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The elderberry (E) and rhubarb (R) strains of cherry leaf roll virus were each purified and separated into middle (M) and bottom (B) nucleoprotein components by sucrose gradient centrifugation followed by near-equilibrium banding in caesium chloride. Heterologous mixtures of the RNAs from the separated components of the two strains were inoculated into a series of test plants. New local lesion types in Nicotiana clevelandii thus induced were passaged three times through local lesions on N. clevelandii to generate pure pseudorecombinant stocks. M-RNA determined serological specificity, the distribution of virus particle components, the type of systemic symptom in N. clevelandii and N. tabacum cv. Samsun and Xanthi, and local and systemic symptoms in Chenopodium amaranticolor and C. quinoa. B-RNA determined the ability to induce systemic symptoms in Gomphrena globosa and the type of local lesion in N. clevelandii. Backcrossing of the pseudorecombinants generated isolates with properties identical to those of the parental strains.
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