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Plant and Cell Physiology Advance Access published online on July 30, 2008
Plant and Cell Physiology, doi:10.1093/pcp/pcn108
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© The Author 2008. Published by Oxford University Press on behalf of Japanese Society of Plant Physiologists. All rights reserved. For Permissions, please e-mail: journals.permissions@oxfordjournals.org

Purification, cloning and functional characterization of an endogenous beta-glucuronidase in Arabidopsis thaliana

Aymerick Eudes1, Grégory Mouille, Johanne Thévenin, Zoran Minic and Lise Jouanin*

INRA, Centre de Versailles, Institut Jean-Pierre Bourgin, Laboratoire de Biologie Cellulaire, 78026 Versailles cedex, France

Corresponding author: Lise Jouanin. INRA, Centre de Versailles, Institut Jean-Pierre Bourgin, Laboratoire de Biologie Cellulaire, 78026 Versailles cedex, France. Tel 33 1 30 83 30 63, Fax 33 1 30 83 30 99, E-mail: jouanin{at}versailles.inra.fr


  Abstract

Beta-Glucuronidase (GUS) activities have been extensively characterized in bacteria, fungi, and animals, and the bacterial enzyme GUSA from Escherichia coli is commonly used as a reporter for gene expression studies in plants. Although endogenous GUS activity has been observed in plants, the nature and function of the enzymes involved remain elusive. Here we report on tissue specific localization, partial purification and identification of AtGUS2, a GUS active under acidic conditions from Arabidopsis thaliana. This enzyme belongs to the GH79 family in the Carbohydrate-Active Enzymes database, which also includes mammalian heparanases that degrade the carbohydrate moieties of cell surface proteoglycans, and fungal enzymes active on arabinogalactan proteins (AGPs). We characterized a knockout insertion line (atgus2-1) and transgenic lines overexpressing AtGUS2 (Pro35S:AtGUS2). Endogenous GUS activity assayed histochemically and biochemically was absent in atgus2-1 tissues and five times higher in Pro35S:AtGUS2 lines. AGPs purified from atgus2-1 and Pro35S:AtGUS2 seedlings showed higher and markedly lower glucuronic acid content, respectively. Our results suggest that endogenous GUS activity influences the sugar composition of the complex polysaccharide chains of AGPs. We also show that transgenics display hypocotyl and root growth defects compared to wild-type plants. Hypocotyl and root lengths are increased in Pro35S:AtGUS2 seedlings, whereas hypocoyl length is reduced in atgus2-1 seedlings. These data are consistent with a role for the carbohydrate moieties of AGPs in cell growth.

Keywords: Arabidopsis - beta-glucuronidase - family 79 glycoside hydrolase - arabinogalactan protein - hypocotyl and root growth

1Present address: University of Florida, Horticultural Sciences Department, P.O. Box 110690, Gainesville, FL, 32611, USA.

(Received July 8, 2008; Accepted July 27, 2008)
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