Analysis of Complex Carbohydrate Composition in Plant Cell Wall Using Fourier Transformed Mid-Infrared Spectroscopy (FT-IR)

Part of the Methods in Molecular Biology book series (MIMB, volume 1588)


Fourier transformed mid-infrared spectroscopy (FTIR) is a powerful tool for compositional analysis of plant cell walls (Acebes et al., Front Plant Sci 5:303, 2014; Badhan et al., Biotechnol Biofuels 7:1–15, 2014; Badhan et al., BioMed Res Int 2015: 562952, 2015; Roach et al., Plant Physiol 156:1351–1363, 2011). The infrared spectrum generates a fingerprint of a sample with absorption peaks corresponding to the frequency of vibrations between the bonds of the atoms making up the material. Here, we describe a method focused on the use of FTIR in combination with principal component analysis (PCA) to characterize the composition of the plant cell wall. This method has been successfully used to study complex enzyme saccharification processes like rumen digestion to identify recalcitrant moieties in low-quality forage which resist rumen digestion (Badhan et al., BioMed Res Int 2015: 562952, 2015), as well as to characterize cell wall mutant lines or transgenic lines expressing exogenous hydrolases (Badhan et al., Biotechnol Biofuels 7:1–15, 2014; Roach et al., Plant Physiol 156:1351–1363, 2011). The FTIR method described here facilitates high-throughput identification of the major compositional differences across a large set of samples in a low cost and nondestructive manner.

Key words

Fourier transformed infrared spectroscopy Plant cell wall Carbohydrate Cellulose Xylan Lignin 



The authors wish to acknowledge the support of Cellulosic Biofuel Network of Agriculture and Agri-Food Canada, Genome Canada, Genome Alberta, and Genome Quebec for their generous support of their research program.


  1. 1.
    Acebes JL, Largo-Gosens A, Hernández-Altamirano M, García-Calvo L, Alonso-Simón A, Álvarez JM (2014) Fourier transform mid infrared spectroscopy applications for monitoring the structural plasticity of plant cell walls. Front Plant Sci 5:303. doi: 10.3389/fpls.2014.00303 PubMedPubMedCentralGoogle Scholar
  2. 2.
    Badhan A, Jin L, Wang Y, Han S, Kowalczys K, Brown DC, Ayala CJ, Latoszek-Green M, Miki B, Tsang A, McAllister T (2014) Expression of a fungal ferulic acid esterase in alfalfa modifies cell wall digestibility. Biotechnol Biofuels 7:1–15CrossRefGoogle Scholar
  3. 3.
    Badhan A, Wang YX, Gruninger R, Patton D, Powlowski J, Tsang A, TA MA (2015) Improvement in saccharification yield of mixed rumen enzymes by identification of recalcitrant cell wall constituents using enzyme fingerprinting. BioMed Res Int 2015:562952. doi: 10.1155/2015/562952 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Roach MJ, Mokshina NY, Badhan A, Snegireva AV, Hobson N, Deyholos MK, Gorshkova TA (2011) Development of cellulosic secondary walls in flax fibers requires β-galactosidase. Plant Physiol 156:1351–1363CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Ribeiro GO, Gruninger RJ, Badhan A, McAllister TA (2016) Mining the rumen for fibrolytic feed enzymes. Anim Front 6:20–26CrossRefGoogle Scholar
  6. 6.
    Badhan A, McAllister TA (2015) Designer plants for biofuel; a review. Curr Metabolomics 4(1):49–55CrossRefGoogle Scholar
  7. 7.
    Badhan AK, Chadha BS, Kaur J, Sonia KG, Saini HS, Bhat MK (2007) Role of transglycosylation products in the expression of multiple xylanases in myceliophthora sp. IMI 387099. Curr Microbiol 54:405–409CrossRefPubMedGoogle Scholar
  8. 8.
    Baker MJ, Trevisan J, Bassan P, Bhargava R, Butler HJ, Dorling KM, Fielden PR, Fogarty SW, Fullwood NJ, Heys KA, Hughes C, Lasch P, Martin-Hirsch PL, Obinaju B, Sockalingum GD, Sulé-Suso J, Strong RJ, Walsh MJ, Wood BR, Gardner P, Martin FL (2014) Using Fourier transform IR spectroscopy to analyze biological materials. Nat Protoc 9:1771–1791CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Allison GG, Thain SC, Morris P, Morris C, Hawkins S, Hauck B, Barraclough T, Yates N, Shield I, Bridgwater AV, Donnison IS (2009) Quantification of hydroxycinnamic acids and lignin in perennial forage and energy grasses by Fourier-transform infrared spectroscopy and partial least squares regression. Bioresour Technol 100:1252–1261CrossRefPubMedGoogle Scholar
  10. 10.
    Carpita NC, McCann MC (2015) Characterizing visible and invisible cell wall mutant phenotypes. J Exp Bot 66:4145–4163CrossRefPubMedGoogle Scholar
  11. 11.
    Chen L, Carpita NC, Reiter WD, Wilson RH, Jeffries C, McCann MC (1998) A rapid method to screen for cell-wall mutants using discriminant analysis of Fourier transform infrared spectra. Plant J 16:385–392CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.Lethbridge Research and Development Centre, Agriculture and Agri-Food CanadaLethbridgeCanada

Personalised recommendations