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Modes of Infection of Barley Protoplasts with Brome Mosaic Virus

Laboratory of Plant Pathology, Faculty of Agriculture, Kyoto University, Kyoto 606, Japan

Mesophyll protoplasts of barley were efficiently infected with brome mosaic virus (BMV) in two different ways under different conditions. Polycation-independent infection occurs at low pH in low ionic strength inoculum whereas polycation-dependent infection occurs at high pH in higher ionic strength inoculum. Optimal conditions at low pH were 20 µg/ml BMV, 0.2 µg/ml poly-L-ornithine and 0.7 M-mannitol in 0.1 mM-potassium citrate buffer, pH 5.0; about 70% of protoplasts were infected. Optimal conditions at high pH were 1 µg/ml BMV, 1 µg/ml poly-L-ornithine and 0.7 M-mannitol in 50 mM-phosphate buffer, pH 8.8; nearly 60% of protoplasts became infected. A brief contact of virus and poly-L-ornithine was essential for infection at high pH whereas it inhibited polycation independent infection at low pH. At high pH, poly-L-ornithine formed aggregates with virus. Infection of protoplasts seemed to be controlled by the strength of the positive charge on virus particles at low pH and that on the virus-poly-L-ornithine complexes at high pH.

Received 5 July 1977; accepted 7 September 1977.