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Infectivity Changes in Preparations from Plants Infected with Tobacco Mosaic Virus

Rothamsted Experimental Station, Harpenden, Herts

Replication of tobacco mosaic virus or its nucleic acid in cell-free preparations from plants has recently been claimed. Some of the evidence supporting such claims has been based on increases in infectivity measured by inoculating assay plants with these preparations or phenol extracts from them (1–5). During attempts to repeat this work it was observed that such increases in infectivity also occurred in conditions precluding the replication of the virus or its nucleic acid.

Cell-free preparations were made from leaves of Nicotiana tabacum, type White Burley, var. Judy's Pride, 5 to 6 days after they were inoculated with tobacco mosaic virus. The leaf tissue was ground with pestle and mortar adding per gram of leaf 1 ml. of buffer containing 0·04 M-sucrose, 0·05 M 2-amino-2-hydroxymethylpropane-1,3-diol (tris) adjusted to pH 7·5 with HCl, and 0·005 M-MgCl₂. The mush was strained through two layers of cheesecloth, and the filtrate centrifuged for 1 hr at 106,000g, or for 3 hr at 42,000g (all operations at 0 to 4°).

^* Present address: Stanford University of Medicine, Palo Alto, California, U.S.A.

Received 28 October 1966; accepted 2 December 1966.