Characterization of Plant Cell Wall Damage-Associated Molecular Patterns Regulating Immune Responses

  • Laura Bacete
  • Hugo Mélida
  • Sivakumar Pattathil
  • Michael G. Hahn
  • Antonio Molina
  • Eva MiedesEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1578)


The plant cell wall is one of the first defensive barriers that pathogens need to overcome to successfully colonize plant tissues. Plant cell wall is considered a dynamic structure that regulates both constitutive and inducible defense mechanisms. The wall is a potential source of a diverse set of Damage-Associated Molecular Patterns (DAMPs), which are signalling molecules that trigger immune responses. However, just a few active wall ligands, such as oligogalacturonic acids (OGs), have been characterized so far. To identify additional wall-derived DAMPs, we obtained different plant wall fractions and tested their capacity to trigger immune responses using a calcium read-out system. To characterize the active DAMPs structures present in these fractions, we applied Glycome Profiling, a technology that uses a large and diverse set of specific monoclonal antibodies against wall carbohydrate ligands. The methods describe here can be used in combination with other biochemical approaches to identify and purify new plant cell wall DAMPs.

Key words

Pectin Hemicellulose Cell wall Arabidopsis Immunity 



Research in the A. M. lab was financially supported by grants BIO2012-32910 and BIO2015-64077-R from Spanish Ministry of Economy and Competitiveness (MINECO), and in the M. G. H. lab (CCRC) by the United States NSF Plant Genome Program (DBI-0421683 and IOS-0923992). L. B. was a PhD fellow (BES-2013-065010) from MINECO and H. M. was supported by an IEF-SignWALLINg-624721 grant from the European Union.


  1. 1.
    Miedes E, Vanholme R, Boerjan W, Molina A (2014) The role of the secondary cell wall in plant resistance to pathogens. Front Plant Sci 5:358. doi: 10.3389/fpls.2014.00358 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Thorsten H (2012) Plant cell wall integrity maintenance as an essential component of biotic stress response mechanisms. Front Plant Sci 23:77. doi: 10.3389/fpls.2012.00077 Google Scholar
  3. 3.
    Thorsten H (2015) The plant cell wall integrity maintenance mechanism—concepts for organization and mode of action. Plant Cell Physiol 56:215–223CrossRefGoogle Scholar
  4. 4.
    Boller T, Felix G (2009) A renaissance of elicitors: perception of microbe-associated molecular patterns and danger signals by pattern-recognition receptors. Annu Rev Plant Biol 60:379–406CrossRefPubMedGoogle Scholar
  5. 5.
    Monaghan J, Zipfel C (2012) Plant pattern recognition receptor complexes at the plasma membrane. Curr Opin Plant Biol 15:349–357CrossRefPubMedGoogle Scholar
  6. 6.
    Miedes E, Lorences EP (2004) Apple (Malus domestica) and tomato (Lycopersicum esculentum) fruits cell-wall hemicelluloses and xyloglucan degradation during Penicillium expansum infection. J Agric Food Chem 52:7957–7963CrossRefPubMedGoogle Scholar
  7. 7.
    Miedes E, Lorences EP (2006) Changes in cell wall pectin and pectinase activity in apple and tomato fruits during Penicillium expansum infection. J Sci Food Agric 86:1359–1364CrossRefGoogle Scholar
  8. 8.
    Knight MR, Campbell AK, Smith SM, Trewavas AJ (1991) Transgenic plant aequorin reports the effects of touch and cold-shock and elicitors on cytoplasmic calcium. Nature 352:524–526CrossRefPubMedGoogle Scholar
  9. 9.
    Ranf S, Gish N, Schäffer M, Illig T, Westphal L, Knirel YA et al (2015) A lectin S-domain receptor kinase mediates lipopolysaccharide sensing in Arabidopsis thaliana. Nat Inmunol 16:426–433CrossRefGoogle Scholar
  10. 10.
    Ranf S, Eschen-Lippold L, Pecher P, Lee J, Scheel D (2011) Interplay between calcium signalling and early signalling elements during defence responses to microbe- or damage-associated molecular patterns. Plant J 68:100–113CrossRefPubMedGoogle Scholar
  11. 11.
    Meng X, Zhang S (2013) MAPK cascades in plant disease resistance signaling. Annu Rev Phytopathol 51:245–266. doi: 10.1146/annurev-phyto-082712-102314 CrossRefPubMedGoogle Scholar
  12. 12.
    Pattathil S, Avci U, Baldwin D, Swennes AG, McGill JA, Popper Z et al (2010) A comprehensive toolkit of plant cell wall glycan- directed monoclonal antibodies. Plant Physiol 153:514–525CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Murashige T, Skoog F (1962) A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  14. 14.
    Pattathil S, Avci U, Miller JS, Hahn MG (2012) Immunological approaches to plant cell wall and biomass characterization: glycome profiling. In: Himmel M (ed) Biomass conversion: methods and protocols. Methods in molecular biology, vol 908. Springer Science+Business Media, LLC, New York, NY, pp 61–72Google Scholar
  15. 15.
    Dubois M, Gilles KA, Hamilton JK, Rebers PA, Smith F (1956) Colorimetric method for the determination of sugars and related substances. Anal Chem 28:350–356CrossRefGoogle Scholar
  16. 16.
    Ranf S, Grimmer J, Poschl Y, Pecher P, Chinchilla D, Scheel D, Lee J (2012) Defense-related calcium signaling mutants uncovered via a quantitative high-throughput screen in Arabidopsis thaliana. Mol Plant 5:115–130CrossRefPubMedGoogle Scholar
  17. 17.
    Mort AJ, Moerschbacher BM, Pierce ML, Maness NO (1991) Problems encountered during the extraction, purification, and chromatography of pectic fragments, and some solutions to them. Carbohydr Res 215:219–227CrossRefGoogle Scholar
  18. 18.
    Coimbra MA, Waldron KW, Selvendran RR (1994) Isolation and characterisation of cell wall polymers from olive pulp (Olea europaea L). Carbohydr Res 252:245–262CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Laura Bacete
    • 1
  • Hugo Mélida
    • 1
  • Sivakumar Pattathil
    • 2
  • Michael G. Hahn
    • 2
  • Antonio Molina
    • 1
  • Eva Miedes
    • 1
    Email author
  1. 1.Centro de Biotecnología y Genómica de Plantas, Universidad Politécnica de Madrid (UPM)—Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria (INIA)Pozuelo de Alarcón (Madrid)Spain
  2. 2.Complex Carbohydrate Research Center (CCRC)University of GeorgiaAthensUSA

Personalised recommendations