The invention relates to the technical field of agrocybe aegerita culture, particularly an agrocybe aegerita culture bag and a preparation method thereof. The culture bag is prepared from the following raw materials in parts by weight: 20-80 parts of polyethylene, 20-40 parts of polypropylene, 8-12 parts of compatilizer, 8-12 parts of toughener, 5-10 parts of heat-resistance modifier, 5-10 parts of impact modifier, 10-20 parts of flame retardant, 1-2 parts of initiator, 1-2 parts of polymerization inhibitor, 0.5-1.5 parts of lubricant and 0.5-1.5 parts of antioxidant. The agrocybe aegerita culture bag has favorable toughness and high-temperature resistance, can be attached to the culture medium at any time, and can not separate. The sterilization temperature can be enhanced to 118 DEG C. The preparation method has the advantages of simple technique, stable quality and high production efficiency, is convenient for operation control, and can implement large-scale industrial production.