Ultrastructure of freeze-substitutedFrankia strain HFPCcI 3, the actinomycete isolated from root nodules ofCasuarina cunninghamiana

Summary Frankia strain HFPCcI 3 is an actinomycete isolated from root nodules ofCasuarina cunninghamiana. In culture it exhibits typicalFrankia morphology and may produce three distinct morphological forms: branching septate hyphae, terminal or intercalary sporangia, and specialized structures termed vesicles which are the purported site of nitrogenase activity. An examination of the ultrastructure of all three morphological forms using both conventional chemical fixation (CF) and quick-freezing followed by freeze-substitution (FS) reveals some interesting differences between the two fixation methods. Unique to FS material are: 1. smooth membrane profiles; 2. lack of mesosomes; 3. lack of discernible nucleoid regions with condensed chromatin; 4. clarity of cytoplasmic elements such as ribosomes and granular bodies; 5. large cytoplasmic tubules in hyphae and young sporangia; 6. outer wall layer not widely separated from the spherical portion of the vesicle, and 7. bundles of microfilaments in vesicles. The quality of preservation after FS appears to be far superior to that obtained with CF. Accordingly the structures observed after FS are thought to represent more faithfully the structure of the living cell.