Agrobacterium tumefaciens-mediated transformation of Cyclamen persicum Mill.

A method for Agrobacterium-mediated transformation of Cyclamen persicum Mill. is reported. Etiolated petiole segments were infected with Agrobacterium tumefaciens strain AGL0 or LBA4404. These strains have a binary vector plasmid, pIG121Hm, that includes the beta-glucuronidase (GUS) gene with an intron as 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 1.0 mg/l thidiazuron, 1.0 mg/l 2,4-dichlorophenoxyacetic acid, 300 mg/l ticarcillin, and 5 mg/l hygromycin or 100 mg/l kanamycin (selection medium) for regeneration. Transformation was confirmed by histochemical assays of GUS activity in plant tissues, and by Southern blot analysis of the GUS gene. Through five experiments, 103 independent GUS-positive plants were obtained from 920 explants. (C) 1999 Elsevier Science Ireland Ltd. All rights reserved.