With orthogonal experiment design,a systematic research of tissue culture and efficient regeneration system of Betula luminifer was carried out.The appropriate medium for differentiation of stem explants was MS+0.50 mg·L^-1 6-BA+0.10 mg·L^-1 TDZ+30 g·L^-1 sugar+5.50 g·L^-1 agar,and the suitable rooting medium was 1/2MS+1 mg·L^-1 IBA+20 g·L^-1 sugar+5.50 g·L^-1 agar.The survival rate reached to 90%.These results could lay a good foundation for multipropagation and genetic transformation of B.luminifera.