A High-Throughput Chemical Screening Method for Inhibitors and Potentiators of Hypersensitive Cell Death Using Suspension Cell Culture of Arabidopsis thaliana
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Chemical biology provides an alternative way to identify genes involved in a particular biological process. It has the potential to overcome issues such as redundancy or lethality often found in genetic approaches, since the chemical compounds can simultaneously target all homologous proteins that function at the same step, and chemicals can be applied conditionally. Even with a variety of genetic approaches, the molecular mechanisms of plant hypersensitive cell death that occurs during disease resistance responses remain unclear. Therefore, application of chemical biology should provide new insights into this phenomenon. Here we describe a high-throughput chemical screening procedure to detect hypersensitive cell death quantitatively, using a suspension cell culture of Arabidopsis thaliana and a well-studied avirulent bacterial pathogen, Pseudomonas syringae pv. tomato DC3000 avrRpm1.
Key wordsHypersensitive response Programmed cell death Disease resistance response Plant activator Evans Blue
Tiadinil was kindly provided by Nihon Nohyaku Co. Ltd. This work was supported by ALCA Grant from the Japan Science and Technology Agency and KAKENHI Grant 25292035 from the Ministry of Education, Culture, Sports, Science and Technology of Japan to Y.N. and in part 15H05959 and 17H06172 to K.S.
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