The invention discloses a culture medium of needle mushrooms and a preparation method thereof, and relates to the technical field of culturing of edible mushrooms. The culture medium is prepared fromthe following raw materials in parts by weight: 20 to 30 parts of peony seed husks, 35 to 45 parts of wood chips, 15 to 25 parts of sweet corn cobs, 6 to 12 parts of sweet potato residues, 5 to 10 parts of grape skin, 3 to 7 parts of vinasse, 5 to 10 parts of bran, 3 to 7 parts of tea residues, 1.6 to 2.4 parts of limes, 0.2 to 1 part of peanut shell based water retaining agent, 0.3 to 0.7 part ofpotassium dihydrogen phosphate and 0.2 to 0.6 part of zinc sulfate. The culture medium of the needle mushrooms prepared by the method disclosed by the invention contains many carbon nitrides and trace elements, so that growth of mycelium and fruiting of the mushrooms can be promoted; by adding the peanut shell based water retaining agent, a wet environment is provided for growth of the needle mushrooms for a long time, influence on growth of the mycelium and a fruiting body by dehydration or lack of water of the culture medium is avoided; moreover, the culture medium is used for culturing theneedle mushrooms, so that the output and quality of the needle mushrooms can be obviously improved; the obtained needle mushrooms are rich in nutrition, and the content of the contained saccharide, crude proteins, amino acid and the like is higher.