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The karyotypes and chromosomal polymorphism of Chironomus pseudothummi were investigated in different parts of its range. It was established that chromosomal variability in the natural populations of this species was represented mainly by the inversion polymorphism of arm G. Only rare and unique inversions were found as heterozygous in arms C, D, and E. In total, the 14 banding sequences of polytene chromosomes form the banding sequences pool of Ch. pseudothummi. Geographic differences in distribution of chromosomal banding sequences throughout the range were established. The presence of banding sequences pstG1 and pstG2 is characteristic of European populations. Banding sequence pstG1 was completely vanished with simultaneous increase in frequency of pstG2 and appearance of new inversion banding sequence pstG3 in Siberian populations. The differences in the set of the rare and unique inversions in arms C, D, and E between west-European and west-Siberian populations were revealed.