14 March 2018

Dual Color Sensors for Simultaneous Analysis of Calcium Signal Dynamics in the Nuclear and Cytoplasmic Compartments of Plant Cells.

Spatiotemporal changes in cellular calcium (Ca2+) concentrations are essential for signal transduction in a wide range of plant cellular processes. In legumes, nuclear and perinuclear-localized Ca2+oscillations have emerged as key signatures preceding downstream symbiotic signaling responses. Förster resonance energy transfer (FRET) yellow-based Ca2+cameleon probes have been successfully exploited to measure the spatiotemporal dynamics of symbiotic Ca2+signaling in legumes. Although providing cellular resolution, these sensors were restricted to measuring Ca2+changes in single subcellular compartments. In this study, we have explored the potential of single fluorescent protein-based Ca2+sensors, the GECOs, for multicolor and simultaneous imaging of the spatiotemporal dynamics of cytoplasmic and nuclear Ca2+signaling in root cells. Single and dual fluorescence nuclear and cytoplasmic-localized GECOs expressed in transgenicMedicago truncatularoots andArabidopsis thalianawere used to successfully monitor Ca2+responses to microbial biotic and abiotic elicitors. InM. truncatula, we demonstrate that GECOs detect symbiosis-related Ca2+spiking variations with higher sensitivity than the yellow FRET-based sensors previously used. Additionally, in bothM. truncatulaandA. thaliana, the dual sensor is now able to resolve in a single root cell the coordinated spatiotemporal dynamics of nuclear and cytoplasmic Ca2+signalingin vivo. The GECO-based sensors presented here therefore represent powerful tools to monitor Ca2+signaling dynamics invivoin response to different stimuli in multi-subcellular compartments of plant cells.

Publisher's Version

JIC Cookie Policy. We use cookies on this site to enhance your user experience. By clicking any link on this page you are giving your consent for us to set cookies. You can find out more about the cookies by clicking here.

Accept cookies