INCUBATION OF BARLEY KERNEL SECTIONS WITH PURIFIED CELL WALL DEGRADING ENZYMES

Abstract

The effects of purified Trichoderma reesei endo-xylanase, XYN II and endoglucanases, EG I and EG II on the cell walls of embedded barley cross sections were studied. The cell walls were stained with Calcofluor which is specific for (1–3)(1–4)-β-D-glucans (mixed-linked β-glucans) in histochemical applications. Whereas endosperm cell walls remained highly fluorescent after incubation with buffer, the aleurone cell walls were only weakly fluorescent. Both EG I and EG II were effective in degrading the outer endosperm cell walls, but had little or no effect on the inner endosperm and aleurone cell walls. Corresponding sections incubated with XYN II showed highly intensive fluorescence of aleurone cell walls suggesting that the surface of alaurone cell walls contains a high level of xylan which is removed by xylanase treatment exposing a layer rich in mixed linked β-glucans. This indicates that arabinoxylans and β-glucans in the aleurone cell walls of barley exist as separate, macroscopic layers. Xylanase did not have any Influence on the microstructure of endosperm cell walls which are rich in mixed linked β-glucans. However, incubation with EG II after xylanase treatment caused the aleurone cell walls to become very thin.