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A genetic transformation system via the pollen tube pathway is presented, plasmid DNA prepared with each of 4 different methods was applied to the surface of an ovary wound site after removal of the style of florets following pollination. Movement of the plasmid DNA indicated plasmid DNA reached the ovules of decapitated florets within about 24 hours after its application to the surface of remaining styles or ovary wound site after pollination. Based on the result of PCR analyses of genomic DNA, 12% to 15% of the plants tested had the 282 bp fragment, the specific portion of the luciferase gene construct into the genome. Southern blotting of genomic DNA from PCR positive plants indicated that the firefly luciferase gene construct may have been incorporated into the genomic DNA of the plants. Some of the PCR positive plants, derived from seeds harvested from florets treated with a DNA solution preparation, still had the 282 bp fragment of the luciferase gene construct in their sexually produced progenies, indicating that transformation via the pollen tube pathway has the potential to produce stably transformed plants.
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