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Stable transformation and regeneration of transgenic plants of Pinus radiata D. Don. Plant Cell Reports 17, 460-468.
| Walter C., Grace, L. J., Wagner, A., White, D. W. R., Walden, A. R., Donaldson, S. S., Hinton, H., Gardner, R. C., Smith, D. R. (1998).
| A biolistic particle delivery system was used to genetically transform embryogenic tissue of Pinus radiata. The introduced DNA contained a uidA reporter gene under the control of either the tandem CaMV 35S or the artificial Emu promoter, and the npt II selectable marker controlled by the CaMV 35S promoter. The average number of stable, geneticin-resistant lines recovered was 0.5 per 200 mg fresh weight bombarded tissue. Expression of the uidA reporter gene was detected histochemically and fluorimetrically in transformed embryogenic tissue and in derived mature somatic embryos and regenerated plants. The integration of uidA and npt II genes into the Pinus radiata genome was demonstrated using PCR amplification of the inserts and Southern hybridisation analysis. The expression of both genes in transformed tissue was confirmed by Northern hybridisation analysis. More than 150 transgenic Pinus radiata plants were produced from 20 independent transformation experiments with four different embryogenic clones.
This is the first published report of stable genetic transformation and regeneration of any pine. Microprojectile bombardment of embryogenic cultures with constructs carrying the nptII gene under the conrolled by the CaMV 35S promoter and the uidA gene under the control of a tandem CaMV 35S or the artificial Emu promoter, followed by selection on geneticin was used to obtain transformed embryogenic material, from which transgenic somatic seedlings were regenerated.
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