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The Infection of Tobacco Protoplasts with Cowpea Chlorotic Mottle Virus and its RNA

John Innes Institute, Colney Lane, Norwich, NOR 70F, U.K.

Tobacco protoplasts were infected with cowpea chlorotic mottle virus and its RNA in the presence of poly-L-ornithine to yield 10⁶ to 10⁷ particles per infected protoplast representing a probable increase in virus concentration of at least 1000-fold by 24 to 72 h after inoculation. The optimum inoculum input for virus was about 0.5 µg/ml and for RNA about 1.2 µg/ml; about 60 and 7%, respectively, of the protoplasts became infected, as judged by fluorescent antibody staining. Data relating to growth curves, inoculum and poly-L-ornithine concentrations, and pH effects are presented along with electron micrographs of sectioned protoplasts.

Received 14 February 1973; accepted 12 March 1973.