A revision of mapping of main and alternative banding sequences in chromosome III (EF) has been made for 14 species of the Chironomus plumosus group. In total, new versions of mapping are presented for 18 banding sequences of arm E and 18 banding sequences of arm F. A new way of tracing the origins of banding sequences in chromosome III of the Ch. plumosus group in comparison with basic banding sequences of the genus Chironomus is suggested. The presented data indicate that h'pluE2 in arm E and p'borF2 in arm F are the closest to banding sequences of Ch. piger Strenzke, 1959 and thus should be considered the most ancient among banding sequences of chromosome III in the Ch. plumosus group. Phylogenetic relationships of banding sequences of chromosome III are discussed.