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Transgenic Paulownia elongata S. Y. Hu plants using biolistic-mediated transformation

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Abstract

A biolistic protocol for the stable genetic transformation of the hardwood tree Paulownia elongata was developed. Leaf explants were bombarded using the PDS-1000/He system with plasmid pBI121. The introduced DNA contained the β-glucuronidase (GUS) reporter gene and neomycin phosphotransferase (nptII) as a selection marker. Transformed calli were induced and selected on medium supplemented with 50 mg L−1 kanamycin, and transgenic plants were regenerated through indirect organogenesis. Complete plants were successfully transferred to soil and established under greenhouse conditions. Different helium pressures and explant positions were used and the transformation frequency was calculated. Optimal conditions for genetic transformation were bombardment of the abaxial leaf surface at a pressure of 450 psi. The integration of the transgenes in the plant genome and their stable expression was demonstrated by fluorometric GUS assay, determination of NPTII activity and PCR analysis. This method allows the production of transgenic trees of P. elongata in a relatively short time.

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Acknowledgments

OACH and ARS wish to thank CONACYT (Consejo Nacional de Ciencia y Tecnologia) for PhD fellowships, Authors are grateful to Aurora Verver y Vargas for her invaluable help with the histological analysis.

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Authors and Affiliations

  1. Departamento de Ciencias Básicas, Centro Universitario de la Ciénega, Universidad de Guadalajara (CUCI-UdeG), Av. Universidad 1115, 47820, Ocotlán, Jalisco, Mexico

    Osvaldo A. Castellanos-Hernández & Araceli Rodríguez-Sahagún

  2. Unidad de Biotecnología Vegetal, Centro de Investigación y Asistencia en Tecnología y Diseño del Estado de Jalisco, A. C. (CIATEJ), Normalistas 800 Colinas de la Normal, 44270, Guadalajara, Jalisco, Mexico

    Benjamín Rodríguez-Garay

  3. Laboratorio Nacional de Genómica para la Biodiversidad, Cinvestav Campus Guanajuato, Km 9.6 Libramiento Norte Carretera Irapuato-León, Apartado Postal 629, CP 36500, Irapuato, Guanajuato, Mexico

    Gustavo J. Acevedo-Hernández & Luis Rafael Herrera-Estrella

  4. Departamento de Ingeniería Genética, Cinvestav Campus Guanajuato, Km 9.6 Libramiento Norte Carretera Irapuato-León, Apartado Postal 629, CP 36500, Irapuato, Guanajuato, Mexico

    José Luis Cabrera-Ponce

Authors
  1. Osvaldo A. Castellanos-Hernández
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  2. Araceli Rodríguez-Sahagún
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  3. Gustavo J. Acevedo-Hernández
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  4. Benjamín Rodríguez-Garay
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  5. José Luis Cabrera-Ponce
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  6. Luis Rafael Herrera-Estrella
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Correspondence to Osvaldo A. Castellanos-Hernández.

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Castellanos-Hernández, O.A., Rodríguez-Sahagún, A., Acevedo-Hernández, G.J. et al. Transgenic Paulownia elongata S. Y. Hu plants using biolistic-mediated transformation. Plant Cell Tiss Organ Cult 99, 175–181 (2009). https://doi.org/10.1007/s11240-009-9590-2

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  • DOI: https://doi.org/10.1007/s11240-009-9590-2

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