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Rapid induction of genomic demethylation and T-DNA gene expression in plant cells by 5-azacytosine derivatives

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Abstract

We have optimized conditions for demethylation of the genome and induction of a silent, hypermethylated T-DNA gene (ipt) by 5-azacytosine (5-azaCyt) derivatives in a suspension culture of tobacco cells. In this system, 5-azacytidine (5-azaC) is more effective in causing genomic demethylation and ipt gene induction than 5-azaCyt or 5-azadeoxycytidine (5-azadC). A single treatment with 2.5 μM 5-azaC resulted in a maximal level of ipt gene induction without inhibiting cell growth. However, we could not reduce the level of genomic methylation below approximately 2/3 of that found in untreated controls, even after extensive 5-azaC treatment. Furthermore, remethylation of the genome occurred after removal of 5-azaC. The use of 5-azaC as an inducer of silent plant genes is discussed, along with differences in the response of plant and animal genomes to demethylating agents.

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Abbreviations

C:

cytidine

Cyt:

cytosine

5-azaCyt:

5-azacytosine

5-azaC:

5-azacytidine

5-azadC:

5-azadeoxycytidine

m5Cyt:

5-methylcytosine

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Authors and Affiliations

  1. Department of Biochemistry, College of Agricultural and Life Sciences, University of Wisconsin-Madison, 53706, Madison, WI, USA

    Manfred Klaas, Manorama C. John, Dring N. Crowell & Richard M. Amasino

Authors
  1. Manfred Klaas
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  2. Manorama C. John
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  3. Dring N. Crowell
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  4. Richard M. Amasino
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Klaas, M., John, M.C., Crowell, D.N. et al. Rapid induction of genomic demethylation and T-DNA gene expression in plant cells by 5-azacytosine derivatives. Plant Mol Biol 12, 413–423 (1989). https://doi.org/10.1007/BF00017581

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  • DOI: https://doi.org/10.1007/BF00017581

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