Noutoshi Yoshiteru, Shirasu Ken
Graduate School of Environmental and Life Science, Okayama University, Okayama, Japan.
Plant Immunity Research Group, RIKEN Center for Sustainable Resource Science, Yokohama, Kanagawa, Japan.
Methods Mol Biol. 2018;1795:39-47. doi: 10.1007/978-1-4939-7874-8_4.
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.
化学生物学提供了一种识别参与特定生物学过程的基因的替代方法。它有潜力克服遗传方法中经常出现的冗余或致死性等问题,因为化学化合物可以同时靶向在同一步骤发挥作用的所有同源蛋白质,并且化学物质可以有条件地应用。即使有多种遗传方法,植物在抗病反应过程中发生的超敏细胞死亡的分子机制仍不清楚。因此,化学生物学的应用应该为这一现象提供新的见解。在这里,我们描述了一种高通量化学筛选程序,使用拟南芥悬浮细胞培养物和一种经过充分研究的无毒细菌病原体——丁香假单胞菌番茄致病变种DC3000 avrRpm1,来定量检测超敏细胞死亡。