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Techniques

Partial Purification and Antiserum Production to the 19 × 50-nm Mushroom Virus Particle. J. W. Moyer, Graduate Assistant, Department of Plant Pathology, The Pennsylvania State University, University Park 16802, Present address of senior author: Department of Plant Pathology, North Carolina State University, Raleigh 27607; S. H. Smith, Professor, Department of Plant Pathology, The Pennsylvania State University, University Park 16802. Phytopathology 66:1260-1261. Accepted for publication 26 March 1976. Copyright © 1976 The American Phytopathological Society, 3340 Pilot Knob Road, St. Paul, MN 55121. All rights reserved.. DOI: 10.1094/Phyto-66-1260.

The 19 × 50-nm virus particles were purified from infected sporophores collected from mushroom beds exhibiting characteristic disease symptoms. Sporophores were frozen in liquid nitrogen and stored at –20 C. The polyhedral virus particles were removed by differential precipitation with 10% polyethylene glycol and 0.6 M NaCl. Sucrose density-gradient centrifugation resulted in one major peak containing the 19 × 50-nm particles which exhibited an ultraviolet absorbance spectrum characteristic of a nucleoprotein with a 260/280 ratio of 1.4. Antiserum prepared against the virus contained a host-specific component which was removed by absorption with an extract from healthy tissue. The remaining antiserum was specific for the 19 × 50-nm virus particles.