Proceedings of the National Academy of Sciences · 1992 · 124 citations · 16 references
AgriculturePlant BiologyPlant SciencesPlant GeneticsPlant VirologyTransgenic CropsAgrobacterium-mediated TransformationPlant Disease ResistancePlant Virus ReplicationPlant Virus ResistancePlant VirusesAgrobacterium TumefaciensAgrobacterium-mediated Transformation ConstructsMolecular Characterization Of Transgenic PlantsResistance Gene-mediated ResistanceTransgenic PlantsTransgenic Virus ResistanceViral Replicase ComplexVirus-resistant Plants
Nicotiana tabacum cv. Turkish Samsun NN plants were transformed with a modified and truncated replicase gene encoded by RNA-2 of cucumber mosaic virus strain Fny. The replicase gene had been modified by deleting a 94-base-pair region spanning nucleotides 1857-1950; the deletion also caused a shift in the open reading frame, resulting in a truncated translation product approximately 75% as large as the full-length protein. Upon transformation via Agrobacterium tumefaciens, transgenic plants were obtained that were resistant to virus disease when challenged with either cucumber mosaic virus virions or RNA at concentrations up to 500 micrograms/ml or 50 micrograms/ml, respectively, the highest concentrations tested. This resistance was absolute, as neither symptoms nor virus could be detected in uninoculated leaves, even after prolonged incubation (120 days after inoculation). These data suggest, therefore, that such a "replicase-mediated" resistance strategy may be applicable to other plant and animal viruses.
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Rapid isolation of high molecular weight plant DNA
Michael G. Murray, William F. Thompson · Nucleic Acids Research · 1980 · 12.4K citations · Full text