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Complete substitution of a secondary cell wall with a primary cell wall in Arabidopsis


The bulk of a plant’s biomass, termed secondary cell walls, accumulates in woody xylem tissues and is largely recalcitrant to biochemical degradation and saccharification1. By contrast, primary cell walls, which are chemically distinct, flexible and generally unlignified2, are easier to deconstruct. Thus, engineering certain primary wall characteristics into xylem secondary walls would be interesting to readily exploit biomass for industrial processing. Here, we demonstrated that by expressing AP2/ERF transcription factors from group IIId and IIIe in xylem fibre cells of mutants lacking secondary walls, we could generate plants with thickened cell wall characteristics of primary cell walls in the place of secondary cell walls. These unique, newly formed walls displayed physicochemical and ultrastructural features consistent with primary walls and had gene expression profiles illustrative of primary wall synthesis. These data indicate that the group IIId and IIIe AP2/ERFs are transcription factors regulating primary cell wall deposition and could form the foundation for exchanging one cell wall type for another in plants.

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Fig. 1: Accumulation of lignin-deficient cell walls by ERF035-VP16 plants.
Fig. 2: ERF035 is sufficient to activate primary cell wall formation.
Fig. 3: Induced cell walls in nst1nst3 NST3pro::ERF035-VP16 plant stems are similar to primary cell walls.
Fig. 4: Typical primary cell wall structures are deposited in induced fibre cell walls of nst1nst3 NST3pro::ERF035-VP16 plants.

Data availability

The microarray data in this study were deposited to the NCBI GEO under the accession number GSE81039. Other data that support the findings of this study are available from the corresponding author upon reasonable request. Figures 1l–o, 2b,d and 3a,b,d–f and Supplementary Figs. 3, 4, 7c, 10, 11, 1418, 21 and 22 have associated raw data.


  1. 1.

    Harris, D. & DeBolt, S. Synthesis, regulation and utilization of lignocellulosic biomass. Plant Biotechnol. J. 8, 244–262 (2010).

    CAS  Article  Google Scholar 

  2. 2.

    Cosgrove, D. J. Re-constructing our models of cellulose and primary cell wall assembly. Curr. Opin. Plant Biol. 22, 122–131 (2014).

    CAS  Article  Google Scholar 

  3. 3.

    Wang, T., McFarlane, H. E. & Persson, S. The impact of abiotic factors on cellulose synthesis. J. Exp. Bot. 67, 543–552 (2016).

    CAS  Article  Google Scholar 

  4. 4.

    Albersheim, P., Darvill, A., Roberts, K., Sederoff, R. & Staehelin, A. Plant Cell Walls: From Chemistry to Biology (Garland Science, New York, 2010).

  5. 5.

    Kumar, M., Campbell, L. & Turner, S. Secondary cell walls: biosynthesis and manipulation. J. Exp. Bot. 67, 515–531 (2016).

    CAS  Article  Google Scholar 

  6. 6.

    Phitsuwan, P., Sakka, K. & Ratanakhanokchai, K. Improvement of lignocellulosic biomass in planta: a review of feedstocks, biomass recalcitrance, and strategic manipulation of ideal plants designed for ethanol production and processability. Biomass Bioenergy 58, 390–405 (2013).

    CAS  Article  Google Scholar 

  7. 7.

    Mottiar, Y., Vanholme, R., Boerjan, W., Ralph, J. & Mansfield, S. D. Designer lignins: harnessing the plasticity of lignification. Curr. Opin. Biotechnol. 37, 190–200 (2016).

    CAS  Article  Google Scholar 

  8. 8.

    Engels, F. M. & Jung, H. G. Alfalfa stem tissues: cell-wall development and lignification. Annals Bot. 82, 561–568 (1998).

    Article  Google Scholar 

  9. 9.

    Müller, S. et al. Cell wall composition of vascular and parenchyma tissues in broccoli stems. J. Sci. Food. Agric. 83, 1289–1292 (2003).

    Article  Google Scholar 

  10. 10.

    Wilson, W. D., Jarvis, M. C. & Duncan, H. J. In-vitro digestibility of kale (Brassica oleracea) secondary xylem and parenchyma cell walls and their polysaccharide components. J. Sci. Food. Agric. 48, 9–14 (1989).

    CAS  Article  Google Scholar 

  11. 11.

    Brereton, N. J. B. et al. Reaction wood—a key cause of variation in cell wall recalcitrance in willow. Biotech. Biofuels 5, 83 (2012).

    CAS  Article  Google Scholar 

  12. 12.

    Mitsuda, N. et al. NAC transcription factors, NST1 and NST3, are key regulators of the formation of secondary walls in woody tissues of Arabidopsis. Plant Cell 19, 270–280 (2007).

    CAS  Article  Google Scholar 

  13. 13.

    Nakano, T., Suzuki, K., Fujimura, T. & Shinshi, H. Genome-wide analysis of the ERF gene family in Arabidopsis and rice. Plant Physiol. 140, 411–432 (2006).

    CAS  Article  Google Scholar 

  14. 14.

    Wilson, K., Long, D., Swinburne, J. & Coupland, G. A dissociation insertion causes a semidominant mutation that increases expression of TINY, an Arabidopsis gene related to APETALA2. Plant Cell 8, 659–671 (1996).

    CAS  Article  Google Scholar 

  15. 15.

    O’Malley, R. C. et al. Cistrome and epicistrome features shape the regulatory DNA landscape. Cell 165, 1280–1292 (2016).

    Article  Google Scholar 

  16. 16.

    Gaudinier, A. et al. Enhanced Y1H assays for Arabidopsis. Nat. Methods 8, 1053–1055 (2011).

    CAS  Article  Google Scholar 

  17. 17.

    Nagata, T. & Takebe, I. Cell wall regeneration and cell division in isolated tobacco mesophyll protoplasts. Planta 92, 301–308 (1970).

    CAS  Article  Google Scholar 

  18. 18.

    Hiratsu, K., Matsui, K., Koyama, T. & Ohme-Takagi, M. Dominant repression of target genes by chimeric repressors that include the EAR motif, a repression domain, in Arabidopsis. Plant J. 34, 733–739 (2003).

    CAS  Article  Google Scholar 

  19. 19.

    Kang, J. S. et al. Salt tolerance of Arabidopsis thaliana requires maturation of N-glycosylated proteins in the Golgi apparatus. Proc. Natl Acad. Sci. USA 105, 5933–5938 (2008).

    CAS  Article  Google Scholar 

  20. 20.

    Scheible, W. R., Eshed, R., Richmond, T., Delmer, D. & Somerville, C. Modifications of cellulose synthase confer resistance to isoxaben and thiazolidinone herbicides in Arabidopsis Ixr1 mutants. Proc. Natl Acad. Sci. USA 98, 10079–10084 (2001).

    CAS  Article  Google Scholar 

  21. 21.

    Lasserre, E., Jobet, E., Llauro, C. & Delseny, M. AtERF38 (At2g35700), an AP2/ERF family transcription factor gene from Arabidopsis thaliana, is expressed in specific cell types of roots, stems and seeds that undergo suberization. Plant Physiol. Biochem. 46, 1051–1061 (2008).

    CAS  Article  Google Scholar 

  22. 22.

    Sakamoto, S. & Mitsuda, N. Reconstitution of a secondary cell wall in a secondary cell wall-deficient Arabidopsis mutant. Plant Cell Physiol. 56, 299–310 (2015).

    CAS  Article  Google Scholar 

  23. 23.

    Pattathil, S. et al. A comprehensive toolkit of plant cell wall glycan-directed monoclonal antibodies. Plant Physiol. 153, 514–525 (2010).

    CAS  Article  Google Scholar 

  24. 24.

    Knox, J. P., Linstead, P. J., King, J., Cooper, C. & Roberts, K. Pectin esterification is spatially regulated both within cell walls and between developing tissues of root apices. Planta 181, 512–521 (1990).

    CAS  Article  Google Scholar 

  25. 25.

    Jones, L., Seymour, G. B. & Knox, J. P. Localization of pectic galactan in tomato cell walls using a monoclonal antibody specific to (1→4)-β-d-galactan. Plant Physiol. 113, 1405–1412 (1997).

    CAS  Article  Google Scholar 

  26. 26.

    Willats, W. G., Marcus, S. E. & Knox, J. P. Generation of a monoclonal antibody specific to (1→5)-α-l-arabinan. Carbohydr. Res. 308, 149–152 (1998).

    CAS  Article  Google Scholar 

  27. 27.

    McCartney, L., Marcus, S. E. & Knox, J. P. Monoclonal antibodies to plant cell wall xylans and arabinoxylans. J. Histochem. Cytochem. 53, 543–546 (2005).

    CAS  Article  Google Scholar 

  28. 28.

    Blake, A. W. et al. Understanding the biological rationale for the diversity of cellulose-directed carbohydrate-binding modules in prokaryotic enzymes. J. Biol. Chem. 281, 29321–29329 (2006).

    CAS  Article  Google Scholar 

  29. 29.

    Anderson, C. T., Carroll, A., Akhmetova, L. & Somerville, C. Real-time imaging of cellulose reorientation during cell wall expansion in Arabidopsis roots. Plant Physiol. 152, 787–796 (2010).

    CAS  Article  Google Scholar 

  30. 30.

    Mitra, P. P. & Loqué, D. Histochemical staining of Arabidopsis thaliana secondary cell wall elements. J. Vis. Exp. 87, 51381 (2014).

    Google Scholar 

  31. 31.

    Zeng, Y., Zhao, S., Yang, S. & Ding, S. Y. Lignin plays a negative role in the biochemical process for producing lignocellulosic biofuels. Curr. Opin. Biotechnol. 27, 38–45 (2014).

    CAS  Article  Google Scholar 

  32. 32.

    Stewart, J. J., Akiyama, T., Chapple, C., Ralph, J. & Mansfield, S. D. The effects on lignin structure of overexpression of ferulate 5-hydroxylase in hybrid poplar. Plant Physiol. 150, 621–635 (2009).

    CAS  Article  Google Scholar 

  33. 33.

    Vanholme, R. et al. Caffeoyl shikimate esterase (CSE) is an enzyme in the lignin biosynthetic pathway in Arabidopsis. Science 341, 1103–1106 (2013).

    CAS  Article  Google Scholar 

  34. 34.

    Wilkerson, C. G. et al. Monolignol ferulate transferase introduces chemically labile linkages into the lignin backbone. Science 344, 90–93 (2014).

    CAS  Article  Google Scholar 

  35. 35.

    Sakamoto, S. et al. Wood reinforcement of poplar by rice NAC transcription factor. Sci. Rep. 6, 19925 (2016).

    CAS  Article  Google Scholar 

  36. 36.

    Storey, J. D. & Tibshirani, R. Statistical significance for genome wide studies. Proc. Natl Acad. Sci. USA 100, 9440–9445 (2003).

    CAS  Article  Google Scholar 

  37. 37.

    Kumar, S., Stecher, G., Li, M., Knyaz, C. & Tamura, K. MEGA X: molecular evolutionary genetics analysis across computing platforms. Mol. Biol. Evol. 35, 1547–1549 (2018).

    Article  Google Scholar 

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We thank A. Hosaka, A. Kuwazawa, F. Tobe, M. Yamada, Y. Sugimoto and Y. Takiguchi for their technical support. This work was supported by the JST ALCA program (grant no. JPMJAL1107) (to N.M.), a postdoctoral fellowship from the German Research Foundation (DFG, project 344523413) (to M.S.), University of Melbourne R@MAP Professorship, an ARC Future Fellowship grant (FT160100218) and a UoM IRRTF RNC grant (501892) (to S.P.) and an NSERC Discovery Grant (to S.D.M.).

Author information




N.M. conceived and executed the project as principal investigator. N.M. and S.S. designed the experimental plans. K.Y. performed the tensile test and microarray data analysis. S.P. and S.D.M. designed the experiment related to CoMPP and cellulose analyses. S.S. performed all of the experiments except for CoMPP, cellulose analyses, TEM imaging and the yeast one-hybrid assay. M.S. performed the CoMPP analysis with the supervision of S.P. M.T.N. observed the root of the 35Spro::ERF035 plant. F.U. and S.D.M. performed the cellulose analyses. T.W., A.-M.B. and A.G. performed the yeast one-hybrid assay and its preparation. S.M.B. and S.P. designed and supervised the yeast one-hybrid assay. K.A. and Y.K. performed the TEM imaging, preparation of the thin section and the analysis of images. S.S., S.P. and N.M. mainly wrote the manuscript, with all other authors contributing to revisions.

Corresponding author

Correspondence to Nobutaka Mitsuda.

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The authors declare no competing interests.

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Supplementary information

Supplementary Information

Supplementary Methods, Supplementary References and Supplementary Figures 1–25.

Reporting Summary

Supplementary Table 1

Over-represented cell-wall-related GO terms in up-regulated genes in nst1 nst3 NST3pro:ERF035-VP16 plants.

Supplementary Table 2

Fold changes of cell wall-related genes in NST3p:ERF035-VP16 nst1 nst3 as per nst1 nst3.

Supplementary Table 3

Fold changes of cell wall-related genes in NST3p:ERF035-VP16 nst1 nst3 as per nst1 nst3 and in wt as per nst1 nst3.

Supplementary Table 4

List of upregulated genes related to cell wall biosynthesis in the nst1 nst3 NST3pro:ERF035-VP16 transgenic plants.

Supplementary Table 5

Over-represented cell-wall-related GO terms in putative target genes of group IIId and IIIe ERFs.

Supplementary Table 6

Primers in this study.

Supplementary Table 7

All transcription factor genes focused in this study.

Supplementary Table 8

Amino acid sequences of conserved domain used for constructing phylogenetic tree shown in Supplementary Fig. 25.

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Sakamoto, S., Somssich, M., Nakata, M.T. et al. Complete substitution of a secondary cell wall with a primary cell wall in Arabidopsis. Nature Plants 4, 777–783 (2018).

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