Using genetically encoded fluorescent indicators in Arabidopsis plants

Abstract

Genetically-encoded fluorescent indicators (GEFIs) are engineered fluorescent proteins containing sensory domains that affect excitation and/or emission in response to specific analytes. In recent years, GEFIs have become a prominent tool in the research of plant signalling pathways as they allow real-time monitoring molecular events by converting them into readable and quantifiable signals. This study describes how plant lines with specific mutations and expressing GEFIs were genotyped, characterized, and employed to detect accumulation of cytosolic CO2 and H2O2 in response to O3 treatment. The gas exchange experiments with GEFI lines revealed that ABAleon, an abscisic acid sensor, affected stomatal responses to high CO2 and O3. It was also demonstrated that HPCA1 which recognizes apoplastic H2O2 in plants is essential for the increase of Ca2+ in the cytosol of plant cells during an exposure to O3.