Abstract

Banana viruses have had a major negative impact on the production of bananas worldwide. Among banana viruses, Banana Bunchy Top Virus (BBTV) is one of the most destructive viral diseases affecting Hill banana (AAB) which is known for its special flavor and long shelf life and are unique to the lower Pulney hills of Tamil Nadu, India. In transgenics, Pathogen Derived Resistance (PDR) approach is one of the potential strategies for developing virus resistance in crops which has been proven to be difficult to obtain by conventional breeding. Here we have reported a reproducible and efficient in planta transformation method via Agrobacterium- mediated genetic transformation in banana. Three-month-old banana suckers were used as explant and Agrobacterium tumefaciens strain LBA4404 harboring the binary vector pCAMBIA1301:bbtv rep was used in the co-cultivation. The presence and integration of the gus and replicase genes into the banana genome were confirmed by GUS histochemical assay, polymerase chain reaction and Southern blot hybridization. These results suggest that the in planta Agrobacterium-mediated transformation protocol developed in this study for stable integration of rep gene of BBTV is efficient and that this transformation system could be widely adopted for routine transformation in banana for transferring genes.

Keywords: Banana, Bunchy top, Decortication, Acetosyringone, GUS histochemical assay, In planta transformation, Replicase gene