Previously we identified a trout myogenic factor related to MyoD. We now present a cDNA encoding a novel trout myogenic factor (TMyoD2) expressed in embryonic muscle. Nucleotide analysis and amino acid comparison showed that this cDNA encodes a MyoD-like protein of 275 amino acids that is distinct but related to TMyoD with 78% identity over the entire length. The protein sequence conservation between TMyoD2 and TMyoD was calculated to be 90% within the basic/ helix-loop-helix domain that is involved in DNA binding and heterooligomerisation. At the nucleotide level, comparison of TMyoD with TMyoD2 reveals that the translated regions are flanked by highly divergent 3' and 5' ends. The substantial differences observed at translated and especially untranslated regions strongly suggest that TMyoD and TMyoD2 mRNA originate from different loci. The TMyoD and TMyoD2 mRNA are likely transcribed from two distinct genes which were duplicated during the tetraploidization of the salmonid genome.