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Whole plant cell wall characterization using solution-state 2D NMR

Nature Protocols volume 7, pages 1579–1589 (2012)Cite this article

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Abstract

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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Figure 1: 2D NMR spectra revealing lignin unit compositions.
Figure 2: 2D NMR spectra revealing lignin interunit distributions, polysaccharide acylation (and polysaccharides).
Figure 3: 2D NMR spectra revealing polysaccharide anomerics.

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Acknowledgements

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).

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Authors and Affiliations

  1. Department of Wood Science, University of British Columbia, Vancouver, Canada

    Shawn D Mansfield

  2. US Department of Energy (DOE) Great Lakes Bioenergy Research Center and Wisconsin Bioenergy Initiative, University of Wisconsin, Madison, Wisconsin, USA

    Hoon Kim, Fachuang Lu & John Ralph

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  1. Shawn D Mansfield
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Contributions

S.D.M., J.R., H.K. and F.L. conceived and designed the experiments. H.K. and F.L. performed the NMR experimental evaluation. J.R. contributed to spectral interpretation. S.D.M. performed the wet chemical evaluation of the cell walls. S.D.M. and J.R. wrote the paper (with input from H.K. and F.L.).

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Correspondence to Shawn D Mansfield or John Ralph.

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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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