The present investigation was carried out at Tissue Culture Laboratory of Breeding Division of Bangladesh Sugarcane Research Institute (BSRI), Ishurdi, Pabna on in vitro shoot regeneration of sugarcane callus during the period of 2011. The regeneration of sugarcane varieties with supplementation of cytokinin (1mg/l of BA) and auxin IBA and IAA 0.25mg/l, 0.50mg/l and 0.75 mg/l respectively, were used in sugarcane on defined MS media for shoot induction. The shortest days to shoot initiation and the highest number of shoot per culture were found in Isd 40. The highest shoot length was observed in Isd 37. The concentration of 1mg/l of BA and 0.50 mg/l of IBA showed the highest number shoots per culture and the highest shoot length. The combination of 1.0 mg/l BA and 0.5 mg/l of IBA produced the maximum number of shoots per culture and shoot length was noticed in Isd 40 and Isd 37 respectively. Between two types of auxin, IBA was found more effective than IAA at different concentrations tested for producing shoots.