The invention relates to the technical field of fungus cultivation, in particular to a sterilization method. The sterilization method of an edible fungus culture medium comprises the following steps: placing the edible fungus culture medium into a sealed container and vacuuming the sealed container; maintaining the temperature of the container at 90 DEG C - 100 DEG C at least 15 minutes; increasing the temperature of the container to 105 DEG C - 110 DEG C and maintaining at least 15 minutes; increasing the temperature of the container to 123 DEG C and maintaining at least 30 minutes. A multi-step sterilization process is provided in the sterilization method of the edible fungus culture medium, sterilization effect is improved through multi-step sterilization and therefore culture medium sterilization is achieved. Every step is carried out in a vacuumed condition, vacuum degree is increased and vacuum effect is improved.