Culture methods and detection of glucanases in suspension cultures of wheat and barley

Abstract

Suspension cultures of Triticum monococcum L., Triticum vulgare Vill. var. Thatcher, Hordeum vulgare L. var. Gateway, and Hordeum vulgare L. var. Gateway mutant yv2 have been established. The cultures were derived from root sections of seedlings and cultured in a denned medium consisting of mineral salts, sucrose, B vitamins, and 2,4-dichlorophenoxyacetic acid, with nitrate and ammonia as the sources of nitrogen. In the early period of the cultures the cell aggregates readily, differentiated to form roots, but this characteristic diminished after several generations of subculture. The cells and medium contained a number of glucanases. The presence of a laminaranase (endo-β-(1 → 3)-D-glucan glucanohydrolase (EC 3.2.1.99)) that did not attack lichenan was established. The culture media of the wheat contained an oligosaccharide which on acid hydrolysis yielded galactose, arabinose, and xylose. Hydrolysis of a cell-wall fraction yielded the same sugars in addition to glucose and mannose.