A preparation method of an oudemansiella raphanipes liquid strain comprises the steps that firstly, a culture medium is prepared, wherein 1 kg of the culture medium serves as a benchmark, the culturemedium contains 300-400g of potato juice, 100-200 g of grape juice, 30-50 g of agar, 0.5-1 g of magnesium sulfate, 3-6 g of paper mulberry fermented powder and balance water; secondly, the culture medium is sterilized, wherein after being filtered, the culture medium obtained in the first step is placed in a sterilization device to be sterilized, and then is cooled after sterilization; thirdly, astrain is inoculated, wherein oudemansiella raphanipes mycelia are inoculated in the culture medium obtained in the second step, and then the culture medium is placed on a shaking bed for oscillationculture; fourthly, fermentation culture is conducted, wherein the liquid strain obtained after oscillation culture in the third step is placed in a fermentation tank for fermentation culture. According to the method, the liquid culture medium prepared from the potato juice, the grape juice, the agar, the magnesium sulfate and the paper mulberry fermented powder is used, the components work together to provide various nutrients for growth of the mycelia, the growth speed of the mycelia can be high, the biomass of the mycelia is 2.3 g/ ml or above, and the mycelia are thick and a little amber.