Study of grain cell wall structures by microscopic analysis with four different staining techniques

Abstract

Four different grain cell wall staining techniques were compared. Two techniques specifically detected arabinoxylan (AX). The first technique used a xylanase probe, while the other one was based on immunolabeling of AX using monoclonal antibodies. The two other staining techniques, one based on Calcofluor and the other on immunolabeling using monoclonal antibodies, stained mixed-linkage β-glucan. Cell walls of wheat, barley, oat and rye grains, differing both in content and location of AX and β-glucan, were examined. The staining methods were complementary to each other in revealing the location and distribution of the major cereal dietary fiber components AX and β-glucan in the different grains. AX was mostly concentrated in nucellar epidermis and aleurone cells, whereas β-glucan was concentrated more in subaleurone cells. Furthermore, in the case of barley and rye, the endosperm cell walls also contained high amounts of β-glucan. Interestingly, β-glucan in rye and barley endosperm cell walls was located adjacent to the cell contents, suggesting that it is not evenly distributed in the endosperm cell walls. The results give new insight into the structure of the cereal dietary fiber complex. Further development of microscopic techniques will help in elucidating the cereal cell wall structure even in more detail.