Abstract: In order to protect the wild resource of Psammosilene tunicoides effectively, tender stems of Ptunicoides were used as explants to study the in vitro rapid propagation technique of Ptunicoides. The results showed that the method of sterilization was that tender stems were rinsed with water, then disinfect with 75% alcohol for 15s and 01% mercuric for 10min. The contamination rate was 361%. Differentiation culture medium was MS + 20mg/L 6BA + 005mg/L TDZ + 005mg/L NAA, the average quantity of differentiation bud was 349, proliferation culture medium was MS + 03mg/L 6BA + 005mg/L TDZ + 001mg/L NAA, the proliferation multiple was 429, Rooting culture medium was 1/2MS + 03mg/L IBA + 01mg/L NAA + 03g/L activated carbon, the rooting rate was 917%, the tissue culture seedlings were transplanted into the matrix (red soil∶humus∶perlite=1∶1∶1), the survival rate was 945%. The technology of tissue culture is hopeful to solve the problem of wild resource shortage and insufficient supply of raw materials in the production.