The visualization of hormonal signaling input and output is key to understanding how multicellular development is regulated. The plant signaling molecule auxin triggers many growth and developmental responses, but current tools lack the sensitivity or precision to visualize these. We developed a set of fluorescent reporters that allow sensitive and semiquantitative readout of auxin responses at cellular resolution in Arabidopsis thaliana. These generic tools are suitable for any transformable plant species.
This is a preview of subscription content, access via your institution
Open Access articles citing this article.
Nature Communications Open Access 15 November 2022
Nature Communications Open Access 09 July 2022
BMC Biology Open Access 24 September 2021
Subscribe to Journal
Get full journal access for 1 year
only $8.25 per issue
All prices are NET prices.
VAT will be added later in the checkout.
Tax calculation will be finalised during checkout.
Get time limited or full article access on ReadCube.
All prices are NET prices.
Lokerse, A.S. & Weijers, D. Curr. Opin. Plant Biol. 12, 520–526 (2009).
Muday, G.K. J. Plant Growth Regul. 20, 226–243 (2001).
Dharmasiri, N., Dharmasiri, S. & Estelle, M. Nature 435, 441–445 (2005).
Kepinski, S. & Leyser, O. Nature 435, 446–451 (2005).
Gray, W.M., Kepinski, S., Rouse, D., Leyser, O. & Estelle, M. Nature 414, 271–276 (2001).
Tan, X. et al. Nature 446, 640–645 (2007).
Ulmasov, T., Hagen, G. & Guilfoyle, T.J. Science 276, 1865–1868 (1997).
Ulmasov, T., Hagen, G. & Guilfoyle, T.J. Proc. Natl. Acad. Sci. USA 96, 5844–5849 (1999).
Wang, R. & Estelle, M. Curr. Opin. Plant Biol. 21, 51–58 (2014).
Ulmasov, T., Murfett, J., Hagen, G. & Guilfoyle, T.J. Plant Cell 9, 1963–1971 (1997).
Friml, J. et al. Nature 426, 147–153 (2003).
Moreno-Risueno, M.A. et al. Science 329, 1306–1311 (2010).
Jones, A.R. et al. Nat. Cell Biol. 11, 78–84 (2009).
Scarpella, E., Marcos, D., Friml, J. & Berleth, T. Genes Dev. 20, 1015–1027 (2006).
Grieneisen, V.A., Xu, J., Marée, A.F.M., Hogeweg, P. & Scheres, B. Nature 449, 1008–1013 (2007).
Boer, D.R. et al. Cell 156, 577–589 (2014).
Hardtke, C.S. et al. Development 131, 1089–1100 (2004).
Perrot-Rechenmann, C. Cold Spring Harb. Perspect. Biol. 2, a001446 (2010).
Brunoud, G. et al. Nature 482, 103–106 (2012).
Völker, A., Stierhof, Y.D. & Jürgens, G. J. Cell Sci. 114, 3001–3012 (2001).
Weijers, D. et al. Development 128, 4289–4299 (2001).
Federici, F., Dupuy, L., Laplaze, L., Heisler, M. & Haseloff, J. Nat. Methods 9, 483–485 (2012).
Wend, S. et al. Sci. Rep. 3, 2052 (2013).
Robert, H.S. et al. Curr. Biol. 23, 2506–2512 (2013).
Vernoux, T. et al. Mol. Syst. Biol. 7, 508 (2011).
De Rybel, B.D. et al. Plant Physiol. 156, 1292–1299 (2011).
Willemsen, V. et al. Plant Cell 15, 612–625 (Humana Press, 2003).
Llavata-Peris, C., Lokerse, A., Möller, B., De Rybel, B. & Weijers, D. in Plant Organogenesis: Methods and Protocols (ed. De Smet, Ive) Ch. 8, 137–148 (Humana Press, 2013).
van den Berg, C., Willemsen, V., Hage, W., Weisbeek, P. & Scheres, B. Nature 378, 62–65 (1995).
Daghma, D.S., Kumlehn, J., Hensel, G., Rutten, T. & Melzer, M. J. Exp. Bot. 63, 6017–6021 (2012).
Hellemans, J., Mortier, G., De Paepe, A., Speleman, F. & Vandesompele, J. Genome Biol. 8, R19 (2007).
We thank T. Laux (Universität Freiburg) for plasmids and B. de Rybel for helpful comments on the manuscript. This work was supported by grants from the European Research Council (ERC; CELLPATTERN; contract number 281573) and the Netherlands Organization for Scientific Research (NWO; ALW-820.02.019) to D.W. and Human Frontier Science Program (HFSP; research grant RGP0054-2013) and Agence Nationale de la Recherche (ANR; AuxiFlo; grant ANR-12-BSV6-0005) to T.V.
The authors declare no competing financial interests.
Integrated supplementary information
Maximal projection of propidium iodine stained root of (a) DR5:: n3eGFP and (b) DR5v2:: n3eGFP reporter lines. Scale bars are 10 µm.
qRT-PCR of serial diluted serial diluted pGIIM/DR5v2::ntdTomato-DR5::n3eGFP plasmid with primers used in this study. Bars indicate standard error from the mean (n=3).
Fluorescent signal intensity of n3xGFP (top row) and ntdTomato (bottom row) in DR5v2::ntdTomato-DR5::n3eGFP root tips following a 12-hour co-treatment of 10 µM NPA and the indicated concentrations of IAA. Detector gain was saturated for each channel separately at the highest signal intensity of the 1000 nM IAA treated root, and all other images were acquired using these same settings. Signal intensity is displayed as a false color scale. Scale bars are 10 µm.
(a) pRPS5a:: DII: Venus and (b) pRPS5a:: mDII: Venus. Scale bars are 10 µm.
Scale bars are 10 µm.
Normalized ntdTomato/n3xVenus signal ratio in nuclei at increasing distance from the QC (see dashed lines in image on the right). Cell 1 corresponds to the first daughter of the initial for each cell file. Red/yellow ratio was set to “1” in cell 1 for each cell file. Bars indicate standard error from the mean (n>30 cell files per tissue). Scale bars are 10 µm.
Supplementary Figures 1–6, Supplementary Table 1 and Supplementary Note 1 (PDF 1731 kb)
Overlay of red and green fluorescence in a root after treatment with 1 μM IAA during 33 minutes (1 frame every 3 minutes). (AVI 8450 kb)
Overlay of red and green fluorescence in a root after treatment with control medium during 33 minutes (1 frame every 3 minutes). (AVI 8450 kb)
About this article
Cite this article
Liao, CY., Smet, W., Brunoud, G. et al. Reporters for sensitive and quantitative measurement of auxin response. Nat Methods 12, 207–210 (2015). https://doi.org/10.1038/nmeth.3279
This article is cited by
Nature Communications (2022)
Nature Reviews Molecular Cell Biology (2022)
Nature Communications (2022)
Nature Plants (2022)
Spatiotemporal plant hormone analysis from cryosections using laser microdissection-liquid chromatography-mass spectrometry
Journal of Plant Research (2022)