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Relationship between allelic state of T-DNA and DNA methylation of chromosomal integration region in transformed Arabidopsis thaliana plants

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Abstract

T-DNA insertions are currently used as a tool to introduce, or knock out, specific genes. The expression of the inserted gene is frequently haphazard and up to now, it was proposed that transgene expression depends on the site of insertion within the genome, as well as the number of copies of the transgene. In this paper, we show that the allelic state of a T-DNA insertion can be at the origin of epigenetic silencing. A T-DNA insertional mutant was characterized to explore the function of AtBP80a′, a vacuolar sorting receptor previously associated with germination. Seeds homozygous for the T-DNA do not germinate, but this can be overcome by a cold treatment and maintained by the following generations. The non-germinating phenotype is only observed in homozygous seed produced by heterozygous plants indicating that it is correlated with the allelic state of the T-DNA in parental lines. Analysis of the region between the T-DNA insertion and the ATG codon of atbp80a′ showed that cytosine methylation is highly enhanced in chromatin containing the T-DNA. Data presented here show that an unpaired DNA region during meiosis could be at the origin of a de novo cytosine methylation mechanism.

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Abbreviations

KANR:

kanamycin resistant

T-DNA:

transfer DNA

VSR:

Vacuolar Sorting Receptor

WT:

wild type

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Authors and Affiliations

  1. Surfaces Cellulaires et Signalisation chez les Végétaux, UMR 5546 CNRS-Université Paul Sabatier, Pôle de Biotechnologie végétale, 24 chemin de Borde-Rouge, BP42617, 31326, Castanet-Tolosan, France

    Frédéric G. Masclaux, Rafael Pont-Lezica & Jean-Philippe Galaud

  2. Laboratoire de Génétique végétale, Université de Genève, Sciences III 30, CH-1211, Quai Ernest-Ansermet, Genève 4, Switzerland

    Frédéric G. Masclaux

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  1. Frédéric G. Masclaux
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  2. Rafael Pont-Lezica
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  3. Jean-Philippe Galaud
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Correspondence to Jean-Philippe Galaud.

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Masclaux, F.G., Pont-Lezica, R. & Galaud, JP. Relationship between allelic state of T-DNA and DNA methylation of chromosomal integration region in transformed Arabidopsis thaliana plants. Plant Mol Biol 58, 295–303 (2005). https://doi.org/10.1007/s11103-005-4808-0

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  • DOI: https://doi.org/10.1007/s11103-005-4808-0

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