Full Length Research Paper

Rapid and efficient production of transgenic East African Highland Banana (Musa spp.) using intercalary meristematic tissues

Leena Tripathi¹*, Jaindra Nath Tripathi¹ and Wilberforce Kateera Tushemereirwe²

¹International Institute of Tropical Agriculture, PO Box 7878, Kampala, Uganda.

²Kawanda Agriculture Research Institute, PO Box 7065, Kampala, Uganda.

*Corresponding author. E-mail: l.tripathi@cgiar.org. Tel: +256 414 285060. Fax: +256 414 285079.

Abbreviations: BAP, 6-benzylaminopurine; EAHBs, East African Highland Bananas; gusA, beta-,glucuronidase gene; MS, Murashige and Skoog; nptII, neomycin phosphotransferase gene.

Accepted 16 April, 2008

East Africa is the largest banana producing and consuming region in Africa.  In particular, the East African Highland Banana serves as the major staple crop of countries like Uganda, but production is constrained by a number of serious pests and diseases.  Banana breeding is a very difficult and slow process, so genetic engineering offers an alternative approach to improvement. A transformation system using intercalary meristematic tissues was developed using Agrobacterium strain EHA105 harboring the binary vector pCAMBIA2301 containing the gusA reporter gene and nptII as selectable marker. In this paper, a new transformation protocol is described that yields kanamycin-resistant, GUS-expressing banana plants from roughly 10% of the initial explants. The resulting fully-rooted transgenic plants do not appear to be chimeras since they can be stably propagated, GUS activity is observed uniformly throughout the plants including the germline cells of the meristem, and PCR and Southern blots indicate stable integration of the genes into the genome.

Key words: Agrobacterium, intercalary meristem, genetic transformation, banana.