Proline Accumulates in Transgenic Red Banana (Musa acuminateColla) Transformed with P5CSF129A Gene

Order of Publishing in Issue: 
9
Volume :8
Issue :4
October, 2014
Page No: 
413-422
Authors: 
Srinath Rao[1*], C.Nagappa[1], P.Surender Reddy[2], S.Anil Kumar[2], P. Hima Kumari[2] and P. B. Kavi Kishor[2]
Address: 
[1]Plant Tissue Culture and Genetic Engineering Laboratory, Department of P.G Studies and Research in Botany, Gulbarga University, Gulbarga 585106, India
Address: 
[2]Department of Genetics, Osmania University, Hyderabad 500 007, India

Agrobacterium tumefaciens mediated genetic transformation in red banana (Musa acuminataColla) is reported in the present communication.Male flower buds and suckers of red banana were transformed using A. tumefaciens strain LBA 4404 with the binary vector pCAMBIA1301, having hygromycin resistance gene as a selectable marker,âglucuronidase (GUS) as a reporter and a mutated version of the gene “1-pyrroline 5-carboxylate synthetase (P5CSF129A) encoding a proline biosynthetic pathway enzyme P5CS (EC 2.7.2.11/ 1.2.1.41) isolated from Vignaaconitifolia. Male flower buds proved to be better explants than suckers for the regeneration and genetic transformation. Green shoots were recovered from A. tumefaciens-transformed explants on selection medium consisting of Murashige and Skoog’s (MS) basal medium supplemented with 5 mg/L 6-benzylaminopurine (BAP), 2 mg/L indole-3-butyric acid (IBA), 250 mg/L cefotaxime and 25 mg/L hygromycin. Shoots were rooted on MS basal medium fortified with 3 mg/L IBA, 250 mg/L cefotaxime and 25 mg/L hygromycin. Transient expression of the GUS was observed in transgenic plants. The integration of the P5CSF129A gene was confirmed by polymerase chain reaction (PCR) and Southern blotting techniques. Transformation frequency was 11.3% and transgenic T0 plants accumulated 2 to 3-folds higher proline when compared to untransformed control plants. This work possibly paves the way for exploiting other genes for engineering salt, drought and temperature stress tolerance in red banana.

Keywords: 
Musa acuminata, P5CSF129A, proline accumulation, genetic transformation.
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