Skip to main content
SpringerLink
Log in

Histochemical Localization of Phenolic Compounds and Reactive Oxygen Species in Eucalypt Microcuttings

  • Protocol
  • First Online:
Plant Secondary Metabolism Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2469))

  • 693 Accesses

Abstract

Among the main features of plant specialized metabolism are cell- and tissue- specific expression and responsiveness to oxidative stress conditions. Although quantitative techniques have significantly improved over time, allowing higher levels of resolution in plant metabolic studies, such analyses are often expensive and/or require relatively large amounts of starting material. The following protocols offer a relatively simple way to survey specialized and related metabolites (total phenolics, flavonoids, lignins), as well as reactive oxygen species (superoxide and hydrogen peroxide), using light and fluorescence microscopy. Also, a step-by-step guide on how to quantify stained areas is provided. Through the association of qualitative and quantitative data, general patterns of molecule distribution across plant tissues may be inferred, allowing for the solving of biological questions, and contributing to hypothesis refinement. Eucalyptus microcuttings were chosen as plant material to exemplify how these protocols can provide useful data for the understanding of complex developmental processes, such as adventitious root formation, which may be influenced by specialized metabolites and redox conditions.

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

Access this chapter

Log in via an institution

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 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
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. Wick MR (2012) Histochemistry as a tool in morphological analysis: a historical review. Ann Diagn Pathol 16:71–78. https://doi.org/10.1016/j.anndiagpath.2011.10.010

    Article  PubMed  Google Scholar 

  2. Coleman R (2000) The impact of histochemistry–a historical perspective. Acta Histochem 102:5–14. https://doi.org/10.1078/0065-1281-00542

    Article  CAS  PubMed  Google Scholar 

  3. Johansen DA (1940) Plant microtechnique. McGraw-Hill, London

    Google Scholar 

  4. Matsuura HN, Malik S, de Costa F et al (2018) Specialized plant metabolism characteristics and impact on target molecule biotechnological production. Mol Biotechnol 60:169–183. https://doi.org/10.1007/s12033-017-0056-1

    Article  CAS  PubMed  Google Scholar 

  5. Nützmann HW, Scazzocchio C, Osbourn A (2018) Metabolic gene clusters in eukaryotes. Annu Rev Genet 52:159–183. https://doi.org/10.1146/annurev-genet-120417-031237

    Article  CAS  PubMed  Google Scholar 

  6. Lattanzio V (2013) Phenolic compounds: introduction. In: Ramawat K, Mérillon JM (eds) Natural products. Springer, Berlin. https://doi.org/10.1007/978-3-642-22144-6_57

    Chapter  Google Scholar 

  7. Pereira DM, Valentão P, Pereira JA et al (2009) Phenolics: from chemistry to biology. Molecules 14:2202–2211. https://doi.org/10.3390/molecules14062202

    Article  CAS  PubMed Central  Google Scholar 

  8. Mittler R, Vanderauwera S, Gollery M et al (2004) Reactive oxygen gene network of plants. Trends Plant Sci 9:490–498. https://doi.org/10.1016/j.tplants.2004.08.009

    Article  CAS  PubMed  Google Scholar 

  9. Mittler R, Vanderauwera S, Suzuki N et al (2011) ROS signaling: the new wave? Trends Plant Sci 16:300–309. https://doi.org/10.1016/j.tplants.2011.03.007

    Article  CAS  PubMed  Google Scholar 

  10. Hidalgo GI, Almajano MP (2017) Red fruits: extraction of antioxidants, phenolic content, and radical scavenging determination: a review. Antioxidants (Basel) 6:7. https://doi.org/10.3390/antiox6010007

    Article  CAS  Google Scholar 

  11. Matsuura HN, Rau MR, Fett-Neto AG (2014) Oxidative stress and production of bioactive monoterpene indole alkaloids: biotechnological implications. Biotechnol Lett 36:191–200. https://doi.org/10.1007/s10529-013-1348-6

    Article  CAS  PubMed  Google Scholar 

  12. McDowell EM, Trump BF (1976) Histologic fixatives suitable for diagnostic light and electron microscopy. Arch Pathol Lab Med 100(8):405–414

    CAS  PubMed  Google Scholar 

  13. Peer WA, Brown D, Tague B et al (2001) Flavonoid accumulation patterns of transparent testa mutants of Arabidopsis. Plant Physiol 126(2):536–548. https://doi.org/10.1104/pp.126.2.536

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  14. Romero-Puertas MC, Rodriguez-Serrano M, Corpas FJ et al (2004) Cadmium-induced subcellular accumulation of O2•– and H2O2 in pea leaves. Plant Cell Environ 27:1122–1134. https://doi.org/10.1111/j.1365-3040.2004.01217.x

    Article  CAS  Google Scholar 

  15. Vilasboa J, da Costa CT, Ransan LG et al (2021) Microcutting redox profile and anatomy in Eucalyptus spp. with distinct adventitious rooting competence. Front. Plant Sci 11:620832. https://doi.org/10.3389/fpls.2020.620832

    Article  Google Scholar 

Download references

Author information

Author notes

  1. Johnatan Vilasboa and Cibele Tesser da Costa contributed equally to this work.

Authors and Affiliations

  1. Plant Physiology Laboratory, Center for Biotechnology and Department of Botany, Federal University of Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil

    Johnatan Vilasboa & Cibele Tesser Da Costa

  2. Plant Anatomy Laboratory, Department of Botany, Federal University of Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brazil

    Jorge Ernesto de Araújo Mariath

  3. Center for Biotechnology and Department of Botany, Plant Physiology Laboratory, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, Brazil

    Arthur Germano Fett-Neto

Authors
  1. Johnatan Vilasboa
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Cibele Tesser Da Costa
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jorge Ernesto de Araújo Mariath
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Arthur Germano Fett-Neto
    View author publications

    You can also search for this author in PubMed Google Scholar

Editor information

Editors and Affiliations

  1. Center for Biotechnology and Department of Botany, Plant Physiology Laboratory, Federal University of Rio Grande do Sul (UFRGS), Porto Alegre, Brazil

    Arthur Germano Fett-Neto

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

Vilasboa, J., Da Costa, C.T., Mariath, J.E.d.A., Fett-Neto, A.G. (2022). Histochemical Localization of Phenolic Compounds and Reactive Oxygen Species in Eucalypt Microcuttings. In: Fett-Neto, A.G. (eds) Plant Secondary Metabolism Engineering. Methods in Molecular Biology, vol 2469. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2185-1_3

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-2185-1_3

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2184-4

  • Online ISBN: 978-1-0716-2185-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation