Abstract: 【Objective】In this study, we aimed to establish an efficient plant regeneration protocol for Salix suchowensis through tissue culture, which would serve as a foundation for developing a genetic transformation system for this woody shrub. 【Method】By using young stems with axillary buds and mature seeds as explants, the influences of different factors on plantlet regeneration were systematically studied, including sterilization methods, types of the basic medium, light conditions, and plant growth regulators. 【Result】The optimal basic medium for S. suchowensis was Murashige and Skoog(MS)medium + sucrose(30 g/L)+ Phytagel(4.4 g/L). When using young stem nodal segments as explants, the best sterilization method was 70% ethanol for 30 sec and then 20% sodium hypochlorite for 10 min, while the optimal medium for axillary buds induction and multiplication was the basic medium supplemented with 6-BA(1 mg/L)+ NAA(0.01 mg/L)and 6-BA(0.5 mg/L)+ NAA(0.05 mg/L), respectively, and the appropriate medium for rooting was the basic medium + IBA(0.2 mg/L)+ NAA(0.03 mg/L). While for mature seeds, the optimal sterilizing treatment was 70% ethanol for 10 sec and then 20% sodium hypochlorite for 2 min; the appropriate medium for callus indution, proliferation and subsequent shoot differentiation was the basic medium supplemented with 6-BA(0.5 mg/L)+NAA(0.3 mg/L), 6-BA(0.5 mg/L)+ NAA(0.5 or 0.8 mg/L), 6-BA(0.5 mg/L)+ TDZ(0.2 mg/L), respectively, and the best rooting medium was the basic medium containing NAA(0.01 mg/L). 【Conclusion】We established a regeneration protocol for S. suchowensis from the axillary buds or callus by using young stem nodal segments and mature seeds as explants respectively.