Abstract: The stem segments from elite Toxicodendron vernicifluum tree were used as explants to establish the technical system of tissue culture. The disinfection and browning protection of stem segments, the medium of axillary bud startup, adventitious buds proliferation and seedlings rooting, and the conditions of plantlet transplanting were studied. It was found that the best disinfection method of explants was to be successively soaked in 75% ethanol for 20 s, 0.05% benzalkonium for 1 min, and 0.1% mercuric chloride for 20 min, and the explants was pollution-free. White medium supplemented with 100 mg/L cysteine could reach the lowest browning rate (1.15%). The optimum medium for axillary bud startup was White + 100 mg/L cysteine + 2.0 mg/L 6-BA + 0.5 mg/L NAA, with a germination rate of 91.67%. The optimum medium for seedling proliferation was MS + 0.5 mg/LZT + 1.0 mg/L 6-BA + 0.1 mg/L TDZ, with a multiplication coefficient over 4.89. Culture medium for seedlings rooting was 1/2 MS + 0.05 mg/L IAA + 0.2 mg/L IBA, with a rooting rate of 98.89% and an average root number per plantlet of 3.98. The survival rate of complete plantlets was more than 80% when transplanted into the medium made up 50% humus and 50% river sand.