An experiment containing shoot regeneration has been conducted at the Laboratory of Breeding Division of Bangladesh Sugarcane Research Institute (BSRI) Ishurdi, Pabna during the period of 2011. The present study was conducted to standardize a suitable protocol of in vitro plant regeneration potentiality of five sugarcane varieties. After shoot proliferation, the mini regenerated shoots were separated aseptically and cultured in MS medium containing five concentrations (viz.1.0, 2.0, 3.0, 4.0 and 5.0mg/l) of NAA for adventitious root induction. The concentration of 5mg/l of NAA produced the minimum (9) days to root initiation the highest number (11.01) of root per culture and the highest number (0.21cm) of root length. All the variety showed the best root forming performance in concentration of 5mg/l of NAA.