Abstract: To establish an effective and rapid propagation method for Dendrobium officinale, sterile stem cuttings from the plant were used as the explants for the study. Effects of cultivation media, growth regulators, sucrose concentrations, and additives on the protocorm induction, proliferation and differentiation, as well as the rooting of the plantlets were investigated in an orthogonal experiment. The results showed that the optimal medium for the protocorm induction consisted of 1/2MS+6-BA4 mg·L^-1+NAA0.1 mg·L^-1; that for the proliferation, MS+30 g·L^-1; that for the differentiation, MS+10 g·L^-1sucrose+10% potato slurry; and, that for the rooting, 1/2MS+NAA1 mg·L^-1+10% potato slurry+10% banana slurry. The method appeared to enable a successful regeneration of D. officinale plantlets through tissue culture propagation.