Abstract: To obtain the system of tissue culture and rapid propagation of Anoectochilus roxburghii from different origins, the tender stem segments of A. roxburghii from different origins were taken as explant, and screened optimum culture medium formula to induce shoot clusters. The results showed that: the optimum initial mediums of A. roxburghii in Fujian, Taiwan and Guangxi was MS+20 mg/L 6-BA+02 mg/L NAA+10 mg/L KT, MS+30 mg/L 6-BA+02 mg/L NAA+10 mg/L KT, and MS+40 mg/L 6-BA+02 mg/L NAA+10 mg/L KT, respectively In addition, the optimum subculture medium of A roxburghii in Fujian, Taiwan and Guangxi was MS+20 mg/L 6-BA+02 mg/L NAA for, MS+30 mg/L 6-BA+03 mg/L NAA for, and MS+20 mg/L 6-BA+02 mg/L NAA, respectively In the stage of seedling cultivation, all plants from different producing areas could be improved growth in different degrees after applying exogenous peptone Additional, the optimum rooting medium A roxburghii in Fujian, Taiwan and Guangxi was 1/2MS+15 mg/L IBA+05 mg/L NAA+20 mg/L ABT, 1/2MS+05 mg/L IBA+05 mg/L NAA, and 1/2MS+05 mg/L IBA+ 10 mg/L NAA+10 mg/L ABT1+ 15 g/L Ac Meanwhile, plantlets from test tube grew well on the growth medium of vermiculitethe and peat soil mixed at the rate of 1∶1, the rooting rate of the A roxburghii reached 90%