Accurate and versatile 3D segmentation of plant tissues at cellular resolution

Abstract

Quantitative analysis of plant and animal morphogenesis requires accurate segmentation of individual cells in volumetric images of growing organs. In the last years, deep learning has provided robust automated algorithms that approach human performance, with applications to bio-image analysis now starting to emerge. Here, we present PlantSeg, a pipeline for volumetric segmentation of plant tissues into cells. PlantSeg employs a convolutional neural network to predict cell boundaries and graph partitioning to segment cells based on the neural network predictions. PlantSeg was trained on 1xed and live plant organs imaged with confocal and light sheet microscopes. PlantSeg delivers accurate results and generalizes well across different tissues, scales, acquisition settings even on non plant samples. We present results of PlantSeg applications in diverse developmental contexts. PlantSeg is free and open-source, with both a command line and a user-friendly graphical interface (https://github.com/hci-unihd/plant-seg).

Data availability

All data used in this study have been deposited in Open Science Framework: https://osf.io/uzq3w/Additionally Arabidopsis 3D Digital Tissue Atlas is available under https://osf.io/fzr56/

The following data sets were generated
The following previously published data sets were used

Article and author information

Author details

  1. Adrian Wolny

    Cell Biology and Biophysics Unit, EMBL, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  2. Lorenzo Cerrone

    Heidelberg Collaboratory for Image Processing, Heidelberg University, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  3. Athul Vijayan

    School of Life Sciences Weihenstephan, Technical University of Munich, Freising, Germany
    Competing interests
    The authors declare that no competing interests exist.
  4. Rachele Tofanelli

    School of Life Sciences Weihenstephan, Technical University of Munich, Freising, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0002-5196-1122
  5. Amaya Vilches Barro

    Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  6. Marion Louveaux

    Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  7. Christian Wenzl

    Department of Stem Cell Biology, Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  8. Sören Strauss

    Department of Comparative Development and Genetics, Max Planck Institute for Plant Breeding Research, Cologne, Germany
    Competing interests
    The authors declare that no competing interests exist.
  9. David Wilson-Sánchez

    Department of Comparative Development and Genetics, Max Planck Institute for Plant Breeding Research, Cologne, Germany
    Competing interests
    The authors declare that no competing interests exist.
  10. Rena Lymbouridou

    Department of Comparative Development and Genetics, Max Planck Institute for Plant Breeding Research, Cologne, Germany
    Competing interests
    The authors declare that no competing interests exist.
  11. Susanne Steigleder

    Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  12. Constantin Pape

    Cell Biology and Biophysics Unit, EMBL, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  13. Alberto Bailoni

    Heidelberg Collaboratory for Image Processing, Heidelberg University, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  14. Salva Duran-Nebreda

    School of Life Sciences, University of Warwick, Coventry, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  15. George Bassel

    School of Life Sciences, University of Warwick, Coventry, United Kingdom
    Competing interests
    The authors declare that no competing interests exist.
  16. Jan U Lohmann

    Department of Stem Cell Biology, Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-3667-187X
  17. Miltos Tsiantis

    Department of Comparative Development and Genetics, Max Planck Institute for Plant Breeding Research, Cologne, Germany
    Competing interests
    The authors declare that no competing interests exist.
  18. Fred Hamprecht

    Department of Stem Cell Biology, Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  19. Kay Schneitz

    School of Life Sciences Weihenstephan, Technical University of Munich, Freising, Germany
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0001-6688-0539
  20. Alexis Maizel

    Department of Stem Cell Biology, Centre for Organismal Studies, Heidelberg University, Heidelberg, Germany
    Competing interests
    The authors declare that no competing interests exist.
  21. Anna Kreshuk

    Cell Biology and Biophysics Unit, EMBL, Heidelberg, Germany
    For correspondence
    anna.kreshuk@embl.de
    Competing interests
    The authors declare that no competing interests exist.
    ORCID icon "This ORCID iD identifies the author of this article:" 0000-0003-1334-6388

Funding

Deutsche Forschungsgemeinschaft (FOR2581)

  • Jan U Lohmann
  • Miltos Tsiantis
  • Fred Hamprecht
  • Kay Schneitz
  • Alexis Maizel
  • Anna Kreshuk

Leverhulme Trust (RPG-2016-049)

  • George Bassel

The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication.

Copyright

© 2020, Wolny et al.

This article is distributed under the terms of the Creative Commons Attribution License permitting unrestricted use and redistribution provided that the original author and source are credited.

Metrics

  • 12,435
    views
  • 1,466
    downloads
  • 178
    citations

Views, downloads and citations are aggregated across all versions of this paper published by eLife.

Download links

A two-part list of links to download the article, or parts of the article, in various formats.

Downloads (link to download the article as PDF)

Open citations (links to open the citations from this article in various online reference manager services)

Cite this article (links to download the citations from this article in formats compatible with various reference manager tools)

  1. Adrian Wolny
  2. Lorenzo Cerrone
  3. Athul Vijayan
  4. Rachele Tofanelli
  5. Amaya Vilches Barro
  6. Marion Louveaux
  7. Christian Wenzl
  8. Sören Strauss
  9. David Wilson-Sánchez
  10. Rena Lymbouridou
  11. Susanne Steigleder
  12. Constantin Pape
  13. Alberto Bailoni
  14. Salva Duran-Nebreda
  15. George Bassel
  16. Jan U Lohmann
  17. Miltos Tsiantis
  18. Fred Hamprecht
  19. Kay Schneitz
  20. Alexis Maizel
  21. Anna Kreshuk
(2020)
Accurate and versatile 3D segmentation of plant tissues at cellular resolution
eLife 9:e57613.
https://doi.org/10.7554/eLife.57613

Share this article

https://doi.org/10.7554/eLife.57613

Further reading

    1. Developmental Biology
    2. Plant Biology
    Shijia Lin, Yiwen Zhang ... Zhaoliang Zhang
    Research Article

    Root-synthesized secondary metabolites are critical quality-conferring compounds of foods, plant-derived medicines, and beverages. However, information at a single-cell level on root-specific secondary metabolism remains largely unexplored. L-Theanine, an important quality component of tea, is primarily synthesized in roots, from which it is then transported to new shoots of tea plant. In this study, we present a single-cell RNA sequencing (scRNA-seq)-derived map for the tea plant root, which enabled cell-type-specific analysis of glutamate and ethylamine (two precursors of theanine biosynthesis) metabolism, and theanine biosynthesis, storage, and transport. Our findings support a model in which the theanine biosynthesis pathway occurs via multicellular compartmentation and does not require high co-expression levels of transcription factors and their target genes within the same cell cluster. This study provides novel insights into theanine metabolism and regulation, at the single-cell level, and offers an example for studying root-specific secondary metabolism in other plant systems.

    1. Plant Biology
    Ann-Kathrin Rößling, Kai Dünser ... Jürgen Kleine-Vehn
    Research Article

    The extracellular matrix plays an integrative role in cellular responses in plants, but its contribution to the signalling of extracellular ligands largely remains to be explored. Rapid alkalinisation factors (RALFs) are extracellular peptide hormones that play pivotal roles in various physiological processes. Here, we address a crucial connection between the de-methylesterification machinery of the cell wall component pectin and RALF1 activity. Pectin is a polysaccharide, contributing to the structural integrity of the cell wall. Our data illustrate that the pharmacological and genetic interference with pectin methyl esterases (PMEs) abolishes RALF1-induced root growth repression. Our data suggest that positively charged RALF1 peptides bind negatively charged, de-methylesterified pectin with high avidity. We illustrate that the RALF1 association with de-methylesterified pectin is required for its FERONIA-dependent perception, contributing to the control of the extracellular matrix and the regulation of plasma membrane dynamics. Notably, this mode of action is independent of the FER-dependent extracellular matrix sensing mechanism provided by FER interaction with the leucine-rich repeat extensin (LRX) proteins. We propose that the methylation status of pectin acts as a contextualizing signalling scaffold for RALF peptides, linking extracellular matrix dynamics to peptide hormone-mediated responses.