2002 Volume 19 Issue 5 Pages 377-382
A successful transformation method was reported on Delphinium spp., using Agrobacterium tumefaciens. Elongated hypocotyl and cotyledon petiole segments were infected with Agrobacterium tumefaciens strain LBA4404, which harbored a binary vector plasmid, pIG121-Hm, which included the β-glucuronidase (GUS) gene (with an intron) as a reporter gene, and the neomycin phosphotransferase II gene and the hygromycin phosphotransferase gene as selection markers. Explants were cultured on Murashige and Skoog medium supplemented with 1.0mgl-1 thidiazuron, 1.0mgl-1 2, 4-dichlorophenoxyacetic acid, 300mgl-1 ticarcillin, and 5mgl-1 hygromycin or 100mgl-1 kanamycin (selection medium) for regeneration. Transformation was confirmed by histochemical assays of GUS activity in plant tissues, and by PCR analysis of the GUS gene. Through four experiments, six independent GUS-positive regenerants were obtained out of 1276 explants.
