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Shank-localized cell wall growth contributes to Arabidopsis root hair elongation

Nature Plants volume 8, pages 1222–1232 (2022)Cite this article

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Abstract

Root hairs are highly elongated tubular extensions of root epidermal cells with a plethora of physiological functions, particularly in establishing the root–rhizosphere interface. Anisotropic expansion of root hairs is generally thought to be exclusively mediated by tip growth—a highly controlled apically localized secretion of cell wall material-enriched vesicles that drives the extension of the apical dome. Here we show that tip growth is not the only mode of root hair elongation. We identified events of substantial shank-localized cell wall expansion along the polar growth axis of Arabidopsis root hairs using morphometric analysis with quantum dots. These regions expanded after in vivo immunolocalization using cell wall-directed antibodies and appeared as distinct bands that were devoid of cell wall labelling. Application of a novel click chemistry-enabled galactose analogue for pulse chase and real-time imaging allowed us to label xyloglucan, a major root hair glycan, and demonstrate its de novo deposition and enzymatic remodelling in these shank regions. Our data reveal a previously unknown aspect of root hair growth in which both tip- and shank-localized dynamic cell wall deposition and remodelling contribute to root hair elongation.

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Fig. 1: Tracking Arabidopsis root hair shank elongation using q-dots.
Fig. 2: Immunolocalization of galactosylated xyloglucan (LM25) and xylan (LM10) in root hairs.
Fig. 3: Metabolic click labelling of xyloglucan in root hairs and roots.
Fig. 4: Bio-orthogonal metabolic chase labelling of growing root hair cell walls.

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All the data supporting the findings of this study are available within the paper and its supplementary information files.

References

  1. Jungk, A. Root hairs and the acquisition of plant nutrients from soil. J. Plant Nutr. Soil Sci. 164, 121–129 (2001).

    Article  CAS  Google Scholar 

  2. Carol, R. J. & Dolan, L. Building a hair: tip growth in Arabidopsis thaliana root hairs. Philos. Trans. R. Soc. Lond. B 357, 815–821 (2002).

    Article  CAS  Google Scholar 

  3. Cormack, R. G. H. Investigations on the development of root hairs. N. Phytol. 34, 30–54 (1935).

    Article  Google Scholar 

  4. Hirano, T. et al. PtdIns(3, 5)P2 mediates root hair shank hardening in Arabidopsis. Nat. Plants 4, 888–897 (2018).

    Article  CAS  PubMed  Google Scholar 

  5. Jones, M. A. et al. The Arabidopsis Rop2 GTPase is a positive regulator of both root hair initiation and tip growth. Plant Cell 14, 763–776 (2002).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Rounds, C. M. & Bezanilla, M. Growth mechanisms in tip-growing plant cells. Annu. Rev. Plant Biol. 64, 243–265 (2013).

    Article  CAS  PubMed  Google Scholar 

  7. Denninger, P. et al. Distinct RopGEFs successively drive polarization and outgrowth of root hairs. Curr. Biol. 29, 1854–1865. e1855 (2019).

    Article  CAS  PubMed  Google Scholar 

  8. Galway, M., Heckman, J. & Schiefelbein, J. Growth and ultrastructure of Arabidopsis root hairs: the rhd3 mutation alters vacuole enlargement and tip growth. Planta 201, 209–218 (1997).

    Article  CAS  PubMed  Google Scholar 

  9. Shaw, S. L., Dumais, J. & Long, S. R. Cell surface expansion in polarly growing root hairs of Medicago truncatula. Plant Physiol. 124, 959–970 (2000).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Dumais, J., Long, S. R. & Shaw, S. L. The mechanics of surface expansion anisotropy in Medicago truncatula root hairs. Plant Physiol. 136, 3266–3275 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. Mravec, J. et al. An oligogalacturonide‐derived molecular probe demonstrates the dynamics of calcium‐mediated pectin complexation in cell walls of tip‐growing structures. Plant J. 91, 534–546 (2017).

    Article  CAS  PubMed  Google Scholar 

  12. Galloway, A. F. et al. Xyloglucan is released by plants and promotes soil particle aggregation. N. Phytol. 217, 1128–1136 (2018).

    Article  CAS  Google Scholar 

  13. Galloway, A. F. et al. A soil-binding polysaccharide complex released from root hairs functions in rhizosheath formation. Preprint at bioRxiv https://doi.org/10.1101/2021.04.15.440065 (2021).

  14. Minnaar, C. & Anderson, B. Using quantum dots as pollen labels to track the fates of individual pollen grains. Methods Ecol. Evol. 10, 604–614 (2019).

    Article  Google Scholar 

  15. Schoenaers, S. et al. The auxin-regulated CrRLK1L kinase ERULUS controls cell wall composition during root hair tip growth. Curr. Biol. 28, 722–732. e726 (2018).

    Article  CAS  PubMed  Google Scholar 

  16. Dünser, K. et al. Extracellular matrix sensing by FERONIA and Leucine‐Rich Repeat Extensins controls vacuolar expansion during cellular elongation in Arabidopsis thaliana. EMBO J. 38, e100353 (2019).

    Article  PubMed  PubMed Central  Google Scholar 

  17. DeVree, B. T. et al. Current and future advances in fluorescence-based visualization of plant cell wall components and cell wall biosynthetic machineries. Biotechnol. Biofuels 14, 1–26 (2021).

    Article  Google Scholar 

  18. Wang, H., Liu, Y., Xu, M. & Cheng, J. Azido-galactose outperforms azido-mannose for metabolic labeling and targeting of hepatocellular carcinoma. Biomater. Sci. 7, 4166–4173 (2019).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Daughtry, J. L., Cao, W., Ye, J. & Baskin, J. M. Clickable galactose analogues for imaging glycans in developing zebrafish. ACS Chem. Biol. 15, 318–324 (2020).

    Article  CAS  PubMed  Google Scholar 

  20. Cavalier, D. M. et al. Disrupting two Arabidopsis thaliana xylosyltransferase genes results in plants deficient in xyloglucan, a major primary cell wall component. Plant Cell 20, 1519–1537 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Dumont, M. et al. Inhibition of fucosylation of cell wall components by 2‐fluoro 2‐deoxy‐l‐fucose induces defects in root cell elongation. Plant J. 84, 1137–1151 (2015).

    Article  CAS  PubMed  Google Scholar 

  22. Albersheim, P., Darvill, A., Roberts, K., Sederoff, R. & Staehelin, A. Plant Cell Walls (Garland Science, 2010).

  23. Anderson, C. T., Wallace, I. S. & Somerville, C. R. Metabolic click-labeling with a fucose analog reveals pectin delivery, architecture, and dynamics in Arabidopsis cell walls. Proc. Natl Acad. Sci. USA 109, 1329–1334 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Herburger, K. et al. Hetero-trans-β-glucanase produces cellulose–xyloglucan covalent bonds in the cell walls of structural plant tissues and is stimulated by expansin. Mol. Plant 13, 1047–1062 (2020).

    Article  CAS  PubMed  Google Scholar 

  25. Vissenberg, K., Fry, S. C., Pauly, M., Höfte, H. & Verbelen, J. P. XTH acts at the microfibril–matrix interface during cell elongation. J. Exp. Bot. 56, 673–683 (2005).

    Article  CAS  PubMed  Google Scholar 

  26. Popper, Z. A. & Fry, S. C. Xyloglucan−pectin linkages are formed intra-protoplasmically, contribute to wall-assembly, and remain stable in the cell wall. Planta 227, 781–794 (2008).

    Article  CAS  PubMed  Google Scholar 

  27. Zabotina, O. A. et al. Mutations in multiple XXT genes of Arabidopsis reveal the complexity of xyloglucan biosynthesis. Plant Physiol. 159, 1367–1384 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Vissenberg, K., Fry, S. C. & Verbelen, J. P. Root hair initiation is coupled to a highly localized increase of xyloglucan endotransglycosylase action in Arabidopsis roots. Plant Physiol. 127, 1125–1135 (2001).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Franková, L. & Fry, S. C. Biochemistry and physiological roles of enzymes that ‘cut and paste’ plant cell-wall polysaccharides. J. Exp. Bot. 64, 3519–3550 (2013).

    Article  PubMed  Google Scholar 

  30. Maris, A., Suslov, D., Fry, S. C., Verbelen, J.-P. & Vissenberg, K. Enzymic characterization of two recombinant xyloglucan endotransglucosylase/hydrolase (XTH) proteins of Arabidopsis and their effect on root growth and cell wall extension. J. Exp. Bot. 60, 3959–3972 (2009).

    Article  CAS  PubMed  Google Scholar 

  31. Herburger, K. et al. Defining natural factors that stimulate and inhibit cellulose: xyloglucan hetero‐transglucosylation. Plant J. 105, 1549–1565 (2021).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  32. Yokoyama, R. et al. Biological implications of the occurrence of 32 members of the XTH (xyloglucan endotransglucosylase/hydrolase) family of proteins in the bryophyte Physcomitrella patens. Plant J. 64, 645–656 (2010).

    Article  CAS  PubMed  Google Scholar 

  33. de Azevedo Souza, C. et al. Cellulose-derived oligomers act as damage-associated molecular patterns and trigger defense-like responses. Plant Physiol. 173, 2383–2398 (2017).

    Article  Google Scholar 

  34. Johnson, J. M. et al. A poly (A) ribonuclease controls the cellotriose-based interaction between Piriformospora indica and its host Arabidopsis. Plant Physiol. 176, 2496–2514 (2018).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Leitner, D. et al. A dynamic model of nutrient uptake by root hairs. N. Phytol. 185, 792–802 (2010).

    Article  CAS  Google Scholar 

  36. Geldner, N. et al. Rapid, combinatorial analysis of membrane compartments in intact plants with a multicolor marker set. Plant J. 59, 169–178 (2009).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Grossmann, G. et al. The RootChip: an integrated microfluidic chip for plant science. Plant Cell 23, 4234–4240 (2011).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Guichard, M., Bertran Garcia de Olalla, E., Stanley, C. E. & Grossmann, G. Microfluidic systems for plant root imaging. Methods Cell. Biol. 160, 381–404 (2020).

    Article  CAS  PubMed  Google Scholar 

  39. Schindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).

    Article  CAS  PubMed  Google Scholar 

  40. Parslow, A., Cardona, A. & Bryson-Richardson, R. J. Sample drift correction following 4D confocal time-lapse imaging. J. Vis. Exp. 86, 51086 (2014).

    Google Scholar 

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

    Article  CAS  PubMed  Google Scholar 

  42. Pedersen, H. L. et al. Versatile high resolution oligosaccharide microarrays for plant glycobiology and cell wall research. J. Biol. Chem. 287, 39429–39438 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Herburger, K. & Holzinger, A. Localization and quantification of callose in the streptophyte green algae Zygnema and Klebsormidium: correlation with desiccation tolerance. Plant Cell Physiol. 56, 2259–2270 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  44. Pauly, M. et al. A xyloglucan-specific endo-β-1, 4-glucanase from Aspergillus aculeatus: expression cloning in yeast, purification and characterization of the recombinant enzyme. Glycobiology 9, 93–100 (1999).

    Article  CAS  PubMed  Google Scholar 

  45. Franková, L. & Fry, S.C. Activity and action of cell-wall transglycanases. Methods Mol. Biol. 2149, 165–192 (2020).

    Article  PubMed  Google Scholar 

  46. Grierson, C. & Schiefelbein, J. Root hairs. Arabidopsis Book 1, e0060 (2002).

    Article  PubMed  PubMed Central  Google Scholar 

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Acknowledgements

This work was supported by the Villum Foundation grants (00023089; ‘TIPorNOT’) to K.H. and (00017489; ‘Instant Architect’) to J.M. S.S. acknowledges the financial support of the Research Foundation Flanders (FWO post-doc fellowship), and K.V. was supported by a University Antwerpen Grant (BOF-DOCPRO4). S. C. Fry (University of Edinburgh, UK) is kindly acknowledged for supplying [3H]XXXGol. Imaging data were in part acquired at the Center for Advanced Bioimaging (CAB) Denmark, University of Copenhagen.

Author information

Author notes

  1. Klaus Herburger

    Present address: Institute of Biological Sciences, University of Rostock, Rostock, Germany

Authors and Affiliations

  1. Section for Plant Glycobiology, Department of Plant and Environmental Sciences, University of Copenhagen, Frederiksberg, Denmark

    Klaus Herburger & Jozef Mravec

  2. Integrated Molecular Plant Physiology Research, Biology Department, University of Antwerp, Antwerp, Belgium

    Sébastjen Schoenaers & Kris Vissenberg

  3. Institut Jean-Pierre Bourgin, INRAE, AgroParisTech, Université Paris-Saclay, Versailles, France

    Sébastjen Schoenaers

  4. Plant Biochemistry and Biotechnology Lab, Department of Agriculture, Hellenic Mediterranean University, Heraklion, Greece

    Kris Vissenberg

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Contributions

J.M. and K.H. conceived the project. K.H. and J.M. designed and planned experiments. K.H., J.M. performed experiments, collected and analysed data. S.S. and K.V. provided expert help in using microfluidic imaging devices, reagents and access to imaging infrastructure at the University of Antwerp, Belgium. K.H. and J.M. wrote the manuscript with input from S.S. and K.V. All authors read, edited and approved the manuscript.

Corresponding authors

Correspondence to Klaus Herburger or Jozef Mravec.

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Nature Plants thanks Masa Sato and the other, anonymous, reviewer(s) for their contribution to the peer review of this work.

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Supplementary information

Supplementary Information

Supplementary Figs. 1–10, legends for Supplementary Movies 1–8.

Reporting Summary

Supplementary Video 1

Time lapse (50 min) of root hair labelled with q-dots (1.5 µg ml−1). Confocal fluorescence images (Z-projections; red) were merged with corresponding images collected in a separate channel. Q-dots sprinkled to the root hair shank do not move, suggesting no shank expansion. Scale bar, 10 µm.

Supplementary Video 2

Time lapse (78 min) of root hair labelled with q-dots (1.5 µg ml−1). Confocal fluorescence (red) and corresponding bright-field images are shown. The nucleus migrating into the growing root hair is highlighted with an asterisk. Root hair shank-located q-dots move towards the polar growth direction, tracking shank expansion. Corresponds to data shown in Fig. 1b. Scale bar, 10 µm.

Supplementary Video 3

Time lapse (49 min) of root hair labelled with q-dots (1.5 µg ml−1). Arrows follow two q-dots that move towards the growth direction. Pale arrows highlight the original locations of the moving q-dots. The left movie shows a magnification of the marked area in the left movie. Corresponds to data shown in Fig. 1c. Scale bar, 10 µm.

Supplementary Video 4

Time lapse (30 min) of root hair with Golgi apparatus marker (Wave18, Got1P homologue). Scale bar, 10 µm.

Supplementary Video 5

Time lapse (60 min) of root hair with ER marker (Wave7Y, YFP-NIP1;1). Scale bar, 10 µm.

Supplementary Video 6

Time lapse (78 min) of root hair ‘click labelled’ with aGal-DBCO-Cy5 (for concentrations of aGal and dye, see Fig. 4b). Confocal fluorescence images (red) are shown. Note emerging unlabelled bands in two different root hairs. Corresponds to data shown in Fig. 4b. Scale bar, 10 µm.

Supplementary Video 7

Time lapse (140 min) of root hairs from xyloglucan-deficient xxt1/2/5 mutant plant labelled with q-dots (1.5 µg ml−1). Scale bar, 10 µm.

Supplementary Video 8

Time lapse (40 min) of root hair growing in presence of 30 mM C4. Corresponds to data in Fig. 4d. Scale bar, 10 µm.

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Herburger, K., Schoenaers, S., Vissenberg, K. et al. Shank-localized cell wall growth contributes to Arabidopsis root hair elongation. Nat. Plants 8, 1222–1232 (2022). https://doi.org/10.1038/s41477-022-01259-y

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