In the last decade, the view of circadian oscillators has expanded from transcriptional feedback to incorporate post-transcriptional, post-translational, metabolic processes and ionic signalling. In plants and animals, there are circadian oscillations in the concentration of cytosolic free Ca2+ ([Ca2+]cyt), though their purpose has not been fully characterized. We investigated whether circadian oscillations of [Ca2+]cyt regulate the circadian oscillator of Arabidopsis thaliana. We report that in Arabidopsis, [Ca2+]cyt circadian oscillations can regulate circadian clock function through the Ca2+-dependent action of CALMODULIN-LIKE24 (CML24). Genetic analyses demonstrate a linkage between CML24 and the circadian oscillator, through pathways involving the circadian oscillator gene TIMING OF CAB2 EXPRESSION1 (TOC1).
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Supported by BBSRC UK research grants BBSRC BB/D010381/1 (A.N.D.), BB/D017904/1 (F.R.) BB/M00113X/1 (H.J.H.) awarded to (A.A.R.W.), Research Studentship (K.H.) and BBSRC Industrial Case (T.H.). A Swiss Science Foundation Award (PBZHP3-123289) and the Isaac Newton Trust Cambridge (M.C.M.R. and S.A.), the National Science Foundation under Grant No. MCB 0817976 (Y-C.T. and J.B.), a Royal Society Grant RG081257 and Corpus Christi College, Cambridge Junior Research Fellowship (M.J.G.), a Cordenadoria de Apoio ao Ensino Superior Brazil studentship (C.T.H.), IEF Marie Curie (Project No. 272186) (M.C.M.R.), a Broodbank Fellowship (M.C.M.R.), a Malaysian Government Studentship (N.I.M-H.). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript. The authors are very grateful to the unnamed laboratories who provided (un)published material for the screen.
The authors declare no competing interests.
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Supplementary Table 1, Supplementary Methods, Supplementary Statistical Parameters, Supplementary References and Supplementary Figures 1–6
Reverse genetic screen of Ca2+ -signalling related mutants.
Circadian period estimates of leaf movement for characterization of the genetic relationship between CML23/CML24 and the clock genes. Relative to Figure 5, 6 and Supplementary Figure 4.
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Martí Ruiz, M.C., Hubbard, K.E., Gardner, M.J. et al. Circadian oscillations of cytosolic free calcium regulate the Arabidopsis circadian clock. Nature Plants 4, 690–698 (2018). https://doi.org/10.1038/s41477-018-0224-8
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