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Standardization in Inoculation Procedure and Effect of a Resistance Gene on Infection of Tomato Protoplasts with Tobacco Mosaic Virus RNA

Institute for Plant Virus Research, Tsukuba Science City, Yatabe, Ibaraki 300-21, Japan

Inoculation at 0 °C improved the efficiency of infecting tomato protoplasts with RNA of tobacco mosaic virus (TMV). Up to 41% infection was obtained when inoculation was at 0 °C with 10 µg/ml RNA in the presence of 1 µg/ml poly-D-lysine in 0.01 M-potassium citrate buffered 0.7 M-mannitol (pH 5.2). The homozygous gene for resistance Tm-1 was expressed in tomato protoplasts inoculated with RNA of TMV-L, a common tomato strain of TMV; no virus progeny were detected by fluorescent antibody staining or infectivity assay. Virus multiplied rapidly in protoplasts from susceptible homozygotes. Protoplasts homozygous for Tm-1 were infected by the RNA of TMV-CH2, a tomato strain which can overcome this resistance in plants. These results resemble those previously reported for inocula using intact virus, and suggest that Tm-1 blocks virus growth after the uncoating stage.

Received 18 December 1978; accepted 21 March 1979.