DESIGN OF MOLECULAR MARKERS FOR THE IDENTIFICATION OF SPECIES OF THE GENUS CHIRONOMUS (DIPTERA, CHIRONOMIDAE)

Accurate identification and differentiation of species of the genus Chironomus based on their morphological features is a difficult problem. Molecular markers enable the unambiguous species identification of the individuals studied irrespective of their sex and stage of the life cycle. The polymerase chain reaction (PCR) with species-specific primers was used to develop molecular markers (amplicons) for the identification of Ch. piger, Ch. dorsalis, and Ch. pseudothummi - species of the genus Chironomus. Nucleotide sequences of the internal transcribed spacer region of the locus coding for ribosomal RNA were used to design species-specific primers for these target species. Each of the species-specific primer pairs yielded species-specific amplicons (molecular markers) only with the DNA of target species - Ch. piger, Ch. dorsalis, and Ch. pseudothummi. The test PCRs with the DNA of eighteen Chironomus species confirmed the species specificity of the designed primers. The molecular markers produced in the PCR with the designed species-specific primers enable the reliable identification of Ch. piger, Ch. dorsalis and Ch. pseudothummi and their differentiation from the other species of the genus Chironomus.