Cell Walls of Germinating Uredospores: II. Carbohydrate Polymers

Polymeric carbohydrates in (14)C-labeled germ tube and uredospore walls of Uromyces phaseoli var. typica were studied by permethylation and by enzymatic hydrolysis. The native structure of the uredospore wall limited the effectiveness of both techniques with this wall, but evidence for two distinct polysaccharides was obtained. A linear (1-->3) glucan, containing minor quantities of (1-->6) linkages, may account for most of the glucose in the uredospore wall. A second uredospore polymer was a glucomannan similar to one reported for other rust fungi in that it consisted of approximately equal numbers of beta(1-->3) and beta(1-->4) mannosidic linkages with glucose as a minor component at the nonreducing end. Branching, most likely by (1-->6) mannose links, was low. In contrast to uredospore wall, considerably more germ tube polysaccharide was accessible to enzymes and to methylation. Methylation studies indicate that (1-->3) glucose and mannose bonds occur predominantly. Evidence from hydrolysis with exo- (beta)-(1-->3) glucanase suggests distinct wall regions of beta(1-->3) glycan, highly branched by (1-->6) bonds, as well as wall regions of a glucomannan with alternating (1-->3) glucose and (1-->3) mannose residues. Polymer heterogeneity was indicated by differences in the proportions of mannose, glucose, and galactose as reducing end groups in different solubility fractions. In germ tube walls, but not in uredospore walls, glucosamine apparently existed as part of chitin polymer as evidenced by the isolation of N,N-diacetylchitobiose from chitinase digestion.