Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Mechanical interactions between tissue layers underlie plant morphogenesis

Nature Plants volume 11, pages 909–923 (2025)Cite this article

Subjects

Abstract

Differential growth between tissues generates mechanical conflicts influencing organogenesis in plants. Here we use the anther, the male floral reproductive organ, as a model system to understand how cell dynamics and tissue-scale mechanics control 3D morphogenesis of a complex shape. Combining deep live-cell imaging, growth analysis, osmotic treatments, genetics and mechanical modelling, we show that localized expansion of internal cells actively drives anther lobe outgrowth, while the epidermis stretches in response. At later stages, mechanical load is transferred to the sub-epidermal layer (endothecium), contributing to proper organ shape. We propose the concept of ‘inflation potential’, encapsulating mechanical and anatomical features causing differential growth. Our data emphasize the active mechanical role of inner tissue in controlling both organ shape acquisition and cell dynamics in outer layers.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on SpringerLink
  • Instant access to the full article PDF.

USD 39.95

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Localized growth in inner tissue underlies lobe formation.
Fig. 2: Gradients of mechanical and anatomical properties between layers can explain faster growth in lobe inner tissue.
Fig. 3: The epidermis in the anther lobe is under local tension.
Fig. 4: Emergence of endothecium reduces tension in the epidermis.

Similar content being viewed by others

Data availability

The data used to quantify growth parameters, models and starting templates with a copy of the MDX software are available to download from the Open Science Framework repository at https://osf.io/h8r76/?view_only=46b882efd7ec4c55ab798f7f992504ab (ref. 72).

Code availability

The Python scripts used to analyse data are available to download from the Open Science Framework repository at https://osf.io/h8r76/?view_only=46b882efd7ec4c55ab798f7f992504ab (ref. 72).

References

  1. Jorgensen, P. & Tyers, M. How cells coordinate growth and division. Curr. Biol. 14, R1014–R1027 (2004).

    CAS  PubMed  Google Scholar 

  2. Vaughan, J. G. The morphology and growth of the vegetative and reproductive apices of Arabidopsis thaliana (L.) Heynh., Capsella bursa-pastoris (L.) Medic. and Anagallis arvensis L. J. Linn. Soc. Lond. Bot. 55, 279–301 (1955).

    Google Scholar 

  3. Dubrovsky, J. G., Doerner, P. W., Colón-Carmona, A. & Rost, T. L. Pericycle cell proliferation and lateral root initiation in Arabidopsis. Plant Physiol. 124, 1648–1657 (2000).

    CAS  PubMed  PubMed Central  Google Scholar 

  4. Rebocho, A. B., Southam, P., Kennaway, J. R., Bangham, J. A. & Coen, E. Generation of shape complexity through tissue conflict resolution. eLife 6, e20156 (2017).

    PubMed  PubMed Central  Google Scholar 

  5. Whitewoods, C. D. et al. Evolution of carnivorous traps from planar leaves through simple shifts in gene expression. Science 367, 91–96 (2020).

    CAS  PubMed  Google Scholar 

  6. Jonsson, K. et al. Mechanochemical feedback mediates tissue bending required for seedling emergence. Curr. Biol. 31, 1154–1164 (2021).

    CAS  PubMed  Google Scholar 

  7. Kelly-Bellow, R. et al. Brassinosteroid coordinates cell layer interactions in plants via cell wall and tissue mechanics. Science 380, 1275–1281 (2023).

    CAS  PubMed  Google Scholar 

  8. Green, P. Phyllotactic patterns: a biophysical mechanism for their origin. Ann. Bot. 77, 515–528 (1996).

    Google Scholar 

  9. Kutschera, U. & Niklas, K. J. The epidermal-growth-control theory of stem elongation: an old and a new perspective. J. Plant Physiol. 164, 1395–1409 (2007).

    CAS  PubMed  Google Scholar 

  10. Kierzkowski, D. et al. Elastic domains regulate growth and organogenesis in the plant shoot apical meristem. Science 335, 1096–1099 (2012).

    CAS  PubMed  Google Scholar 

  11. Cosgrove, D. J. Plant cell wall extensibility: connecting plant cell growth with cell wall structure, mechanics, and the action of wall-modifying enzymes. J. Exp. Bot. 67, 463–476 (2016).

    CAS  PubMed  Google Scholar 

  12. Beauzamy, L., Louveaux, M., Hamant, O. & Boudaoud, A. Mechanically, the shoot apical meristem of Arabidopsis behaves like a shell inflated by a pressure of about 1 MPa. Front. Plant Sci. 6, 1038 (2015).

    PubMed  PubMed Central  Google Scholar 

  13. Selker, J. M. L., Steucek, G. L. & Green, P. B. Biophysical mechanisms for morphogenetic progressions at the shoot apex. Dev. Biol. 153, 29–43 (1992).

    CAS  PubMed  Google Scholar 

  14. Peaucelle, A. et al. Pectin-induced changes in cell wall mechanics underlie organ initiation in Arabidopsis. Curr. Biol. 21, 1720–1726 (2011).

    CAS  PubMed  Google Scholar 

  15. Silveira, S. R., Le Gloanec, C., Gómez-Felipe, A., Routier-Kierzkowska, A.-L. & Kierzkowski, D. Live-imaging provides an atlas of cellular growth dynamics in the stamen. Plant Physiol. 188, 769–781 (2022).

    CAS  PubMed  Google Scholar 

  16. Sanders, P. M. et al. Anther developmental defects in Arabidopsis thaliana male-sterile mutants. Sex. Plant Reprod. 11, 297–322 (1999).

    CAS  Google Scholar 

  17. Ma, H. Molecular genetic analyses of microsporogenesis and microgametogenesis in flowering plants. Annu. Rev. Plant Biol. 56, 393–434 (2005).

    CAS  PubMed  Google Scholar 

  18. Green, P. G. Mechanism for plant cellular morphogenesis. Science 138, 1404–1405 (1962).

    CAS  PubMed  Google Scholar 

  19. Cosgrove, D. J. Wall extensibility: its nature, measurement and relationship to plant cell growth. New Phytol. 124, 1–23 (1993).

    CAS  PubMed  Google Scholar 

  20. Niklas, K. J. Plant Biomechanics: An Engineering Approach to Plant Form and Function (Univ. Chicago Press, 1992).

  21. Bassel, G. W. et al. Mechanical constraints imposed by 3D cellular geometry and arrangement modulate growth patterns in the Arabidopsis embryo. Proc. Natl Acad. Sci. USA 111, 8685–8690 (2014).

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Wang, C. X., Wang, L. & Thomas, C. R. Modelling the mechanical properties of single suspension-cultured tomato cells. Ann. Bot. 93, 443–453 (2004).

    CAS  PubMed  PubMed Central  Google Scholar 

  23. Weber, A. et al. Measuring the mechanical properties of plant cells by combining micro-indentation with osmotic treatments. J. Exp. Bot. 66, 3229–3241 (2015).

    CAS  PubMed  PubMed Central  Google Scholar 

  24. Yu, J., Zhang, Y. & Cosgrove, D. J. The nonlinear mechanics of highly extensible plant epidermal cell walls. Proc. Natl Acad. Sci. USA 121, e231639612 (2024).

    Google Scholar 

  25. Flügge, W. Stresses in Shells 2nd edn (Springer, 1973).

  26. Venkova, L. et al. A mechano-osmotic feedback couples cell volume to the rate of cell deformation. eLife 11, e72381 (2022).

    CAS  PubMed  PubMed Central  Google Scholar 

  27. Bonazzi, D. et al. Symmetry breaking in spore germination relies on an interplay between polar cap stability and spore wall mechanics. Dev. Cell 28, 534–546 (2014).

    CAS  PubMed  Google Scholar 

  28. Misra, G., Rojas, E. R., Gopinathan, A. & Huang, K. C. Mechanical consequences of cell-wall turnover in the elongation of a Gram-positive bacterium. Biophys. J. 104, 2342–2352 (2013).

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Hong, L. et al. Variable cell growth yields reproducible organ development through spatiotemporal averaging. Dev. Cell 38, 15–32 (2016).

    CAS  PubMed  Google Scholar 

  30. Mosca, G., Sapala, A., Strauss, S., Routier-Kierzkowska, A.-L. & Smith, R. S. On the micro-indentation of plant cells in a tissue context. Phys. Biol. 14, 015003 (2017).

    PubMed  Google Scholar 

  31. Hofhuis, H. et al. Morphomechanical innovation drives explosive seed dispersal. Cell 166, 222–233 (2016).

    CAS  PubMed  PubMed Central  Google Scholar 

  32. Hoermayer, L. et al. Mechanical forces in plant tissue matrix orient cell divisions via microtubule stabilization. Dev. Cell 59, 1333–1344.e4 (2024).

    CAS  PubMed  Google Scholar 

  33. Yang, W. C., Ye, D., Xu, J. & Sundaresan, V. The SPOROCYTELESS gene of Arabidopsis is required for initiation of sporogenesis and encodes a novel nuclear protein. Genes Dev. 13, 2108–2117 (1999).

    CAS  PubMed  PubMed Central  Google Scholar 

  34. Verger, S., Long, Y., Boudaoud, A. & Hamant, O. A tension–adhesion feedback loop in plant epidermis. eLife 7, e34460 (2018).

    PubMed  PubMed Central  Google Scholar 

  35. Peters, W. S. & Tomos, A. D. The history of tissue tension. Ann Bot. 77, 657–665 (1996).

    CAS  PubMed  Google Scholar 

  36. Coen, E. & Cosgrove, D. J. The mechanics of plant morphogenesis. Science 379, eade8055 (2023).

    CAS  PubMed  Google Scholar 

  37. Kutschera, U. Determination of the longitudinal tissue stresses in the growing and non-growing regions of sunflower hypocotyls. J. Plant Physiol. 138, 460–465 (1991).

    Google Scholar 

  38. Hejnowicz, Z. & Sievers, A. Tissue stresses in organs of herbaceous plants: I. Poisson ratios of tissues and their role in determination of the stresses. J. Exp. Bot. 46, 1035–1043 (1995).

    CAS  Google Scholar 

  39. Bozorg, B., Krupinski, P. & Jönsson, H. Stress and strain provide positional and directional cues in development. PLoS Comput. Biol. 10, e1003410 (2014).

    PubMed  PubMed Central  Google Scholar 

  40. Ali, O., Oliveri, H., Traas, J. & Godin, C. Simulating turgor-induced stress patterns in multilayered plant tissues. Bull. Math. Biol. 81, 3362–3384 (2019).

    PubMed  Google Scholar 

  41. Dumais, J. & Steele, C. R. New evidence for the role of mechanical forces in the shoot apical meristem. J. Plant Growth Regul. 19, 7–18 (2000).

    CAS  PubMed  Google Scholar 

  42. Hamant, O. et al. Developmental patterning by mechanical signals in Arabidopsis. Science 322, 1650–1655 (2008).

    CAS  PubMed  Google Scholar 

  43. Hamant, O. & Moulia, B. How do plants read their own shapes? New Phytol. 212, 333–337 (2016).

    CAS  PubMed  Google Scholar 

  44. Creff, A. et al. Evidence that endosperm turgor pressure both promotes and restricts seed growth and size. Nat. Commun. 14, 67 (2023).

    CAS  PubMed  PubMed Central  Google Scholar 

  45. Cosgrove, D. J. Structure and growth of plant cell walls. Nat. Rev. Mol. Cell Biol. 25, 340–358 (2024).

    CAS  PubMed  Google Scholar 

  46. Hord, C. L. H., Chen, C., DeYoung, B. J., Clark, S. E. & Ma, H. The BAM1/BAM2 receptor-like kinases are important regulators of Arabidopsis early anther development. Plant Cell 18, 1667–1680 (2006).

    CAS  PubMed  PubMed Central  Google Scholar 

  47. Fridman, Y. et al. The root meristem is shaped by brassinosteroid control of cell geometry. Nat. Plants 7, 1475–1484 (2021).

    CAS  PubMed  PubMed Central  Google Scholar 

  48. Zheng, Y. et al. Auxin guides germ-cell specification in Arabidopsis anthers. Proc. Natl Acad. Sci. USA 118, e2101492118 (2021).

    CAS  PubMed  PubMed Central  Google Scholar 

  49. Boudaoud, A. An introduction to the mechanics of morphogenesis for plant biologists. Trends Plant Sci. 15, 353–360 (2010).

    CAS  PubMed  Google Scholar 

  50. Oliveri, H., Traas, J., Godin, C. & Ali, O. Regulation of plant cell wall stiffness by mechanical stress: a mesoscale physical model. J. Math. Biol. 78, 625–653 (2019).

    PubMed  Google Scholar 

  51. Collinet, C. & Lecuit, T. Programmed and self-organized flow of information during morphogenesis. Nat. Rev. Mol. Cell Biol. 22, 245–265 (2021).

    CAS  PubMed  Google Scholar 

  52. Zhao, F. et al. Microtubule-mediated wall anisotropy contributes to leaf blade flattening. Curr. Biol. 30, 3972–3985 (2020).

    CAS  PubMed  PubMed Central  Google Scholar 

  53. Bauer, A. et al. Spatiotemporally distinct responses to mechanical forces shape the developing seed of Arabidopsis. EMBO J. 43, 1–26 (2024).

    Google Scholar 

  54. Villeneuve, C. et al. Mechanical forces across compartments coordinate cell shape and fate transitions to generate tissue architecture. Nat. Cell Biol. 26, 207–218 (2024).

    CAS  PubMed  PubMed Central  Google Scholar 

  55. Dumortier, J. G. et al. Hydraulic fracturing and active coarsening position the lumen of the mouse blastocyst. Science 365, 465–468 (2019).

    CAS  PubMed  Google Scholar 

  56. Wang, H. et al. Molecular regulation of oil gland development and biosynthesis of essential oils in Citrus spp. Science 383, 659–666 (2024).

    CAS  PubMed  Google Scholar 

  57. McCartney, B. & Dudin, O. Cellularization across eukaryotes: conserved mechanisms and novel strategies. Curr. Opin. Cell Biol. 80, 102157 (2023).

    CAS  PubMed  Google Scholar 

  58. Yang, W. et al. Regulation of meristem morphogenesis by cell wall synthases in Arabidopsis. Curr. Biol. 26, 1404–1415 (2016).

    CAS  PubMed  PubMed Central  Google Scholar 

  59. Murashige, T. & Skoog, F. A revised medium for rapid growth and bio assays with tobacco tissue cultures. Physiol. Plant. 15, 473–497 (1962).

  60. Wang, B., Bauer, A., Gómez-Felipe, A., Silveira, S. R. & Kierzkowski, D. Confocal live imaging of reproductive organs development in Arabidopsis. Bio Protoc. 15, e5177 (2025).

    Google Scholar 

  61. Wojciechowska, N. et al. Autophagy counteracts instantaneous cell death during seasonal senescence of the fine roots and leaves in Populus trichocarpa. BMC Plant Biol. 18, 260 (2018).

    CAS  PubMed  PubMed Central  Google Scholar 

  62. Barbier de Reuille, P. et al. MorphoGraphX: a platform for quantifying morphogenesis in 4D. eLife 6, 05864 (2015).

    Google Scholar 

  63. Strauss, S. et al. Using positional information to provide context for biological image analysis with MorphoGraphX 2.0. eLife 11, e72601 (2022).

    CAS  PubMed  PubMed Central  Google Scholar 

  64. Schindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 28, 676–682 (2012).

    Google Scholar 

  65. Mann, H. B. & Whitney, D. R. On a test of whether one of two random variables is stochastically larger than the other. Ann. Math. Stat. 18, 50–60 (1947).

    Google Scholar 

  66. Wagner, C. H. Simpson's paradox in real life. Am. Stat. 36, 46–48 (1982).

    Google Scholar 

  67. Sapala, A. et al. Why plants make puzzle cells, and how their shape emerges. eLife 27, e32794 (2018).

    Google Scholar 

  68. Natonik-Białoń, S. et al. Deformation of a cell monolayer due to osmotic treatment: a case study of onion scale epidermis. Botany 98, 21–36 (2020).

    Google Scholar 

  69. Geuzaine, C. & Remacle, J. F. Gmsh: a three-dimensional finite element mesh generator with built-in pre-and post-processing facilities. Int. J. Numer. Meth. Eng. 79, 1309–1331 (2009).

    Google Scholar 

  70. Mosca, G. et al. in Mathematical Modelling in Plant Biology (Morris, R. J.) 107–138 (Springer, 2018).

  71. Majda, M. et al. Elongation of wood fibers combines features of diffuse and tip growth. New Phytol. 232, 673–691 (2021).

    CAS  PubMed  Google Scholar 

  72. Kierzkowski, D. & Silveira, S. R. Mechanical interactions between tissue layers. OSF https://osf.io/h8r76/?view_only=46b882efd7ec4c55ab798f7f992504ab (2024).

Download references

Acknowledgements

We thank N. Boodhoo and T. Stan for help with image segmentations; S. Strauss for help with MorphoGraphX 2.0; G. Mosca and B. Lane for help with MorphoDynamX; K. Jonsson for seeds of qua2-1 mutant crossed with membrane marker line; and K. Jonsson and A. Bauer for critical reading of the paper. This work was supported by a New Frontiers in Research Fund Exploration grant (no. NFRFE-2018-00953) from the Government of Canada (D.K., A.-L.R.-K. and F.P.G.), Natural Sciences and Engineering Research Council of Canada Discovery grant RGPIN-2018-05762 (A.-L.R.-K.), Natural Sciences and Engineering Research Council of Canada Discovery grant RGPIN-2018-04897 (D.K.), Natural Sciences and Engineering Research Council of Canada Discovery grant RGPIN-2019-07072 (F.P.G.), Human Frontier Science Program research grant RGY0077/2021 (A.-L.R.-K.), and a Biotechnological and Biological Sciences Research Council Institute Strategic Programme Grant to the John Innes Centre BB/X01102X/1 (R.S.S.). This research was also supported in part by grant NSF PHY-2309135 to the Kavli Institute for Theoretical Physics (KITP) (D.K. and A.-L.R.-K.), and Centre SÈVE - strategic regroupement of the Fonds de Recherche du Québec Nature et Technologies (D.K. and A.-L.R.-K.). Confocal imaging was conducted using the instrument supported by Canada Foundation for Innovation grants (37805) (D.K.).

Author information

Author notes

  1. These authors contributed equally: Sylvia R. Silveira, Loann Collet.

Authors and Affiliations

  1. Institut de Recherche en Biologie Végétale, Département de Sciences Biologiques, Université de Montréal, Montréal, Quebec, Canada

    Sylvia R. Silveira, Loann Collet, Sahil M. Haque, Luc Lapierre, Anne-Lise Routier-Kierzkowska & Daniel Kierzkowski

  2. Department of General Botany, Institute of Experimental Biology, Faculty of Biology, Adam Mickiewicz University, Poznań, Poland

    Agnieszka Bagniewska-Zadworna

  3. Department of Computational and Systems Biology, John Innes Centre, Norwich, UK

    Richard S. Smith

  4. Laboratory of Multiscale Mechanics (LM2), Department of Mechanical Engineering, Polytechnique Montréal, Montréal, Quebec, Canada

    Frederick P. Gosselin

Authors
  1. Sylvia R. Silveira
    View author publications

    Search author on:PubMed Google Scholar

  2. Loann Collet
    View author publications

    Search author on:PubMed Google Scholar

  3. Sahil M. Haque
    View author publications

    Search author on:PubMed Google Scholar

  4. Luc Lapierre
    View author publications

    Search author on:PubMed Google Scholar

  5. Agnieszka Bagniewska-Zadworna
    View author publications

    Search author on:PubMed Google Scholar

  6. Richard S. Smith
    View author publications

    Search author on:PubMed Google Scholar

  7. Frederick P. Gosselin
    View author publications

    Search author on:PubMed Google Scholar

  8. Anne-Lise Routier-Kierzkowska
    View author publications

    Search author on:PubMed Google Scholar

  9. Daniel Kierzkowski
    View author publications

    Search author on:PubMed Google Scholar

Contributions

S.R.S., L.C., A.-L.R.-K. and D.K. conceived and designed the experiments. S.R.S., L.C., S.M.H., A.-L.R.-K. and D.K. developed methods. L.C., F.P.G., R.S.S. and A.-L.R.-K. conceived and designed the computational models. S.R.S., A.B.-Z., L.C. and D.K. performed experiments and extracted data with help from L.L. and S.M.H.; S.R.S., L.C., A.-L.R.-K. and D.K. analysed the data. A.-L.R.-K. and D.K. supervised the project. S.R.S., L.C., A.-L.R.-K. and D.K. wrote the manuscript with input from all other authors. A.-L.R.-K., D.K., F.P.G. and R.S.S. provided funding. All authors reviewed the manuscript.

Corresponding authors

Correspondence to Anne-Lise Routier-Kierzkowska or Daniel Kierzkowski.

Ethics declarations

Competing interests

The authors declare no competing interests.

Peer review

Peer review information

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

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Extended data

Extended Data Fig. 1 Localized growth in inner tissue underlies lobe formation.

a, Quantification of cell area increase per anther region (n represent cell number for consecutive measurement for each sample from top to bottom: 5, 17, 9, 25, 15, 42, 21, 62; 8, 15, 11, 17, 17, 27, 24, 22; 12, 14, 23, 28, 43, 62; and 14, 11, 17, 17, 25, 20). b, Quantification of cell volume increase per anther region (n represent cell number for consecutive measurement for each sample from top to bottom: 5, 4, 14, 19, 10, 8, 27, 29, 14, 25, 42, 34, 21, 48, 62, 34; 8, 6, 14, 12, 11, 13, 17, 17, 16, 26, 28, 25, 24, 43, 22, 5; 14, 13, 24, 31, 28, 53, 45, 75, 62, 124, 66, 80; and 14, 10, 13, 23, 17, 26, 16, 46, 25, 41, 19, 38). Each of the four time-lapse series that were normalized and pooled in the boxplots shown in Fig. 1. In box plots, the lines represent the median and dots represent the mean values, the edges of the boxes define interquartile ranges, the whiskers represent 90% confidence intervals. For each time-point different letters indicate statistical significance determined by two-sided Mann Whitney U test (P < 0.05).

Extended Data Fig. 2 Cube of cube model details.

a to c, Cross-section of the model template of the cube of cubes. a, Outer layer of the tissue with constant properties. It is of total size Lout = Uoutlout where Uout is the number of cells along the side of the cubic tissue and lout the side of those cells. b, Inner tissue of total size Lin = (Uout-2)lout. It is made of UinxUinxUin cells of size lin = Lin/Uin. c, Assembled outer layer and inner tissue. d, Detailed view of Fig. 2b. Color scale: Trace of the Green-Lagrangian strain tensor Tr(ε) per element. (e and f) For each parameter Et, s, and P, areal strains (in %) of anticlinal walls within inner e and outer f cells are plotted against the ratio of inflation potential in inner vs outer cells.

Extended Data Fig. 3 Measurements of cell wall thickness and Incipient plasmolysis analysis in ~4 days old anthers.

a, Representative transmission electron microscopy micrograph of an anther lobe at a stage before lobation. Brackets indicate the layers considered for measurements as shown in Fig. 2e, epidermis (magenta) and inner tissue (green). Insets show segmented lines used to quantify cell wall thickness shown in Fig. 2f. b, Confocal images of anthers, before and after osmotic treatment with solutions of increasing NaCl concentration. The top row shows epidermis while the bottom row shows digital cross sections in the inner tissue. Arrow heads indicate the interstitial spaces created at incipient plasmolysis. Scale bars, 2.5 μm (a), 10 μm (b).

Extended Data Fig. 4 Obtaining the starting mesh for stiffness reverse-engineering.

a, 2D meshes extracted from the top-to-bottom and bottom-to-top confocal microscopy scans of the same plasmolyzed sample Cell junctions are highlighted in red. We compute the transformation from one 3D point cloud to the other, M b, Starting mesh and transformed mesh obtained by applying half the transformation M1/2 to the former. The heatmap of volume increase of each cell from the plasmolyzed experimental state to the turgid. c, Pooling of cells into regions. Regions are based on layer, connectivity and volume increase displayed in b. Scale bars, 5 μm.

Extended Data Fig. 5 Convergence of reverse engineering and significance of results.

a, Convergence of the volume distribution of all cells of each sample over the reverse engineering steps. b, For each sample: final volume increase distributions from the plasmolyzed state to the turgid state, simulated and experimental. c, Guessing of the next stiffness of a given cell. A linear trend is drawn from the two previous states (En-2,Vn-2) and (En-1,Vn-1), and the intersection between this line and the V = Vexp horizontal line yields the new cell stiffness EGuess. d, Stiffness values after the 20th reverse engineering step per region and stiffness ratios between regions of interest. An inner/outer gradient can be observed for each sample in the locule region.

Extended Data Fig. 6 Lobe development is abolished in the absence of localized internal growth in the locule.

a, Digital cross sections of a time-lapse series of the developing spl-1 anther. b, Heat maps of volume increase in anther epidermis and inner tissue. c-d, Quantification of volume increase per anther region in the epidermis (n = 16, 23, 64, 92 cells in lobe, 26, 48, 128, 182 in connective, at consecutive time points and inner tissue (n = 10, 27, 80, 73 cells in lobe, and 42, 66, 163, 109 in the connective tissue; 4 independent time-lapse series). In box plots, the lines represent the median and dots represent the mean values, the edges of the boxes define interquartile ranges, the whiskers represent 90% confidence intervals. No statistical significance was found by two-sided Mann Whitney U test (P < 0.01). Mann-Whitney U test, P < 0.01 was detected Scale bars, 20 μm (a and b).

Extended Data Fig. 7 The epidermis in the anther lobe is under local tension.

a, Digital longitudinal cross sections of the anther lobe (top) and tip of connective (bottom) for wild type (left) and spl/nzz (right). Arrows indicate intercellular spaces. b-c, Quantification of ratio between cell growth within the organ plane and growth in cell thickness, per anther region in the epidermis in wild type (b) and spl-1 (c) (n = 13, 49, 77, 134, 175 cell in lobe epidermis, 32, 81, 132, 172, 195 cells in connective epidermis in wild type, 16, 22, 63, 92, 103 cells in lobe epidermis, 26, 47, 127, 182, 205 cells in connective epidermis of the mutant at consecutive time points; 4 independent time-lapse series). In box plots, the lines represent the median and dots represent the mean values, the edges of the boxes define interquartile ranges, the whiskers represent 90% confidence intervals. Asterisks indicate statistical significance determined by two-sided Mann Whitney U test (P < 0.01). d, Confocal images of dissected qua2-1 mutant anther. Insets show details of mid connective (left), tip of connective (top), and lobe (right). Arrows indicate points of cell detachments in the epidermis. Scale bars, 10 μm (a) 20 μm (d).

Extended Data Fig. 8 Cylindrical Voronoï construction and simulation.

a, Construction of the cylindrical epidermis. b, 3D view of the cylindrical epidermis. c, 2D simplified schematic of the spring network used to place the Voronoï diagram control points within the epidermis. d, Symmetries of the cloud point and Voronoï diagram generation. e, Final Voronoï diagram. f, 3D view of a cylindrical Voronoï model. g, Side view of the model. Continuous red line indicates immobile vertices along the z axis. Dotted red line indicates the mobile plane that will adjust the axial force the model is subjected to. dn is the current displacement of that plane. h, Estimation of the next displacement dn based on the previous steps (dn-2, Fn-2) and (dn-1, Fn-1) shown as green dots and the goal force of 0 N. The new state (dn, Fn) is represented as the red dot.

Extended Data Fig. 9 Construction of the flat twin model for the epidermis.

a, Left side: Initial states of the voronoï in ring model and of the corresponding flat twin. Right side: Final states after pressurization. b, Superpositions of the initial and final simulation states of the flat twin model in the xz and yz planes respectively. The cell walls marked in red remain flat due to boundary conditions. The same walls in the yz plane are also moved to match the stretching of the epidermis. c, Tissue circumferential tension for varying inner over outer inflation potential ratios plotted against the organ radius before pressurization. d, Ratio of the tissue circumferential tension over the tissue vertical tension, plotted against the organ radius before pressurization. e, Pressurized voronoï in ring models for different inner inflation potentials and tissue sizes. Color scales: Trace of the Green-Lagrangian strain tensor per element (a and e).

Extended Data Fig. 10 Emergence of endothecium reduces tension in the epidermis.

(a) Representative digital cross-sections of the anther lobe region in WT, bam1bam2, and spl-1 from around 3 to 13 days*. Asterisk indicates estimated days after initiation. (b) Quantification of cell area per lobe region in WT anthers (n = 35, 36, 35, 53, 31, 70, 18, 20 cells in epidermis; 10, 18, 34, 22, 50, 15, 15 cells in endothecium; 59, 41, 19, 24, 13, 26, 19, 14 cells in inner tissue at consecutive time points; 5 independent inflorescences). (c) Example of transmission electron microscopy micrograph of an anther lobe after ‘butterfly’ shape acquisition. Brackets indicate the layers considered for measurements. (d) Quantification of cell wall thickness in different layers of the lobe (n = 24 walls in epidermis, 24 walls in endothecium, and 38 walls in inner tissue; 3 independent anthers). (e) Quantification of volume increase per region in 12 h interval (n = 88, 78 cells in epidermis, 36 cells in endothecium, and 42, 54 cells in inner tissue at consecutive time points; 4 independent time lapse series). In box plots, the lines represent the median and dots represent the mean values, the edges of the boxes define interquartile ranges, the whiskers represent 90% confidence intervals. For each time-point different letters indicate statistical significance determined by two-sided Mann Whitney U test (p < 0.01). Scale bar, 40 μm (a) 2.5 μm (b).

Supplementary information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Silveira, S.R., Collet, L., Haque, S.M. et al. Mechanical interactions between tissue layers underlie plant morphogenesis. Nat. Plants 11, 909–923 (2025). https://doi.org/10.1038/s41477-025-01944-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Version of record:

  • Issue date:

  • DOI: https://doi.org/10.1038/s41477-025-01944-8

Search

Advanced search

Quick links

Nature Briefing

Sign up for the Nature Briefing newsletter — what matters in science, free to your inbox daily.

Get the most important science stories of the day, free in your inbox. Sign up for Nature Briefing