Academic Bibliography

 Search 200 years of publications by Ghent University researchers.
Advanced search
1 file | 444.86 KB
Add to list
  1. Search results
  2. T-DNA transfer and T-DNA integration ...

T-DNA transfer and T-DNA integration efficiencies upon Arabidopsis thaliana root explant cocultivation and floral dip transformation

Rim Ghedira (UGent) , Sylvie De Buck (UGent) , Frédéric Van Ex, Geert Angenon and Anna Depicker (UGent)
(2013) PLANTA. 238(6). p.1025-1037
Author
Organization
Abstract
T-DNA transfer and integration frequencies during Agrobacterium-mediated root explant cocultivation and floral dip transformations of Arabidopsis thaliana were analyzed with and without selection for transformation-competent cells. Based on the presence or absence of CRE recombinase activity without or with the CRE T-DNA being integrated, transient expression versus stable transformation was differentiated. During root explant cocultivation, continuous light enhanced the number of plant cells competent for interaction with Agrobacterium and thus the number of transient gene expression events. However, in transformation competent plant cells, continuous light did not further enhance cotransfer or cointegration frequencies. Upon selection for root transformants expressing a first T-DNA, 43-69 % of these transformants showed cotransfer of another non-selected T-DNA in two different light regimes. However, integration of the non-selected cotransferred T-DNA occurred only in 19-46 % of these transformants, indicating that T-DNA integration in regenerating root cells limits the transformation frequencies. After floral dip transformation, transient T-DNA expression without integration could not be detected, while stable T-DNA transformation occurred in 0.5-1.3 % of the T1 seedlings. Upon selection for floral dip transformants with a first T-DNA, 8-34 % of the transformants showed cotransfer of the other non-selected T-DNA and in 93-100 % of them, the T-DNA was also integrated. Therefore, a productive interaction between the agrobacteria and the female gametophyte, rather than the T-DNA integration process, restricts the floral dip transformation frequencies.
Keywords
SELECTION, AGROBACTERIUM-MEDIATED TRANSFORMATION, RECOMBINATION, LIGHT, TUMEFACIENS, HIGH-FREQUENCY, PLANT-CELLS, VACUUM-INFILTRATION, Transgene expression, Transformation efficiency, CRE recombinase, TARGET, GENETIC-TRANSFORMATION, Agrobacterium

Downloads

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

Citation

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

MLA
Ghedira, Rim, et al. “T-DNA Transfer and T-DNA Integration Efficiencies upon Arabidopsis Thaliana Root Explant Cocultivation and Floral Dip Transformation.” PLANTA, vol. 238, no. 6, 2013, pp. 1025–37, doi:10.1007/s00425-013-1948-3.
APA
Ghedira, R., De Buck, S., Van Ex, F., Angenon, G., & Depicker, A. (2013). T-DNA transfer and T-DNA integration efficiencies upon Arabidopsis thaliana root explant cocultivation and floral dip transformation. PLANTA, 238(6), 1025–1037. https://doi.org/10.1007/s00425-013-1948-3
Chicago author-date
Ghedira, Rim, Sylvie De Buck, Frédéric Van Ex, Geert Angenon, and Anna Depicker. 2013. “T-DNA Transfer and T-DNA Integration Efficiencies upon Arabidopsis Thaliana Root Explant Cocultivation and Floral Dip Transformation.” PLANTA 238 (6): 1025–37. https://doi.org/10.1007/s00425-013-1948-3.
Chicago author-date (all authors)
Ghedira, Rim, Sylvie De Buck, Frédéric Van Ex, Geert Angenon, and Anna Depicker. 2013. “T-DNA Transfer and T-DNA Integration Efficiencies upon Arabidopsis Thaliana Root Explant Cocultivation and Floral Dip Transformation.” PLANTA 238 (6): 1025–1037. doi:10.1007/s00425-013-1948-3.
Vancouver
1.
Ghedira R, De Buck S, Van Ex F, Angenon G, Depicker A. T-DNA transfer and T-DNA integration efficiencies upon Arabidopsis thaliana root explant cocultivation and floral dip transformation. PLANTA. 2013;238(6):1025–37.
IEEE
[1]
R. Ghedira, S. De Buck, F. Van Ex, G. Angenon, and A. Depicker, “T-DNA transfer and T-DNA integration efficiencies upon Arabidopsis thaliana root explant cocultivation and floral dip transformation,” PLANTA, vol. 238, no. 6, pp. 1025–1037, 2013.
@article{4213826,
  abstract     = {{T-DNA transfer and integration frequencies during Agrobacterium-mediated root explant cocultivation and floral dip transformations of Arabidopsis thaliana were analyzed with and without selection for transformation-competent cells. Based on the presence or absence of CRE recombinase activity without or with the CRE T-DNA being integrated, transient expression versus stable transformation was differentiated. During root explant cocultivation, continuous light enhanced the number of plant cells competent for interaction with Agrobacterium and thus the number of transient gene expression events. However, in transformation competent plant cells, continuous light did not further enhance cotransfer or cointegration frequencies. Upon selection for root transformants expressing a first T-DNA, 43-69 % of these transformants showed cotransfer of another non-selected T-DNA in two different light regimes. However, integration of the non-selected cotransferred T-DNA occurred only in 19-46 % of these transformants, indicating that T-DNA integration in regenerating root cells limits the transformation frequencies. After floral dip transformation, transient T-DNA expression without integration could not be detected, while stable T-DNA transformation occurred in 0.5-1.3 % of the T1 seedlings. Upon selection for floral dip transformants with a first T-DNA, 8-34 % of the transformants showed cotransfer of the other non-selected T-DNA and in 93-100 % of them, the T-DNA was also integrated. Therefore, a productive interaction between the agrobacteria and the female gametophyte, rather than the T-DNA integration process, restricts the floral dip transformation frequencies.}},
  author       = {{Ghedira, Rim and De Buck, Sylvie and Van Ex, Frédéric and Angenon, Geert and Depicker, Anna}},
  issn         = {{0032-0935}},
  journal      = {{PLANTA}},
  keywords     = {{SELECTION,AGROBACTERIUM-MEDIATED TRANSFORMATION,RECOMBINATION,LIGHT,TUMEFACIENS,HIGH-FREQUENCY,PLANT-CELLS,VACUUM-INFILTRATION,Transgene expression,Transformation efficiency,CRE recombinase,TARGET,GENETIC-TRANSFORMATION,Agrobacterium}},
  language     = {{eng}},
  number       = {{6}},
  pages        = {{1025--1037}},
  title        = {{T-DNA transfer and T-DNA integration efficiencies upon Arabidopsis thaliana root explant cocultivation and floral dip transformation}},
  url          = {{http://doi.org/10.1007/s00425-013-1948-3}},
  volume       = {{238}},
  year         = {{2013}},
}
Altmetric
View in Altmetric
Web of Science
Times cited: