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A Novelgene Transformation Technique for Farmer's Preferred Cassava Cultivar (Nwibibi) from Nigeria

2 Abstract: Cassava is an important food security crop in the tropics and subtropics. Nutritionally, the low protein content of cassava, vitamins and minerals is a source of concern for people whose diet is based mainly on this plant. Biotechnology has been identified as a powerful tool for genetic transformation. Agrobacterium tumefaciens, a ubiquitous soil borne pathogen employs a highly evolved and still incompletely understood gene transfer and integration system that appears optimized for efficient nuclear targeting and integration. A bacterial strain LBA 4404 containing a binary vector pB 2300 with npt11 gene as selectable marker and a green fluorescent protein (GFP) as a tracker was used for the experiments. Compared with the previously transformed model cultivar (60444), Nwibibi tissues were more amenable to the antibiotic cefotaxime and responded optimally to tyrosine. Subsequent selection of transformed tissues with cefotaxime and paramomycin resulted in the recovery of antibiotic-resistant, GFP-expressing lines of friable embryogenic callus, from which embryos and subsequent plants were regenerated. The positive response of this local Nigerian farmer preferred cassava cultivar to Agrobacterium transformation therefore supports the assertion that plants do not lack the biological capacity to respond to essential triggers of integrative transformation or have cellular mechanisms preventing integrative transformation. Abbreviations: BAP-benzylaminopurine; Cef.-cefotaxime; DKW-DKW basal salt mixture; FEC-friable embryogenic callus; GD-Gresshoff and Doy; Paro.-paramomycin; P-picloram; MS-Murashige and Skoog.