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Protoplasma

, Volume 249, Issue 1, pp 53–63 | Cite as

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

  • Juraj Bleho
  • Bohuš Obert
  • Tomáš Takáč
  • Beáta Petrovská
  • Claudia Heym
  • Diedrik Menzel
  • Jozef ŠamajEmail author
Original Paper

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.

Keywords

Agrobacterium rhizogenes Agrobacterium tumefaciens Endoplasmic reticulum Flax GFP Non-regenerable transformation Stable transformation 

Notes

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.

Conflict of interest

The authors declare that they have no conflict of interest.

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Juraj Bleho
    • 3
  • Bohuš Obert
    • 3
  • Tomáš Takáč
    • 1
  • Beáta Petrovská
    • 4
  • Claudia Heym
    • 2
  • Diedrik Menzel
    • 2
  • Jozef Šamaj
    • 1
    Email author
  1. 1.Centre of the Region Haná for Biotechnological and Agricultural Research, Department of Cell Biology, Faculty of SciencePalacký UniversityOlomoucCzech Republic
  2. 2.Institute of Cellular and Molecular BotanyUniversity of BonnBonnGermany
  3. 3.Institute of Plant Genetics and BiotechnologySlovak Academy of SciencesNitraSlovak Republic
  4. 4.Institute of Experimental BotanyAcademy of Sciences of the Czech RepublicSokolovská 6OlomoucCzech Republic

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