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      Spatiotemporal coordination of cell division and growth during organ morphogenesis

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          Abstract

          A developing plant organ exhibits complex spatiotemporal patterns of growth, cell division, cell size, cell shape, and organ shape. Explaining these patterns presents a challenge because of their dynamics and cross-correlations, which can make it difficult to disentangle causes from effects. To address these problems, we used live imaging to determine the spatiotemporal patterns of leaf growth and division in different genetic and tissue contexts. In the simplifying background of the speechless ( spch) mutant, which lacks stomatal lineages, the epidermal cell layer exhibits defined patterns of division, cell size, cell shape, and growth along the proximodistal and mediolateral axes. The patterns and correlations are distinctive from those observed in the connected subepidermal layer and also different from the epidermal layer of wild type. Through computational modelling we show that the results can be accounted for by a dual control model in which spatiotemporal control operates on both growth and cell division, with cross-connections between them. The interactions between resulting growth and division patterns lead to a dynamic distributions of cell sizes and shapes within a deforming leaf. By modulating parameters of the model, we illustrate how phenotypes with correlated changes in cell size, cell number, and organ size may be generated. The model thus provides an integrated view of growth and division that can act as a framework for further experimental study.

          Author summary

          Organ morphogenesis involves two coordinated processes: growth of tissue and increase in cell number through cell division. Both processes have been analysed individually in many systems and shown to exhibit complex patterns in space and time. However, it is unclear how these patterns of growth and cell division are coordinated in a growing leaf that is undergoing shape changes. We have addressed this problem using live imaging to track growth and cell division in the developing leaf of the mustard plant Arabidopsis thaliana. Using subsequent computational modelling, we propose an integrated model of leaf growth and cell division, which generates dynamic distributions of cell size and shape in different tissue layers, closely matching those observed experimentally. A key aspect of the model is dual control of spatiotemporal patterns of growth and cell division parameters. By modulating parameters in the model, we illustrate how phenotypes may correlate with changes in cell size, cell number, and organ size.

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          Most cited references59

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          TOR, a Central Controller of Cell Growth

          Cell, 103(2), 253-262
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            High-resolution whole-mount imaging of three-dimensional tissue organization and gene expression enables the study of Phloem development and structure in Arabidopsis.

            Currently, examination of the cellular structure of plant organs and the gene expression therein largely relies on the production of tissue sections. Here, we present a staining technique that can be used to image entire plant organs using confocal laser scanning microscopy. This technique produces high-resolution images that allow three-dimensional reconstruction of the cellular organization of plant organs. Importantly, three-dimensional domains of gene expression can be analyzed with single-cell precision. We used this technique for a detailed examination of phloem cells in the wild type and mutants. We were also able to recognize phloem sieve elements and their differentiation state in any tissue type and visualize the structure of sieve plates. We show that in the altered phloem development mutant, a hybrid cell type with phloem and xylem characteristics develops from initially normally differentiated protophloem cells. The simplicity of sieve element data collection allows for the statistical analysis of structural parameters of sieve plates, essential for the calculation of phloem conductivity. Taken together, this technique significantly improves the speed and accuracy of the investigation of plant growth and development.
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              Transcription factor control of asymmetric cell divisions that establish the stomatal lineage.

              The establishment of new cell lineages during development often requires a symmetry-breaking event. An asymmetric division in the epidermis of plants initiates a lineage that ultimately produces stomatal guard cells. Stomata are pores in the epidermis that serve as the main conduits for gas exchange between plants and the atmosphere; they are critical for photosynthesis and exert a major influence on global carbon and water cycles. Recent studies implicated intercellular signalling in preventing the inappropriate production of stomatal complexes. Genes required to make stomata, however, remained elusive. Here we report the identification of a gene, SPEECHLESS (SPCH), encoding a basic helix-loop-helix (bHLH) transcription factor that is necessary and sufficient for the asymmetric divisions that establish the stomatal lineage in Arabidopsis thaliana. We demonstrate that SPCH and two paralogues are successively required for the initiation, proliferation and terminal differentiation of cells in the stomatal lineage. The stomatal bHLHs define a molecular pathway sufficient to create one of the key cell types in plants. Similar molecules and regulatory mechanisms are used during muscle and neural development, highlighting a conserved use of closely related bHLHs for cell fate specification and differentiation.
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                Author and article information

                Contributors
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: InvestigationRole: MethodologyRole: ResourcesRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: MethodologyRole: ResourcesRole: SoftwareRole: ValidationRole: VisualizationRole: Writing – original draftRole: Writing – review & editing
                Role: ConceptualizationRole: Data curationRole: Formal analysisRole: MethodologyRole: ResourcesRole: SoftwareRole: ValidationRole: VisualizationRole: Writing – review & editing
                Role: Data curationRole: ResourcesRole: Software
                Role: ConceptualizationRole: InvestigationRole: MethodologyRole: ResourcesRole: Writing – review & editing
                Role: Formal analysisRole: InvestigationRole: Writing – review & editing
                Role: InvestigationRole: Writing – review & editing
                Role: Formal analysisRole: ValidationRole: Writing – review & editing
                Role: ResourcesRole: Writing – review & editing
                Role: Writing – review & editing
                Role: Writing – review & editing
                Role: ConceptualizationRole: Supervision
                Role: ConceptualizationRole: Funding acquisitionRole: MethodologyRole: Project administrationRole: SupervisionRole: Writing – original draftRole: Writing – review & editing
                Role: Academic Editor
                Journal
                PLoS Biol
                PLoS Biol
                plos
                plosbiol
                PLoS Biology
                Public Library of Science (San Francisco, CA USA )
                1544-9173
                1545-7885
                1 November 2018
                November 2018
                1 November 2018
                : 16
                : 11
                : e2005952
                Affiliations
                [1 ] Department of Cell and Developmental Biology, John Innes Centre, Norwich, England, United Kingdom
                [2 ] School of Computational Sciences, University of East Anglia, Norwich, England, United Kingdom
                [3 ] Department of Computational and Systems Biology, John Innes Centre, Norwich, England, United Kingdom
                University of California San Diego, United States of America
                Author notes

                The authors have declared that no competing interests exist.

                [¤a]

                Current address: School of Computational Sciences, University of East Anglia, Norwich, England, United Kingdom

                [¤b]

                Current address: UMR LEPSE, Montpellier SupAgro/INRA, Montpellier, France

                [¤c]

                Current address: Institute of Plant Sciences, University of Bern, Bern, Switzerland

                [¤d]

                Current address: Department of Agricultural and Environmental Sciences—Production, Landscape, Agroenergy, University of Milan, Milan, Italy

                [¤e]

                Current address: MRC-Laboratory of Molecular Biology, Cambridge Biomedical Campus, Francis Crick Avenue, Cambridge, England, United Kingdom

                Author information
                http://orcid.org/0000-0003-1271-6412
                Article
                pbio.2005952
                10.1371/journal.pbio.2005952
                6211367
                30383040
                88404253-94ae-4c1c-bf42-953052e004d6
                © 2018 Fox et al

                This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 8 March 2018
                : 20 September 2018
                Page count
                Figures: 10, Tables: 1, Pages: 48
                Funding
                Biotechnology and Biological Sciences Research Council (grant number BB/P013511/1; BB/J004588/1). The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Biotechnology and Biological Sciences Research Council (grant number BB/M023117/1, BB/F005997/1, and BB/L008920/1). Received by EC. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. ERC (grant number Carnomorph, 323028). Received by EC. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Biotechnology and Biological Sciences Research Council (grant number BB/F005555/1). Received by AB. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Marie Skłodowska-Curie actions (grant number HEMOTIONAL, 329784). Received by FP. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. Consejo Nacional de Ciencia y Tecnología. Received by YSC. The funder had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Biology and Life Sciences
                Cell Biology
                Cell Processes
                Cell Cycle and Cell Division
                Biology and Life Sciences
                Anatomy
                Integumentary System
                Skin
                Epidermis
                Medicine and Health Sciences
                Anatomy
                Integumentary System
                Skin
                Epidermis
                Biology and Life Sciences
                Plant Science
                Plant Anatomy
                Leaves
                Research and Analysis Methods
                Bioassays and Physiological Analysis
                Cell Analysis
                Cell Division Analysis
                Research and Analysis Methods
                Animal Studies
                Experimental Organism Systems
                Model Organisms
                Arabidopsis Thaliana
                Research and Analysis Methods
                Model Organisms
                Arabidopsis Thaliana
                Biology and Life Sciences
                Organisms
                Eukaryota
                Plants
                Brassica
                Arabidopsis Thaliana
                Research and Analysis Methods
                Animal Studies
                Experimental Organism Systems
                Plant and Algal Models
                Arabidopsis Thaliana
                Biology and Life Sciences
                Cell Biology
                Cell Physiology
                Cell Polarity
                Computer and Information Sciences
                Network Analysis
                Regulatory Networks
                Biology and Life Sciences
                Organisms
                Eukaryota
                Plants
                Seedlings
                Custom metadata
                Images and models software from this study are available at  https://figshare.com/s/b14c8e6cb1fc5135dd87. Models and GFtbox modeling software are also available at  http://cmpdartsvr3.cmp.uea.ac.uk/wiki/BanghamLab/index.php/Software.

                Life sciences
                Life sciences

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