Levée V, Lelu M-A, Jouanin L, Cornu D, Pilate G
Station d'Amélioration des Arbres Forestiers, INRA-CRO, F-45160 Ardon, France Fax No.: 33-02-38-41-78-79, , , , , , FR.
Laboratoire de Biologie Cellulaire et Moléculaire, INRA, Route de St-Cyr, F-78000 Versailles, France, , , , , , FR.
Plant Cell Rep. 1997 Jul;16(10):680-685. doi: 10.1007/s002990050301.
A transformation procedure was developed for hybrid larch embryogenic tissue using Agrobacterium tumefaciens. The cocultivation procedure yielded one to two transformation events per 100 cocultivated masses. The addition of 100 µM coniferyl alcohol increased the yield. This improved procedure was successfully applied to three other genotypes. After 3 months on selective medium, the transgenic tissue remained embryogenic, which allowed production of transgenic plants in the greenhouse. Stable integration of the transgene was confirmed by PCR and Southern hybridisation on transformed tissues and acclimatised plants.
利用根癌农杆菌开发了一种用于杂种落叶松胚性组织的转化程序。共培养程序每100个共培养团块产生1至2个转化事件。添加100 µM松柏醇可提高产量。这一改进程序已成功应用于其他三种基因型。在选择培养基上培养3个月后,转基因组织仍保持胚性,这使得能够在温室中培育出转基因植株。通过对转化组织和驯化植株进行PCR和Southern杂交,证实了转基因的稳定整合。