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Appearance of Tobacco Mosaic Virus in Thin Section Using Different Staining Procedures

Department of Botany, University of Nottingham and College of Agriculture, Cracow, Poland

Tomato fruit infected with the vulgare strain of tobacco mosaic virus were used for the isolation of protoplasts infected with tobacco mosaic virus (Gregory & Cocking, 1965). Protoplasts were fixed in glutaraldehyde, post-fixed in osmium tetroxide and embedded in a butyl methacrylate+styrene mixture (70:30, v/v) as previously described (Mohr & Cocking, 1968). Two different staining procedures were employed. The first (method A) involved incubation of protoplasts during dehydration in a freshly prepared, Millipore-filtered 1% solution of uranyl acetate in absolute ethanol for 1 hr at 18°. In the second procedure (method B) incubation was for 15 hr at 25° in a freshly prepared, Millipore-filtered 1% solution of uranyl acetate in 70% ethanol. With both staining procedures sections were post-stained for hr with lead citrate (Reynolds, 1963). Electron micrographs were made with an A.E.I. EM6B at 60 kv using a 25 µ objective aperture.

Received 2 November 1967; accepted 10 December 1967.