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Determining the polysaccharide composition of plant cell walls
Filomena A Pettolino1, Cherie Walsh2, Geoffrey B Fincher3 & Antony Bacic2
1Commonwealth Scientific and Industrial Research Organization (CSIRO) Plant Industry, Canberra, Australia. 2Australian Research Council (ARC) Centre for Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Melbourne, Australia. 3ARC Centre for Excellence in Plant Cell Walls, School of Agriculture, Food and Wine, University of Adelaide, Glen Osmond, Australia. Correspondence should be addressed to A.B. ([email protected]).
Published online 2 August 2012; doi:10.1038/nprot.2012.081
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 chromatographymass 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.
2012 Nature America, Inc. All rights reserved.
INTRODUCTIONBackgroundThe composition of cell walls varies depending on the function of the tissue and the species of plant. Primary cell walls surround expanding cells in young tissues, whereas thickened secondary cell walls confer mechanical strength after cell growth has ceased in more mature tissues. In most tissues, cell walls have to be sufficiently porous to allow the movement of molecules between cells (apoplastic transport), but those of some water-conducting cells of the vasculature need to be waterproof. Cell walls are also involved in cell adhesion, cell-cell communication and defense responses1,2.
Polysaccharides make up about 90% of the dry weight of primary cell walls and about 60% of the dry weight of lignified secondary cell walls1. Water may constitute about 60% (wt/vol) of primary cell walls, but is essentially...