The population structure of endemic species Sergentia baicalensis (Diptera, Chironomidae) from Lake Baikal was studied using the first subunit of the cytochrome C oxidase mitochondrial gene (Col). Two populations inhabiting different basins of this lake, the southern-central and northern, were detected. It was confirmed that the divergence time of this species was dated to Late Miocene (9.53 ± 3.9 Mya), during the period when geographically separated basins existed in the Baikal rift zone.