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Agrobacterium-mediated genetic transformation and regeneration of transgenic plants from cotyledon explants of groundnut (Arachis hypogaea L.) via somatic embryogenesis

Venkatachalam, P and Geetha, N and Khandelwal, Abha and Shaila, MS and Sita, Lakshmi G (2000) Agrobacterium-mediated genetic transformation and regeneration of transgenic plants from cotyledon explants of groundnut (Arachis hypogaea L.) via somatic embryogenesis. In: Current Science, 78 (9). pp. 1130-1136.

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Abstract

An efficient transformation protocol was developed for groundnut (Arachis hypogaea L.) plants. Precultured cotyledons were co-cultured with Agrobacterium tumefaciens strain LBA 4404 harbouring the binary vector pBI121 containing the uidA (GUS) and nptII genes for 2 days and cultured on an embryo induction medium containing 0.5 mg/l NAA, 5.0 mg/l BAP, 75 mg/ml kanamycin and 300 mg/ml cefotaxime. The putatively transformed embryos were transferred to the medium with reduced kanamycin (50 mg/ml) for further development. Prolific shoots developed from these embryos on a MS medium containing 0.5 mg/l BAP and 50 mg/ml kanamycin with a transformation efficiency of 47%. The elongated kanamycin-resistant shoots were subsequently rooted on the MS medium supplemented with 1.0 mg/l IBA. The transgenic plants were later established in plastic cups. A strong GUS activity was detected in the putatively transformed plants by histochemical assay. Transformation was confirmed by PCR analyses. Integration of T-DNA into nuclear genome of transgenic plants was further confirmed by Southern hybridization with nptII gene probe. A large number of transgenic plants were obtained in this study. This protocol allows effective transformation and quick regeneration via embryogenesis.

Item Type: Journal Article
Additional Information: Copyright for this article belongs to Indian Academy of Sciences.
Department/Centre: Division of Biological Sciences > Microbiology & Cell Biology
Date Deposited: 07 Oct 2004
Last Modified: 19 Sep 2010 04:15
URI: http://eprints.iisc.ernet.in/id/eprint/1715

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