Genetic transformation in commercial Tasmanian cultivars of opium poppy, Papaver somniferum, and movement of transgenicpollen in the field

Functional Plant Biology : FPB
Julie A ChittyPhilip J Larkin

Abstract

We report a new transformation protocol for the pharmaceutically important opium poppy, Papaver somniferum L.; the protocol allows transformation for the first time of high yielding commercial cultivars. The method involves Agrobacterium tumefaciens infection of hypocotyl explants, followed by the production of antibiotic- or herbicide-resistant embryogenic callus and the subsequent induction of somatic embryos and plants. Key elements of the improvement are the use of buffering agents to stabilise medium pH and bottom-cooling of the cultures. Transformation was verified by PCR and Southern blot hybridisation. Transcription of transgenes was confirmed by RT-PCR and product sequencing. Expression of transgenes was detected by histochemical GUS staining, phosphinothricin acetyltransferase (PAT) enzyme assays for bar and pat genes, and western analysis of transgenic sunflower seed albumin protein. Expression of various transgenes was detected in stem, leaf, seed, capsule and latex. The pat gene was demonstrated to be stably inherited to the T2 generation and to confer phosphinothricin (PPT) herbicide resistance. Most T0 plants showed normal morphology, were self-fertile and the transgenes displayed the expected Mendelian segregati...Continue Reading

Citations

Nov 16, 2004·Nature Biotechnology·Robert S AllenPhilip J Larkin
May 15, 2008·The Plant Journal : for Cell and Molecular Biology·Peter J Facchini, Vincenzo De Luca
Dec 7, 2007·Plant Cell Reports·Peter J FacchiniVincent Blanche
Nov 22, 2011·The Plant Journal : for Cell and Molecular Biology·Champa P Wijekoon, Peter J Facchini
Jul 1, 2005·Functional Plant Biology : FPB·Ani BarbulovaMaurizio Chiurazzi

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