Abstract: Top cut pruning was made on an American sycamore crown, and segments from new sprout were collected as tissue culture initial explants. The results indicated that as for shoot segments induction buds the optimal medium was MS+NAA (0.1mg/L)+6-BA (1.0mg/L)+sucrose (30g/L), and DCR+NAA (0.1mg/L)+6-BA (1.0mg/L)+KT (0.3mg/L)+sucrose (30g/L) was the optimal medium for induction buds proliferation, on which induction buds grew well and coefficient of multiplication reached 6.4 times. We developed two ways to get adventitious buds to root. One was that adventitious buds were transferred on the DCR medium with 150mg/L R. The other, on the DCR medium with 60mg/L R about for 15d and then, transferred on a garniture-free DCR. By the two ways, both of the rooting percentages were up to 100%.