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Pine (Pinus radiata).

Jan Grant1, Tracy Dale, Pauline Cooper

  • 1New Zealand Institute for Crop and Food Research Ltd., Christchurch, New Zealand.

Methods in Molecular Biology (Clifton, N.J.)
|October 13, 2006
PubMed
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This study introduces a new method for transforming Monterey pine (Pinus radiata) using cotyledon explants, offering year-round seed availability and high regeneration rates for improved conifer genetic modification.

Area of Science:

  • Plant Biotechnology
  • Forest Genetics
  • Molecular Biology

Background:

  • Somatic embryogenesis is a common method for conifer transformation.
  • Organogenesis offers an alternative route for plant genetic engineering.

Purpose of the Study:

  • To describe a novel organogenic cotyledon explant method for Pinus radiata transformation.
  • To highlight the advantages of this method over traditional approaches.

Main Methods:

  • Utilized mature cotyledon explants from Pinus radiata seeds.
  • Employed Agrobacterium tumefaciens for gene transfer.
  • Optimized cotyledon survival, Agrobacterium strain, and selection pressure.

Main Results:

  • Achieved a transformation efficiency of 1.7% for regenerated shoots.

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  • Demonstrated year-round seed availability and regeneration of over 80% of genotypes.
  • Observed predominantly single-copy transgene integrations, typical of Agrobacterium-mediated methods.

    • Conclusions:

    • Organogenic cotyledon explants provide a viable and efficient alternative for Pinus radiata transformation.
    • This method offers practical advantages including consistent seed supply and broad genotypic regeneration.
    • Key factors for successful transformation include explant viability, bacterial strain selection, and appropriate selection pressure.