Protoplasts isolated from suspension-cultured tobacco cells were inoculated with tobacco mosaic virus (TMV) using a procedure similar to that for tobacco mesophyll protoplasts. Fifty to 70% of protoplasts were infected, as determined by staining with fluorescent TMV antibody. The formation of progeny virus particles was substantiated by electron microscopy of sectioned protoplasts, and the virus yield was assessed by assaying the infectivity of protoplast extracts. The course of TMV multiplication was studied by following the incorporation of [H]uridine into virus RNA. The results indicated that infection in this system is synchronous and that the rate of virus multiplication is comparable to that in mesophyll protoplasts. A system of undifferentiated, growing plant cells was thus established for one-step growth of TMV.


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