This study was determined optimal condition as culture medium, pH, carbohydrate source and growth regulators through axillary buds cultures of Salix gracilistyla. S. gracilistyla shoots were obtained from axillary bud on 1/2MS basal medium after 2 weeks of culture. Highest shoot growth was showed in the MS medium(GI= 2.43). Shoot growth was highest in 9% sucrose compared to other concentration. BAP affects axillary shoot multiplication in of S. gracilistyla. Among nutrients, Ca2+ was not affected shoot number of S. gracilistyla. However, NH4+, NO3 -, and K+ is greatly influenced shoot number. However, Ca2+ was affected in shoot length of S. gracilistyla. Feeding of Mg2+ was not affected in shoot length. Root number is greatly influenced by NH4 +, NO3 -, and K+, however NH4 + and K+ was not affected root number of S. gracilistyla. NH4 + was affected root length, however Mg4 + was not affected root length. Treatment of 0.1mg/L IBA promoted in vitro rooting. In vitro rooted plantlets were successfully transferred to pots for 4weeks hardening process. Later, these plants were transferred to soil with above 90% survival rate in BS+SD(2:1). These results could be easily adopted for commercial large scale cultivation.