Department of Biological Sciences, The University of Newcastle, 2308, New South Wales, Australia.
Plant Cell Rep. 1992 Apr;11(3):113-7. doi: 10.1007/BF00232161.
Fertile transgenic plants of the annual pasture legume Medicago truncatula were obtained by Agrobacterium-mediated transformation, utilising a disarmed Ti plasmid and a binary vector containing the kanamycin resistance gene under the control of the cauliflower mosaic virus 35S promoter. Factors contributing to the result included an improved plant regeneration protocol and the use of explants from a plant identified as possessing high regeneration capability from tissue culture. Genes present on the T-DNA of the Ri plasmid had a negative effect on somatic embryogenesis. Only tissue inoculated with Agrobacterium strains containing a disarmed Ti plasmid lacking the T-DNA region or a Ri plasmid with an inactivated rol A gene regenerated transgenic plants. Fertile transgenic plants were only obtained with disarmed A. tumefaciens, and the introduced NPT II gene was transmitted to R1 progeny.
通过根癌农杆菌介导的转化，利用无毒性 Ti 质粒和含有卡那霉素抗性基因的二元载体（该基因受花椰菜花叶病毒 35S 启动子的控制），获得了一年生牧草豆科植物蒺藜苜蓿的可育转基因植株。促成这一结果的因素包括改进的植物再生方案，以及利用鉴定出的在组织培养中具有高再生能力的外植体。Ri 质粒 T-DNA 上的基因对体细胞胚胎发生有负面影响。只有用含有无毒性 Ti 质粒（缺失 T-DNA 区域）或 Ri 质粒（rol A 基因失活）的根癌农杆菌菌株接种的组织才能再生出转基因植株。只有无毒性的根癌农杆菌才能获得可育的转基因植株，并且导入的 NPT II 基因被传递到 R1 后代。
