Efficient transformation of Medicago truncatula cv. Jemalong using the hypervirulent Agrobacterium tumefaciens strain AGL1
Efficient transformation of Medicago truncatula cv. Jemalong using the hypervirulent Agrobacterium tumefaciens strain AGL1
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DOI：
10.1007/s00299-003-0649-y
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发表时间：
2003-08-01
影响因子：
6.2
通讯作者：
Barker, DG
中科院分区：
文献类型：
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作者：
Chabaud, M;de Carvalho-Niebel, F;Barker, DG
研究了农杆菌转化模式豆科牧草紫花苜蓿的效率。对菌株LBA 4404、C58pMP90、C58pGV2260和AGL1进行了Jemlong(2ha)基因分型。以携带启动子-GUS/GFP报告基因融合基因和nptII基因为选择标记的双载体用于植物的体外转化/再生。解除武装的超强毒株AGL1(Ti质粒TiBo542)的转化效率最高，其外植体形成卡那霉素抗性愈伤组织的比例是其他三个菌株的两倍。此外，我们能够减少使用AGL1进行植株再生所需的时间，24%的外植体在培养4-5个月内就产生了抗KM的转基因植株。分析了植物中的转基因表达，发现转基因在T-1后代中是保守的。
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.