Truffles are ectomycorrhizal fungi that belong to the genus Tuber. They exhibit symbiotic relationships, particularly with oak (Quercus spp.) and hazel (Corylus spp.) trees. We performed an inoculation using a spore suspension to synthesize mycorrhizae between European truffles, Tuber borchii and Tuber melanosporum, and an indigenous oak species, Quercus acutissima. This resulted in the formation of mycorrhizae within 2 months after inoculation. Despite having the same host plant, differences in features were observed between Tuber species, including color and mantle type. These results indicate that Q. acutissima is a suitable host plant for truffle cultivation in Korea and provide a better understanding of the mycorrhization of T. borchii and T. melanosporum.