Electron Microscopic Detection of Novel, Coiled Viruslike Particles Associated with Graft-Inoculation of Prunus Species.

Coiled, viruslike particles (spirions) were detected by electron microscopic examination of crude extracts from flowers, leaves, and/or roots of infected Prunus avium, P. mume, and P. serrulata. The particles were observed in ultrathin sections of lower epidermis, palisade, and spongy mesophyll cells of leaves of P. avium and P. mume. Spring (March and April) appears to be optimal for detection of the particles in both screenhouse and field-grown plants. The particles were successfully graft-transmitted to P. armeniaca (cvs. Luizet and Tilton), P. avium (cvs. Bing, F12/1, Mazzard, and Sam), P. mahaleb, and P. persica (cv. Elberta). Individual spirions measured 132 × 34 nm. The particles appear to be coiled forms of a filamentous virus. Filaments extending from some coiled particles were approximately 13 nm wide, with striations at a pitch of 3.24 nm. No disease symptoms or cytopathological abnormalities were associated with the presence of the particles in the Prunus species studied. The particles were not detected in virus-free control plants.

Improved staining boxes for fast, uniform staining of ultrathin sections on grids.

A standard LKB (LKB-Produkter Ab. S-161, 25 Brommma 1, Sweden) grid storage box is converted into several grid staining boxes by sawing the body of the box into segments along rows of its grid storage cavities. The staining boxes can be cut out to any required size or shape. The polymethyacrylate storage box cover is discarded. Covers for the staining boxes are cut from thin sheet vinyl, which is more chemically resistant than polymethyacrylate. Corresponding 2 mm diameter holes are drilled through the vinyl covers and the bottoms of the grid storage cavities of the staining boxes to convert the storage cavities into staining chambers. For staining, the covers are tied to the boxes with sewing thread and the assembled units are put into vials. The separate staining chambers prevent intermingling of and mechanical damage to grids during the staining procedure. Ultrathin sections are more cleanly and uniformly stained in bulk by the use of these staining boxes than they are when stained individually by a standard method.