The invention discloses a preparation method of a mushroom preservative film, and belongs to the field of food fresh keeping. The method comprises the following steps: with polylactic acid, citric acid ester compounds and natural antibacterial plant essential oil as raw materials, preparing a composite film through a solvent membrane method; carrying out thermal plastic package on the composite film according to the packaging amount, so as to prepare a bag-shaped packaging bag; putting the clean fresh mushroom into the packaging bag, wherein the mushroom loading amount is 50-80% of the volume of the packaging bag; filling in fresh air and then sealing; and storing in a refrigerator with the temperature being 6+/-1 DEG C. The preservation method is simple in process, and easy to operate; the used materials can achieve biodegradation, and reduce environmental pollution caused by package; due to addition of the natural antibacterial plant essential oil, according to the preservative film used in the preservation method, the mechanical property of the preservation film is improved, the breathability of the preservation film is increased and the preservative film is relatively suitable for modified atmosphere packaging of the mushroom; meanwhile, the preservative film also has certain bacteriostatic action, and is capable of restraining growth and reproduction of microorganisms; and the preservation method is capable of effectively prolonging the storage life of the mushroom.