The invention discloses a preparation method of a potato culture medium. The preparation method comprises the following steps: putting sawdust, sand, peat, vermiculite and rural soil in a dryer and drying for 15-25 minutes in a drying environment of 50-70 DEG C; adding the dried sawdust, sand, peat, vermiculite and rural soil in a mixer and mixing for 5-8 minutes at a mixing speed of 30-60 r/minute to obtain a mixture a; adding a potash fertilizer in the mixture a, wherein the stirring speed is 60-80 r/minute, and the stirring and mixing time is 1-3 minutes; sequentially adding a nitrogen fertilizer, a phosphorus fertilizer, ferrous sulfate, a zinc fertilizer, a magnesium fertilizer, sodium molybdate, chicken manures and cow manures in the mixture a according to the process of adding the potash fertilizer, wherein the stirring speed is 60-80 r/minute, and the stirring and mixing time is 1-3 minutes. The preparation method of the potato culture medium, disclosed by the invention, has the benefits that the culture medium prepared by the preparation method is specially used for potato cultivation, soil is loose, the water-retaining property is good, an organic fertilizer is also contained, wastes are utilized, the cost is reduced, and the yield of last potatoes is high; the preparation method is less in process, fungus is also introduced into the culture medium, so that the use ofthe fertilizers can be greatly reduced.