植物过敏反应抗病反应过程中环磷酸鸟苷的测量

Measurement of Cyclic GMP During Plant Hypersensitive Disease Resistance Response.

作者信息

Chen Jian, Bellin Diana, Vandelle Elodie

机构信息

Department of Biological Sciences, University of South Carolina, Columbia, SC, USA.

Department of Biotechnology, University of Verona, Verona, Italy.

出版信息

Methods Mol Biol. 2018;1743:143-151. doi: 10.1007/978-1-4939-7668-3_13.

DOI:10.1007/978-1-4939-7668-3_13

PMID:29332293

Abstract

Cyclic guanosine-3',5'-monophosphate (cGMP) is recognized as an important second messenger in plants, mediating intracellular signal in important physiological processes, including the hypersensitive disease resistance response induced by avirulent pathogens. In this context, the analysis of cGMP levels in infected plants requires an accurate and specific detection method allowing its quantification. Here, we describe an assay based on the Alphascreen technology, developed for animal cells and further adapted and optimized for the detection of cGMP in plants. The method is applied for the measurement of cGMP in Arabidopsis thaliana plants challenged with an avirulent strain of Pseudomonas syringae pv. tomato. This protocol includes the extraction of cGMP, the assay procedure and the calculation of cGMP concentration.

摘要

环鸟苷-3',5'-单磷酸（cGMP）被认为是植物中一种重要的第二信使，在包括无毒病原体诱导的过敏抗病反应在内的重要生理过程中介导细胞内信号。在此背景下，分析受感染植物中的cGMP水平需要一种准确且特异的检测方法以实现其定量。在这里，我们描述了一种基于AlphaScreen技术的检测方法，该技术最初是为动物细胞开发的，后来经过改进和优化用于检测植物中的cGMP。该方法用于测量用丁香假单胞菌番茄致病变种无毒菌株攻击的拟南芥植物中的cGMP。本方案包括cGMP的提取、检测程序以及cGMP浓度的计算。