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Methotrexate is a new selectable marker for tobacco immature pollen transformation

  • Genetic Transformation and Hybridization
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Abstract

We report here a new selectable marker for tobacco immature pollen transformation based on the expression of dihydrofolate reductase (dhfr) gene which confers resistance to methotrexate (Mtx). Two immature pollen transformation approaches, i.e., male germ line transformation and particle bombardment of embryogenic mid-bicellular pollen have been used for the production of stable transgenic tobacco plants. In the first method, two methotrexate-resistant plants were selected from a total of 7161 seeds recovered after transformation experiments. In the second method, four methotrexate-resistant plants were obtained from 29 bombardments using 3.7×105 pollen grains per bombardment. Southern analysis confirmed the transgenic nature of T0 and T1 candidate transgenic plants, and a genetic analysis showed that the transgenes are transmitted to subsequent generations.

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Acknowledgements

The authors would like to thank Dr J Greilhuber and Dr EM Temsch (Botany Institute, University of Vienna) for technical support and kind help to perform the Flow cytometry analysis. The authors are also very thankful to Dr B Reiss (Max-Planck Institute für Züchtungsforschung, Köln, Germany) for providing the pMN125 plasmid. This study was funded by the grant of the European Union QLK-CT-2000-00365.

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Authors and Affiliations

  1. Max F. Perutz Laboratories at the Vienna Biocenter, Department of Genetics, Vienna University, Dr. Bohrgasse 9/4, A-1030, Vienna, Austria

    Tatiana Aionesei, Julia Hosp, Viktor Voronin, Erwin Heberle-Bors & Alisher Touraev

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  1. Tatiana Aionesei
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  2. Julia Hosp
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  3. Viktor Voronin
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  4. Erwin Heberle-Bors
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  5. Alisher Touraev
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Correspondence to Alisher Touraev.

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Communicated by W. Harwood

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Aionesei, T., Hosp, J., Voronin, V. et al. Methotrexate is a new selectable marker for tobacco immature pollen transformation. Plant Cell Rep 25, 410–416 (2006). https://doi.org/10.1007/s00299-005-0067-4

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  • DOI: https://doi.org/10.1007/s00299-005-0067-4

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