Quantification of Extracellular ATP in Plant Suspension Cell Cultures

Extracellular ATP functions as an important signaling molecule in both plants and animals. In plants, ATP is released in the extracellular region of cells in response to environmental perturbations, such as herbivory, cellular damage, or other abiotic and biotic stimuli, which is then perceived by the purinoceptor P2K1 as a damaged-self signal for activation of defense responses. Given its involvement in various physiological processes, quantification of extracellular ATP is important for further understanding of its molecular function. In this chapter, we describe a method for the accurate and reliable determination of extracellular ATP concentrations in plant cell culture media based on the luciferase-luciferin reaction, using either end-point or real-time detection assays. The protocol can be easily performed with any luminometer within 1 h after sample collection. Although we use Arabidopsis suspension cells, the protocol described can be optimized for any cell type.