Improved callogenesis and plant regeneration from immature male flowers of East African highland banana cv. “Nakitembe” (AAA-EA)

Abstract

Application of biotechnological tools in breeding, germplasm conservation and propagation of the East African bananas (Musa sp.) is limited by the crop’s recalcitrance to somatic embryogenesis. This study was undertaken to establish an efficient callus induction and plant regeneration protocol from immature male flowers in the most commercial and farmer preferred banana cultivar “Nakitembe”. Embryogenic callusing was improved from 2.9% on conventional MA1 culture medium to 4.2% with 500 mg L-1 L-Glutamine, 500 mg L-1 L-Proline and 300 mg L-1 casein hydrolysate supplements. A combination of the three amino acids with Chui N6 salts yielded the highest callusing of 10.2%. Cell suspensions were developed and maintained on conventional MS based MA2 medium. Subsequently, the highest efficiency of embryos germination (up to 80%) was achieved on MA4 medium, supplemented with 2.25 mg L-1 BAP and 0.2 mg L-1 IAA. The developed protocol has been successfully applied in Agrobacterium mediated genetic transformations of this variety.