Station d'Amélioration des Arbres Forestiers, Institut National de la Recherche Agronomique, F-45160, Ardon, Olivet, France.
Plant Cell Rep. 1992 Apr;11(3):137-41. doi: 10.1007/BF00232166.
Leaf or stem explants of a hybrid poplar clone (Populus tremula X Populus alba), sensitive to Agrobacterium tumefaciens, were co-cultivated either by an octopine or a nopaline disarmed A. tumefaciens modified strain. Transformed poplar shoots were readily regenerated from explants. The protocol was improved using the nopaline disarmed strain C58/pMP90 with the binary vector pBI121. This protocol was then used to test three other vectors. The first one, possessing a nptII gene fused to the CaMV 19S promoter, permitted regeneration of transformed shoots in presence of 50 to 100 mg/l kanamycin. The two other vectors carried an additional nptII gene under the control of the CaMV 35S or CaMV 35S promoter with a double enhancer sequence (CaMV 70). CaMV 70 promoter provided consistently higher level of gene expression than the other promoters in both callus and leaf tissues.
杨树无性系(欧洲山杨×银白杨)叶片或茎段外植体对根癌农杆菌敏感,与八氢番茄红素合酶或 nopalin 去激活的根癌农杆菌修饰菌株共培养。转化的杨树芽从外植体中很容易再生。使用带有二元载体 pBI121 的 nopalin 去激活菌株 C58/pMP90 改进了该方案。然后,该方案用于测试另外三个载体。第一个载体,将 nptII 基因与 CaMV 19S 启动子融合,在 50 至 100mg/L 卡那霉素存在的情况下,允许转化芽的再生。另外两个载体在 CaMV 35S 或 CaMV 35S 启动子的控制下携带一个额外的 nptII 基因,带有双增强子序列(CaMV 70)。CaMV 70 启动子在愈伤组织和叶片组织中均提供了比其他启动子更高水平的基因表达。