Among the Palearctic species, butterflies of the genus Melanargia are known for their black and white wing patterns. The morphological character polarization of this genus is full of varying combinations of subgenus, species complex, and subspecies status. Its taxonomy is open to debate, especially in species and subspecies categories, with definitions mostly based on wing color. In recent years, cryptic species, phenotypically masked species, and species with intense intraspecific variation have been identified through the determination of lineages under the leadership of molecular systematics. The mtCOI gene, which is especially described as a species signature, is an important DNA barcode used for Lepidoptera. In the presented study, the mtCOI gene sequence of the populations of Melanargia larissa, M.grumi, M.hylata, M.syriaca, and M.russiae species in the southeastern Anatolia region was determined for the first time. To determine the boundaries of these species, gene characterization and genetic distances were carried out according to the Kimura-2 Parameter, and putative species analyses were carried out by the ABGD method. Trees were constructed with Maximum likelihood and Bayesian inference algorithms to determine the phylogenetic relationships between species of the genus. In light of these analyses, it has been shown that the genetic distance of morphological species M. larissa, M. grumi, M.hylata, and M. syriaca is not at the species level and that M.larissa maintains its species status according to the principle of priority. In addition, the M.russiae population presented in this study forms a monophyletic clade with other populations of the same species in the phylogenetic tree, proving that this taxon is a stable species.
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