Molecular identification of genus Duttaphrynus from Punjab, Pakistan

The Indus valley toad and common Asian toad are widely distributed toads in Pakistan. There is doubt in the taxonomic position of species within the genusDuttaphrynusin Pakistan as most of the species identified on morphology. Previously,Bufo melanostictus hazarensisidentified on morphology but during the present study, it is confirmed asDuttaphrynus melanostictus-based COI sequences (MK941836). The interspecific divergence betweenDuttaphrynus stomaticusandD. melanostictuswas 16%. The intraspecific divergence ofD. stomaticus(MK947909.1) was ranging from 0% to 1% while the intraspecific divergence ofD. melanostictus(MK941836) was high ranging from 10% to 11%. Overall, genetic variation between the species of genusDuttaphrynusbased on p-distance was 14%. In our recommendation, a large-scale molecular identification of amphibians should take into consideration for exact species identification to report any new species from Pakistan.