ER disruption and GFP degradation during non-regenerable transformation of flax with Agrobacterium tumefaciens.
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.
Institute of Plant Genetics and Biotechnology, Slovak Academy of Sciences, Nitra, Slovak Republic.
Juraj
J
Bleho
Bohuš
B
Obert
Tomáš
T
Takáč
Beáta
B
Petrovská
Claudia
C
Heym
Diedrik
D
Menzel
Jozef
J
Samaj
D016960:Agrobacterium tumefaciens / Q000235:genetics / Q000378:metabolism*; D003470:Culture Media / Q000378:metabolism; D024881:Drug Resistance, Bacterial; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000378:metabolism*; D019597:Flax / Q000166:cytology / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism*; D018517:Plant Roots / Q000235:genetics / Q000378:metabolism; D018520:Plant Shoots / Q000254:growth & development / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000166:cytology / Q000235:genetics / Q000254:growth & development / Q000378:metabolism; D013997:Time Factors; D046509:Tissue Culture Techniques; D014170:Transformation, Genetic; D019076:Transgenes
D003470:Culture Media; D049452:Green Fluorescent Proteins
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