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Abstract

How biological systems generate reproducible patterns with high precision is a central question in science1. The shoot apical meristem (SAM), a specialized tissue producing plant aerial organs, is a developmental system of choice to address this question. Organs are periodically initiated at the SAM at specific spatial positions and this spatiotemporal pattern defines phyllotaxis. Accumulation of the plant hormone auxin triggers organ initiation2,3,4,5, whereas auxin depletion around organs generates inhibitory fields that are thought to be sufficient to maintain these patterns and their dynamics4,6,7,8,9,10,11,12,13. Here we show that another type of hormone-based inhibitory fields, generated directly downstream of auxin by intercellular movement of the cytokinin signalling inhibitor ARABIDOPSIS HISTIDINE PHOSPHOTRANSFER PROTEIN 6 (AHP6)14, is involved in regulating phyllotactic patterns. We demonstrate that AHP6-based fields establish patterns of cytokinin signalling in the meristem that contribute to the robustness of phyllotaxis by imposing a temporal sequence on organ initiation. Our findings indicate that not one but two distinct hormone-based fields may be required for achieving temporal precision during formation of reiterative structures at the SAM, thus indicating an original mechanism for providing robustness to a dynamic developmental system.

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Acknowledgements

We thank M. Heisler, B. Müller and D. Weijers for sharing materials; A. Miyawaki for VENUS; D. Mast for help with plant analysis; C. Chamot and C. Lionnet (PLATIM) for help with confocal microscopy and ImageJ (C. Chamot); S. Chamot for help with live imaging; C. Gauthier and A. Laugraud (PRABI, Université Lyon I) for help with statistical analyses; Xavier Jaurand (Pi2) for help with the SEM; Y. Couder, S. Douady, M. Bennett and M.-A. Félix for their insights and support; Y. Jaillais, O. Hamant and C. Scutt for comments on the manuscript. T.V. was supported by HFSPO CDA 0047/2007 (Human Frontier Science Program Organization) and ANR-07-JCJC-0115 (Agence National de la Recherche) grants; R.D., F.P. and T.V. by the ANR-12-BSV6-0005 grant (AuxiFlo); J.L., J.T., C.G., Y.H. and T.V. by a transnational EraSysBio Grant (iSAM); F.B. by a predoctoral grant of the French Ministry of Research; and Y.R. by a CJS grant from INRA.

Author information

Author notes

    • Fabrice Besnard
    • , Valérie Morin
    • , Etienne Farcot
    •  & Anthony Bishopp

    Present addresses: IBENS, ENS, 75005 Paris, France (F.B.); UMR CNRS 5534, Université Claude Bernard Lyon I, Bâtiment Gregor Mendel, 16 rue Raphaël Dubois, 69622 Villeurbanne, France (V.M.); University of Nottingham, University Park, Nottingham NG7 2RD, UK (E.F); University of Nottingham, Sutton Bonington LE12 5RD, UK (A.Bi.).

    • Valérie Morin
    • , Benjamin Marteaux
    •  & Géraldine Brunoud

    These authors contributed equally to this work.

Affiliations

  1. Laboratoire de Reproduction et Développement des Plantes, CNRS, INRA, ENS Lyon, UCBL, Université de Lyon, 69364 Lyon, France

    • Fabrice Besnard
    • , Valérie Morin
    • , Benjamin Marteaux
    • , Géraldine Brunoud
    • , Pierre Chambrier
    • , Frédérique Rozier
    • , Vincent Mirabet
    • , Jonathan Legrand
    • , Stéphanie Lainé
    • , Coralie Cellier
    • , Pradeep Das
    • , Arezki Boudaoud
    • , Jan Traas
    •  & Teva Vernoux
  2. Virtual Plants INRIA/CIRAD/INRA Project Team, UMR AGAP, Institut de Biologie Computationelle, 34095 Montpellier, France

    • Yassin Refahi
    • , Jonathan Legrand
    • , Etienne Farcot
    • , Christophe Godin
    •  & Yann Guédon
  3. Laboratoire Joliot-Curie, CNRS, ENS Lyon, Université de Lyon, 69364 Lyon, France

    • Vincent Mirabet
    • , Jonathan Legrand
    • , Pradeep Das
    •  & Arezki Boudaoud
  4. Laboratoire Physiologie Cellulaire et Végétale, CEA, CNRS, INRA, UJF, 38041 Grenoble, France

    • Emmanuel Thévenon
    • , Renaud Dumas
    •  & François Parcy
  5. Institute of Biotechnology/Department of Biosciences, University of Helsinki, FIN-00014, Finland

    • Anthony Bishopp
    •  & Ykä Helariutta

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Contributions

F.B. and T.V. conceived and designed the experiments. F.B., V.Mo., B.M., G.B., P.C., F.R., J.L., S.L., C.C., E.T., P.D. and T.V. performed the experiments. Y.R., E.F., C.G., F.B., G.B., T.V. and Y.G. performed the mathematical analysis of phyllotaxis. V.Mi., G.B. and A.Bo. analysed the auxin transport network. R.D., F.P., A.Bi., Y.H. and J.T. provided reagents/materials. F.B. and T.V. analysed the data with inputs from all the authors. F.B. and T.V. wrote the paper. All authors discussed the results and commented on the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Teva Vernoux.

Supplementary information

PDF files

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    Supplementary Information

    This file contains Supplementary Figures 1-15 and Supplementary Tables 1-3.

Videos

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    Live-imaging of DR5::VENUS and pAHP6::GFP expression

    The expression of DR5::VENUS (Magenta) and pAHP6::GFP (Green) was followed over time in the shoot apical meristem using confocal microscopy.

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DOI

https://doi.org/10.1038/nature12791

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