At present sugarcane variety Co 86032 is a ruling variety in peninsular zone of India. The present investigation was carried out with different types of explants for callus induction viz., direct leaf whorl, tissue culture leaf, shoot tip and shoot base. Better callus induction was obtained with direct leaf whorl as an explant. Explants of 2-2.5 cm size were taken for callus induction on MS media containing different concentrations of 2,4-D (1mg to 5mg). Maximum callus formation (80-85%) was observed in media containing both 3mg L-1 and 4mg L-1 of 2,4-D. A significant increase in the rate of callus initiation and proliferation was observed in cultures incubated in dark as compared to light grown culture. Two different media were used for shoot initiation, first media contain 1.0mg L-1 BAP, 0.5mg L-1 NAA, 20g L-1sucrose and 7g L-1agar, whereas second media contain 1.0mg L-1 BAP, 0.5mg L-1 IBA, 20g L-1 sucrose and 7 g L-1 agar. Second media proved better for shoot initiation. For multiplication of plantlets 1.0mg L-1 BAP , 0.25mg L-1 GA3, 20g L-1 sucrose and 7g L-1 agar was used. For root development 2.0mg L-1 NAA was used. This is first attempt of application of BAP with GA3 in multiplication media in variety CO 86032 and its gives encouraging result in short time period. The above developed protocol may be useful for genetic transformation and somaclonal variation studies in sugarcane variety CO 86032.