Chabaud M, de Carvalho-Niebel F, Barker D G
Laboratoire des Interactions Plantes-Microorganismes, INRA-CNRS, UMR215, BP 27, 31326 Castanet Tolosan Cedex, France.
Plant Cell Rep. 2003 Aug;22(1):46-51. doi: 10.1007/s00299-003-0649-y. Epub 2003 Jun 24.
The efficiency of Agrobacterium tumefaciens transformation of the model legume Medicago truncatula cv. Jemalong (genotype 2HA) was evaluated for strains LBA 4404, C58pMP90, C58pGV2260 and AGL1. Binary vectors carrying promoter- gus/ gfp reporter gene fusions and the nptII gene as selectable marker were used for plant in vitro transformation/regeneration. The highest transformation efficiency was obtained with the disarmed hypervirulent strain AGL1 (Ti plasmid TiBo542), for which the percentage of explants forming kanamycin (Km)-resistant calli was double that obtained with each of the other three strains. In addition, we were able to reduce the time necessary for plant regeneration using AGL1, with 24% of the explants generating Km-resistant transgenic plantlets within only 4-5 months of culture. Transgene expression in planta was analysed and found to be conserved in the T(1) descendents.
评估了根癌农杆菌菌株LBA 4404、C58pMP90、C58pGV2260和AGL1对豆科模式植物蒺藜苜蓿品种Jemalong（基因型2HA）的转化效率。携带启动子 - gus/gfp报告基因融合体和nptII基因作为选择标记的双元载体用于植物的体外转化/再生。无毒性的超强毒株AGL1（Ti质粒TiBo542）获得了最高的转化效率，形成卡那霉素（Km）抗性愈伤组织的外植体百分比是其他三种菌株中任何一种的两倍。此外，我们能够缩短使用AGL1进行植物再生所需的时间，24%的外植体在仅4 - 5个月的培养期内就产生了Km抗性转基因植株。对植物中的转基因表达进行了分析，发现其在T(1)代后代中得以保留。
