The invention relates to a method for isolating edible fungus strains. The method comprises the following steps: performing exposure of a fresh aseptic tissue of a fruiting body, cutting off a block from the surface of the aseptic tissue with a sharp tip, inserting the shape tip obliquely into the tissue, performing isolation with the shape tip, allowing a slope in a test tube to be downward, pulling out a tampon, clamping the tissue block with tweezers, burning the tissue block over flame lightly and to and fro, putting the burned tissue block into the test tube, covering the test tube with the tampon, tilting the test tube finally, allowing an isolation culture medium to face downwards, shaking the tissue block gently so as to make the tissue block slide directly from the mouth of the test tube to the bottom of the test tube containing the culture medium and be stuck between the culture medium and the test tube, and then putting the test tube at a temperature of 22-28 DEG C, whereingermination of hyphae occurs in 3 days, and transfer of the strains can be conducted in 8-12 days. According to the isolating method, operation can be conducted in an open environment without specialequipment, and in addition, alcohol and other disinfectants are not needed for the surface treatment, and direct clamping with the tweezers and even burning over the flame of an alcohol lamp are conducted, so that the surface dust and hybrid bacteria are burned by the flame; the tissue block does not pass over the culture medium, so that the contamination rate is reduced greatly.