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JXB Advance Access published online on November 1, 2005
Journal of Experimental Botany, doi:10.1093/jxb/eri314
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Published by Oxford University Press [2005] on behalf of the Society for Experimental Biology.
Received April 4, 2005
Accepted September 15, 2005

RESEARCH PAPER

Biochemical and immunohistochemical analysis of pectic polysaccharides in the cell walls of Arabidopsis mutant QUASIMODO 1 suspension-cultured cells: implications for cell adhesion

Edouard Leboeuf 1, Fabienne Guillon 2, Séverine Thoiron 3, and Marc Lahaye 2*

1 INRA-Biopolymères, Interactions, Assemblages, BP 71627, F-44316 Nantes Cedex 3, France; Université de Nantes, Groupe de Physiologie et Pathologie Végétales, Faculté des Sciences et Techniques, 2 rue de la Houssinière, BP 92208, F-44322, Nantes Cedex 3, France
2 INRA-Biopolymères, Interactions, Assemblages, BP 71627, F-44316 Nantes Cedex 3, France
3 Université de Nantes, Groupe de Physiologie et Pathologie Végétales, Faculté des Sciences et Techniques, 2 rue de la Houssinière, BP 92208, F-44322, Nantes Cedex 3, France

* To whom correspondence should be addressed.
Marc Lahaye, E-mail: lahaye{at}nantes.inra.fr


  Abstract

Mutation in the Arabidopsis thaliana QUASIMODO 1 gene (QUA1), which encodes a putative glycosyltransferase, reduces cell wall pectin content and cell adhesion. Suspension-cultured calli were generated from roots of wild-type (wt) and qua1-1 A. thaliana plants. The altered cell adhesion phenotype of the qua1-1 plant was also found with its suspension-cultured calli. Cell walls of both wt and qua1-1 calli were analysed by chemical, enzymatic and immunohistochemical techniques in order to assess the role of pectic polysaccharides in the mutant phenotype. Compared with the wt, qua1-1 calli cell walls contained more arabinose (23.6 versus 21.6 mol%), rhamnose (3.1 versus 2.7 mol%), and fucose (1.4 versus 1.2 mol%) and less uronic acid (24.2 versus 27.6 mol%), and they were less methyl-esterified (DM: 22.9% versus 30.3%). When sequential pectin extraction of calli cell walls was performed, qua1-1 water-soluble and chelator-soluble extracts contained more arabinose and less uronic acid than wt. Water-soluble pectins were less methyl-esterified in qua1-1 than in wt. Chelator-soluble pectins were more acetyl-esterified in qua1-1. Differences in the cell wall chemistry of wt and mutant calli were supported by a reduction in JIM7 labelling (methyl-esterified homogalacturonan) of the whole wall in small cells and particularly by a reduced labelling with 2F4 (calcium-associated homogalacturonan) in the middle lamella at tricellular junctions of large qua1-1 cells. Differences in the oligosaccharide profile obtained after endopolygalacturonase degradation of alkali extracts from qua1-1 and wt calli indicated variations in the structure of covalently bonded homogalacturonan. About 29% more extracellular polymers rich in pectins were recovered from the calli culture medium of qua1-1 compared with wt. These results show that perturbation of QUASIMODO 1-1 gene expression in calli resulted in alterations of homogalacturonan content and cell wall location. The consequences of these structural variations are discussed with regard to plant cell adhesion.

Keywords: Arabidopsis thaliana; cell adhesion; cell culture; cell wall; pectin.
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