Visualization of barley beta-glucan degrading isozymes after gel isoelectric focusing
S Menteur1, L Jestin, T Risacher
1Laboratoire Orge-Triticale, INRA - Station d'Amélioration des Plantes, Domaine de Crouelle, Clermont-Ferrand, France.
Abstract:
A simple method to study the polymorphism of barley (Hordeum vulgare L.) beta-(1-3,1-4)-glucanases (specific enzymes of barley beta-glucan hydrolysis) is described. Proteins of a crude extract of germinated barley kernels were separated in an immobilized pH gradient in two pH ranges (pH 3-10.5 and pH 4-7). beta-glucanases were visualized by contact printing with a polyacrylamide gel containing beta-glucan or lichenan. Patterns of beta-glucanases were revealed by staining with Congo Red with resultant clear zones on a stained background. Various conditions of germination, extraction and visualization were investigated. New isozyme bands could be detected and their nature and origin discussed.
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