Preparation method of moelleriella protoplast

The invention relates to a preparation method of moelleriella protoplast. Healthy and strong mycelia on a PDA thallus culture medium are inoculated into a PDB liquid culture medium; shake cultivationis performed for 36 to 54h at 150 rpm and 28 DEG C; then, the mycelia are filtered by four layers of sterile gauzes; a sterile mortar is used for grinding the mycelia; the ground mycelia are put intothe PDB culture medium to be subjected to shake cultivation for 16 h at 100 rpm and 28 DEG C. Then, the mycelia are filtered out by the four layers of sterile gauzes, then, the mycelia are transferredinto a sterile triangular bottle by a sterile medicine spoon; the mycelia is pretreated for 30 minutes by 10 ml of 0 to 0.08 percent beta-mercaptoethanol; 20 to 26.6 mg of fungal wall degrading enzymes (enzyme liquid pH is 5.6 to 6.2) are used for performing enzymolysis for 2.4 to 3.3h at 24 to 30 DEG C; when 0.4 to 0.7 mol/L of solution is used as an osmotic pressure stabilizing agent, the protoplast quantity reaches 2.2