Production of biosafe transgenic potato plants with coat protein gene for potato virus Y [electronic resource]

Includes references.

Most techniques of Agrobacterium-mediated transformation described in the literature used antibiotic resi.llllnce genes as selectable marker genes. Theoretically, risks of horizontal gene flow of antibiotic resisrance genes from transgenic plants to enteric bacteria are expected. We describe here the possibi/itv of producing transgenic potato plants harbouring the coat protein gene of potato virus Y (CP-PVY). conferring resistance against potyvirus Y without antibiotic resistance genes. To achieve this goal, transformation conditions of different potato explants. using Agrobacterium rumefaciens strain EHA105 harbouring neomycin phosphotransferase II gene (nptII), which 'render plants kanamyein resistance were optimized. The optimized conditions were followed typically to transform new explants with Agrobacterium tumefaciens strain EHA]05 harbouring inrron-conraining reporter gene, B-glucuronidase (GUS), and CP-PVY gene. i.e. only eukaryotic organisms will be able to process this reporter gene. No selective pressure of anribiotic was applied in rhis srage. Putative transfomwnts regenerated from explants on regeneration medium-free of anribiorics were screened using GUS fluorescence assay and cOllfirmed by polymersase chain reaction (PCR). GUS and PCR positive plants were further confirmed .!iJr the integration and expression of CP-PVY gene using reverse transcriptase PCR (RT-PCR). The transformants "'ere mass micropropagated and microtubers were produced in vitro. The obrained results indicared rhal it is possible to avoid the use of antibiotic and herbicide resistance genes as selectable markers and consequently avoid environmental risks, which may develop in the future due to the use 01' anlibiotics resistance genes, as selectable markers genes. Moreover, this svstem may be useful for Ihe transformation of crops known to be recalcitrant for in vitro regeneration, as the omitting oj' antibiotic from the regeneration medium enhances the percentage of shoot recovery. Further studies are going on to evaluate the efficacy of the transgenic potato clone for resistance to potato virus Y isolates under Egyptian conditions.

Summary in Arabic.

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