Characterization of the ethanol-inducible alc gene-expression system in Arabidopsis thaliana

Authors

  • Hairul A. Roslan,

    1. Donnan Laboratories and Life Sciences Building, School of Biological Sciences, University of Liverpool, L69 7ZD, UK, and
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    • Present address: Plant Molecular Biotechnology Laboratory, Faculty of Resource Science and Technology, Universiti Malaysia Sarawak, Jalan Dato’ Musa, 94300 Kota Samarahan Sarawak, Malaysia.

  • Michael G. Salter,

    1. Donnan Laboratories and Life Sciences Building, School of Biological Sciences, University of Liverpool, L69 7ZD, UK, and
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    • Present address: Department of Biology, Adrian Building, University of Leicester, Leicester LE2 7RH, UK.

  • Chris D. Wood,

    1. Donnan Laboratories and Life Sciences Building, School of Biological Sciences, University of Liverpool, L69 7ZD, UK, and
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    • §

      Present address: Department of Physiological Sciences, Medical School, Framlington Place, University of Newcastle, Newcastle-upon-Tyne NE2 4HH, UK.

  • Michael R. H. White,

    1. Donnan Laboratories and Life Sciences Building, School of Biological Sciences, University of Liverpool, L69 7ZD, UK, and
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  • Kevan P. Croft,

    1. Donnan Laboratories and Life Sciences Building, School of Biological Sciences, University of Liverpool, L69 7ZD, UK, and
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  • Frances Robson,

    1. John Innes Centre, Norwich Research Park, Colney Lane, Norwich NR4 7UH, UK
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  • George Coupland,

    1. John Innes Centre, Norwich Research Park, Colney Lane, Norwich NR4 7UH, UK
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  • John Doonan,

    1. John Innes Centre, Norwich Research Park, Colney Lane, Norwich NR4 7UH, UK
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  • Patrick Laufs,

    1. John Innes Centre, Norwich Research Park, Colney Lane, Norwich NR4 7UH, UK
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  • A. Brian Tomsett,

    1. Donnan Laboratories and Life Sciences Building, School of Biological Sciences, University of Liverpool, L69 7ZD, UK, and
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  • Mark X. Caddick

    Corresponding author
    1. Donnan Laboratories and Life Sciences Building, School of Biological Sciences, University of Liverpool, L69 7ZD, UK, and
      For correspondence (fax
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For correspondence (fax +44 151 794 3655; email caddick@liverpool.ac.uk).

Summary

Controlled expression of transgenes in plants is key to the characterization of gene function and the regulated manipulation of growth and development. The alc gene-expression system, derived from the filamentous fungus Aspergillus nidulans, has previously been used successfully in both tobacco and potato, and has potential for use in agriculture. Its value to fundamental research is largely dependent on its utility in Arabidopsis thaliana. We have undertaken a detailed function analysis of the alc regulon in A. thaliana. By linking the alcA promoter to β-glucuronidase (GUS), luciferase (LUC) and green fluorescent protein (GFP) genes, we demonstrate that alcR-mediated expression occurs throughout the plant in a highly responsive manner. Induction occurs within one hour and is dose-dependent, with negligible activity in the absence of the exogenous inducer for soil-grown plants. Direct application of ethanol or exposure of whole plants to ethanol vapour are equally effective means of induction. Maximal expression using soil-grown plants occurred after 5 days of induction. In the majority of transgenics, expression is tightly regulated and reversible. We describe optimal strategies for utilizing the alc system in A. thaliana.

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