. The article presents data on the new location of scorpions of the genus Mesobuthus in the Orenburg Nature Reserve. The aim of the work is to determine the species on the basis of the identifi ed morphological features of populations living in the Southern Urals (the territory of the Orenburg Nature Reserve, Orenburg region, Kuvandyksky Mountains. the district, the vicinity of Aituar village) and phylogenetic relationships with populations living on the territory of the Lower Volga region (Krasnoarmeysky district of the Saratov region, Kamyshinsky district of the Volgograd region, Akhtubinsky district of the Astrakhan region) based on a comparison of nucleotide sequences of COI gene fragments. DNA was isolated from metasomal segments and sequenced using the Sanger reaction. Multiple analysis was performed using the tools of the MEGA X software package. The data obtained by the authors in the course of their work contribute to the development of ideas about phylogenetic relationships between populations of scorpions of the genus Mesobuthus living in the Palearctic in the territories of the Sakhar-Gobi and Scythian steppe regions. In the course of the study, specifi c morphological features were identifi ed for the Orenburg scorpion population, such as: the number of pectin teeth, the color of the chelicerae of the shell and tergite; the shape of the telson. Thus, the data obtained during the multiple analysis of the decoded DNA sequences confi rm the presence of closely related links between the Lower Volga and South Ural scorpion populations. The distinctive features identifi ed during morphological analysis are of interest for further comparative morphological studies in the framework of studying the phylogeny of the genus Mesobuthus.