The invention discloses a cordyceps militaris culture medium. The culture medium adopts water as a solvent. The obtained cordyceps militaris culture medium is prepared from, by weight, 30-70 parts ofmain materials, 5-20 parts of lepidopteron powder, 3-12 parts of glucose, 2-8 parts of peptone, 1-5 parts of corn starch, 0.5-5 parts of ammonia citrate, 0.5-5 parts of magnesium sulfate, 0.5-5 partsof monopotassium phosphate, 7-12 parts of hydroxyethyl cellulose and 15-30 parts of traditional Chinese medicine additives. The traditional Chinese medicine additives include, by weight, 40-55% of tribulusterrestris, 20-40% of aloe leaves and 5-25% of astragalus membranaceus. The invention further discloses a preparation method of the cordyceps militaris culture medium. The cordyceps militaris culture medium obtained according to the preparation method can effectively restrain infectious microbe growth, accelerate hypha forming, enhance cordyceps militaris sporocarp oxidation resistance and have the wide application prospect.