Summary
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.
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Abbreviations
- NPT II:
-
neomycin phosphotransferase II
- Nos:
-
nopaline synthase
- GUS:
-
β-glucuronidase
- CaMV:
-
cauliflower mosaic virus
- 2,4-D:
-
2,4 dichlorophenoxyacetic acid
- PMSF:
-
phenylmethylsulfonyl fluoride
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Communicated by J. M. Widholm
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Thomas, J.C., Guiltinan, M.J., Bustos, S. et al. Carrot (Daucus carota) hypocotyl transformation usingAgrobacterium tumefaciens . Plant Cell Reports 8, 354–357 (1989). https://doi.org/10.1007/BF00716672
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DOI: https://doi.org/10.1007/BF00716672