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Procedures for obtaining sectional views of fungal fructifications by scanning electron microscopy
Abstract:
Procedures for sectioning fungal fructifications in host tissues or on artificial media are described, which allow observation of internal structures by scanning electron microscopy. Perithecia of Ceratocystis fimbriata and Phyllachora graminis, and telia of Puccinia xanthii showed excellent preservation of exposed structures in sections which were osmium-coated before being dried. While similar preservation was obtained in sectioned acervuli of Lecanosticta acicola and Marssonia juglandis and in pycnidia of Dothiorella ribis and Phomopsis occulta, the mucilaginous substances produced in these fructifications precluded observation of conidiophores. Extraction of mucilage from these sections was accomplished by periodic acid and dilute KOH treatments, followed by an osmium-coating procedure. In such preparations, mucilage was removed, internal structures were preserved, and pertinent characteristics of conidiogenous cells were resolved.
Insights
This study presents improved methods for sectioning fungal structures for scanning electron microscopy. These techniques enhance the visualization of internal fungal morphology, aiding in taxonomic and pathological studies.
Area of Science:
- Mycology
- Plant Pathology
- Microscopy
Background:
- Accurate visualization of fungal structures is crucial for identification and understanding disease mechanisms.
- Traditional sectioning methods can be hindered by fungal mucilage and tissue complexity.
Purpose of the Study:
- To develop and refine sectioning techniques for observing internal fungal fructifications using scanning electron microscopy (SEM).
- To overcome limitations posed by mucilaginous substances in certain fungal species.
Main Methods:
- Fungal fructifications were sectioned from host tissues and artificial media.
- Osmium coating was applied to dried sections for SEM observation.
- Periodic acid and dilute KOH treatments were used to extract mucilage.
Main Results:
- Excellent preservation of internal structures was achieved for various fungal types (e.g., perithecia, telia) after osmium coating.
- Mucilage extraction successfully revealed conidiophores in previously obscured structures (acervuli, pycnidia).
- SEM imaging allowed detailed resolution of conidiogenous cells.
Conclusions:
- The described sectioning and mucilage extraction methods significantly improve SEM visualization of fungal internal morphology.
- These techniques are valuable for resolving fine details of fungal reproductive structures, aiding in fungal taxonomy and pathology.