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

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    0379091 - ÚEB 2013 RIV AT eng J - Journal Article
    Bleho, J. - Obert, B. - Takáč, T. - Petrovská, Beáta - Heym, C. - Menzel, D. - Šamaj, J.
    ER disruption and GFP degradation during non-regenerable transformation of flax with Agrobacterium tumefaciens.
    Protoplasma. Roč. 249, č. 1 (2012), s. 53-63. ISSN 0033-183X. E-ISSN 1615-6102
    Grant - others:GA MŠk(CZ) ED0007/01/01
    Institutional research plan: CEZ:AV0Z50380511
    Keywords : Agrobacterium rhizogenes * Agrobacterium tumefaciens * Endoplasmic reticulum
    Subject RIV: EB - Genetics ; Molecular Biology
    Impact factor: 2.855, year: 2012

    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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