Carrot (Daucus carota) hypocotyl transformation usingAgrobacterium tumefaciens.
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.
Department of Biochemistry, University of Arizona, 85721, Tucson, AZ, USA.
J C
JC
Thomas
M J
MJ
Guiltinan
S
S
Bustos
T
T
Thomas
C
C
Nessler