Carrot (Daucus carota) hypocotyl transformation usingAgrobacterium tumefaciens
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Daucus carota hypocotyl sections were transformed withAgrobacterium tumefaciens LBA4404 containing CaMV 35S promoter, β-glucuronidase coding sequence and the nopaline synthase (Nos) poly adenylation sequences in Bin 19. Sliced sterile seedling hypocotyl segments were preincubated for 2 days, co-cultivated withAgrobacterium for an additional 2 days, and then transferred to medium containing 100ug/ml of kanamycin and 400ug/ml carbenicillin. In 6 weeks kanamycin resistant calli were obtained in 5.8% of the explants from one variety. Calli were subcultured on solid medium, and in 4 weeks introduced into suspension culture. NPTII and Southern blot analysis confirmed that three selected lines were transformed with 1-3 copies of the GUSII construction. GUS activity in transformants was 5 to 250 fold over background.
author list (cited authors)
Thomas, J. C., Guiltinan, M. J., Bustos, S., Thomas, T., & Nessler, C.