Plant Cell Reports

, Volume 14, Issue 7, pp 450–454

Efficient Agrobacterium-mediated transformation of Arabidopsis thaliana using the bar gene as selectable marker

  • Kazuhito Akama
  • Holger Puchta
  • Barbara Hohn
Article

DOI: 10.1007/BF00234053

Cite this article as:
Akama, K., Puchta, H. & Hohn, B. Plant Cell Reports (1995) 14: 450. doi:10.1007/BF00234053

Summary

We have established an efficient Agrobacterium-mediated transformation procedure for Arabidopsis thaliana genotype C24 using the chimeric bialaphos resistance gene (bar) coding for phosphinothricin acetyltransferase (PAT). Hypocotyl explants from young seedlings cocultivated with agrobacteria carrying a bar gene were selected on shoot-inducing media containing different concentrations of phosphinothricin (PPT) which is an active component of bialaphos. We found that 20 mg/l of PPT completely inhibited the control explants from growing whereas the explants transformed with the bar gene gave rise to multiple shoots resistant to PPT after 3 weeks under the same selection conditions. The transformation system could also be applied to root explants. Resulting plantlets could produce viable seeds in vitro within 3 months after preparation of the explants. The stable inheritance of the resistance trait, the integration and expression of the bar gene in the progeny were confirmed by genetic tests, Southern analysis and PAT enzyme assay, respectively. In addition, the mature plants in soil showed tolerance to the herbicide Basta.

Key words

Arabidopsis thaliana Transformation Phosphinothricin Bar Basta resistance 

Abbreviations

bar

bialaphos resistance gene

CIM

callus-inducing medium

DTNB

5,5′-dithiobis(2-nitrobenzoic acid)

GM

germination medium

HPT

hygromycin phosphotransferase

MS

Murashige and Skoog salts

NPTII

neomycin phosphotransferase II

PAT

phosphinothricin acetyltransferase

PPT

phosphinothricin

SIM

shoot-inducing medium

Copyright information

© Springer-Verlag 1995

Authors and Affiliations

  • Kazuhito Akama
    • 1
  • Holger Puchta
    • 1
  • Barbara Hohn
    • 1
  1. 1.Friedrich-Mieseher-InstitutBaselSwitzerland
  2. 2.Department of BiologyShimane UniversityMatsueJapan

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