Research Article

Infection of Barley Protoplasts with Barley Stripe Mosaic Virus Detected and Assayed by Immunoperoxidase

Journal of General Virology 1982; 58(2):323 · https://doi.org/10.1099/0022-1317-58-2-323

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Abstract

The kinetics of infection of barley protoplasts with barley stripe mosaic virus (BSMV) was followed by enzyme-linked immunosorbent assay (ELISA) using the double-sandwich procedure. Infected protoplasts were identified by an immunoperoxidase method taking precautions to inhibit endogenous peroxidase. About 70% of the protoplasts were infected with BSMV when the inoculum contained 50 µg/ml purified BSMV in 10 mM-potassium citrate buffer pH 4.7 containing 0.65 M-mannitol and 5 µg/ml poly-L-ornithine. It was estimated by ELISA that about 1400 to 1600 virus particles were adsorbed per protoplast. Following an apparent latent period, virus increased linearly between about 14 and 48 h after inoculation. After 48 h the virus concentration reached between 1.62 x 106 and 2.0 x 106 particles per protoplast.