STABLE TRANSFORMATION AND REGENERATION OF TRANSGENIC PLANTS OF PINUS-RADIATA DON,D

A biolistic particle delivery system was used to genetically transform embryogenic tissue of Pinus radiata. The introduced DNA contained a u idA reporter gene under the control of either the tandem CaMV 35S or t he artificial Emu promoter, and the npt II selectable marker controlle d by the CaMV 35S promoter. The average number of stable, geneticin-re sistant lines recovered was 0.5 per 200 mg fresh weight bombarded tiss ue. 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 N orthern hybridisation analysis. More than 150 transgenic Pinus radiata plants were produced from 20 independent transformation experiments w ith four different embryogenic clones.