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ER disruption and GFP degradation during non-regenerable transformation of flax with Agrobacterium tumefaciens

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Abstract

Flax is considered as plant species susceptible to Agrobacterium-mediated genetic transformation. In this study, stability of flax transformation by Agrobacterium rhizogenes versus Agrobacterium tumefaciens was tested by using combined selection for antibiotic resistance and visual selection of green fluorescent protein (GFP)-fusion reporter targeted to the endoplasmic reticulum (ER). Transformation with A. rhizogenes was stable for over 2 years, whereas transformation by A. tumefaciens resulted in non-regenerable stable transformation which was restricted solely to transgenic callus and lasted only 6–8 weeks. However, shoots regenerated from this callus appeared to be non-transgenic. Importantly, callus and root cells stably transformed with A. rhizogenes showed typical regular organization and dynamics of ER as visualized by GFP-ER marker. On the other hand, callus cells transformed with A. tumefaciens showed disintegrated ER structure and impaired dynamics which was accompanied with developmental degradation of GFP. Consequently, shoots which regenerated from such callus were all non-transgenic. Possible reasons for this non-regenerable flax transformation by A. tumefaciens are discussed.

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Acknowledgments

We thank Dr. Bekir Ülker (IZMB Bonn) for critical reading of the manuscript and useful suggestions. This work was supported by Grant No. ED0007/01/01 Centre of the Region Haná for Biotechnological and Agricultural Research.

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The authors declare that they have no conflict of interest.

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Authors and Affiliations

  1. Centre of the Region Haná for Biotechnological and Agricultural Research, Department of Cell Biology, Faculty of Science, Palacký University, Šlechtitelů 11, 783 71, Olomouc, Czech Republic

    Tomáš Takáč & Jozef Šamaj

  2. Institute of Cellular and Molecular Botany, University of Bonn, Kirschallee 1, 53115, Bonn, Germany

    Claudia Heym & Diedrik Menzel

  3. Institute of Plant Genetics and Biotechnology, Slovak Academy of Sciences, Akademická 2, SK-950 07, Nitra, Slovak Republic

    Juraj Bleho & Bohuš Obert

  4. Institute of Experimental Botany, Academy of Sciences of the Czech Republic, Sokolovská 6, 772 00, Olomouc, Czech Republic

    Beáta Petrovská

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  1. Juraj Bleho
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Correspondence to Jozef Šamaj.

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Bleho, J., Obert, B., Takáč, T. et al. ER disruption and GFP degradation during non-regenerable transformation of flax with Agrobacterium tumefaciens . Protoplasma 249, 53–63 (2012). https://doi.org/10.1007/s00709-010-0261-2

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  • DOI: https://doi.org/10.1007/s00709-010-0261-2

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