Abstract
Understanding the biological background of strigolactone (SL) structural diversity and the SL signaling pathway at molecular level requires quantitative and sensitive tools that precisely determine SL dynamics. Such biosensors may be also very helpful in screening for SL analogs and mimics with defined biological functions.
Recently, the genetically encoded, ratiometric sensor StrigoQuant was developed and allowed the quantification of the activity of a wide concentration range of SLs. StrigoQuant can be used for studies on the biosynthesis, function and signal transduction of this hormone class.
Here, we provide a comprehensive protocol for establishing the use of StrigoQuant in Arabidopsis protoplasts. We first describe the generation and transformation of the protoplasts with StrigoQuant and detail the application of the synthetic SL analogue GR24. We then show the recording of the luminescence signal and how the obtained data are processed and used to assess/determine SL perception.
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Acknowledgments
This work was supported by the baseline funding and Competitive Research Grant 4 (CRG4) given to S.A. from King Abdullah University of Science and Technology and Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) under Germany’s Excellence Strategy (CEPLAS—EXC-2048/—Project ID 390686111) to M.D.Z. and the iGRAD Plant (IRTG 1525) to R.O.F. and M.D.Z. We thank Maximilian Augustin and Hannes Beyer for preliminary work on the sensors.
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Braguy, J., Samodelov, S.L., Andres, J., Ochoa-Fernandez, R., Al-Babili, S., Zurbriggen, M.D. (2021). A Protoplast-Based Bioassay to Quantify Strigolactone Activity in Arabidopsis Using StrigoQuant. In: Prandi, C., Cardinale, F. (eds) Strigolactones. Methods in Molecular Biology, vol 2309. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1429-7_16
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DOI: https://doi.org/10.1007/978-1-0716-1429-7_16
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