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http://nopr.niscpr.res.in/handle/123456789/19879| Title: | Agrobacterium-mediated Transformation of Lucerne (Medicago sativa Linn.): Optimizing Biological and Physical Parameters |
| Authors: | Kumar, Suresh Bhat, Vishnu Bhat, B V Gupta, M G |
| Keywords: | Lucerne;Agrobacterium;Genetic transformation;Forage quality improvement |
| Issue Date: | Jul-2002 |
| Publisher: | NISCAIR-CSIR, India |
| Abstract: | An optimized protocol for the transformation of Indian cultivars of lucerne (Medicago sativa Linn.) using Agrobacterium tumefaciens 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 in 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. tumefaciens strain LBA-4404 harbouring plasmid pCAMBIA-1301. Cotyledonary explant was found to be more responsive for transformation. Bacterial density of 5×109 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. |
| Page(s): | 298-300 |
| ISSN: | 0975-0967 (Online); 0972-5849 (Print) |
| Appears in Collections: | IJBT Vol.01(3) [July 2002] |
Files in This Item:
| File | Description | Size | Format | |
|---|---|---|---|---|
| IJBT 1(3) 298-300.pdf | 759.59 kB | Adobe PDF | View/Open |
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