A novel sensor to map auxin response and distribution at high spatio-temporal resolution

Abstract

Auxin is a key plant morphogenetic signal1 but tools to analyse dynamically its distribution and signalling during development are still limited. Auxin perception directly triggers the degradation of Aux/IAA repressor proteins2,3,4,5,6. Here we describe a novel Aux/IAA-based auxin signalling sensor termed DII-VENUS that was engineered in the model plant Arabidopsis thaliana. The VENUS fast maturing form of yellow fluorescent protein7 was fused in-frame to the Aux/IAA auxin-interaction domain (termed domain II; DII)5 and expressed under a constitutive promoter. We initially show that DII-VENUS abundance is dependent on auxin, its TIR1/AFBs co-receptors4,5,6,8 and proteasome activities. Next, we demonstrate that DII-VENUS provides a map of relative auxin distribution at cellular resolution in different tissues. DII-VENUS is also rapidly degraded in response to auxin and we used it to visualize dynamic changes in cellular auxin distribution successfully during two developmental responses, the root gravitropic response and lateral organ production at the shoot apex. Our results illustrate the value of developing response input sensors such as DII-VENUS to provide high-resolution spatio-temporal information about hormone distribution and response during plant growth and development.

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Figure 1: DII-VENUS degradation is dependent on auxin, TIR1/AFBs and proteasome activity.
Figure 2: DII-VENUS provides a sensor to map auxin distribution in plant tissues.
Figure 3: DII-VENUS monitors changes in auxin response and distribution at high temporal resolution.
Figure 4: DII-VENUS allows visualization of changes in auxin distribution during development.

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Acknowledgements

We thank A. Erktan and C. Cellier for help with marker expression analysis; J. Neve, A. Miyawaki, M. Heisler and M. Estelle for providing the 35S::Flag-TIR1 line, VENUS complementary DNA, DR5::VENUS plasmids and TIR/AFB GUS lines, respectively; the PLATIM for access to confocal microscopes; F. Parcy, O. Hamant, A. Boudaoud and P. Das for discussions. T.V. was supported by the Human Frontier Science Program Organization (CDA 0047/2007 HFSPO) and the Agence National de la Recherche (ANR-07-JCJC-0115 and EraSysBio+ iSAM). D.M.W., A.L. and M.J.B. acknowledge the support of the Biotechnology and Biological Sciences Research Council (BBSRC) and Engineering and Physical Sciences Research Council (EPSRC) funding to the Centre for Plant Integrative Biology (CPIB); BBSRC grants BB/F013981/1 and BB/F007418/1 to S.K.; BBSRC Professorial Research Fellowship funding to D.M.W. & M.J.B.; and Belgian Scientific policy (BELSPO contract BARN) to A.L., T.B. and M.J.B.

Author information

T.V. designed the DII-VENUS tool. G.B., M.O. and T.V. engineered and characterized DII-VENUS transgenic lines. D.M.W., G.B., A.L. and V.M. quantified the spatial and temporal dynamics of DII-VENUS. A.H.B. did the pull-down assay. T.V. and M.J.B. designed the experiments with the help of T.B., S.K. and J.T. T.V. and M.J.B. analysed the data and wrote the paper. All authors discussed the results and commented on the manuscript.

Correspondence to Teva Vernoux.

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Supplementary Information

The file contains Supplementary Text, Supplementary References, Supplementary Figures 1-10 with legends and Supplementary Table 1. (PDF 2832 kb)

Supplementary Movie 1

This file shows DII-VENUS degradation upon auxin treatment. (MOV 4466 kb)

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Brunoud, G., Wells, D., Oliva, M. et al. A novel sensor to map auxin response and distribution at high spatio-temporal resolution. Nature 482, 103–106 (2012). https://doi.org/10.1038/nature10791

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