Abstract
Isolate No.30 of bean yellow mosaic virus (BYMV) was purified from broad bean leaves by extraction with 0.1M Tris-HCl buffer, pH7.0, containing 0.05M EDTA and 1% 2-mercaptoethanol; repeated clarification with carbon tetrachloride and precipitation with 4% polyethylene glycol (#6, 000) followed by differential centrifugation and sucrose density-gradient centrifugation. ISCO fractionator scanning patterns indicated a high degree of purity but serological results revealed the presence of some host contaminates. The yield of virus was about 2mg/100g of broad bean leaves.
The titer of the antiserum against isolate No.30 was 1/2048 by ring precipitin tests. Although the intact virus particles could not diffuse in agar gel plates, a clear precipitin line was observed when 0.5% SDS (or LIS) was added to the agar or virus suspension. Isolate No.30 was compared serologically with a necrotic strain and a chlorotic spot strain of BYMV in agar gel diffusion tests. Spur reactions were observed between isolate No.30 and these strains.
