Abstract
This chapter describes procedures to find small molecules that bind to abscisic acid (ABA) receptors and improve plant tolerance to water stress. Abscisic acid promotes the interaction between ABA receptors and protein phosphatase 2Cs (PP2Cs), which are negative regulators of ABA signaling. This receptor-mediated ABA-dependent inhibition of PP2C activity is required for ABA action in vivo. ABA agonists can be identified by high-throughput screening for molecules that promote agonist-induced ABA receptor–PP2C interactions using yeast two-hybrid assays. In addition to in vitro assays, an in vivo test to evaluate ABA agonist activity has been devised in which an ABA-inducible GUS reporter strain is used to evaluate the effect of each ABA agonist. The effects of ABA agonists can also be determined using thermal imaging analyses and a water loss assay of stomata. Finally, the ability of each ABA agonist to confer drought stress tolerance upon plants grown in soil is determined. These methods can be used to identify various ABA agonists that protect plants against water stress.
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Acknowledgments
The authors would like to thank Dr. Andrew Defries and Dr. Sang-Youl Park, Center for Plant Cell Biology, Department of Botany and Plant Sciences, UC Riverside, for supporting ABA agonist screening. This work was supported in part by the Japanese Society for the Promotion of Science (JSPS) KAKENHI (17H05009 to M.O.), PRESTO JST (JPMJPR15Q5 to M.O.), and NSF (IOS1656890 to S.R.C.).
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Okamoto, M., Cutler, S.R. (2018). Chemical Control of ABA Receptors to Enable Plant Protection Against Water Stress. In: Fauser, F., Jonikas, M. (eds) Plant Chemical Genomics. Methods in Molecular Biology, vol 1795. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7874-8_11
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DOI: https://doi.org/10.1007/978-1-4939-7874-8_11
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