Agrobacterium-mediated gene transfer in brinjal Kumari Anita, Gambhir Geetika, Srivastava D.K.* Department of Biotechnology, Dr. Y.S. Parmar University of Horticulture and Forestry, Solan-173230, India *Email: dksuhf89@gmail.com
Online published on 29 March, 2016. Abstract Plant regeneration studies were carried out using two types of explants viz cotyledon and hypocotyl in brinjal. The cotyledon explants showed high frequency of shoot regeneration (77.77%) on MS medium supplemented with 2.5 mg/l Kn and 0.4 mg/l IAA as compared to hypocotyl (50%) on MS medium supplemented with 2.5 mg/l BAP and 0.5 mg/l IAA. MS medium supplemented with 0.10 mg/l IAA was found to be best for root regeneration (81.81%). The brinjal plantlets were able to regenerate within 6–7 weeks. Regenerated plantlets were acclimatized. For genetic transformation, disarmed Agrobacterium tumefaciens LBA 4404 strain containing a reporter β-glucuronidase (gus) gene in binary vector pBI 121 system along with kanamycin resistance gene (npt-II) for selection in both bacteria and plant was used for co-cultivation experiment to transfer gus and npt-II genes in brinjal cells. After cocultivation only the transformed cells were able to grow on selective shoot regeneration medium containing kanamycin and cefotaxime, whereas control explants died on the selective medium. Transformation experiment could be scored as early as 4 weeks after selection. Most of the calli obtained on the selective medium were GUS positive. Putative transgenic shoots were obtained, which were able to grow on the selective medium containing 50 mg/l kanamycin. Top Keywords Brinjal, plant regeneration, genetic transformation, binary vector. Top |
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