Caucasia is a global biodiversity hotspot, rich in amphibians, including several endemic species. We sequenced samples from Parsley frogs (genus Pelodytes) across their Anatolian range to generate a barcode reference database and to assess patterns of genetic diversity in the species. Different species delimitation methods (ABGD, ASAP, GMYC and PTP) were applied to assess species diversity in the genus Pelodytes based on published and newly obtained mtDNA sequences. A majority of the species delimitation tests (ABGD, GMYC and ASAP) recovered four taxonomic units corresponding to currently accepted taxonomy (P. atlanticus, P. caucasicus, P. ibericus and P. punctatus). PTP, on the other hand, recovered only two taxonomic units, one combining the three Iberian taxa (P. atlanticus, P. ibericus, and P. punctatus), and the other, P. caucasicus. In Anatolia, individuals from Giresun and Trabzon were found to be genetically closer to each other compared to those from Rize and Artvin, based on genetic distances and phylogenetic and haplotype network analyses.