Abstract: GM256 (Malus domestica Borkh) was a cold- hardy and dwarfing apple rootstock, having high commercial value in production. But its stem cutting was difficult to root, hampered its use in production. In order to propagate it rapidly, tissue culture method was employed. The effects of basal medium, the kinds and concentrations of phytohormones on proliferation and growth of in vitro shoots were examined. The effect of dark duration on in vitro shoots rooting was investigated. The results showed that the optimal basal medium for proliferation was QL. In vitro shoots growth for proliferation and elongation was best on QL with 0.5 mg/L BA and 0.05 mg/L IBA. Dark culture significantly improved rooting percentage and root number per plantlet than direct light. Rooting percentage and root number was no significant different between 5 days and 10 days in the dark. But rooting plantlets obtained from 5 initial dark days was healthier and stronger than that 10 initial dark days, transplanting survival rate of healthy plantlets was above 85% . The optimal proliferation medium and optimal rooting condition obtained in this study could be used for large scale in vitro propagation in factory of GM256.