The tissue culture conditions of fast-growing clone K2 and fodder-type clone K3 of Robinia pseudoacacia L. tetraploid were investigated using shoot sprouting in hydroponic growth chamber as explants. After disinfected by 70% ethanol then sterilized with NaClO solution containing 0.5% available chlorine, the explants should be initially cultured with MS medium (containing 30 g·L^-1 sucrose and 6 g·L^-1 agar, pH 5.8), and survival rate of K2 and K3 explants all reaches 90% in initial culture medium. The best proliferation medium of K2 adventitious bud is MS medium adding 0.3 mg·L^-1 6-BA and 0.05 mg·L^-1 NAA, with the highest proliferation coefficient of 5.29,and that of K3 is MS medium adding 1.0 mg·L^-1 6-BA and 0.10 mg·L^-1 NAA, with the highest proliferation coefficient of 3.36. The optimized rooting medium of K2 and K3 plantlets is 1/2MS medium (containing 30 g·L^-1 sucrose and 6 g·L^-1 agar, pH 5.8) adding 0.1 mg·L^-1 IBA and 0.1 mg·L^-1 NAA, respectively. After training,K2 and K3 plantlets are transplanted by two-step transplanting method, all of survival rate reach 98%.