The invention discloses an acer palmatum Sangokaku tissue culture propagation process which comprises the following steps: S1, explant sterilization; S2, inoculation and propagation, namely inoculating a sterilized stem section with a bud to a culture medium for growth, culturing for 2-3 weeks, and inducing the germination of an axillary bud; S3, enrichment subculture, namely shearing the germinated axillary bud, inoculating on a subculture medium, culturing for 3-5 weeks, and inducing the generation of caespitose buds; S4 rooting culture, namely cutting adventitious buds positioned on the caespitose buds, inoculating on a rooting culture medium, culturing for 2-3 weeks, and inducing the generation of a root. The acer palmatum Sangokaku tissue culture propagation process disclosed by the invention can be used for building a Sangokaku tissue culture propagation system, and realizing the breakthrough of the Sangokaku tissue culture propagation system, is low in production cost, high in propagation coefficient, short in propagation period, an effective way for the fast propagation of acer palmatum Sangokaku, and capable of preventing the virus accumulation, and achieving the easiness for domesticated seedling survival, and by using the process, the issue culture seedlings with consistent inheritable characters and the sterile acer palmatum Sangokaku seedlings can be relatively easily obtained.