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Agrobacterium-mediated transformation of embryogenic tissues of hybrid firs (Abies spp.) and regeneration of transgenic emblings

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Abstract

A genetic transformation system has been developed for selected embryogenic cell lines of hybrids Abies alba × A. cephalonica (cell lines AC2, AC78) and Abies alba × A. numidica (cell line AN72) using Agrobacterium tumefaciens. The cell lines were derived from immature or mature zygotic embryos on DCR medium containing BA (1 mg l−1). The T-DNA of plant transformation vector contained the β-glucuronidase reporter gene under the control of double dCaMV 35S promoter and the neomycin phosphotransferase selection marker gene driven by the nos promoter. The regeneration of putative transformed tissues started approximately 1 week after transfer to the selection medium containing 10 mg geneticin l−1. GUS activity was detected in most of the geneticin-resistant sub-lines AN72, AC2 and AC78, and the transgenic nature of embryogenic cell lines was confirmed by PCR approach. Plantlet regeneration from PCR-positive embryogenic tissues has been obtained as well. The presence of both gus and nptII genes was confirmed in 11 out of 36 analysed emblings.

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Acknowledgements

This work was funded by the Slovak Grant Agency VEGA, projects No. 2-5022-25 and No. 2-0011-08.

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Authors and Affiliations

  1. Institute of Plant Genetics and Biotechnology, Slovak Academy of Sciences, Akademická 2, P.O. Box 39A, 950 07, Nitra, Slovak Republic

    Terézia Salaj, Jana Moravčíková, Božena Vooková & Ján Salaj

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  1. Terézia Salaj
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  2. Jana Moravčíková
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  3. Božena Vooková
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  4. Ján Salaj
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Correspondence to Terézia Salaj.

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Salaj, T., Moravčíková, J., Vooková, B. et al. Agrobacterium-mediated transformation of embryogenic tissues of hybrid firs (Abies spp.) and regeneration of transgenic emblings. Biotechnol Lett 31, 647–652 (2009). https://doi.org/10.1007/s10529-009-9923-6

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  • DOI: https://doi.org/10.1007/s10529-009-9923-6

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