The invention relates to a method for preserving strains of ectomycorrhizal fungi. The method is characterized by comprising the following steps of: a, placing spores or sporocarp tissues of ectomycorrhizal fungi in an agar culture medium to culture mycelium; b, inoculating the agar mycelium into a liquid culture medium to culture; c, homogenizing the cultured mycelium through a homogenizer, and inoculating into the liquid culture medium to culture once again until the mycelium accords with the requirement; and d, transferring the cultured mycelium pellet into a screw cuvette or is sealed andpreserved together with a culture container. Various ectomycorrhizal fungi strain preservation experiments prove that: the method is easy and efficient in operation, has high efficiency, is suitable for popularization and application in preservation of massive ectomycorrhizal fungi strains, and has remarkable economical benefit.