Advertisement

Planta

, Volume 247, Issue 4, pp 887–897 | Cite as

The effect of altered lignin composition on mechanical properties of CINNAMYL ALCOHOL DEHYDROGENASE (CAD) deficient poplars

  • Merve Özparpucu
  • Notburga Gierlinger
  • Ingo Burgert
  • Rebecca Van Acker
  • Ruben Vanholme
  • Wout Boerjan
  • Gilles Pilate
  • Annabelle Déjardin
  • Markus Rüggeberg
Original Article

Abstract

Main conclusion

CAD-deficient poplars enabled studying the influence of altered lignin composition on mechanical properties. Severe alterations in lignin composition did not influence the mechanical properties.

Wood represents a hierarchical fiber-composite material with excellent mechanical properties. Despite its wide use and versatility, its mechanical behavior has not been entirely understood. It has especially been challenging to unravel the mechanical function of the cell wall matrix. Lignin engineering has been a useful tool to increase the knowledge on the mechanical function of lignin as it allows for modifications of lignin content and composition and the subsequent studying of the mechanical properties of these transgenics. Hereby, in most cases, both lignin composition and content are altered and the specific influence of lignin composition has hardly been revealed. Here, we have performed a comprehensive micromechanical, structural, and spectroscopic analysis on xylem strips of transgenic poplar plants, which are downregulated for cinnamyl alcohol dehydrogenase (CAD) by a hairpin-RNA-mediated silencing approach. All parameters were evaluated on the same samples. Raman microscopy revealed that the lignin of the hpCAD poplars was significantly enriched in aldehydes and reduced in the (relative) amount of G-units. FTIR spectra indicated pronounced changes in lignin composition, whereas lignin content was not significantly changed between WT and the hpCAD poplars. Microfibril angles were in the range of 18°–24° and were not significantly different between WT and transgenics. No significant changes were observed in mechanical properties, such as tensile stiffness, ultimate stress, and yield stress. The specific findings on hpCAD poplar allowed studying the specific influence of lignin composition on mechanics. It can be concluded that the changes in lignin composition in hpCAD poplars did not affect the micromechanical tensile properties.

Keywords

Plant cell wall Cell wall mechanics Genetic modification Lignin composition 

Notes

Acknowledgements

This work was supported by Grants from the Agency for Innovation by Science and Technology (IWT) through the SBO project BIOLEUM (Grant no. 130039) and the SBO-FISH project ARBOREF (Grant no. 140894), and the European Framework Project MultiBioPro (project number: 311804). R. V. is indebted to the Research Foundation Flanders for a postdoctoral fellowship. N. G. acknowledges funding by the Austrian Science Fund (START-project SURFINPLANT Y-728-316) and the European community (ERC-consolidator grant SCATAPNUT 681885).

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

425_2017_2828_MOESM1_ESM.pdf (419 kb)
Supplementary material 1 (PDF 419 kb)

References

  1. Adapa PK, Karunakaran C, Tabil LG, Schoenau GJ (2009) Potential applications of infrared and Raman spectromicroscopy for agricultural biomass. Agri Eng Int CIGR J XI:1081Google Scholar
  2. Agarwal UP (1999) An overview of Raman spectroscopy as applied to lignocellulosic materials. In: Argyropoulos DS (ed) Advances in lignocellulosics characterization. Tappi Press, pp 201–225Google Scholar
  3. Agarwal UP, Terashima N (2003) FT-Raman study of dehydrogenation polymer (DHP) lignins. In: Proceedings of 12th International Symposium Wood Pulping Chemistry, Department of Forest Ecology and Management, University of Wisconsin Madison, WI, pp 123–126Google Scholar
  4. Agarwal UP, Ralph SA, Atalla RH (1997) FT Raman spectroscopic study of softwood lignin. In: Proceedings of 9th international symposium on wood and pulping chemistry (ISWPC), Montreal, pp 8-1Google Scholar
  5. Agarwal UP, McSweeny JD, Ralph SA (2011) FT–Raman investigation of milled-wood lignins: softwood, hardwood, and chemically modified black spruce lignins. J Wood Chem Technol 31:324–344CrossRefGoogle Scholar
  6. Awad H, Herbette S, Brunel N, Tixier A, Pilate G, Cochard H, Badel E (2012) No trade-off between hydraulic and mechanical properties in several transgenic poplars modified for lignins metabolism. Environ Exp Bot 77:185–195CrossRefGoogle Scholar
  7. Baucher M, Chabbert B, Pilate G, Van Doorsselaere J, Tollier M-T, Petit-Conil M, Cornu D, Monties B, Van Montagu M, Inze D (1996) Red xylem and higher lignin extractability by down-regulating a cinnamyl alcohol dehydrogenase in poplar. Plant Physiol 112:1479–1490CrossRefPubMedPubMedCentralGoogle Scholar
  8. Baucher M, Monties B, Montagu MV, Boerjan W (1998) Biosynthesis and genetic engineering of lignin. Crit Rev Plant Sci 17:125–197CrossRefGoogle Scholar
  9. Bjurhager I, Olsson A-M, Zhang B, Gerber L, Kumar M, Berglund LA, Burgert I, Br Sundberg, Salmén L (2010) Ultrastructure and mechanical properties of Populus wood with reduced lignin content caused by transgenic down-regulation of cinnamate 4-hydroxylase. Biomacromol 11:2359–2365CrossRefGoogle Scholar
  10. Boerjan W, Ralph J, Baucher M (2003) Lignin biosynthesis. Ann Rev Plant Biol 54:519–546CrossRefGoogle Scholar
  11. Bonawitz ND, Chapple C (2010) The genetics of lignin biosynthesis: connecting genotype to phenotype. Annu Rev Genet 44:337–363CrossRefPubMedGoogle Scholar
  12. Burgert I, Frühmann K, Keckes J, Fratzl P, Stanzl-Tschegg SE (2003) Microtensile testing of wood fibers combined with video extensometry for efficient strain detection. Holzforschung 57:661–664CrossRefGoogle Scholar
  13. Cave ID (1968) The anisotropic elasticity of the plant cell wall. Wood Sci Technol 2(4):268–278CrossRefGoogle Scholar
  14. Chen F, Dixon RA (2007) Lignin modification improves fermentable sugar yields for biofuel production. Nat Biotechnol 25:759–761CrossRefPubMedGoogle Scholar
  15. Cosgrove DC, Jarvis M (2012) Comparative structure and biomechanics of plant primary and secondary cell walls. Front Plant Sci 3:204CrossRefPubMedPubMedCentralGoogle Scholar
  16. Donaldson L (2008) Microfibril angle: measurement, variation and relationships–A review. Iawa J 29:345CrossRefGoogle Scholar
  17. Faix O (1991) Classification of lignins from different botanical origins by FT-IR spectroscopy. Holzforschung-Int J Biol Chem Phys Technol Wood 45:21–28Google Scholar
  18. Forbes JC, Watson D (1992) Plants in agriculture. Cambridge University Press, CambridgeGoogle Scholar
  19. Gierlinger N (2014) Revealing changes in molecular composition of plant cell walls on the micron-level by Raman mapping and vertex component analysis (VCA). Front Plant Sci 5:306CrossRefPubMedPubMedCentralGoogle Scholar
  20. Gierlinger N, Keplinger T, Harrington M (2012) Imaging of plant cell walls by confocal Raman microscopy. Nat Protoc 7:1694–1708CrossRefPubMedGoogle Scholar
  21. Gierlinger N, Keplinger T, Harrington M, Schwanninger M (2013) Raman imaging of lignocellulosic feedstock. In: Ven Tvd, Kadla J (eds) Cellulose–biomass conversion. INTECH, pp 159–192Google Scholar
  22. Gorisek Z, Torelli N, Vilhar B, Grill D, Guttenberger H (1999) Microfibril angle in juvenile, adult and compression wood of spruce and silver fir. PHYTON-HORN 39:129–132Google Scholar
  23. Halpin C, Knight ME, Foxon GA, Campbell MM, Boudet AM, Boon JJ, Chabbert B, Tollier MT, Schuch W (1994) Manipulation of lignin quality by downregulation of cinnamyl alcohol dehydrogenase. Plant J 6:339–350CrossRefGoogle Scholar
  24. Hepworth D, Vincent J (1998) The mechanical properties of xylem tissue from tobacco plants (Nicotiana tabacum ‘Samsun’). Ann Bot 81:751–759CrossRefGoogle Scholar
  25. Horvath B, Peralta P, Peszlen I, Divos F, Kasal B, LaiGeng L (2010) Elastic modulus of transgenic aspen. Wood Res (Bratisl) 55:1–10Google Scholar
  26. Hu W-J, Harding SA, Popko JL, Ralph J, Stokke DD, Tsai C-J, Chiang VL (1999) Repression of lignin biosynthesis promotes cellulose accumulation and growth in transgenic trees. Nat Biotechnol 17:808CrossRefPubMedGoogle Scholar
  27. Jin K, Qin Z, Buehler MJ (2015) Molecular deformation mechanisms of the wood cell wall material. J Mech Behav Biomed Mat 42:198–206CrossRefGoogle Scholar
  28. Kačuráková M, Belton PS, Wilson RH, Hirsch J, Ebringerová A (1998) Hydration properties of xylan-type structures: an FTIR study of xylooligosaccharides. J Sci Food Agric 77:38–44CrossRefGoogle Scholar
  29. Köhler L, Spatz H-C (2002) Micromechanics of plant tissues beyond the linear-elastic range. Planta 215:33–40.  https://doi.org/10.1007/s00425-001-0718-9 CrossRefPubMedGoogle Scholar
  30. Lapierre C, Pollet B, Petit-Conil M, Toval G, Romero J, Pilate G, Leplé J-C, Boerjan W, Ferret V, De Nadai V (1999) Structural alterations of lignins in transgenic poplars with depressed cinnamyl alcohol dehydrogenase or caffeic acid O-methyltransferase activity have an opposite impact on the efficiency of industrial kraft pulping. Plant Physiol 119:153–164CrossRefPubMedPubMedCentralGoogle Scholar
  31. Larsen KL, Barsberg S (2010) Theoretical and Raman spectroscopic studies of phenolic lignin model monomers. J Phys Chem B 114:8009–8021CrossRefPubMedGoogle Scholar
  32. Liu B, Wang P, Kim JI, Zhang D, Xia Y, Chapple C, Cheng J-X (2015) Vibrational fingerprint mapping reveals spatial distribution of functional groups of lignin in plant cell wall. Anal Chem 87:9436–9442CrossRefPubMedGoogle Scholar
  33. Mackenzie-Helnwein P, Müllner HW, Eberhardsteiner J, Mang HA (2005) Analysis of layered wooden shells using an orthotropic elasto-plastic model for multi-axial loading of clear spruce wood. Comput Methods Appl Mech Eng 194:2661–2685.  https://doi.org/10.1016/j.cma.2004.07.051 CrossRefGoogle Scholar
  34. Özparpucu M, Rüggeberg M, Gierlinger N, Cesarino I, Vanholme R, Boerjan W, Burgert I (2017) Unravelling the impact of lignin on cell wall mechanics—a comprehensive study on young poplar trees downregulated for cinnamyl alcohol dehyrogenase (CAD). Plant J 91:480–490CrossRefPubMedGoogle Scholar
  35. Pilate G, Guiney E, Holt K, Petit-Conil M, Lapierre C, Leplé J-C, Pollet B, Mila I, Webster EA, Marstorp HG (2002) Field and pulping performances of transgenic trees with altered lignification. Nat Biotechnol 20:607–612CrossRefPubMedGoogle Scholar
  36. Rowell RM (2012) Handbook of wood chemistry and wood composites. CRC Press, Boca RatonCrossRefGoogle Scholar
  37. Rüggeberg M, Speck T, Paris O, Lapierre C, Pollet B, Koch G, Burgert I (2008) Stiffness gradients in vascular bundles of the palm Washingtonia robusta. Proceed R Soc B Biol Sci 275:2221–2229CrossRefGoogle Scholar
  38. Rüggeberg M, Saxe F, Metzger TH, Sundberg B, Fratzl P, Burgert I (2013) Enhanced cellulose orientation analysis in complex model plant tissues. J Struct Biol 183:419–428CrossRefPubMedGoogle Scholar
  39. Salmén L, Burgert I (2009) Cell wall features with regard to mechanical performance. A review COST Action E35 2004–2008: Wood machining–micromechanics and fracture. Holzforschung 63:121–129CrossRefGoogle Scholar
  40. Schwanninger M, Rodrigues J, Pereira H, Hinterstoisser B (2004) Effects of short-time vibratory ball milling on the shape of FT-IR spectra of wood and cellulose. Vib Spectrosc 36:23–40CrossRefGoogle Scholar
  41. Speck T, Burgert I (2011) Plant stems: functional design and mechanics. Annu Rev Mater Res 41:169–193CrossRefGoogle Scholar
  42. Sun L, Varanasi P, Yang F, Loqué D, Simmons BA, Singh S (2012) Rapid determination of syringyl: Guaiacyl ratios using FT-Raman spectroscopy. Biotechnol Bioeng 109:647–656CrossRefPubMedGoogle Scholar
  43. Van Acker R, Leplé J-C, Aerts D, Storme V, Goeminne G, Ivens B, Légée F, Lapierre C, Piens K, Van Montagu MC (2014) Improved saccharification and ethanol yield from field-grown transgenic poplar deficient in cinnamoyl-CoA reductase. Proc Natl Acad Sci 111:845–850CrossRefPubMedGoogle Scholar
  44. Van Acker R, Déjardin A, Desmet S, Vanholme R, Morreel K, Laurans F, Kim H, Santoro N, Foster C, Goeminne G, Légée F, Lapierre C, Pilate G, Ralph J, Boerjan W (2017) Different metabolic routes for coniferaldehyde and sinapaldehyde with CINNAMYL ALCOHOL DEHYDROGENASE1 deficiency. Plant Physiol 175(3):1018–1039.  https://doi.org/10.1104/pp.17.00834 CrossRefPubMedPubMedCentralGoogle Scholar
  45. Vanholme R, Morreel K, Ralph J, Boerjan W (2008) Lignin engineering. Curr Opin Plant Biol 11:278–285.  https://doi.org/10.1016/j.pbi.2008.03.005 CrossRefPubMedGoogle Scholar
  46. Vargas L, Cesarino I, Vanholme R, Voorend W, Saleme MdLS, Morreel K, Boerjan W (2016) Improving total saccharification yield of Arabidopsis plants by vessel-specific complementation of caffeoyl shikimate esterase (cse) mutants. Biotechnol Biofuels 9:139CrossRefPubMedPubMedCentralGoogle Scholar
  47. Voelker SL, Lachenbruch B, Meinzer FC, Strauss SH (2011) Reduced wood stiffness and strength, and altered stem form, in young antisense 4CL transgenic poplars with reduced lignin contents. New Phytol 189:1096–1109.  https://doi.org/10.1111/j.1469-8137.2010.03572.x CrossRefPubMedGoogle Scholar
  48. Zobel BJ, Van Buijtenen JP (2012) Wood variation: its causes and control. Springer Science & Business Media, BerlinGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2017

Authors and Affiliations

  • Merve Özparpucu
    • 1
  • Notburga Gierlinger
    • 3
  • Ingo Burgert
    • 1
    • 2
  • Rebecca Van Acker
    • 4
    • 5
  • Ruben Vanholme
    • 4
    • 5
  • Wout Boerjan
    • 4
    • 5
  • Gilles Pilate
    • 6
  • Annabelle Déjardin
    • 6
  • Markus Rüggeberg
    • 1
    • 2
  1. 1.Institute for Building Materials (IfB)ETH ZurichZurichSwitzerland
  2. 2.Laboratory of Applied Wood MaterialsEMPADübendorfSwitzerland
  3. 3.Institute for BiophysicsUniversity of Natural Resources and Life Sciences ViennaViennaAustria
  4. 4.Department of Plant Biotechnology and BioinformaticsGhent UniversityGhentBelgium
  5. 5.VIB Center for Plant Systems BiologyGhentBelgium
  6. 6.AGPF, INRAOrléansFrance

Personalised recommendations