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Plant Physiol, July 2001, Vol. 126, pp. 981-992

The Experimental Herbicide CGA 325'615 Inhibits Synthesis of Crystalline Cellulose and Causes Accumulation of Non-Crystalline beta -1,4-Glucan Associated with CesA Protein1

Liangcai Peng,2 Fan Xiang, Eric Roberts,3 Yasushi Kawagoe,4 L. Carl Greve, Klaus Kreuz, and Deborah P. Delmer*

Section of Plant Biology (L.P., E.R., Y.K., D.P.D.), Molecular Structure Facility (F.X.), and Department of Pomology (L.C.G.), University of California, One Shields Avenue, Davis, California 95616; and Novartis Crop Protection, CH-4002, Basel, Switzerland (K.K.)

Developing cotton (Gossypium hirsutum) fibers, cultured in vitro with their associated ovules, were used to compare the effects of two herbicides that inhibit cellulose synthesis: 2,6-dichlorobenzonitrile (DCB) and an experimental thiatriazine-based herbicide, CGA 325'615. CGA 325'615 in nanomolar concentrations or DCB in micromolar concentrations causes inhibition of synthesis of crystalline cellulose. Unlike DCB, CGA 325'615 also causes concomitant accumulation of non-crystalline beta -1,4-glucan that can be at least partially solubilized from fiber walls with ammonium oxalate. The unusual solubility of this accumulated glucan may be explained by its strong association with protein. Treatment of the glucan fraction with protease changes its size distribution and leads to precipitation of the glucan. Treatment of the glucan fraction with cellulase digests the glucan and also releases protein that has been characterized as GhCesA-1 and GhCesA-2---proteins that are believed to represent the catalytic subunit of cellulose synthase. The fact that cellulase treatment is required to release this protein indicates an extremely tight association of the glucan with the CesA proteins. In addition, CGA 325'615, but not DCB, also causes accumulation of CesA protein and a membrane-associated cellulase in the membrane fraction of fibers. In addition to the effects of CGA 325'615 on levels of both of these proteins, the level of both also shows coordinate regulation during fiber development, further suggesting they are both important for cellulose synthesis. The accumulation of non-crystalline glucan caused by CGA 325'615 mimics the phenotype of the cellulose-deficient rsw1 mutant of Arabidopsis that also accumulates an apparently similar glucan (T. Arioli, L. Peng, A.S. Betzner, J. Burn, W. Wittke, W. Herth, C. Camilleri, H. Hofte, J. Plazinski, R. Birch et al. [1998] Science 279: 717).

1 This work was supported by Novartis Crop Protection and by the U.S. Department of Energy (grant no. DE-FG-03-963-ER 20238 to D.P.D.).

2 Present address: Plant Gene Expression Center, 800 Buchanan Street, Albany, CA 94710-1198.

3 Present address: Biological Sciences Department, University of Rhode Island, 10 Ranger Road, Kingston, RI 02881.

4 Present address: National Institute of Agrobiological Resources, Kannondai 2-1-2, Tsukuba, Ibaraki, Japan 305-8602.

* Corresponding author; e-mail dpdelmer{at}ucdavis.edu; fax 530-752-5410.

© 2001 American Society of Plant Physiologists


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