AUTHOR=Weigand Chrystle , Kim Su-Hwa , Brown Elizabeth , Medina Emily , Mares Moises , Miller Gad , Harper Jeffrey F. , Choi Won-Gyu TITLE=A Ratiometric Calcium Reporter CGf Reveals Calcium Dynamics Both in the Single Cell and Whole Plant Levels Under Heat Stress JOURNAL=Frontiers in Plant Science VOLUME=12 YEAR=2021 URL=https://www.frontiersin.org/journals/plant-science/articles/10.3389/fpls.2021.777975 DOI=10.3389/fpls.2021.777975 ISSN=1664-462X ABSTRACT=

Land plants evolved to quickly sense and adapt to temperature changes, such as hot days and cold nights. Given that calcium (Ca²⁺) signaling networks are implicated in most abiotic stress responses, heat-triggered changes in cytosolic Ca²⁺ were investigated in Arabidopsis leaves and pollen. Plants were engineered with a reporter called CGf, a ratiometric, genetically encoded Ca²⁺ reporter with an mCherry reference domain fused to an intensiometric Ca²⁺ reporter GCaMP6f. Relative changes in [Ca²⁺]_cyt were estimated based on CGf’s apparent K_D around 220 nM. The ratiometric output provided an opportunity to compare Ca²⁺ dynamics between different tissues, cell types, or subcellular locations. In leaves, CGf detected heat-triggered cytosolic Ca²⁺ signals, comprised of three different signatures showing similarly rapid rates of Ca²⁺ influx followed by differing rates of efflux (50% durations ranging from 5 to 19 min). These heat-triggered Ca²⁺ signals were approximately 1.5-fold greater in magnitude than blue light-triggered signals in the same leaves. In contrast, growing pollen tubes showed two different heat-triggered responses. Exposure to heat caused tip-focused steady growth [Ca²⁺]_cyt oscillations to shift to a pattern characteristic of a growth arrest (22%), or an almost undetectable [Ca²⁺]_cyt (78%). Together, these contrasting examples of heat-triggered Ca²⁺ responses in leaves and pollen highlight the diversity of Ca²⁺ signals in plants, inviting speculations about their differing kinetic features and biological functions.