Abstract: In order to obtain the appropriate growth regulators for flowering in vitro of Dendrobium huoshanense, laboratory experiments were conducted to investigate the collaboration effects of growth regulators (6-BA, 2,4-D, NAA, Ad, PP₃₃₃ and TDZ) on the flowering in vitro of D. huoshanense. The results showed that growth regulator combination increased significantly the flower bud of D. huoshanense, and the optimum treatment was 0.5 mg/L 6-BA 0.2 mg/L 2,4-D, with the 35% flower bud induction rate, 20% flower rate and 8.9% normal flowers in 120 days. The next best treatment was 0.5 mg/L 6-BA 0.2 mg/L NAA, with the 24.4% flower bud induction rate, 12.8% flower rate and 5.0% normal flowers.