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Radioautographic visualization of beta glucans formed by pea pisum sativum cultivar alaska membranes from udp glucose



Radioautographic visualization of beta glucans formed by pea pisum sativum cultivar alaska membranes from udp glucose



Canadian Journal of Botany 61(4): 1266-1275



Radioautographic experiments were carried out using pea stem slices to determine the site of glucose incorporation from UDP-glucose. Cut or damaged pea stem cells were the only cells to incorporate [3H]glucose from UDP-[3H]glucose. The product formed at 20 .mu.M UDP-glucose was observed in EM thin sections in patches on the plasma membrane and the cell wall. The product formed at 5 mM UDP-glucose occurred in fibrillar bundles that stretched between the plasma membrane and the cell wall. This periplasmic material fluoresced when stained with aniline blue. Experiments in which slices were subjected to sequential incubations in radioactive 5 mM UDP-glucose followed by unlabeled 5 mM UDP-glucose, or incubations in the reverse order, indicated that incorporation of [3H]glucose into products insoluble in chloroform:methanol:water or hot water occurs at the plasma membrane, and radioactivity is displaced from the membrane by subsequent incubations. A similar experiment, in which slices were first incubated in radioactive 20 .mu.M UDP-glucose followed by unlabeled 5 mM UDP-glucose, indicated that the synthesis of fibrillar material from 5 mM UDP-glucose displaces the labeled product that had been formed from 20 .mu.m UDP-glucose. Only cut or damaged pea stem cells utilize UDP-glucose and the plamsa membrane enzymes that incorporate [3H]glucose from 20 or 5 mM UDP-[3H]glucose are in close physical proximity.

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