Stem sections from shoot cultures maintained in vitro were used to produce transgenic plants of the potato, Solanum tuberosum L. cv. 'Russet Burbank'. Stem internode pieces inoculated with Agrobacterium tumefaciens containing coat protein genes from potato virus X and potato virus Y, produced shoots with a frequency of 60% in the absence of selection and 10% on medium containing 100 mg/l kanamycin monosulfate. Regenerated shoots were assayed for kanamycin resistance by placing stem segments on callus induction medium containing an increased level of kanamycin. Of a total 255 regenerated shoots, 47 (18%) were kanamycin resistant. Of the kanamycin resistant shoots, 25 (53%) expressed the PVX or PVY coat protein genes as assayed by enzyme-linked immunosorbent assay or Western immunoblot analysis.
Russet Burbank potato was transformed with plant expression vectors containing the potato leafroll luteovirus (PLRV) coat protein (CP) gene. Transgenic potato lines contained a gene expression cassette with two copies of a PLRV CP gene in which the nucleotide sequence was modified to improve expression of the gene. In addition, the two copies of the PLRV CP gene were each driven by a different promoter. Field test screening for PLRV resistance identified 15 lines which showed moderate resistance to PLRV infection and virus titer build-up and a longer incubation period for systemic infection. By conducting field resistance assays during a period when the vector of PLRV was not present, it was possible to test whether the observed resistance was sufficient to restrict aphid transmission of PLRV in a field test. Two years of field testing demonstrated that PLRV-spread from an infected plant to adjacent healthy plants of the same line was severely restricted in nearly all the transgenic lines in the field. These lines have useful resistance to PLRV and could aid in managing PLRV disease in Russet Burbank potato.
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