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Treatment of Agrobacterium or leaf disks with 5-azacytidine increases transgene expression in tobacco

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Abstract

We have studied the effect of the demethylating agent azacytidine (azaC) on expression of a β-glucuronidase (GUS) gene transferred to tobacco leaf disks by Agrobacterium-mediated transformation. In a system where no selection was performed, where shoot formation was partially repressed, and where Agrobacterium does not express the GUS gene, we were able to follow the early events of transient and stable expression. Two days after inoculation, 8% of the cells expressed GUS but this proportion rapidly decreased to near zero in the following week. Treatment of leaf disks with azaC just after transformation retarded this inactivation to some extent, while treatment of Agrobacterium prior to transformation increased the frequency of transient expression. Three weeks after inoculation the number of GUS-expressing cells increased 4- to 6-fold in the leaf disks treated with azaC and in the leaf disks transformed with azaC-treated bacteria, while the control remained low. These data suggest that DNA methylation is involved in transgene inactivation and that a large number of silent but potentially active transgenes become integrated.

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Author notes

  1. Gorm Palmgren

    Present address: Max Planck Institut für Züchtungsforschung, Carl von Linné Weg 10, D-5000, Köln 30, FRG

Authors and Affiliations

  1. Institute of Plant Physiology, University of Copenhagen, Øster Farimagsgade 2A, DK-1353, Copenhagen K, Denmark

    Gorm Palmgren & Ole Mattson

  2. Maribo Seed Biotechnology, Danisco A/S, 1 Langebrogade, P.O. Box 17, DK-1001, Copenhagen K, Denmark

    Finn Thyge Okkels

Authors
  1. Gorm Palmgren
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  2. Ole Mattson
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  3. Finn Thyge Okkels
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Palmgren, G., Mattson, O. & Okkels, F.T. Treatment of Agrobacterium or leaf disks with 5-azacytidine increases transgene expression in tobacco. Plant Mol Biol 21, 429–435 (1993). https://doi.org/10.1007/BF00028801

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  • DOI: https://doi.org/10.1007/BF00028801

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