Plant Molecular & Cellular Genetics & Centre for Plant Molecular Biology, Bose Institute, P1/12, C.I.T. Scheme VII-M, 700 054, Calcutta, India.
Plant Cell Rep. 1996 Nov;16(1-2):32-7. doi: 10.1007/BF01275444.
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 26(0)C. 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.
三种鹰嘴豆 ICCV-1、ICCV-6 和一种本地品种（Desi）通过丛生芽再生被测试进行植物再生。从成熟种子中取出胚轴，丢弃根分生组织和茎尖。这些外植体在含有 MS 大量元素盐、4X MS 微量元素盐、I35 维生素、3.0 mg/1 BAP、0.004 mg/1 NAA、3%（w/v）蔗糖的培养基上培养，并在 26(0)C 下孵育。用携带 uidA 和 nptII 基因的二元载体 pBI121 的根癌农杆菌菌株 LBA4404 转化外植体。用卡那霉素反复选择丛生芽。选择的卡那霉素抗性芽在补充有 0.05 mg/1 113A 的 MS 培养基上生根。假定的转化体经 GUS 组织化学染色呈阳性。此外，nptll 测定法证实了 nptII 在卡那霉素抗性植物中的表达。转基因植物被转移到土壤中并在温室中生长。
