Abstract: This study aims to develop a rapid propagation method for the cassava cultivar ‘Huanan 9’ using an apical bud and stem segment with one axillary bud as explants. The MS basal culture medium was modified,and a self-created basal culture medium（named ZCM）was designed to conduct experiments in parallel with the MS medium.Cultivate and induce sterile seedlings in MS and ZCM media, thus different concentrations 6-BA and IBA were added to MS or ZCM medium to study the effects of different concentration ratios of plant-growth regulator on the subculture proliferation, and different concentrations NAA and IBA were added to MS or ZCM medium to study the effects of rooting. Screening suitable proliferation and rooting medium formulas.And explore the effect of seedling hardening methods on the survival rate of tissue cultured rooting seedlings after transplantation.The results indicate that both MS and ZCM media can effectively induce germination and growth of the plants, achieving a sterile seedling induction rate of 66.7% for stem segment and 46.7% for apical bud,stem segments with one axillary bud are better explants.The optimal medium for proliferation culture was identified as MS or ZCM,the proliferation coefficients are 3.5 and 3.4, respectively. MS or ZCM were proved suitable for rooting,achieving a rooting rate of 100%.The tissue-cultured seedlings developed numerous and long roots. After removal from the tissue culture bottle, the rooted seedlings were cultured with a nutrient solution outside the bottle, which improved the survival rate during transplantation.