zenodo.12594174Keywords:
Mitochondrial DNA, phylogenetic analysis, genetic divergence, species identification, COIAbstract
The herpetofauna diversity of Pakistan is underestimated due to the country's lack of molecular-based identification. Field surveys were conducted from August 2018 through July 2022 to collect as many as possible specimens from Punjab, Pakistan. A total of 21 species were collected and initially identified by morphological characteristics. The three gene fragments in four amphibian species and seven reptile species were successfully amplified and sequenced. A total of 18 DNA sequences of 11 species representing nine genera and five families were deposited in GenBank, and accession numbers were obtained. Furthermore, phylogenetic analysis was performed through the Neighbor-joining method using 100 bootstrap pseudo-replicates in MEGA X. Closely related toad species, namely Duttaphrynus melanostictus and Duttaphrynus stomatitis, were clearly separated in the tree inferred from Cytb gene sequences. Similarly, conspecific sequences were analyzed for multiple individuals of Platyceps rhodorachis clustered together in the tree inferred from COI gene sequences. In our findings, 16S rRNA appears to be more reliable in identifying amphibian species, while COI has a better success rate in reptile species identification. In our recommendations, molecular-based identification of herpetofauna is necessary nationwide to document any new subspecies.
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