Abstract: The paper aims to construct a highly efficient asexual system of Ipomoea aquatic, which is the main cultivated vegetable in Chonqing. The young stems were cultured on MS medium with 6-BA, IAA, NAA and 2, 4-D of different concentrations. The results showed that, lots of callus were induced in the medium of 1/2 MS+ 6-BA 0.5 mg/L+ 2,4-D 1.5 mg/L, and the inducement ratio was up to 93.3%. In seedling subculture, the differentiation medium of callus induction was 1/2MS+ 6-BA 0.5 mg/L, and the differentiation frequency was 92% . The medium of 1/2 MS + 0.9 mg/L NAA was suitable for water spinach root growth. The substrate composition of peat, mushroom dregs and vermiculite (1:1:1) was suitable for aseptic seedlings growth, with shoot length of 11.43 cm and the survival rate of 98%.