Molecular phylogeny and cryptic species identification of the genus Triplophysa in the freshwater of Malakand division, Pakistan

Triplophysa is an ecologically and commercially important genus of freshwater fish consisting of cryptic species with uncertain taxonomic status. The current study was the first report on the molecular phylogeny of the genus Triplophysa in the freshwater of Malakand Division, Pakistan. The mitochondrial gene cytochrome c oxidase subunit I (COI) was used for the identification of cryptic species in the genus Triplophysa and for finding out the molecular phylogeny of the identified species. Fish specimens were collected, and DNA was extracted, then amplified using genes specific primers. The PCR results were sequenced, and consensus sequences were created using BioEdit software. The consensus nucleotide sequences of each Triplophysa species were BLAST, and each query sample showed similarities to gene bank sequences. The three species of Triplophysa (Triplophysa microps, Triplophysa naziri, and Triplophysa choprai) were identified, and their phylogenetic analysis was carried out using MEGA software. The mitochondrial gene sequences of the three recognized species (T. naziri, T. microps, and T. choprai) have been uploaded for the first time to the GenBank database. These findings will be an addition to the indigenous ichthyofauna of Pakistan and will act as a reference library for future studies.