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Land plants evolved to quickly sense and adapt to temperature changes, such as hot days and cold nights. Given that calcium (Ca 2+ ) signaling networks are implicated in most abiotic stress responses, heat-triggered changes in cytosolic Ca 2+  were investigated in  Arabidopsis  leaves and pollen. Plants were engineered with a reporter called CGf, a ratiometric, genetically encoded Ca 2+  reporter with an m  C  herry reference domain fused to an intensiometric Ca 2+  reporter   G  CaMP6  f  . Relative changes in [Ca 2+ ] cyt  were estimated based on CGf’s apparent  K  D  around 220 nM. The ratiometric output provided an opportunity to compare Ca 2+  dynamics between different tissues, cell types, or subcellular locations. In leaves, CGf detected heat-triggered cytosolic Ca 2+  signals, comprised of three different signatures showing similarly rapid rates of Ca 2+  influx followed by differing rates of efflux (50% durations ranging from 5 to 19 min). These heat-triggered Ca 2+  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 [Ca 2+ ] cyt  oscillations to shift to a pattern characteristic of a growth arrest (22%), or an almost undetectable [Ca 2+ ] cyt  (78%). Together, these contrasting examples of heat-triggered Ca 2+  responses in leaves and pollen highlight the diversity of Ca 2+  signals in plants, inviting speculations about their differing kinetic features and biological functions.

A Ratiometric Calcium Reporter CGf Reveals Calcium Dynamics Both in the Single Cell and Whole Plant Levels Under Heat Stress