Abstract: The in vitro culture of Limnophila sessiliflora was established by using the nodal stem segments from submerged branch as explants.The method of surface sterilization of explants,the effects of different plant growth regulators on the multiplication and rooting were investigated.The results showed that 15% sterile explants could be obtained by twice surface treatment with 0.1% HgCl2 for 4 min with an interval of 4 h.When cultured on a 1/2 MS multiplication medium supplemented with 2.0 mg·L^-1 of 6-BA and 0.1-0.2 mg·L^-1 of NAA for 35 d,the test-tube seedling exhibited a multiplication coefficient higher than 30.8.For optimal rooting,induction on a 1/2 MS rooting medium supplemented with 0.3 mg·L^-1 of 6-BA and 0.5 mg·L^-1 of NAA for 28 d could produce rooted plantlets with 3-5 lateral shoots,and each plantlet generated 4.8 roots on average.Rooted plantlets were acclimatized and transferred into outdoor aquatic environment with 100% survival ratio.