Cytochemical study of the yeast and mycelial cell walls ofParacoccidioides brasiliensis

Cell wall structure and macromolecular organization of the various growth forms ofParacoccidioides brasiliensis were investigated using chemical, enzymatic, and cytochemical methods. The wall of yeast-like cells was not sensitive to periodic acid and β(1 → 3) glucanase treatments, but bound calcofluor suggesting that it is composed of an α(1 → 3) glucan and of chitin. The fibrillar outer layer of bud cell initials and of abscision areas, were characterized by the presence of a strongly periodic acid- and concanavalin A-reactive substance that was sensitive to the lytic action of protease, presumably composed of mannan and proteins. The outermost layer of the mycelium was sensitive to protease, and periodic acid and peanut agglutinin positive indicating that it was composed of galactomannan and protein. The inner layer is mainly composed of a β(1 → 3), (1 → 6) glucan (sensitive to snail enzyme but not to an exo β(1 → 3) glucanase), and chitin. Septa were brightly fluorescent with calcofluor. Our results are compared with models previously proposed by other authors.