Abstract: In order to resolve the insufficient quantity of high quality seeding of Trichosanthes kirilowii, virusfree, higher seed setting and strong sprouts were selected as explants in tissue culture. In this paper, the technology of tissue culture was studied, including the selection and sterile techniques of the explants, different explant callus induction and buds induction and proliferation, root induction of tube plantlets of T. kirilowii. The main results showed that: buds germinated from the roots of female T. kirilowii between April and May were suitable for tissue culture explants. Explants were immersed in 75% ethanol for 30 s, washed by sterile water three times, then immersed in 0.31% ClO2 for 5 min and washed by sterile water 4-5 times again had a better result. The MS medium with different plant growth regulators for buds induction was MS 1 mg/L 6-BA 0.3 mg/L NAA, MS 0.3 mg/L 6-BA 0.1 mg/L NAA and MS 0.4 mg/L 6-BA 0.2 mg/L NAA were suitable for propagation, MS 0.1 mg/L IBA and MS 0.1 mg/L NAA were suitable for roots induction in T. kirilowii tissue culture.