Advanced search
1 file | 111.89 KB

Agrobacterium tumefaciens transformation and cotransformation frequencies of Arabidopsis thaliana root explants and tobacco protoplasts

Sylvie De Buck (UGent) , Anni Jacobs (UGent) , Marc Van Montagu (UGent) and Anna Depicker (UGent)
Author
Organization
Abstract
In view of the recent finding that different T-DNAs tend to ligate and integrate as repeats at single chromosomal positions, the frequency of transformation and cotransformation was determined during cocultivation of Arabidopsis thaliana root explants and Nicotiana tabacum protoplasts with two Agrobacterium strains. The transformation frequency of unselected A, thaliana shoots was lower than 1% whereas that of cocultivated tobacco protoplasts was approximately 18%, The cotransformation frequencies, defined as the frequencies with which cells transformed with a first T-DNA contained a second unselected T-DNA, were approximately 40% reproducible, irrespective of the selection, the transformation frequency, and the plant system used. Extrapolation of these results suggests that at least two independently transferred T-DNAs were present in 64% of the transformed plant cells, Molecular analysis of cocultivated N. tabacum shoots regenerated on nonselective medium showed that only a few transformants had a silenced (2/46) or truncated (1/46) T-DNA, Therefore, most integrated T-DNAs expressed their selectable or screenable markers in primary transgenic plants. Remarkably, 10 to 30% of the selected A. thaliana shoots or progenies lost the T-DNA marker they were selected on. As these regenerants contained the unselected T-DNA with a high frequency (17%), these selected plants might result from the expression of unstable, transiently expressed T-DNAs, In conclusion, a significant part of the T-DNAs is lost from the transformed cells.
Keywords
SEQUENCES, VECTORS, SELECTION, COMPETENCE, gene silencing, T-DNA transfer, transgene expression, MEDIATED GENE-TRANSFER, T-DNA, PLANT-CELLS, TRANSGENE EXPRESSION, SINGLE-COPY, INTEGRATION

Downloads

  • (...).pdf
    • full text
    • |
    • UGent only
    • |
    • PDF
    • |
    • 111.89 KB

Citation

Please use this url to cite or link to this publication:

Chicago
De Buck, Sylvie, Anni Jacobs, Marc Van Montagu, and Anna Depicker. 1998. “Agrobacterium Tumefaciens Transformation and Cotransformation Frequencies of Arabidopsis Thaliana Root Explants and Tobacco Protoplasts.” Molecular Plant-microbe Interactions 11 (6): 449–457.
APA
De Buck, S., Jacobs, A., Van Montagu, M., & Depicker, A. (1998). Agrobacterium tumefaciens transformation and cotransformation frequencies of Arabidopsis thaliana root explants and tobacco protoplasts. MOLECULAR PLANT-MICROBE INTERACTIONS, 11(6), 449–457.
Vancouver
1.
De Buck S, Jacobs A, Van Montagu M, Depicker A. Agrobacterium tumefaciens transformation and cotransformation frequencies of Arabidopsis thaliana root explants and tobacco protoplasts. MOLECULAR PLANT-MICROBE INTERACTIONS. 1998;11(6):449–57.
MLA
De Buck, Sylvie, Anni Jacobs, Marc Van Montagu, et al. “Agrobacterium Tumefaciens Transformation and Cotransformation Frequencies of Arabidopsis Thaliana Root Explants and Tobacco Protoplasts.” MOLECULAR PLANT-MICROBE INTERACTIONS 11.6 (1998): 449–457. Print.
@article{178119,
  abstract     = {In view of the recent finding that different T-DNAs tend to ligate and integrate as repeats at single chromosomal positions, the frequency of transformation and cotransformation was determined during cocultivation of Arabidopsis thaliana root explants and Nicotiana tabacum protoplasts with two Agrobacterium strains. The transformation frequency of unselected A, thaliana shoots was lower than 1\% whereas that of cocultivated tobacco protoplasts was approximately 18\%, The cotransformation frequencies, defined as the frequencies with which cells transformed with a first T-DNA contained a second unselected T-DNA, were approximately 40\% reproducible, irrespective of the selection, the transformation frequency, and the plant system used. Extrapolation of these results suggests that at least two independently transferred T-DNAs were present in 64\% of the transformed plant cells, Molecular analysis of cocultivated N. tabacum shoots regenerated on nonselective medium showed that only a few transformants had a silenced (2/46) or truncated (1/46) T-DNA, Therefore, most integrated T-DNAs expressed their selectable or screenable markers in primary transgenic plants. Remarkably, 10 to 30\% of the selected A. thaliana shoots or progenies lost the T-DNA marker they were selected on. As these regenerants contained the unselected T-DNA with a high frequency (17\%), these selected plants might result from the expression of unstable, transiently expressed T-DNAs, In conclusion, a significant part of the T-DNAs is lost from the transformed cells.},
  author       = {De Buck, Sylvie and Jacobs, Anni and Van Montagu, Marc and Depicker, Anna},
  issn         = {0894-0282},
  journal      = {MOLECULAR PLANT-MICROBE INTERACTIONS},
  language     = {eng},
  number       = {6},
  pages        = {449--457},
  title        = {Agrobacterium tumefaciens transformation and cotransformation frequencies of Arabidopsis thaliana root explants and tobacco protoplasts},
  url          = {http://dx.doi.org/10.1094/MPMI.1998.11.6.449},
  volume       = {11},
  year         = {1998},
}

Altmetric
View in Altmetric
Web of Science
Times cited: