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

Osvaldo A. Castellanos-Hernández, Araceli Rodríguez-Sahagún, Gustavo J. Acevedo-Hernández, Benjamín Rodríguez-Garay, José Luis Cabrera-Ponce, Luis Rafael Herrera-Estrella

Research output: Contribution to journalArticle

7 Scopus citations

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.

Original languageEnglish
Pages (from-to)175-181
Number of pages7
JournalPlant Cell, Tissue and Organ Culture
Volume99
Issue number2
DOIs
StatePublished - Sep 2009

Keywords

  • Biolistics
  • NptII
  • Organogenesis
  • Transformation
  • Tree

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    Castellanos-Hernández, O. A., Rodríguez-Sahagún, A., Acevedo-Hernández, G. J., Rodríguez-Garay, B., Cabrera-Ponce, J. L., & Herrera-Estrella, L. R. (2009). Transgenic Paulownia elongata S. Y. Hu plants using biolistic-mediated transformation. Plant Cell, Tissue and Organ Culture, 99(2), 175-181. https://doi.org/10.1007/s11240-009-9590-2