Title: Agrobacterium-mediated Transformation of Lucerne (Medicago sativa Linn.): Optimizing Biological and Physical Parameters
Abstract: An optimized protocol for the transformation of Indian cultivars of lucerne (Medicago sativa Linn.) using Agrobacterium tlllnejaciefls has been developed. It involved a selectable marker gene encoding hygromycin phosphotransferase (hpt II) which confers resistance to the antibiotic hygromycin, together with a reporter gene (uid A) encoding P-D glucuronidase (GUS); cotyledonary and hypocotyl explants from ill vitro grown 5 days -old seedlings raised from mature seeds of genotype L-2 and IL-75 as target tissues, and co-cultivation of explants with A. tumejaciells strain LBA-4404 harbouring plasmid pCAMBIA-1301. Cotyledonary explant was found to be more responsive for transformation. Bacterial density of 5xl0 9 cells/ml and co cultivation for three days were found to be most effective for stable GUS expression in 91 % of the hygromycin-resistant calli.
Publication Year: 2002
Publication Date: 2002-01-01
Language: en
Type: article
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Cited By Count: 3
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