The effects of different factors, such as different combinations of plant growth regulators, position of leaves on seedling and GA3 concentrations, on shoot regeneration from leaves of hybrid Broussonetia papyrifera were studied. The results indicated that the optimum medium for callus induction and shoot regeneration was MS+6-BA 2.0 mg/L+NAA 0.1 mg/L, the callus induction rate could reach 100% and the frequency of adventitious bud regeneration was 91.7%. The best leaves for adventitious bud regeneration were the first three top leaves of a shoot. When MS medium was supplemented with 0.5 mg/L GA3, the shoot propagation coefficient reached 8.22 and the length of shoots increased.When low concentration of 6-BA (0.3 mg/L) was supplemented to MS medium, the length of shoots was 3.89 cm, which was significantly increased, and the adventitious bud could be used for rooting culture directly. When NAA(1.0 mg/L) was added in MS medium, the rooting time was only 9 days and rooting rate reached 86.7%.