Cellulose is synthesized by Cellulose Synthase A proteins at the plasma membrane using the substrate UDP glucose. Herein, we provide a detailed method for measuring the incorporation of radiolabeled glucose into the cellulose fraction of the cell wall. In this method Arabidopsis seedlings are treated for 2 h with a cellulose biosynthesis inhibitor in the presence of radiolabeled glucose, and are subsequently boiled in acetic-nitric acid to solubilize non-cellulosic material. The radiolabeled glucose detected in the insoluble fraction indicates the amount of cellulose synthesized during the experimental timeframe. The short-term nature of this method is a useful tool in determining if inhibition of cellulose biosynthesis is the herbicides primary mode of action.
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This work was supported by the U. S. Department of Energy (grant no. DOE–FOA 10–0000368 to S. D. and C. B.), the U. S. Department of Agriculture (Hatch Act grant to S. D. and J. S.)
Brabham, Chad; Stork, Jozsef; and DeBolt, Seth, "[14C] Glucose Cell Wall Incorporation Assay for the Estimation of Cellulose Biosynthesis" (2015). Horticulture Faculty Publications. 38.