IN VITRO REGENERATION PROTOCOLS FROM IMMATURE MALE FLOWER BUDS OF BANANA CV. KARPURA CHAKKARAKELI (AAB) AND ASSESSMENT OF GENETIC FIDELITY WITH SSR & ISSR MARKERS M. Viswanath 1* B. V. K. Bhagavan2 K. Ravindra Kumar3 P. Subbaramamma4 M. GirijaRani5 *1 Ph.D Scholar, Department of Fruit Science, College of Horticulture, V.R. Gudem, 2 Principal scientist & Head, 3Scientist, 4Associate professor, 5Senior Scientist & Head Dr. Y.S.R. Horticultural University, Venkataramannagudem – 534101, Andhra Pradesh, India. *Corresponding author’s email:viswahorti123@gmail.com 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. Plate 1. Different stages of in vitro regeneration in banana cv. Karpura Chakkarakeli (AAB). a Mother plant bearing fruits and male flower bud. b Excised male flower bud. c Reduced male floral bud (6-7cm). d A group of male flowers (hands) used for culture. e Floral explants initiated on MS basal medium supplemented with 1 mgL-1 TDZ + 0.5 mgL-1 IAA + 2 CH. f Clusters of Cauliflower-like-bodies (CLBs). g CLBs converting to shoots. h In vitro shoot clusters i Plantlets ready for hardening. Secondary hardened plantlets. . ur s Plate 2. Morphogenic responses from the immature male flower explants of banana cv Karpura Chakkarakeli (AAB) in T14 treatment A. Responded explant (7 DAI) B. Green colo swollen base (15 DAI) C. Perianth necrosis (20 DAI) D. Formation of Cauliflower-like-bodie (CLBs) (30 DAI) E. Formation of shoot buds (60 DAI) F. Shoot clusters(90 DAI). Murashige and Skoog’s (MS) media supplemented with 1.0 mgL-1 TDZ + 0.5 mgL-1 IAA + 2 mgL-1 casein hydrolysate 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 amplified products were 100% monomorphic across all the regenerated plants and mother plant. RESULTS & DISCUSSION MS media supplemented with 1.0 mgL-1 TDZ + 0.5 mgL-1 IAA + 2 mgL-1 CH in 0.25% gel-rite was found optimum for mass production of this commercially important banana cv. Karpura Chakkarakeli (AAB) from IMF buds. CONCLUSION Amplification with SSR & ISSR primers A) Ma-SSR-230 B) UBC 868 300 bp B ABSTRACT INTRODUCTION Karpura Chakkarakeli (AAB), the choicest table cultivar of banana belongs to Mysore subgroup commercially grown in the coastal districts of Andhra Pradesh. Due to its highly recalcitrant nature, excessive phenol compounds, biotic contamination and poor multiplication rate causing the major hindrances for its commercial multiplication through shoot tip explants. Keeping in this view, this study was aimed to examine the potential of male flower explants as an alternate source material for culture establishment, morphogenetic responses and direct organogenesis with different combinations and concentrations of plant