Production of antiserum to a non-structural potyviral protein and its use to detect narcissus yellow stripe and other potyviruses.
A protein, of apparent molecular weight 72,000, was purified from experimentally infected narcissus plants with yellow stripe symptoms utilising SDS-polyacrylamide gel electrophoresis. This protein was excised from the gels and used to prepare antiserum, which reacted specifically with cytoplasmic cylindrical inclusions in ultra-thin sections of virus-infected cells and, in immunoblots, with the 72 kDa protein in preparations containing cytoplasmic inclusions. The antiserum reacted in ELISA with leaf extracts from yellow stripe diseased plants of four narcissus cultivars but not with extracts from comparable symptomless plants. In tests with extracts of plants infected with seven definitive potyviruses, reactions were obtained with bean yellow mosaic and iris mild mosaic viruses. Virus-specific reactions in dot-blot ELISA were dependent on the presence of Tween 20 in the extraction buffer. In contrast, an antiserum to the putative cytoplasmic inclusion protein of alstroemeria mosaic virus reacted only with SDS-treated leaf extracts of infected plants. In limited tests, the method of purifying cytoplasmic inclusion protein was successfully applied to four definitive potyviruses, suggesting that it may be generally applicable to potyviruses and of use for preparing antisera when purification of virus particles is difficult.
Scottish Crop Research Institute, Invergowrie, Dundee, U.K.
W P
WP
Mowat
S
S
Dawson
G H
GH
Duncan
D000914:Antibodies, Viral; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D007106:Immune Sera; D015151:Immunoblotting; D007181:Inclusion Bodies, Viral / Q000032:analysis*; D010942:Plant Viruses / Q000032:analysis* / Q000302:isolation & purification; D010944:Plants / Q000382:microbiology*; D014764:Viral Proteins / Q000032:analysis / Q000276:immunology*
D000914:Antibodies, Viral; D007106:Immune Sera; D014764:Viral Proteins