Purification and serology of a Japanese isolate of barley yellow dwarf virus.

Abstract

A Japanese isolate of barley yellow dwarf virus (BYDV) was purified from infected plants by an improved procedure. The procedure included extraction of the virus by grinding the frozen plant tissues in a meat grinder followed by regrinding in phosphate buffer with motar and pestle for extended periods (more than 2hr) in room temperature, clarification of the sap with chloroform, concentration of the virus by polyethylene glycol, two cycles of differential centrifugation and a sucrose density gradient centrifugation. The average yield of the virus was 44μg per 100g tissues. The Japanese isolate had particle diameter of 27.3nm (2% PTA, pH 5.0) and UV absorbance spectrum with an average A260/A280 ratio of 1.71. In ELISA using the double antibody sandwich method, the virus reacted only with homologous antiserum but not with antiserum to potato leafroll virus (PLRV), and vice versa. The virus, however, reacted slightly with heterologous antisera to other luteoviruses such as PLRV, soybean dwarf and beet western yellows viruses in agar gel double diffusion tests.