Recent advances in nuclear magnetic resonance (NMR) technology have made it possible to rapidly screen plant material and discern whole cell wall information without the need to deconstruct and fractionate the plant cell wall. This approach can be used to improve our understanding of the biology of cell wall structure and biosynthesis, and as a tool to select plant material for the most appropriate industrial applications. This is particularly true in an era when renewable materials are vital to the emerging bio-based economies. This protocol describes procedures for (i) the preparation and extraction of a biological plant tissue, (ii) solubilization strategies for plant material of varying composition and (iii) 2D NMR acquisition (for typically 15 min–5 h) and integration methods used to elucidate lignin subunit composition and lignin interunit linkage distribution, as well as cell wall polysaccharide profiling. Furthermore, we present data that demonstrate the utility of this new NMR whole cell wall characterization procedure with a variety of degradative methods traditionally used for cell wall compositional analysis.
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We gratefully acknowledge funding from the Natural Sciences and Engineering Research Council of Canada′s Discovery Program held by S.D.M.; H.K., F.L., S.D.M. and J.R. were funded in part by the DOE Great Lakes Bioenergy Research Center (DOE Office of Science BER DE-FC02-07ER64494).
The authors declare no competing financial interests.
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Mansfield, S., Kim, H., Lu, F. et al. Whole plant cell wall characterization using solution-state 2D NMR. Nat Protoc 7, 1579–1589 (2012). https://doi.org/10.1038/nprot.2012.064
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