An efficient in vitro propagation protocol has been developed for the first time on direct differentiation of shoot buds from immature male flower buds of banana cv. Karpura Chakkarakeli (AAB). MS media supplemented with 1.0 mgL-1 TDZ + 0.5 mgL-1 IAA + 2 mgL-1 CH was found optimal for per -cent cauliflower-like bodies (CLBs) formation (75.04%), maximum number of CLBs (15.01) explant-1 and minimum number of days (43.54d) taken for CLBs induction. The highest per cent (75.37%) and maximum number (12.78) of shoots proliferated explant-1 within 120 days after culture initiation. The regenerated plantlets rooted better in half MS media supplemented with 0.5 mgL-1 IBA and hardened with 97% survival rate. SSR & ISSR markers were employed to assess the genetic stability of in vitro regenerated and multiplied plantlets of banana cv. Karpura Chakkarakeli (AAB) recorded 100% monomorphic across all the regenerated plants with the mother plant.