Abstract
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.
Document Type
Article
Publication Date
9-20-2015
Digital Object Identifier (DOI)
https://doi.org/10.21769/BioProtoc.1589
Funding Information
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.)
Repository Citation
Brabham, Chad; Stork, Jozsef; and DeBolt, Seth, "[14C] Glucose Cell Wall Incorporation Assay for the Estimation of Cellulose Biosynthesis" (2015). Horticulture Faculty Publications. 38.
https://uknowledge.uky.edu/horticulture_facpub/38
Notes/Citation Information
Published in Bio-protocol, v. 5, issue 18, p. 1-6.
Copyright © 2015 The Authors; exclusive licensee Bio-protocol LLC.
The publisher has granted the permission for posting the article here.