Ultrastructural comparisons of fungus hyphal cells using frozen-etched replicas and thin sections of the fungus Pyrenochaeta terrestris

Abstract

The ultrastucture of P. terrestris hyphal cells was investigated to compare frozen-etched replicas with chemically fixed thin sections. The fungus used in this study uses glycerol as a sole carbon source and survives the freezing procedures necessary for freeze-etching; thus frozen-etched replicas reflect the living state. Frozen-etched membrane systems have particles of various sizes and concentrations and have a smooth appearance as contrasted to chemically fixed membrane systems, which have particles difficult to distinguish and somewhat irregular membrane systems. Frozen-etched cell walls are seen to contain particles, and microfibrillar orientation is evident in older cell walls, whereas substructure is not evident in chemically fixed cell walls, although secretion products of the fungus accumulate on cell surfaces. Chemically fixed ground cytoplasm has ribosomes and areas of high-and low-electron scattering which are not seen with freeze-etching. Cells fixed in glutaraldehyde-acrolein-OsO4 more nearly resemble frozen-etched cells than cells fixed in potassium permanganate.