The interface between fungal hyphae and orchid protocorm cells

Seeds of the orchids Platanthera hyperborea, Spiranthes lacera, and Spiranthes sinensis were germinated in vitro in the presence of compatible fungal species and the resulting colonized protocorms were studied by light microscopy, transmission electron microscopy, and colloidal-gold affinity techniques. Protocorm cells in early stages of colonization contained coils of fungal hyphae (pelotons) separated from host cell cytoplasm by the host plasma membrane and interfacial matrix material. Best cell walls were labelled by the colloidal gold - cellobiohydralase I (CBH-I) complex to detect cellulose and, particularly over the middle lamella, by antibodies that bind to pectins (JIM 5 and JIM 7). A polyclonal antibody that binds to beta-1,3-glucans labelled the fungal cell wall heavily. None of the probes, however, labelled the interfacial matrix between the wall of active fungal hyphae and the surrounding plasma membrane. In contrast, the interfacial matrix material that ensheathed collapsing hyphae showed labelling after treatment with JIM 5, the polyclonal antibody, and the CBH-I complex. Labelling of host cell walls and fungal walls was similar to that described for early stages.