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

Chrystle Weigand, Su Hwa Kim, Elizabeth Brown, Emily Medina, Moises Mares, Gad Miller, Jeffrey F. Harper, Won Gyu Choi

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

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 KD 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.

Original languageEnglish
Article number777975
JournalFrontiers in Plant Science
Volume12
DOIs
StatePublished - 17 Dec 2021

Bibliographical note

Publisher Copyright:
Copyright © 2021 Weigand, Kim, Brown, Medina, Mares, Miller, Harper and Choi.

Funding

This work was supported by the United States Department of Agriculture (HATCH grant no. NEV00384 to JH), the National Science Foundation (IOS grant no. 1656774 to JH, and MCB grant no. 2016143 to W-GC), and US-Israel Binational Agricultural Research and Development Fund (BARD grant no. IS-4652-13 to JH and GM).

FundersFunder number
National Science Foundation
U.S. Department of AgricultureNEV00384
United States - Israel Binational Agricultural Research and Development Fund
Illinois Ornithology Society1656774, 2016143
BARDIS-4652-13

    Keywords

    • calcium
    • heat stress
    • mCherry fused GCaMP6f
    • pollen tube imaging
    • ratiometric calcium reporter CGf
    • single cell imaging
    • whole rosette imaging

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