5 kinds of proliferation medium formulations,4 kinds of rooting medium formulations,3 kinds of substrate compositions were used to research the technology of multiplication,rooting and transplanting technology of Prunus humilis tissue culture seedlings.The results showed that the multiplication effect was the best in the medium of MS + 6BA 1.0 mg/L + NAA 0.2 mg/L.The rooting effect was the best in the medium of 1/2MS + NAA 0.2 mg/L.The effect of using sand: peat: garden soil=1:1:1 as substrate was the best.This provided the references for the establishment of cultural system of in vitro Prunus humilis.