Isolation and characterization of a novel mycovirus, PeSV, in Pleurotus eryngii and the development of a diagnostic system for it.
A novel mycovirus was isolated from a cultivated edible mushroom, Pleurotus eryngii, with severe epidemic symptoms. Purification of the virus was carried out by a sequential procedure of polyethylene glycol precipitation, differential centrifugation, and equilibrium centrifugation in a CsCl gradient. Nuclease digestion assay and protein analysis revealed that the virus consisted of a single-stranded RNA (ssRNA) genome of 7.8 kbp which was encapsulated by a coat protein of 22 kDa. Transmission electron microscope showed that it was spherical with a diameter of 31 nm. Since there was neither a previous report on discovery of a virus in P. eryngii, nor known mushroom viruses with similar characteristics, we concluded that this is a novel virus and thus have named it as P. e ryngii Spherical Virus (PeSV). Because of a diagnostic test would be helpful in preventing the PeSV-related disease outbreaks, we developed a triple antibody sandwich-ELISA (TAS-ELISA) system using anti-PeSV mouse monoclonal and anti-PeSV rabbit polyclonal antibodies. The TAS-ELISA system successfully detected less than 0.5 microg of the virus particles in 1 g diseased mushroom tissue collected from various commercial farms.
Ro
Hyeon-Su
HS
Department of Microbiology and Research Institute of Life Sciences, GyeongSang National University, Chinju, Korea.
Kang
Eon-Ju
EJ
Yu
Jae-San
JS
Lee
Tae-Soo
TS
Lee
Chang-Won
CW
Lee
Hyun-Sook
HS
D002499:Centrifugation, Density Gradient / Q000379:methods*; D011232:Chemical Precipitation; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D010935:Plant Diseases / Q000821:virology*; D020076:Pleurotus / Q000821:virology*; D012328:RNA Viruses / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*