The experiment was conducted at the Laboratory of Breeding Division of Bangladesh Sugarcane Research Institute (BSRI), Ishurdi, Pabna during the period of 2011 for the effect of BA and NNA for shoot induction in vitro culture of sugarcane. The regeneration of sugarcane varieties with supplementation of 1mg/l of BA and different concentration of NNA (0.5, 1.0 and 2.0 mg/l) were used in this experiment for shoot induction. In varietal response to application of BA and NNA, among five varieties, the early days to shoot initiation were found in Isd 2-54 and Isd 40. The highest number of shoots per culture and the highest shoot length were observed in Isd 40 and Isd 37 respectively.The effect of concentrations of BA and NAA, 1.0 mg/l BA and 0.5 mg/l of NAA were showed the highest number of shoot per culture and the highest shoot length. Regarding interaction effect of varieties and the doses of BA and NNA, 1 mg/l of BA and 0.50 mg/l of NAA produced the highest number of shoots per culture and shoot length in Isd 40 and Isd 37 respectively.