Fine structure of the extracellular sheath and cell walls inOphiostoma novo-ulmigrowing on various substrates

Abstract

The presence of microfilamentous-like structures of tubular appearance (MFS) in cell walls and extracellular sheath material (ES) in a number of isolates of Ophiostoma novo-ulmi Brasier grown on various substrates and following various treatments is reported. Standard fixation or high-pressure freezing methods were used, and cytochemical tests were carried out to detect fungal and host wall components and, in some cases, fungal DNA. In some cases, serial 0.2-μm-thick sections were examined at 120 kV and tilted to obtain stereoscopic images. Whether the fungal cell walls were thick and composed of an outer opaque and inner more electron-lucent layers, or thin and barely perceptible, MFS were observed to extend from the cell cytoplasm as parallel structures across the walls into the surrounding medium, including host cell components in infected elm tissues. MFS were associated (in samples from inoculated trees) with cleavage and desquamation of fungal walls. ES and MFS did not label for cellulose or chitin, but generally labelled slightly for β-(1-3)-glucan and mannose, and strongly for galactose. Only the lucent, inner fungal wall layer labelled for chitin and cellulose. DNA labelling was confined to nuclei and mitochondria in fungal cells from cultures on agar medium; in cells from cultures on millipore membranes, it was pronounced over imprecisely delimited cell regions. The possible ontogeny of MFS components and their importance are discussed. Key words: chitin, Dutch elm disease, fungal fimbriae, fungal walls, gold-complexed probes, microfilamentous structures (MFS).