Skip to main content

Measuring Circadian Oscillations of Cytosolic-Free Calcium in Arabidopsis thaliana

  • Protocol
  • First Online:
Plant Circadian Networks

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1158))

Abstract

Circadian oscillations of cytosolic-free calcium can be measured in plants by observing luminescence of the bioluminescent calcium binding protein aequorin. Here we describe the use of intensified photon-counting CCD cameras to measure circadian oscillations in aequorin bioluminescence from Arabidopsis thaliana.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Subscribe and save

Springer+ Basic
$34.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or eBook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 89.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 119.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

Similar content being viewed by others

References

  1. Dodd AN, Salathia N, Hall A et al (2005) Plant circadian clocks increase photosynthesis, growth, survival, and competitive advantage. Science 309:630–633

    Article  CAS  PubMed  Google Scholar 

  2. Johnson CH, Knight MR, Kondo T et al (1995) Circadian oscillations of cytosolic and chloroplastic free calcium in plants. Science 269:1863–1865

    Article  CAS  PubMed  Google Scholar 

  3. Dalchau N, Hubbard KE, Robertson FC et al (2010) Correct biological timing in Arabidopsis requires multiple light-signaling pathways. Proc Natl Acad Sci U S A 107:13171–13176

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Xu X, Hotta CT, Dodd AN et al (2007) Distinct light and clock modulation of cytosolic free Ca2+ oscillations and rhythmic CHLOROPHYLL A/B BINDING PROTEIN2 promoter activity in Arabidopsis. Plant cell 19:3474–3490

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  5. Dodd AN, Gardner MJ, Hotta CT et al (2007) The Arabidopsis circadian clock incorporates a cADPR-based feedback loop. Science 318:1789–1792

    Article  CAS  PubMed  Google Scholar 

  6. Sai J, Johnson CH (1999) Different circadian oscillators control Ca2+ fluxes and lhcb gene expression. Proc Natl Acad Sci U S A 96:11659–11663

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  7. Dodd AN, Kudla J, Sanders D (2010) The language of calcium signaling. Annu Rev Plant Biol 61:593–620

    Article  CAS  PubMed  Google Scholar 

  8. McAinsh M, Ng CKY (2005) Measurement of cytosolic-free Ca2+ in plant tissue. In: Lambert DG (ed) Calcium signalling protocols, methods in molecular biology. Humana, New York, pp 289–302

    Chapter  Google Scholar 

  9. Knight MR, Campbell AK, Smith SM, Trewavas AJ (1991) Transgenic plant aequorin reports the effects of touch and cold-shock and elicitors on cytoplasmic calcium. Nature 352:524–526

    Article  CAS  PubMed  Google Scholar 

  10. Gould PD, Diaz P, Hogben C et al (2009) Delayed fluorescence as a universal tool for the measurement of circadian rhythms in higher plants. Pant J 58:893–901

    CAS  Google Scholar 

  11. Plautz JD, Straume M, Stanewsky R et al (1997) Quantitative analysis of Drosophila period gene transcription in living animals. J Biol Rhythms 12:204–217

    Article  CAS  PubMed  Google Scholar 

  12. Love J, Dodd AN, Webb AAR (2004) Circadian and diurnal calcium oscillations encode photoperiodic information in Arabidopsis. Plant Cell 16:956–966

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  13. Haydon MJ, Bell LJ, Webb AAR (2011) Interactions between plant circadian clocks and solute transport. J Exp Bot 62:2333–2348

    Article  CAS  PubMed  Google Scholar 

  14. Cobbold PH, Rink TJ (1987) Fluorescence and bioluminescence measurement of cytoplasmic free calcium. Biochem J 248:313–328

    CAS  PubMed Central  PubMed  Google Scholar 

  15. Fricker MD, Plieth C, Knight H et al (1999) Fluorescence and luminescence techniques to probe ion activities in living plant cells. In: Mason WT (ed) Fluorescent and luminescent probes for biological activity, 2nd edn. Academic Press, San Diego, CA, pp 569–596

    Chapter  Google Scholar 

  16. Tracy FE, Gilliham M, Dodd AN et al (2008) NaCl-induced changes in cytosolic free Ca2+ in Arabidopsis thaliana are heterogeneous and modified by external ionic composition. Plant Cell Environ 31:1063–1073

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Alex A. R. Webb .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2014 Springer Science+Business Media New York

About this protocol

Cite this protocol

Hearn, T.J., Webb, A.A.R. (2014). Measuring Circadian Oscillations of Cytosolic-Free Calcium in Arabidopsis thaliana . In: Staiger, D. (eds) Plant Circadian Networks. Methods in Molecular Biology, vol 1158. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-0700-7_15

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-0700-7_15

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-0699-4

  • Online ISBN: 978-1-4939-0700-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics