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A comprehensive fluorescent sensor for spatiotemporal cell cycle analysis in Arabidopsis

Nature Plants volume 6, pages 1330–1334 (2020)Cite this article

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Abstract

Assessing cell proliferation dynamics is crucial to understand the spatiotemporal control of organogenesis. Here we have generated a versatile fluorescent sensor, PlaCCI (plant cell cycle indicator) on the basis of the expression of CDT1a-CFP, H3.1-mCherry and CYCB1;1-YFP, that identifies cell cycle phases in Arabidopsis thaliana. This tool works in a variety of organs, and all markers and the antibiotic resistance are expressed from a single cassette, facilitating the selection in mutant backgrounds. We also show the robustness of PlaCCI line in live-imaging experiments to follow and quantify cell cycle phase progression.

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Fig. 1: Development of PlaCCI.
Fig. 2: Use of PlaCCI reporter in various experimental settings.

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Acknowledgements

We thank the Confocal Microscopy and the Cytometry services of CBMSO for support, E. Martinez-Salas for comments, S. Herrero-Desvoyes for help with cell size measurements, E. Caro for the terminator modules for GoldenBraid constructs, V. Mora-Gil for technical assistance and members of the laboratory for continuous feedback. Research in our laboratory is supported by grant nos. BIO2017-92329-EXP and RTI2018-094793-B-I00 from MICIU, and ERC-2018-AdG_833617 from European Union, and by institutional grants from Banco de Santander and Fundación Ramon Areces to the Centro de Biología Molecular Severo Ochoa.

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Authors and Affiliations

  1. Centro de Biologia Molecular Severo Ochoa, CSIC-UAM, Cantoblanco, Madrid, Spain

    Bénédicte Desvoyes, Ainhoa Arana-Echarri, María D. Barea & Crisanto Gutierrez

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  1. Bénédicte Desvoyes
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  2. Ainhoa Arana-Echarri
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  3. María D. Barea
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  4. Crisanto Gutierrez
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Contributions

B.D. and C.G designed this work. A.A.-E. and M.D.B. participated in the initial cloning steps. B.D. performed all the experiments. B.D. and C.G. wrote the manuscript.

Corresponding authors

Correspondence to Bénédicte Desvoyes or Crisanto Gutierrez.

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The authors declare no competing interests.

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Peer review information Nature Plants thanks Lieven De Veylder 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. 1–6 and Table 1.

Reporting Summary

Supplementary Video 1

Time lapse (7 h) of the root meristem of a PlaCCI line showing the switching on/off of various cell cycle markers.

Supplementary Data 1

Source data for Supplementary Fig. 4.

Supplementary Data 2

Source data for Supplementary Fig. 5a.

Supplementary Data 3

Source data for Supplementary Fig. 5b.

Source data

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Desvoyes, B., Arana-Echarri, A., Barea, M.D. et al. A comprehensive fluorescent sensor for spatiotemporal cell cycle analysis in Arabidopsis. Nat. Plants 6, 1330–1334 (2020). https://doi.org/10.1038/s41477-020-00770-4

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