DNA Barcoding of Tree Frogs: Testing the Existence of Two Mitochondrial Lineages of Hyla savignyi

DNA barcoding is a popular approach that can aid in identifying species and has led to the discovery of many new amphibian species. In this study, we first performed DNA barcoding for two Hylidae family representatives (Hyla orientalis and Hyla savignyi) in the Anatolia region of Turkiye. Five species delineation tests (ABGD, GMYC, PTP, RESL, and statistical parsimony analysis) were applied, and four of them indicated that three mitochondrial lineages of Hylidae are present in Anatolia. Intraspecific genetic distances (K2P) ranged from 0.0014 to 0.0045, while the interspecific genetic distances ranged from 0.0753 to 0.1933. According to the tree topologies obtained from maximum likelihood (ML) within H. savignyi from Anatolia, two lineages can be differentiated, and further investigation is warranted as to whether these lineages represent distinct species.