A Robust and Sensitive Synthetic Sensor to Monitor the Transcriptional Output of the Cytokinin Signaling Network in Planta
Abstract
Cytokinins are classic plant hormones that orchestrate plant growth, development, and physiology. They affect gene expression in target cells by activating a multistep phosphorelay network. Type-B response regulators, acting as transcriptional activators, mediate the final step in the signaling cascade. Previously, we have introduced a synthetic reporter, Two Component signaling Sensor (TCS)::green fluorescent protein (GFP), which reflects the transcriptional activity of type-B response regulators. TCS::GFP was instrumental in uncovering roles of cytokinin and deepening our understanding of existing functions. However, TCS-mediated expression of reporters is weak in some developmental contexts where cytokinin signaling has a documented role, such as in the shoot apical meristem or in the vasculature of Arabidopsis (Arabidopsis thaliana). We also observed that GFP expression becomes rapidly silenced in TCS::GFP transgenic plants. Here, we present an improved version of the reporter, TCS new (TCSn), which, compared with TCS, is more sensitive to phosphorelay signaling in Arabidopsis and maize (Zea mays) cellular assays while retaining its specificity. Transgenic Arabidopsis TCSn::GFP plants exhibit strong and dynamic GFP expression patterns consistent with known cytokinin functions. In addition, GFP expression has been stable over generations, allowing for crosses with different genetic backgrounds. Thus, TCSn represents a significant improvement to report the transcriptional output profile of phosphorelay signaling networks in Arabidopsis, maize, and likely other plants that display common response regulator DNA-binding specificities.
Additional Information
© 2013 American Society of Plant Biologists. Published online before print January 2013. Received November 27, 2012. Accepted January 24, 2013. Published January 25, 2013. This work was supported a Syngenta Fellowship from the Zurich-Basel Plant Science Center, University of Zurich (to B.M.) and a grant from the Swiss National Science Foundation (to B.M.). We thank Ueli Grossniklaus (University of Zurich) for helpful discussions, financial support, and critical reading of the manuscript; Marco Celio (University of Fribourg), Jean-Pierre Métraux (University of Fribourg), and Jen Sheen (Harvard Medical School) for financial support; Tatsuo Kakimoto (Osaka University) for ahk mutant seeds; and Eva Benková (Institute of Science and Technology Austria), Ari Pekka Mähönen (University of Helsinki), and Jan Hejátko (Masaryk University) for testing TCSn::GFP.Attached Files
Published - 1066.full.pdf
Supplemental Material - 211763suppdata.xls
Supplemental Material - Plant_Physiol.__161_3__1066-75,_Figures.ppt
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Additional details
- PMCID
- PMC3585579
- Eprint ID
- 38281
- Resolver ID
- CaltechAUTHORS:20130506-082723428
- University of Zurich
- Swiss National Science Foundation (SNSF)
- University of Fribourg
- Harvard Medical School
- Created
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2013-05-06Created from EPrint's datestamp field
- Updated
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2023-06-02Created from EPrint's last_modified field