In order to establish an efficient plant regeneration system,young stem segments of Dendrobium officinale Kimura et Migo were used as explants,and the effects of plant growth regulators on the protocorm-like body（PLB） induction,proliferation and differentiation were studied. The results showed that the optimum medium for PLB induction was MS + 6-BA 1.0 mg/L + IBA 1.0 mg/L + mashed potato 60 g/L + acticarbon 0.5 g/L + sucrose 25 g/L + agar 6.5 g/L,and the PLB induction rate was 97.78% after cultured for 45 d. The optimum medium for PLB proliferation and differentiation was MS + 6-BA 2.5 mg/L + IBA 1.0 mg/L,the proliferation multiple of PLB reached 7.94 after cultured for 30 d,and the induction rate of cluster bud was 97.33% after cultured for 60 d. The optimum rooting medium was MS + NAA 0.5 mg/L,and the rooting rate was 100% after cultured for 30 d.