Sonication assisted Agrobacterium transformation of banana cv. Neypoovan shoot tips with GUS reporter gene

Abstract

In the present study, Agrobacterium-mediated transformation was performed with shoot tips of banana cultivar, Ney Poovan (AB). The plant expression vector pBI 121, harboring reporter gene, (β-glucuronidase and plant selection marker), kanamycin, was used for co-cultivation. The apical meristems were sonicated and vacuum infiltered with Agrobacterium and co-cultured on liquid MS basal medium supplemented with 100 μM acetosyringone for 72 h. The explants were then cultured on solid shoot induction medium [SIM-MS salts, B5 vitamins, 3% sucrose, 400 mg L"1 of cefotaxime and 100 mg L"1 of kanamycin and 5 mg L"1 of BAP], which showed a regeneration percentage that ranged from 20% to 40%. Among different time intervals tested, sonication assisted Agrobacterium transformation for 8 min was found to be highly efficient in Ney Poovan with a maximum of 56% GUS positive shoots. The uniform GUS staining of transformed plant confirmed the presence and integration of the GUS gene into the banana genome. Polymerase chain reaction confirmed the presence of 500 bp amplified product of partial GUS gene in transformed plants. Thus an efficient protocol for Agrobacterium- mediated transformation of banana cv Ney Poovan was standardized which may possibly be useful for future studies on genetic modification and development of transgenics using novel genes