MAMP-triggered Medium Alkalinization of Plant Cell Cultures

• Published in: Bio-protocol Vol. 10; no. 8; p. e3588
• Main Authors: Fiorin, Gabriel L, Sánchez-Vallet, Andrea, Thomma, Bart P H J, Pereira, Gonçalo A G, Teixeira, Paulo J P L
• Format: Journal Article
• Language: English
• Published: United States Bio-Protocol 20.04.2020; Bio-protocol LLC
• Subjects: Biology; Methods; Host-microbe interactions; Elicitor; Phytopathology; Effector Biology; Plant immunity
• ISSN: 2331-8325; 2331-8325
• DOI: 10.21769/BioProtoc.3588

Plants recognize a wide variety of microbial molecules to detect and respond to potential invaders. Recognition of Microbe-Associated Molecular Patterns (MAMPs) by cell surface receptors initiate a cascade of biochemical responses that include, among others, ion fluxes across the plasma membrane. A consequence of such event is a decrease in the concentration of extracellular H ions, which can be experimentally detected in plant cell suspensions as a shift in the pH of the medium. Thus, similarly to reactive oxygen species (ROS) accumulation, phosphorylation of MAP kinases and induction of defense-related genes, MAMP-induced medium alkalinization can be used as a proxy for the activation of plant immune responses. Here, we describe a detailed protocol for the measurement of medium alkalinization of tobacco BY-2 cell suspensions upon treatment with two different MAMPs: chitohexamers derived from fungal cell walls (NAG6; N-acetylglucosamine) and the flagellin epitope flg22, found in the bacterial flagellum. This method provides a reliable and fast platform to access MAMP-Triggered Immunity (MTI) in tobacco cell suspensions and can be easily adapted to other plant species as well as to other MAMPs.