A High-Throughput Chemical Screening Method for Inhibitors and Potentiators of Hypersensitive Cell Death Using Suspension Cell Culture of Arabidopsis thaliana

Methods Mol Biol. 2018:1795:39-47. doi: 10.1007/978-1-4939-7874-8_4.

Abstract

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.

Keywords: Disease resistance response; Evans Blue; Hypersensitive response; Plant activator; Programmed cell death.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / chemistry*
  • Arabidopsis / genetics
  • Arabidopsis / metabolism
  • Cell Culture Techniques
  • Cell Death / drug effects
  • Disease Resistance / genetics
  • Drug Discovery* / methods
  • High-Throughput Screening Assays*
  • Phytochemicals / chemistry*
  • Phytochemicals / pharmacology
  • Plant Cells / drug effects
  • Plant Cells / metabolism
  • Plant Diseases / genetics
  • Plant Diseases / microbiology

Substances

  • Phytochemicals