Trichoderma spp. cause large crop losses of the cultivated shiitake mushroom, Lentinula edodes. We bred several shiitake strains that are resistant to Trichoderma spp. using di-mon mating to establish a useful method for controlling the greenmold disease. We examined the competitive ability of L. edodes against Trichoderma spp. using a dual culture system to select resistant strains. By screening Trichoderma-resistant strains, we found that among 11 parental strains, 4 strains, including KFRI 36, were confirmed resistant strains. They showed especially strong resistance to T. harzianum, which formed deadlock after mycelial contact and then invaded into the territory of T. harzianum. KFRI 171 also showed resistance to T. atroviride strains. Among 13 strains, which were made by hybridization of shiitake strains, 5 were confirmed to be resistant to Trichoderma, including KFRI 58-1. Their resistance was not correlated to the resistant activity of their parents’ strains. Two strains lose resistance and two strains acquire resistance compared to their parents’ strains. In SEM observation, the mycelium of L. edodes at the interaction zone of Lentinula-Trichoderma was rugged and swollen by T. harzianum.