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Agrobacterium-mediated genetic transformation of groundnut (arachis hypogaea L.): An assessment of factors affecting regeneration of transgenic plants

Venkatachalam, Perumal and Geetha, Natesan and Jayabalan, Narayanasamipillai and Saravanababu, * and Sita, Lakshmi (1998) Agrobacterium-mediated genetic transformation of groundnut (arachis hypogaea L.): An assessment of factors affecting regeneration of transgenic plants. In: Journal of Plant Research, 111 (4). pp. 565-572.

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Abstract

Transgenic groundnut (Arachis hypogaea L.) plants were produced efficiently by inoculating different explants with Agrobacterium tumefaciens strain LBA4404 harbouring a binary vector pBI121 containing uidA (GUS) and nptll (neomycin phosphotransferase) genes. Genetic transformation frequency was found to be high with cotyledonary node explants followed by 4 d cocultivation. This method required 3 days of precultivation period before cocultivation with Agrobacterium. A concentration of 75 mg/l kanamycin sulfate was added to regeneration medium in order to select transformed shoots. Shoot regeneration occurred within 4 weeks; excised shoots were rooted on MS medium containing 50 mg/l kanamycin sulfate before transferring to soil. The expression of GUS gene (uidA gene) in the regenerated plants was verified by histochemical and fluorimetric assays. The presence of uidA and nptll genes in the putative transgenic lines was confirmed by PCR analysis. Insertion of the nptll gene in the nuclear genome of transgenic plants was verified by genomic Southern hybridization analysis. Factors affecting transformation efficiency are discussed.

Item Type: Journal Article
Additional Information: Copyright of this article belongs to Springer.
Keywords: Agrobacterium tumefaciens; Arachis hypogaea; gene expression; polymerase chain reaction; Southern hybridization; transformation.
Department/Centre: Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited: 16 Feb 2009 11:33
Last Modified: 19 Sep 2010 04:59
URI: http://eprints.iisc.ernet.in/id/eprint/17901

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