Reference Report for CAIN769091588
| Title: | Purification and serology of soybean dwarf virus |
| Authors: | Tamada, T., Kojima, M. |
| Source: | Phytopath. Z. 1976, 85(3):237-250 |
| Abstract: | Soybean dwarf virus (SDV), a member of potato leafroll virus group, consists of 2 strains differing in host range and symptoms: dwarfing strain (SDV-DS) and yellowing strain (SDV-Y). Infectivity of virus was assayed by insect injection and membrane feeding methods, and the sensitivity of both methods was similar. Tests with several buffers and pH values showed that phosphate buffer containing EDTA, pH 7.4, was useful for extraction of virus and maintaining its infectivity. Concentrated virus preparations remained infectious for at least 4 mo. at 4.degree. C and for at least 20 days at 15.degree. C. The thermal inactivation point of both strains was 45-50.degree. C. The virus was purified from soybean plants by maceration with high salt concentration buffer, clarification with chloroform and n-butanol and fluorocarbon, precipitation with polyethylene glycol, differential centrifugation and density-gradient centrifugation. The average yield of virus (per kilogram of source tissue) was 369 .mu.g and 166 .mu.g, respectively, for SDV-DS and SDV-Y. The A 260/280 ratio for SDV-DS was 1.96 and that for SDV-Y was 1.90. The virus particles were isometric, about 25 nm in diameter in negatively stained preparations. No differences in sedimentation rate between the 2 isolates were detected in sucrose density-gradient centrifugation tests. Antisera against SDV-DS and SDV-Y, prepared using purified virus, had titers of 2048 and 4096, respectively, in ring precipitin tests. In agar gel-diffusion tests, a single precipitin band was produced between the virus antigens and their homologous or heterologous antisera. In infectivity neutralization tests, antiserum against either isolate completely neutralizied infectivity of the other isolate |
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