Skip to main content
SpringerLink
Log in
Book cover

Plant Proteases and Plant Cell Death pp 185–192Cite as

Measuring and Perturbing Ferroptosis in Plants

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2447))

Abstract

Ferroptosis is an oxidative iron-dependent cell death that was recently described in vertebrates, invertebrates, fungi, plants, and bacteria. In plants, ferroptosis has been reported in response to heat shock in roots of 6-day-old Arabidopsis thaliana seedlings. Generally, all biochemical and morphological ferroptosis hallmarks are conserved between animals and plants. Here, we describe a protocol to induce and quantify ferroptosis in plants based on the analysis of dead cells with a Sytox Green stain. Furthermore, heat shock induced cell death is prevented by using specific ferroptosis inhibitors.

Key words

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

Protocol
EUR   44.95
Price includes VAT (Finland)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
EUR   99.99
Price includes VAT (Finland)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
EUR   131.99
Price includes VAT (Finland)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
EUR   175.99
Price includes VAT (Finland)
  • 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

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Dixon SJ, Lemberg KM, Lamprecht MR et al (2012) Ferroptosis: an iron-dependent form of non-apoptotic cell death. Cell 149:1060–1072. https://doi.org/10.1016/j.cell.2012.03.042

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Han C, Liu Y, Dai R et al (2020) Ferroptosis and its potential role in human diseases. Front Pharmacol 11:239. https://doi.org/10.3389/fphar.2020.00239

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Conrad M, Kagan VE, Bayir H et al (2018) Regulation of lipid peroxidation and ferroptosis in diverse species. Genes Dev 32:602–619. https://doi.org/10.1101/gad.314674.118

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  4. Shen Q, Liang M, Yang F et al (2020) Ferroptosis contributes to developmental cell death in rice blast. New Phytol 227(6):1831–1846. https://doi.org/10.1111/nph.16636

    Article  CAS  PubMed  Google Scholar 

  5. Distéfano AM, López GA, Setzes N et al (2020) Ferroptosis in plants: triggers, proposed mechanisms and the role of iron in modulating cell death. J Exp Bot 72(6):2125–2135. https://doi.org/10.1093/jxb/eraa425

    Article  CAS  Google Scholar 

  6. Shen X, Ma R, Huang Y et al (2020) Nano-decocted ferrous polysulfide coordinates ferroptosis-like death in bacteria for anti-infection therapy. Nano Today 35:100981. https://doi.org/10.1016/j.nantod.2020.100981

    Article  CAS  Google Scholar 

  7. Stockwell BR, Jiang X (2020) The chemistry and biology of ferroptosis. Cell Chem Biol 27:365–375. https://doi.org/10.1016/j.chembiol.2020.03.013

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Friedmann Angeli JP, Schneider M, Proneth B et al (2014) Inactivation of the ferroptosis regulator Gpx4 triggers acute renal failure in mice. Nat Cell Biol 16:1180–1191. https://doi.org/10.1038/ncb3064

    Article  CAS  PubMed  Google Scholar 

  9. Kagan VE, Mao G, Qu F et al (2017) Oxidized arachidonic/adrenic phosphatidylethanolamines navigate cells to ferroptosis. Nat Chem Biol 13:81–90. https://doi.org/10.1038/nchembio.2238

    Article  CAS  PubMed  Google Scholar 

  10. Stoyanovsky DA, Tyurina YY, Shrivastava I et al (2019) Iron catalysis of lipid peroxidation in ferroptosis: regulated enzymatic or random free radical reaction? Free Radic Biol Med 133:153–161. https://doi.org/10.1016/j.freeradbiomed.2018.09.008

    Article  CAS  PubMed  Google Scholar 

  11. Distéfano AM, Martin MV, Córdoba JP et al (2017) Heat stress induces ferroptosis-like cell death in plants. J Cell Biol 216:463–476. https://doi.org/10.1083/jcb.201605110

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  12. Hajdinák P, Czobor Á, Szarka A (2019) The potential role of acrolein in plant ferroptosis-like cell death. PLoS One 14:e0227278. https://doi.org/10.1371/journal.pone.0227278

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Yang WS, Sri Ramaratnam R, Welsch ME et al (2014) Regulation of ferroptotic cancer cell death by GPX4. Cell 156:317–331. https://doi.org/10.1016/j.cell.2013.12.010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Dangol S, Chen Y, Hwang BK, Jwa N-S (2019) Iron- and reactive oxygen species-dependent ferroptotic cell death in rice-magnaportheoryzae interactions. Plant Cell 31:189–209. https://doi.org/10.1105/tpc.18.00535

    Article  CAS  PubMed  Google Scholar 

  15. Macharia M, Das PP, Naqvi NI, Wong S-M (2020) iTRAQ-based quantitative proteomics reveals a ferroptosis-like programmed cell death in plants infected by a highly virulent tobacco mosaic virus mutant 24A+UPD. Phytopathol Res 2:1. https://doi.org/10.1186/s42483-019-0043-5

    Article  Google Scholar 

  16. Tsuboyama S, Nonaka S, Ezura H, Kodama Y (2018) Improved G-AgarTrap: a highly efficient transformation method for intact gemmalings of the liverwort Marchantiapolymorpha. Sci Rep 8:10800. https://doi.org/10.1038/s41598-018-28947-0

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Müller M, Schmidt W (2004) Environmentally induced plasticity of root hair development in Arabidopsis. Plant Physiol 134:409–419. https://doi.org/10.1104/pp.103.029066

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Vissenberg K, Claeijs N, Balcerowicz D, Schoenaers S (2020) Hormonal regulation of root hair growth and responses to the environment in Arabidopsis. J Exp Bot 71:2412–2427. https://doi.org/10.1093/jxb/eraa048

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Kacprzyk J, McCabe PF (2015) A root hair assay to expedite cell death research. Methods Mol Biol Clifton NJ 1242:73–82. https://doi.org/10.1007/978-1-4939-1902-4_7

    Article  CAS  Google Scholar 

  20. Vacca RA, Valenti D, Bobba A et al (2006) Cytochrome C is released in a reactive oxygen species-dependent manner and is degraded via caspase-like proteases in tobacco bright-yellow 2 cells en route to heat shock-induced cell death. Plant Physiol 141:208–219. https://doi.org/10.1104/pp.106.078683

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Marsoni M, Cantara C, De Pinto MC et al (2010) Exploring the soluble proteome of tobacco bright yellow-2 cells at the switch towards different cell fates in response to heat shocks. Plant Cell Environ 33:1161–1175. https://doi.org/10.1111/j.1365-3040.2010.02137.x

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We acknowledge the International Centre for Genetic Engineering and Biotechnology (ICGEB) and The Argentinean Agency for the promotion of Science and Technology (ANPCyT) for financial support (ICGB-CRP/19/020 grant to GCP; ANPCyT PICTs 2017-00201 and 2016-00110 to GCP; PICT 2016-00382 to AMD). FM is ANPCyT Post-Doctoral fellow; AMD, EZ, and GCP are CONICET researchers.

Contribution agreement: Conceptualization: AMD, FM, EZ, and GCP; Funding Acquisition: GCP, EZ, and AMD; Project Administration: GCP and AMD; Supervision: AMD and GCP; Writing: AMD, FM, and GCP; Writing—Review & Editing: AMD, FM, EZ, and GCP.

Author information

Authors and Affiliations

  1. Instituto de Investigaciones Biológicas, Universidad Nacional de Mar del Plata, CONICET, Mar del Plata, Argentina

    Ayelen M. Distéfano, Fernanda Marchetti, Eduardo Zabaleta & Gabriela C. Pagnussat

Authors
  1. Ayelen M. Distéfano
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Fernanda Marchetti
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Eduardo Zabaleta
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Gabriela C. Pagnussat
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Ayelen M. Distéfano .

Editor information

Editors and Affiliations

  1. Chemistry and Chemical Technology, University of Ljubljana, Ljubljana, Slovenia

    Marina Klemenčič

  2. VIB-UGent Cntr for Plant Systems Biology, Ghent University, Ghent, Belgium

    Simon Stael

  3. ZEA-3 Analytik, Forschungszentrum Juelich, Juelich, Germany

    Pitter F. Huesgen

Rights and permissions

Reprints and permissions

Copyright information

© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Distéfano, A.M., Marchetti, F., Zabaleta, E., Pagnussat, G.C. (2022). Measuring and Perturbing Ferroptosis in Plants. In: Klemenčič, M., Stael, S., Huesgen, P.F. (eds) Plant Proteases and Plant Cell Death. Methods in Molecular Biology, vol 2447. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2079-3_15

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-2079-3_15

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2078-6

  • Online ISBN: 978-1-0716-2079-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation