The invention relates to a mushroom secondary punching cultivation method. The method includes the specific steps that firstly, fungus bags are prepared according to a traditional method; then hairy fungus inoculation and chamfer transfer are conducted, secondary punching is conducted, punching positions are located at mycelium growth positions, punching is conducted in a shape like a Chinese character 'pin', the intervals between holes are 5-8 cm, no overlapping exists during secondary punching, the fungus bags are pierced, but mycelia are not punctured, and illumination intensity, the temperature of 12-18 DEG C and the humidity of 80% to 90% in a mushroom producing chamber need to be kept after the fungus bags are punched; after secondary punching, part of the mycelia continue to grow and change colors, part of the mycelia start to be twisted, fungus buds are formed, and at the moment, formation of mushroom fruiting bodies can be promoted through a wood beating mushroom inducing method; mushrooms can be harvested in real time 40 days later. The novel mushroom cultivation method is provided and is simple, feasible and easy to operate; through two times of punching for exhausting, the mycelia can be promoted to grow, change colors and produce the mushrooms rapidly, the growth period of the mushrooms is greatly shortened, and the production efficiency of the mushrooms is improved.