Particle Bombardment-mediated Co-transformation of Chitinase and β-1, 3 Glucanase Genes in Banana

An efficient transformation system for Pisang Rastali (AAB) was developed using an optimised particle bombardment device, improved antibiotic selections and co-transformations procedure and a simple protocol for regeneration of transformants. Recent progress with advanced in vitro cultures of banana such as establishment of highly regenerable tiny single meristem buds opened the opportunity for the production of disease tolerant transgenic bananas. Chitinase and glucanase the important disease tolerant genes were successfully transformed into banana together with gfp and gusA genes as reporter gene. Five different treatments using different chitinase and glucanase genes inserted singly or in combination were carried out and resulted in earlier detection of transient expression of gfp and gusA genes. Proliferating single buds were selected on geneticin G-418 (solid-liquid-solid medium) to produce a number of putatively transformed bananas. Molecular analyses such as Polymerase Chain Reactions (PCR) and Southern blot was performed to confirm the integration and expression of the introduced genes in genome. Protein assay of extract from the transgenic plantlets showed an increased in chitinase and β-1,3-glucanase enzymes activity over the untransformed plantlets. The present of particle bombardment-mediated transformation system reported here is suitable for using tiny single meristem buds to obtain fungal disease tolerant or resistant banana through genetic engineering.