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Licensed Unlicensed Requires Authentication Published by De Gruyter March 30, 2022

XET activity determination in powdered wood samples as an indicator of tension wood, tested on juvenile Populus x euramericana exposed to severe long-term static bending

  • Miloš Prokopijević , Jasna Simonović Radosavljević , Dragica Spasojević , Katarina Vojisavljević , Ksenija Radotić and Aleksandra Lj Mitrović ORCID logo EMAIL logo
From the journal Holzforschung


Leaning stems of woody plants form reaction wood, in hardwood trees termed tension wood (TW). Typical TW fibers, gelatinous fibers (G-fibers), are characterized by an inner gelatinous cell wall layer (G-layer). Xyloglucan endotransglycosylases (XETs) was proposed as the essential enzyme in cell wall modifications in TW, by making xyloglucan (XG) cross-links between G- and S2-layers in G-fibers, and thus maintaining their contact. The determination of TW presence in a sample is of great importance for the forest products industry, biofuel production, and tree physiology studies. However, TW is not easy to detect visually. The colorimetric assay for XET activity determination as an indicator of TW presence in a sample was tested on powdered stem segments of juvenile Populus x euramericana trees exposed to severe long-term static bending. In parallel, histochemical and ultrastructural characterization of stem samples of bent and control trees was performed. The tested colorimetric assay for XET activity determination could be suggested as a useful and easily applicable tool for fast screening of powdered wood samples for the presence of TW.

Corresponding author: Aleksandra Lj Mitrović, Institute for Multidisciplinary Research, University of Belgrade, Kneza Višeslava 1, 11030, Belgrade, Serbia; and Center for Green Technologies, Institute for Multidisciplinary Research, University of Belgrade, Kneza Višeslava 1, 11030, Belgrade, Serbia, E-mail:
Miloš Prokopijević and Jasna Simonović Radosavljević contributed equally to this work.

Funding source: Ministry of Education, Science and Technological Development of the Republic of Serbia

Award Identifier / Grant number: 451-03-68/2022-14/200053


The authors wish to thank the Institute of Lowland Forestry and Environment, University of Novi Sad for providing Populus x euramericana cl. NS 11-8 cuttings. We thank Dr Vladimir Farkaš, Slovak Akademy os Sciences, Bratislava, for providing XG and XGOs.

  1. Author contributions: KR and ALjM – research design; JSR, DS and ALjM – sample collection and processing; MP, DS, JSR – XET extraction and activity determination and results interpretation; KV – SEM. ALjM – histochemical and structural analysis; ALjM – writing of the manuscript. All authors read and approved the manuscript.

  2. Research funding: This work was supported by the Ministry of Education, Science and Technological Development of the Republic of Serbia (contract no. 451-03-68/2022-14/200053).

  3. Conflict of interest statement: The authors declare that they have no conflicts of interest regarding this article.


Alméras, T. and Clair, B. (2016). Critical review on the mechanisms of maturation stress generation in trees. J. R. Soc. Interface 13: 20160550, in Google Scholar PubMed PubMed Central

Badia, M.A., Mothe, F., Constant, T., and Nepveu, G. (2005). Assessment of tension wood detection based on shiny appearance for three poplar cultivars. Ann. For. Sci. 62: 43–49, in Google Scholar

Bourquin, V., Nyshikubo, N., Abe, H., Brumer, H., Denman, S., Eklund, M., Christiernin, M., Teer, T.T., Sundberg, B., and Mellerowicz, E.J. (2002). Xyloglucan endotransglycosylases have a function during the formation of secondary cell walls of vascular tissues. Plant Cell 14: 3073–3088, in Google Scholar PubMed PubMed Central

Bowling, A.J. and Vaughn, K.C. (2008). Immunocytochemical characterization of tension wood: Gelatinous fibers contain more than just cellulose. Am. J. Bot. 95: 655–663, in Google Scholar PubMed

Clair, B., Thibaut, B., and Sugiyama, J. (2005). On the detachment of gelatinous layer in tension wood fibre. J. Wood Sci. 51: 218–221, in Google Scholar

Donaldson, L.A. and Singh, A.P. (2016). Reaction wood. In: Kim, Y.S., et al.. (Eds.), Secondary xylem biology. Elsevier, Amsterdam, pp. 93–110.10.1016/B978-0-12-802185-9.00006-1Search in Google Scholar

Farkaš, V., Ait-Mohand, F., and Stratilová, E. (2005). Sensitive detection of transglycosylating activity of xyloglucan endotransglycosylase/hydrolase (XTH) after isoelectric focusing in polyacrylamide gels. Plant Physiol. Biochem. 43: 431–435.10.1016/j.plaphy.2005.03.006Search in Google Scholar PubMed

Han, Y., Zhu, Q., Zhang, Z., Meng, K., Hou, Y., Ban, Q., Suo, J., and Rao, J. (2015). Analysis of xyloglucan endotransglycosylase/hydrolase (XTH) genes and diverse roles of isoenzymes during persimmon fruit development and postharvest softening. PLoS One 10: 1–20, in Google Scholar PubMed PubMed Central

Henriksson, H., Denman, S.E., Campuzano, I.D., Ademark, P., Master, E.R., Teeri, T.T., and Brumer, H.3rd (2003). N-linked glycosylation of native and recombinant cauliflower xyloglucan endotransglycosylase 16A. Biochem. J. 375: 61–73, in Google Scholar PubMed PubMed Central

Jensen, W.A. (1962). Botanical histochemistry: Principles and practice. W.H. Freeman and Company, San Francisco, California.Search in Google Scholar

Kaku, T., Serada, S., Baba, K., Tanaka, F., and Hayashi, T. (2009). Proteomic analysis of the G-layer in poplar tension wood. J. Wood Sci. 55: 250–257, in Google Scholar

Kaur, G.A. (2019). Review of XET Enzymes, Current applications and future trends. Int. J. Latest Technol. Eng. Manag. Appl. Sci. VIII: 52–55.Search in Google Scholar

Maeglin, R.R. (1987). Juvenile wood, tension wood, and growth stress effects on processing hardwoods. In: Applying the latest research to hardwood problems. Proceedings of the 15th annual hardwood symposium of the Hardwood Research Council. Hardwood Research Council, Memphis, TN, pp. 100–108.Search in Google Scholar

Nyshikubo, N., Awano, T., Banasiak, A., Bourquin, V., Ibatullin, F., Funada, R., Brumer, H., Teeri, T.T., Hayashi, T., Sundberg, B., et al.. (2007). Xyloglucan endo-transglycosylase (XET) functions in gelatinous layers of tension wood fibers in poplar--a glimpse into the mechanism of the balancing act of trees. Plant Cell Physiol 48: 843–855.10.1093/pcp/pcm055Search in Google Scholar PubMed

Nyshikubo, N., Takahashi, J., Roos, A.A., Derba-Maceluch, M., Piens, K., Brumer, H., Teeri, T.T., Stålbrand, H., and Mellerowicz, E.J. (2011). Xyloglucan endo-transglycosylase-mediated xyloglucan rearrangements in developing wood of hybrid aspen. Plant Physiol. 155: 399–413.10.1104/pp.110.166934Search in Google Scholar PubMed PubMed Central

Perre, P., Tuan Dinh, A., Assor, C., Frank, X., and Pilate, G. (2013). Stiffness of normal, opposite, and tension poplar wood determined using micro-samples in the three material directions. Wood Sci. Technol. 47: 481–498, in Google Scholar

Roussel, J.R. and Clair, B. (2015). Evidence of the late lignification of the G-layer in Simarouba tension wood, to assist understanding how non-G-layer species produce tensile stress. Tree Physiol. 35: 1366–1377, in Google Scholar PubMed

Ruelle, J. (2014). Morphology, anatomy and ultrastructure of reaction wood. In: Gardiner, B., et al.. (Eds.), The biology of reaction wood. Springer series in wood science. Springer, Berlin, pp. 13–35.10.1007/978-3-642-10814-3_2Search in Google Scholar

Sulová, Z., Lednická, M., and Farkaš, V. (1995). A colorimetric assay for xyloglucan-endotransglycosylase from germinating seeds. Anal. Biochem. 229: 80–85.10.1006/abio.1995.1381Search in Google Scholar PubMed

Timell, T.E. (1969). The chemical composition of tension wood. Svensk Papperstidning 72: 173–181.Search in Google Scholar

Received: 2021-11-08
Accepted: 2022-02-24
Published Online: 2022-03-30
Published in Print: 2022-07-26

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