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

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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.

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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

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Author notes

  1. Kazuhito Akama

    Present address: Department of Biology, Shimane University, 690, Matsue, Japan

Authors and Affiliations

  1. Friedrich-Mieseher-Institut, P.O. Box 2543, CH-4002, Basel, Switzerland

    Kazuhito Akama, Holger Puchta & Barbara Hohn

Authors
  1. Kazuhito Akama
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  2. Holger Puchta
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  3. Barbara Hohn
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Communicated by I. Potrykus

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Akama, K., Puchta, H. & Hohn, B. Efficient Agrobacterium-mediated transformation of Arabidopsis thaliana using the bar gene as selectable marker. Plant Cell Reports 14, 450–454 (1995). https://doi.org/10.1007/BF00234053

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  • DOI: https://doi.org/10.1007/BF00234053

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