The invention provides an edible mushroom culture medium which comprises, by weight, 30-50 parts of biogas residues and biogas slurry, 30-50 parts of crop straws, 10-20 parts of grape pips and peels and a sterilizing agent with 0.5-2% of the volume-weight ratio. Hydrogen peroxide is adopted for the sterilizing agent, and the invention further provides a preparation method of the culture medium. The edible mushroom culture medium has the advantages that the grape pipes and peels comprise a large quantity of polyphenol substances and a large amount of linoleic acid and further have a certain bacteriostatic effect, so that more rich nutrition is provided for the culture medium, and the bacteriostatic effect enables the prepared culture medium not to be prone to being infected with infectious microbes; due to the fact that 35% of the hydrogen peroxide is selected and used for the sterilizing agent, the sterilizing effect is remarkable, in addition, due to the selection and use of the grape pips and peels, the culture medium is little infected with bacteria, in the process of high temperature sterilization, the culture medium is volatilized to generate water and oxygen, and no residue and contamination exist; a large amount of agricultural and stock farming waste such as the straws and the biogas residues and biogas slurry are further selected and used for the culture medium, so that reusing of the waste is achieved, and the culture medium meets the requirements of environmental protection and circular economy.