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

J Exp Bot. 2005 Dec;56(422):3171-82. doi: 10.1093/jxb/eri314. Epub 2005 Nov 1.

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.

MeSH terms

  • Alkalies
  • Arabidopsis / chemistry*
  • Arabidopsis / cytology
  • Arabidopsis / growth & development
  • Arabidopsis Proteins / genetics*
  • Calcium / analysis
  • Cell Adhesion
  • Cell Wall / chemistry
  • Cell Wall / ultrastructure
  • Cells, Cultured
  • Chelating Agents
  • Culture Media / chemistry
  • Immunohistochemistry
  • Intercellular Junctions / chemistry
  • Mutation
  • Pectins / analysis*
  • Pectins / chemistry*
  • Polysaccharides / analysis

Substances

  • Alkalies
  • Arabidopsis Proteins
  • Chelating Agents
  • Culture Media
  • Polysaccharides
  • Pectins
  • Calcium
  • polygalacturonic acid