Light and electron microscope studies of host-parasite relations in a mycoparasite

Light microscope studies of the mycoparasite Piptocephalis virginiana revealed that the cylindrical spores of the parasite became spherical upon germination and produced 1-4 germ tubes. Generally t"l.vO germ tubes were produced by each spore. When this parasite was inoculated on its potential hosts, Choanephora cucurbitarum and Phascolomyces articulosus, the germ tube nearest to the host hypha continued to grow and made contact with the host hypha. The tip of the parasite's germ tube became swollen to form a distinct appressorium. Up to this stage the behavior of the parasite was similar regardless of the nature of the host. In the compatible host-parasite combination, the parasite penetrated the host, established a nutritional relationship and continued to grow to cover the host completely with its buff colored spores in 3-4 days. In the incompatible host-parasite combination, the parasite penetrated the host but its further advance was arrested. As a result of failure to establish a nutritional relationship with the resistant host, the parasite made further attempts to penetrate the host at different sites producing multiple infections. In the absence of nutrition the parasite weakened and the host outgrew the parasite completely. In the presence of a non-host species, Linderina pennispora the parasite continued to grow across the non-host 1).yp_hae vlithout establishing an initial contact. Germination studies showed that the parasite germinated equally well in the presence of host and non-host species. Further electron microscope studies revealed that the host-parasite interaction between P. virginiana and its host, C. cucurbi tarum, was compatible when the host hyphae were young slender, with a thin cell wall of one layer. The parasite appeared to penetrate mechanically by pushing the host-cell wall inward. The host plasma membrane invaginated along the involuted cell wall. The older hyphae of C. cucurbitarum possessed two distinct layers of cell wall and-showed an incompatible interaction when challenged vlith the parasite. At the point of contact, the outer layer of the host-cell wall dissolved, probably by enzymatic digestion, and the inner layer became thickened and developed a papilla as a result of its response to the parasite. The haustoria of the parasite in the old hyphae were always surrounded by a thick, well developed sheath, whereas the haustoria of the same age in the young host mycelium were devoid of a sheath during early stages of infection. Instead, they were in direct contact with the host protoplast. The incompatible interaction between a resistant host, P. articulosus and the parasite showed similar results as with the old hyphae of C. cucurbitarum. The cell wall of P. articulosus appeared thick-with two or more layers even in the 18-22 h-old hyphae. No contact or interaction was established between the parasite and the non-host L. pennispora. The role of cell wall in the resistance mechanism is discussed.

Investigations on the host-parasite interface of a mycoparasite system /

The cell wall composition of Choanephora cucur - bitarum and the host-parasite interface, after infection with Piptocephalis virginiana , were examined in detail. The cell walls of C_. cucurbitarum were determined to be composed of chitin (17%), chitosan (28.4%), neutral sugars (7.2%),uronic acid (2.4%), proteins (8.2%) and lipids (13.8%). The structure of hyphal walls investigated by electron microscopy of shadowed replicas before and after alkali-acid hydrolysis, showed two distinct regions: microfibrillar and amorphous. The microfibrils which were composed of mainly chitin, were organized into two distinct layers: an outer, thicker layer of randomly orientated microfibrils and an inner, thin layer of parallel microfibrils.Electronmicrographs of the host-parasite interface of C_. cucurbitarum and the mycoparasite , P_. virginiana , 30 h following inoculation, showed that the sheath zone has a similar electron density to that of the host cell wall. The sheath was not present around the young (18 h old) haustorium. High-resolution autoradiographs of infected host hyphae showed that radioactive N-acetyl-D-glucosamine , a precursor of chitin, was incorporated preferentially in the host cell wall and sheath zone. Cell fractionation of label fed hyphae showed that 84% of the label was present in the cell wall and specifically in the chitin portion of the wall. The antifungal antibiotic, Polyoxin D, a specific inhibitor of the enzyme, chitin synthetase, suppressed the incorporation of the label in the cell wall and sheath zone and resulted in a decrease in electron density of the developing sheath. The significance of these results is discussed in the light of host resistance.