Abstract: In order to establish a rapid propagation system for Dendrobium cariniferum Rchb. f., the immature seeds were selected as explants to study the suitable medium for for tissue culture, and mediums aseptic seed germination. Meanwhile, the appropriate multiplication conditions of protocorm and buds were selected subculture of protocom, and the rooting medium and transplanting matrix were also screened. The results showed that the medium of MS+6-BA 1.0 mg/L+NAA 0.1 mg/L+ sucrose 20 g/L had a significant effect on seedlings germination and protocorm induction of D. cariniferum. The medium of MS+6-BA 1.0 mg/L+NAA 0.1 mg/L+sucrose 20 g/L+activated carbon 1.0 g/L was suitable for protocorm proliferation and shoot induction, and the propagation coefficient reached 5.8 times after cultured 90 d. While the optimal medium for rooting was MS+6-BA 1.0 mg/L+KT 0.5 mg/L+NAA 0.2 mg/L+sucrose 30 g/L+banana mud 100 g/L+activated carbon 1.0 g/L, the rooting rate reached 100% and the roots had higher quality and quantity after cultured 90 d. The optimum time for the transplantation of root regeneration plants was March and April in Guilin area, and sphagna was suitable as media for transplanting and domestication, and the rooted plants had a survival rate of 76.7%.