Efficient transgenic plant regeneration throughAgrobacterium-mediated transformation of Chickpea (Cicer arietinum L.) |
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| Authors: | Sanchayita Kar Tony M Johnson Pritilata Nayak S K Sen |
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| Institution: | (1) Plant Molecular & Cellular Genetics & Centre for Plant Molecular Biology, Bose Institute, P1/12, C.I.T. Scheme VII-M, 700 054 Calcutta, India;(2) Present address: Department of Biochemistry & Molecular Biology, Mississipi State University, Box 9650, MS 39762, USA;(3) Present address: Department of Biochemistry, College of Biological Sciences, 140 Gortner Laboratory, 1479 Gortner Avenue, 55108-1022 St. Paul, MN, USA |
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| Abstract: | Three genotypes of chickpea ICCV-1, ICCV-6 and a Desi (local) variety were tested for plant regeneration through multiple shoot production. The embryo axis was removed from mature seeds, the root meristem and the shoot apex were discarded. These explants were cultured on medium containing MS macro salts, 4X MS micro salts, I35 vitamins, 3.0 mg/1 BAP, 0.004 mg/1 NAA, 3% (w/v) sucrose and incubated at 260C. The explants were transformed withAgrobacterium tumefaciens strain LBA4404 with binary vector pBI121 containing theuidA andnptIl genes. Multiple shoots were repeatedly selected with kanamycin. The selected kanamycin resistant shoots were rooted on MS medium supplemented with 0.05 mg/1 113A. The presumptive transformants histochemically stained positive for GUS. Additionally, nptll assay confirmed the expression ofnptII in kanamycin resistant plants. Transgenic plants were transferred to soil and grown in the green house.Abbreviations BAP
6-benzylamino purine
- 2,4-D
2,4dichlorophenoxy acetic acid
- IAA
Indole acetic acid
- IBA
Indole butaric acid
- NAA
Naphthalene acetic acid |
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