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Vacuum infiltration of Petunia hybrida pollen with Agrobacterium tumefaciens to achieve plant transformation

  • GENETIC TRANSFORMATION AND HYBRIDIZATION
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Abstract

 Genetic transformation of Petunia hybrida with a reporter gene and selectable marker gene (35S-bar) was achieved in similar frequencies by pollinating flowers with pollen vacuum-infiltrated with Agrobacterium tumefaciens or applying a drop of Agrobacterium suspension to the stigma immediately prior to pollination. Nine percent of the T1, and 5% of the T2 progeny germinated in nutrient medium with 3 mgl/l BastaR. Polymerase chain reaction assays indicated that of the BastaR-resistant plants, 66% of the T1 plants, and 61% of the T2 plants harboured the GUS gene. Histochemical assays showed that 10% of the putatively transformed T1 plants and 5% of their progeny expressed GUS in leaf tissue, pistils and young anthers. Southern hybridization confirmed genomic integration of the bar gene in one to three places in selected T1 and T2 progeny.

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Received: 12 March 1999 / Revision received: 1 October 1999 / Accepted: 20 October 1999

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Tjokrokusumo, D., Heinrich, T., Wylie, S. et al. Vacuum infiltration of Petunia hybrida pollen with Agrobacterium tumefaciens to achieve plant transformation. Plant Cell Reports 19, 792–797 (2000). https://doi.org/10.1007/s002990050009

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  • DOI: https://doi.org/10.1007/s002990050009

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