Background. The formation of the subdivided genetic pattern is key to the successful adaptation of population. Consequently work on the conservation and restoration of protected steppe snail Helicopsis striata (Müller 1774) should be based on the analysis microprotrusions differentiation of population gene pools. Materials and methods. The genetic structure of six populations of H. striata was studied in a variety conditions of the Southern Mid-Russian Upland. In each population collected snails from five locations, located at an equal distance from each other (150-200 m). As the genetic markers used six polymorphic loci of allozymes (polyacrylamide gel electrophoresis) and DNA loci (ISSR-PCR). Results. In general, on the Southern Mid-Russian Upland H. striata is characterized by a different degree of the spatial differentiation population gene pools (for example, according to the ISSR-loci 0,078 ≤ Фst ≤ 0,251). In comparison with the other investigated species of land snail in the study area H. striata is distinguished by the high degree of а differentiation population gene pools. It is shown that populations with a high degree of genetic structure found in the biotopes with different levels of the landscape fragmentation. Conclusion. Based on these results it can be stated that an important role in shaping the populations genetic structure in H. striata is not so much the level of diversity environmental conditions, as the degree of migratory activity individuals, which was the highest in genetically rich populations of the basin Seversky Donets river. At the same time the biology of this snail decrease the speed of its active and passive migration, which probably leads to increased isolation of populations. The highest probability of extinction, in our opinion, are isolated populations of H. striata with low genetic variability.