During investigating of shiitake mushroom competitors, 289 isolates of Trichoderma spp. were collected from shiitake mushroom farms in different districts and the Forest Mushroom Research Center of Korea, among which 29 representative strains were selected. Based on the DNA sequences of the rpb2 and tef1 genes and the ITS rDNA, and their morphological characteristics, they were identified as T. atroviride, T. citrinoviride, T. harzianum, T. longibrachiatum, and two undescribed species, Trichoderma spp. 1 and 2, which are considered to be the candidate of new species. Competition tests between Lentinula edodes (Sanjo302) and the Trichoderma species indicated that the six species of Trichoderma were significantly different from each other in terms of their ability to invade the mycelial blocks of shiitake. In both of dual cultures on potato dextrose agar and sawdust media, Trichoderma spp. 1 and 2 strongly invaded the mycelial blocks of shiitake. Our results suggest that the two Trichoderma species may cause potentially serious economic losses in shiitake cultivation of Korea.