Abstract: For germplasm conservation and rapid propagation of Euphorbia cotinifolia, the stem-segment with axillary buds，stem tips and leaflet taken as explants to be studied for tissue culture system. The results showed that, the optimizing sterilization method for stem-segment with axillary buds in the second ten days of June was that explants could be treated with 0.1% HgCl2 for 5 min. The percentage of sterile explant could arrive at 79.5%. It was suggested that MS+KT0.5mg/L+IBA0.5 mg/L to be desirable treatment for subculture and multiplication, with 2.45 times multiplication rate after a 20-25 days cultivation. The media with MS+IBA0.5mg/L+KT(0.5~1.0)mg/L and 1/2MS+IBA (0.1~0.5)mg/L could be used for the rooting induction . The rooting rate was 91.14% . The average number of rootlet for the test-tube plantlets was 6.49~13.03.