Protoplasts from a cell culture line were successfully infected with barley yellow dwarf virus. Both purified virions and extracted RNA were shown to be infectious using a polyethylene glycol inoculation procedure. Up to 20% of the protoplasts contained viral antigens as judged by immunofluorescence assay. ELISA analysis showed that virus antigen expression was both dose- and time-dependent. Synthesis of new viral RNAs was confirmed by Northern blot hybridization. Studies of the expression of this economically important aphid-borne, phloem-limited virus should be greatly facilitated using this graminaceous protoplast system. It is the first reproducible protoplast infection system for this virus which can not be transmitted mechanically at the whole plant level.


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