Abstract
Intracellular crystalline viral aggregates of soil-borne wheat mosaic, wheat streak mosaic, barley stripe mosaic, and cowpea mosaic viruses (CPMV) were stabilized when tissue was fixed in glutaraldehyde after equilibration in cold 0.1 M phosphate—citrate, pH 7.3, buffered 0 to 25%/25% (w/v) glycerol/sucrose gradients. Large intranuclear and cytoplasmic CPMV and CPMV capsid protein crystals were common. Tobacco mosaic virus inclusions were not preserved under the same conditions. No membrane blackening resulted after postfixation osmication unless glycerol and sucrose were removed by washing. Cell organelles and cytoplasm of all tested tissues fixed in glutaraldehyde in 25%, in 0 to 25/25, or lower glycerol/sucrose gradients appeared as well preserved as that obtained by prior chilling of intact tissue, but, CPMV crystals were destroyed in lower concentrations of glycerol and sucrose. The method may be useful for the ultrastructural investigation of other difficult-to-fix or labile plant virus inclusions.
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