In Vitro Cellular & Developmental Biology - Plant

, Volume 41, Issue 6, pp 725–730

Stable genetic transformation of embryogenic cultures of Abies nordmanniana (nordmann fir) and regeneration of transgenic plants

Authors

    • New Zealand Forest Research Institute Ltd
    • Tissue Culture Laboratory, Botanic Garden and Museum, Natural History Museum of DenmarkUniversity of Copenhagen
  • Julia A. Charity
    • New Zealand Forest Research Institute Ltd
  • Lynette J. Grace
    • New Zealand Forest Research Institute Ltd
  • Michel M. M. H. Kristensen
    • Tissue Culture Laboratory, Botanic Garden and Museum, Natural History Museum of DenmarkUniversity of Copenhagen
  • Peter Krogstrup
    • Tissue Culture Laboratory, Botanic Garden and Museum, Natural History Museum of DenmarkUniversity of Copenhagen
  • Christian Walter
    • New Zealand Forest Research Institute Ltd
Article

DOI: 10.1079/IVP2005704

Cite this article as:
Find, J.I., Charity, J.A., Grace, L.J. et al. In Vitro Cell.Dev.Biol.-Plant (2005) 41: 725. doi:10.1079/IVP2005704

Summary

Stable genetic transformation of embryogenic cultures of Abies nordmanniana (Nordmann fir or Caucasian fir) was achieved using the Biolistic® transformation technology, followed by regeneration of transgenic plants. Selection of the transgenic tissue was based on the antibiotic resistance induced by the neomycin phosphotransferase II gene (npt II), in combination with the antibiotic geneticin. Six transclones were recovered from a total of 215 bombardments. Expression of the β-glucuronidase gene (uidA) was confirmed by histochemical analysis, and expression of npt II was verified by quantification of NPTII protein by enzyme linked immunosorbent assay (ELISA). Both genes were still expressed in the embryogenic tissue after 5 yr of in vitro culture and in mature somatic embryos and plants produced from these cultures. The integration of npt II was confirmed by Southern hybridization in embryogenic tissue after 5 yr of culture. After 5 yr of growth, uidA was still expressed in needles from the transformed trees.

Key words

Nordmann firAbies nordmannianaBiolistic® transformationgenetic engineeringgene expression
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Copyright information

© Society for In Vitro Biology 2005