High-frequency generation of transgenic tobacco plants after modified leaf disk cocultivation withAgrobacterium tumefaciens Academic Article uri icon


  • A modified protocol for the Agrobacterium tumefaciens-mediated transformation of tobacco (Nicotina tabacum L.) leaf disks was developed for greater recovery of transgenic plants. Modifications include transformation of Agrobacterium by a freeze-thaw procedure, initial cocultivation of leaf disks and Agrobacterium under vacuum, subsequent growth with nurse cells for one week, rooting of shoots in medium lacking carbenicillin, longer, growth in rooting medium, and a shortened "hardening" step. By this procedure, an average of 1.3 kanamycin-resistant calli were obtained per leaf disk, and 38% of, the callus cultures used were regenerated to produce 133 independently transformed tobacco plants. 1990 Kluwer Academic Publishers.

published proceedings

  • Plant Molecular Biology Reporter

altmetric score

  • 3

author list (cited authors)

  • Burow, M. D., Chlan, C. A., Sen, P., Lisca, A., & Murai, N.

citation count

  • 87

complete list of authors

  • Burow, Mark D||Chlan, Caryl A||Sen, Partha||Lisca, Andreana||Murai, Norimoto

publication date

  • May 1990