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Published February 13, 2019 | Supplemental Material + Published
Journal Article Open

Calcium signals are necessary to establish auxin transporter polarity in a plant stem cell niche

Abstract

In plants mechanical signals pattern morphogenesis through the polar transport of the hormone auxin and through regulation of interphase microtubule (MT) orientation. To date, the mechanisms by which such signals induce changes in cell polarity remain unknown. Through a combination of time-lapse imaging, and chemical and mechanical perturbations, we show that mechanical stimulation of the SAM causes transient changes in cytoplasmic calcium ion concentration (Ca^(2+)) and that transient Ca^(2+) response is required for downstream changes in PIN-FORMED 1 (PIN1) polarity. We also find that dynamic changes in Ca^(2+) occur during development of the SAM and this Ca^(2+) response is required for changes in PIN1 polarity, though not sufficient. In contrast, we find that Ca^(2+) is not necessary for the response of MTs to mechanical perturbations revealing that Ca^(2+) specifically acts downstream of mechanics to regulate PIN1 polarity response.

Additional Information

© 2019 The Author(s). This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/. Received 08 June 2018; Accepted 15 January 2019; Published 13 February 2019. Code availability: The corresponding open-source Python code, previously introduced in Refs. 56,57 on natural-cubic-smoothing-splines can be found at https://github.com/eldad-a/natural-cubic-smoothing-splines. All other code, including the programming code for quantitative analysis of the meristem calcium oscillations and waves upon mechanical perturbation, are available from the corresponding authors upon request. Data availability: All data are available from the corresponding author upon request, including all of the microscope time lapse files. The source data underlying Supplementary Figs. 1a, b, 4c, 5a–f, 7e–j, 8a, b, 10, 11i–l, 12k–l and Supplementary Tables 1 and 2 are provided as a Source Data file. We are grateful to Dr. Arun Sampathkumar for support in establishing the calcium confocal imaging process, Dr. Hanako Yashiro for the support in intact plant imaging and Dr. Ivo Grosse for the useful discussions and input on statistical analysis. We thank Dr. Francesca Peri for kindly providing the DNA for GCaMP6f and Dr. Rainer Waadt for R-GECO1 constructs and advice. We also thank Dr. Changfu Yao and Meyerowitz and Heisler lab members for suggestions and discussions. The authors' work was funded by the Howard Hughes Medical Institute and by NASA grant NNX17AD53G to E.M.M., by the European Research Council (ERC) grant 261081 to M.G.H., in part by NIH grant GM060396 to J.I.S., Marie Skłodowska-Curie Action fellowship to P.D-.S., and a California Institute of Technology Biology and Biological Engineering Fellowship and a Zuckerman STEM Leadership Program postdoctoral scholarship to E.A. Author Contributions: T.L., A.Y., N.B., J.I.S, M.G.H., and E.M.M. conceived the experiments. T.L., N.B., A.Y., P.D-.S. performed experiments. A.A. analyzed Ca^(2+) wave imaging data. E.A. analyzed Ca^(2+) oscillation patterns. P.D-.S. invented, built, and provided the indentation and compression devices. P.T.T. contributed reagents. T.L., A.Y., N.B., M.H., and E.M.M. wrote the manuscript. All authors read and approved the manuscript. The authors declare no competing interests.

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Published - s41467-019-08575-6.pdf

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Created:
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