, Volume 219, Issue 2, pp 338–345 | Cite as

Detection in situ and characterization of lignin in the G-layer of tension wood fibres of Populus deltoides

  • Jean-Paul Joseleau
  • Takanori Imai
  • Katsushi Kuroda
  • Katia Ruel
Original Article


The occurrence of lignin in the additional gelatinous (G-) layer that differentiates in the secondary wall of hardwoods during tension wood formation has long been debated. In the present work, the ultrastructural distribution of lignin in the cell walls of normal and tension wood fibres from poplar (Populus deltoides Bartr. ex Marshall) was investigated by transmission electron microscopy using cryo-fixation–freeze-substitution in association with immunogold probes directed against typical structural motifs of lignin. The specificity of the immunological probes for condensed and non-condensed guaiacyl and syringyl interunit linkages of lignin, and their high sensitivity, allowed detection of lignin epitopes of definite chemical structures in the G-layer of tension wood fibres. Semi-quantitative distribution of the corresponding epitopes revealed the abundance of syringyl units in the G-layer. Predominating non-condensed lignin sub-structures appeared to be embedded in the crystalline cellulose matrix prevailing in the G-layer. The endwise mode of polymerization that is known to lead to these types of lignin structures appears consistent with such an organized cellulose environment. Immunochemical labelling provides the first visualization in planta of lignin structures within the G-layer of tension wood. The patterns of distribution of syringyl epitopes indicate that syringyl lignin is deposited more intensely in the later phase of fibre secondary wall assembly. The data also illustrate that syringyl lignin synthesis in tension wood fibres is under specific spatial and temporal regulation targeted differentially throughout cell wall layers.


G-layer Lignin Lignin immunolabelling Populus Syringyl unit Tension wood 



Gelatinous layer


Guaiacyl monomeric unit


Periodic acid–thiocarbohydrazide–silver proteinate


Syringyl monomeric unit



The authors thank Guillaume Chantre (AFOCELL, Nungis, France) for the gift of the sample of poplar, clone Raspalje. Some of the results were acquired in the framework of European program AIR 3 CT 94-2065.


  1. Aoyama W, Matsumura A, Tsutsumi Y, Nishita (2001) Lignification and peroxidase in tension wood of Eucalyptus viminalis seedlings. J Wood Sci 47:419–424Google Scholar
  2. Araki N, Fujita M, Saiki H, Harada H (1982) Transition of the fiber wall from normal wood to tension wood in Robinia pseudoacacia L. and Populus euroamericana Gunii. Mokuzai Gakkaishi 28:267–273Google Scholar
  3. Baillères H, Castan M, Monties B, Pollet B, Lapierre C (1997) Lignin structure in Buxus sempervirens reaction wood. Phytochemistry 44:35–39CrossRefGoogle Scholar
  4. Besombes S, Robert D, Utille J-P, Taravel F-R, Mazeau K (2003) Molecular modelling of syringyl and p-hydroxyphenyl β-O-4 dimers. Comparative study of the computed experimental conformational properties of lignin β-O-4 model compounds. J Agric Food Chem 51:34–42CrossRefPubMedGoogle Scholar
  5. Blanchette RA, Obst JR, Timell TE (1994) Biodegradation of compression wood and tension wood by white and brown rot fungi. Holzforschung 48:34–42Google Scholar
  6. Dixon RA, Chen F, Guo D, Parvathi K (2001) The biosynthesis of monolignols: a “metabolic grid”, or independent pathways to guaiacyl and syringyl units? Phytochemistry 57:1069–1084CrossRefPubMedGoogle Scholar
  7. Donaldson LA (2001) Lignification and lignin topochemistry—an ultrastructural view. Phytochemistry 57:859–873CrossRefPubMedGoogle Scholar
  8. Furuya N, Tatahashi S, Miyasaki M (1970) The chemical composition of the gelatinous layer from the tension wood of Populus euro-americana. Mokuzai Gakkaishi 16:26–30Google Scholar
  9. Gindl W (2002) Comparing mechanical properties of normal and compression wood in Norway spruce: the role of lignin in compression parallel to grain. Holzforschung 56:395–401Google Scholar
  10. Grünwald C, Ruel K, Kim YS, Schmitt U (2002) On the cytochemistry of cell wall formation in poplar trees. Plant Biol 4:13–21Google Scholar
  11. Hawkins S, Boudet A (2003) ‘Defence lignin’ and hydroxycinnamyl alcohol dehydrogenase activities in wounded Eucalyptus gunnii. For Pathol 33:339–352Google Scholar
  12. Houtman CJ, Atalla RH (1995) Cellulose–lignin interactions. A computational study. Plant Physiol 107:977–984PubMedGoogle Scholar
  13. Hu W-J, Harding SA, Lung J, Popko JL, Stokke DD, Tsai CJ, Chiang VL (1999). Repression of lignin biosynthesis promotes cellulose accumulation and growth in transgenic trees. Nature Biotechnol 17:808–812CrossRefGoogle Scholar
  14. Imai T, Ruel K, Pilate G, Leple J-C, Joseleau J-P (2002) Influence of environment or genetic factors on the formation of tension wood. Abstract Journées Réseau Français des Parois, Reims May 2002, p 13Google Scholar
  15. Joseleau J-P, Ruel K (1997) Study of lignification by non invasive techniques in growing maize internodes—an investigation by Fourier transform infrared, cross-polarisation-magic angle spinning 13C-nuclear magnetic resonance spectroscopy. Plant Physiol 114:1123–1133CrossRefPubMedGoogle Scholar
  16. Joseleau J-P, Petit-Conil M, Jouanin L, DeChoudens C, Chantre G, Sollier JN, Ruel K (1999) Immunological characterization of residual lignins in pulps from poplar clones and from a genetically modified variety. 10th international symposium on wood pulping chemistry, Yokohama, Japan, vol II, pp 190–193Google Scholar
  17. Kuo CM, Timell TE (1969) Isolation and characterization of a galactan from tension wood of American beech (Fagus grandifolia Ehrl.) Svensk Papperstidn 72:703–708Google Scholar
  18. Lange BM, Lapierre C, Sandermann (1995) Elicitor-induced spruce stress lignin. Structural similarity to developmental lignins. Plant Physiol 108:1277–1287PubMedGoogle Scholar
  19. Lapierre C (1993) Application of new methods for the investigation of lignin structure. In: Jung HG, Buxton DR, Hatfield RD, Ralph J (eds) Forage cell wall structure and digestibility. American Society of Agronomy Inc, Madison, pp 133–166Google Scholar
  20. Maier-Maercker U, Koch W (1986) Delignification of subsidiary and guard cell walls by SO2 and probable implication on the humidity response of Picea abies (L) Karst. Eur J For Pathol 16:342–351Google Scholar
  21. Nevell TP (1963) Degradation of cellulose—28. Oxidation. In: Wistler RL (ed) Methods in carbohydrate chemistry, vol III. Academic Press, NY, pp 164–284Google Scholar
  22. Nicholson RL, Hammerschmidt R (1992) Phenolic compounds and their role in disease resistance. Annu Rev Phytopathol 30:113–157Google Scholar
  23. Norberg PH, Meier H (1966) Physical and chemical properties of the gelatinous layer in tension wood fibers of aspen (Populus tremula L.). Holzforschung 20:174–178Google Scholar
  24. Parham RA, Côté WA (1971) Distribution of lignin in normal and compression wood of Pinus taeda. Wood Sci Technol 5:49–54Google Scholar
  25. Robards AW (1966) The application of the modified sine rule to tension wood production in the stem of crack willow (Salix fragilis L). Ann Bot 30:513–523Google Scholar
  26. Roussel MR, Lim C (1995) Dynamic model of lignin growing in restricted spaces. Macromolecules 28:370–376Google Scholar
  27. Ruel K, Barnoud F (1978) Détermination quantitative du bois de tension par une méthode analytique chimique: validité du critère galactose. Holzforschung 32:149–156Google Scholar
  28. Ruel K, Barnoud F, Goring DAI (1979) Ultrastructural lamellation in the S2 layer of two hardwoods and a reed. Cell Chem Technol 13:429–432Google Scholar
  29. Ruel K, Faix O, Joseleau J-P (1994) New immunogold probes for studying the distribution of the different lignin types during plant cell wall biogenesis. J Trace Microprobe Tech 12:2247–265Google Scholar
  30. Ruel K, Burlat V, Joseleau J-P (1999) Relationship between ultrastructural topochemistry of lignin and wood properties. IAWA J 20:203–211Google Scholar
  31. Ruel K, Chabannes M, Boudet A-M, Legrand M, Joseleau J-P (2001) Reassessment of quantitative changes in lignification of transgenic tobacco plants and their impact on cell wall assembly. Phytochemistry 57:875–882CrossRefPubMedGoogle Scholar
  32. Ruel K, Faix O, Kuroda K-I Joseleau J-P (2004) A polyclonal antibody directed against syringyl propane epitopes of native lignins. C R Acad Sci (in press)Google Scholar
  33. Sandermann HE, Heller W, Langebartels C (1998) Ozone: an abiotic elicitor of plant defence reactions. Trends Plant Sci 3:47–50CrossRefGoogle Scholar
  34. Scurfield G (1971) Histochemistry of reaction wood cell walls in two species of Eucalyptus and in Tristinia conferta R. BR. Aust J Bot 20:9–26Google Scholar
  35. Scurfield G, Wardrop AB (1963) The nature of reaction wood. VII. Lignification in reaction wood. Aust J Bot 11 107–116Google Scholar
  36. Southerton SG, Deverall BJ (1990) Histochemical and chemical for lignin accumulation during the expression of resistance to leaf rust fungi in wheat. Physiol Mol Plant Pathol 36:483–494Google Scholar
  37. Thiery JP (1967) Mise en évidence des polysaccharides sur coupes fines en microscopie électronique. J Microsc 6:987–1017Google Scholar
  38. Timell TE (1969) The chemical composition of tension wood. Svensk Papperstidn 72:173–178Google Scholar
  39. Vance CP, Kirk TK, Sherwood RT (1980) Lignification as a mechanism of disease resistance. Annu Rev Phytopathol 18:259–288Google Scholar
  40. Wada M, Okano T, Sugiyama J, Horii F (1995) Characterization of tension and normally lignified wood cellulose in Populus maximowiczii. Cellulose 2:223–233Google Scholar
  41. Wardrop AB, Dadswell HE (1955) Nature of reaction wood. IV. Variations in cell wall organization of tension-wood fibres. Aust J Bot 3:177–189Google Scholar
  42. Yamamoto H (1998) Generation process of growth stresses in cell walls: role of lignin deposition and cellulose micro-fibril during cell wall maturation. Wood Sci Technol 32:171–182Google Scholar
  43. Yamamoto H, Okuyama T, Yoshida M (1993) Generation of a process of growth stresses in cell walls. V. Model of tensile stress generation in gelatinous fibers. Mokuzai Gakkaishi 39:118–125Google Scholar
  44. Yamauchi K, Yasuda S, Hamada K, Tsutsumi Y, Fukushima K (2003) Multiform biosynthetic pathway of syringyl lignin in angiosperms. Planta 216:496–501PubMedGoogle Scholar
  45. Yoshida M, Ohta H, Okuyama T (2002) Tensile growth stress and lignin distribution in the cell walls of black locust (Robinia pseudoacacia). J Wood Sci 48:99–105Google Scholar
  46. Yoshida S, Tanahashi M, Shigematsu M, Shinoda Y (1994) Effect of reaction medium on dehydrogenative polymerisation of sinapyl alcohol. Mokuzai Gakkaishi 40:974–979Google Scholar

Copyright information

© Springer-Verlag 2004

Authors and Affiliations

  • Jean-Paul Joseleau
    • 1
  • Takanori Imai
    • 1
    • 2
  • Katsushi Kuroda
    • 1
    • 3
  • Katia Ruel
    • 1
  1. 1.Centre de Recherche sur les Macromolécules VégétalesCERMAV-CNRSGrenoble Cedex 9France
  2. 2.School of Bioagricultural SciencesNagoya UniversityNagoyaJapan
  3. 3.National Institute of Agrobiological SciencesIbarakiJapan

Personalised recommendations