Green mold is a serious disease of the cultivated mushroom causing losses in production of economical importance. In the present study, digestion of a Th444 amplicon with endonuclease BseGI was useful to discriminate Trichoderma aggressivum f. aggressivum (T.a.f.a) from the T. aggressivum f. europeanum (T.a.f.e.). The informative restriction fragments of 260 and 300 bp were revealed in the corresponding reference strains T.a.f.a. and T.a.f.e. The 300-bp marker was found in all 28 Polish mushroom isolates tested.