Purification and serology of beet mosaic virus.

Abstract

A purification procedure for beet mosaic virus (BMV) was improved. The infected swiss chard (Beta vulgaris var. cicla L.) leaves were homogenized with 0.5M borate buffer, pH 8.0, containing 0.05M ethylenediaminetetraacetic acid and 0.1% thioglycolic acid. After clarification with 1% Triton X-100, the virus was concentrated by precipitation with 4% polyethylene glycol (MW 6, 000) (PEG), and centrifugation utilizing 30% sucrose cushion containing 4% PEG, and further purified by rate zonal sucrose density-gradient centrifugation. The ultraviolet absorption spectrum of the purified preparation with a maximum at 260nm and a minimum at 243nm was typical for the potyviruses. The yield of purified virus was about 0.5∼0.7mg per 100g of infected swiss chard leaves. The titer of the antiserum against BMV was 1/512 by ring interface precipitin test. In sodium laurylsulfate-immunodiffusion test, this antiserum reacted not only with purified BMV but also with crude sap of BMV-infected leaves. No reactions were detected with some other potyviruses including turnip mosaic virus, bean yellow mosaic virus, lettuce mosaic virus and potato virus Y.