Determining the polysaccharide composition of plant cell walls

• DOI: 10.1038/nprot.2012.081
• Publication Date: 2012-08-03T15:24:33Z
• Authors: Filomena A Pettolino; Cherie Walsh; Geoffrey B Fincher; Antony Bacic
• Keywords: 0303 health sciences; 03 medical and health sciences; Cell Wall; Polysaccharides; Analytic Sample Preparation Methods; Plants; 540; Methylation; Gas Chromatography-Mass Spectrometry

The plant cell wall is a chemically complex structure composed mostly of polysaccharides. Detailed analyses of these cell wall polysaccharides are essential for our understanding of plant development and for our use of plant biomass (largely wall material) in the food, agriculture, fabric, timber, biofuel and biocomposite industries. We present analytical techniques not only to define the fine chemical structures of individual cell wall polysaccharides but also to estimate the overall polysaccharide composition of cell wall preparations. The procedure covers the preparation of cell walls, together with gas chromatography-mass spectrometry (GC-MS)-based methods, for both the analysis of monosaccharides as their volatile alditol acetate derivatives and for methylation analysis to determine linkage positions between monosaccharide residues as their volatile partially methylated alditol acetate derivatives. Analysis time will vary depending on both the method used and the tissue type, and ranges from 2 d for a simple neutral sugar composition to 2 weeks for a carboxyl reduction/methylation linkage analysis.