Skip to main content

Cell Proliferation Analysis Using EdU Labeling in Whole Plant and Histological Samples of Arabidopsis

  • Protocol
Plant Cell Division

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1370))

Abstract

The ability to analyze cell division in both spatial and temporal dimensions within an organism is a key requirement in developmental biology. Specialized cell types within individual organs, such as those within shoot and root apical meristems, have often been identified by differences in their rates of proliferation prior to the characterization of distinguishing molecular markers. Replication-dependent labeling of DNA is a widely used method for assaying cell proliferation. The earliest approaches used radioactive labeling with tritiated thymidine, which were later followed by immunodetection of bromodeoxyuridine (BrdU). A major advance in DNA labeling came with the use of 5-ethynyl-2′deoxyuridine (EdU) which has proven to have multiple advantages over BrdU. Here we describe the methodology for analyzing EdU labeling and retention in whole plants and histological sections of Arabidopsis.

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

Access this chapter

eBook
USD 39.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 54.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 54.99
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. Clowes FAL (1956) Localization of nucleic acid synthesis in root meristems. J Exp Bot 7:307–312

    Article  CAS  Google Scholar 

  2. Clowes FAL (1956) Nucleic acids in root apical meristems of Zea. New Phytol 55:29–34

    Article  CAS  Google Scholar 

  3. Booth A, Cutter EG, Postlethwait SN, Voeller BR, Wareing PF (1959) Use of radioactive tracers in studies of the shoot apex. Nature 183:1580–1582

    Article  Google Scholar 

  4. Gratzner HG (1982) Monoclonal antibody to 5-bromo- and 5-iododeoxyuridine: a new reagent for detection of DNA replication. Science 218:474–475

    Article  CAS  PubMed  Google Scholar 

  5. Gratzner HG, Leif RC, Ingram DJ, Castro A (1975) Use of antibody specific for bromodeoxyuridine for immunofluorescent determination of DNA-replication in single cells and chromosomes. Exp Cell Res 95:88–94

    Article  CAS  PubMed  Google Scholar 

  6. Salic A, Mitchison TJ (2008) A chemical method for fast and sensitive detection of DNA synthesis in vivo. Proc Natl Acad Sci U S A 105:2415–2420

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  7. Kotogany E, Dudits D, Horvath GV, Ayaydin F (2010) A rapid and robust assay for detection of S-phase cell cycle progression in plant cells and tissues by using ethynyl deoxyuridine. Plant Methods 6:5

    Article  PubMed Central  PubMed  Google Scholar 

  8. Perilli S, Perez-Perez JM, Di Mambro R, Peris CL, Diaz-Trivino S, Del Bianco M, Pierdonati E, Moubayidin L, Cruz-Ramirez A, Costantino P, Scheres B, Sabatini S (2013) RETINOBLASTOMA-RELATED protein stimulates cell differentiation in the Arabidopsis root meristem by interacting with cytokinin signaling. Plant Cell 25:4469–4478

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  9. Vanstraelen M, Baloban M, Da Ines O, Cultrone A, Lammens T, Boudolf V, Brown SC, De Veylder L, Mergaert P, Kondorosi E (2009) APC/C-CCS52A complexes control meristem maintenance in the Arabidopsis root. Proc Natl Acad Sci U S A 106:11806–11811

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  10. Schubert I, Schubert V, Fuchs J (2011) No evidence for “break-induced replication” in a higher plant - but break-induced conversion may occur. Front Plant Sci 2:8

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  11. She W, Grimanelli D, Rutowicz K, Whitehead MW, Puzio M, Kotlinski M, Jerzmanowski A, Baroux C (2013) Chromatin reprogramming during the somatic-to-reproductive cell fate transition in plants. Development 140:4008–4019

    Article  CAS  PubMed  Google Scholar 

  12. Zhu Y, Weng M, Yang Y, Zhang C, Li Z, Shen WH, Dong A (2011) Arabidopsis homologues of the histone chaperone ASF1 are crucial for chromatin replication and cell proliferation in plant development. Plant J 66:443–455

    Article  CAS  PubMed  Google Scholar 

  13. Ichihashi Y, Kawade K, Usami T, Horiguchi G, Takahashi T, Tsukaya H (2011) Key proliferative activity in the junction between the leaf blade and leaf petiole of Arabidopsis. Plant Physiol 157:1151–1162

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  14. Kakar K, Zhang H, Scheres B, Dhonukshe P (2013) CLASP-mediated cortical microtubule organization guides PIN polarization axis. Nature 495:529–533

    Article  CAS  PubMed  Google Scholar 

  15. Kelliher T, Walbot V (2011) Emergence and patterning of the five cell types of the Zea mays anther locule. Dev Biol 350:32–49

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  16. Xiong Y, McCormack M, Li L, Hall Q, Xiang C, Sheen J (2013) Glucose-TOR signalling reprograms the transcriptome and activates meristems. Nature 496:181–186

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  17. Xu D, Huang W, Li Y, Wang H, Huang H, Cui X (2012) Elongator complex is critical for cell cycle progression and leaf patterning in Arabidopsis. Plant J 69:792–808

    Article  CAS  PubMed  Google Scholar 

  18. Neef AB, Luedtke NW (2011) Dynamic metabolic labeling of DNA in vivo with arabinosyl nucleosides. Proc Natl Acad Sci U S A 108:20404–20409

    Article  PubMed Central  CAS  PubMed  Google Scholar 

  19. Cruz-Ramirez A, Diaz-Trivino S, Wachsman G, Du Y, Arteaga-Vazquez M, Zhang H, Benjamins R, Blilou I, Neef AB, Chandler V, Scheres B (2013) A SCARECROW-RETINOBLASTOMA protein network controls protective quiescence in the Arabidopsis root stem cell organizer. PLoS Biol 11:e1001724

    Article  PubMed Central  PubMed  Google Scholar 

  20. Kerk NM, Ceserani T, Tausta SL, Sussex IM, Nelson TM (2003) Laser capture microdissection of cells from plant tissues. Plant Physiol 132:27–35

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to J. Matthew Watson or Karel Riha .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2016 Springer Science+Business Media New York

About this protocol

Cite this protocol

Kazda, A., Akimcheva, S., Watson, J.M., Riha, K. (2016). Cell Proliferation Analysis Using EdU Labeling in Whole Plant and Histological Samples of Arabidopsis. In: Caillaud, MC. (eds) Plant Cell Division. Methods in Molecular Biology, vol 1370. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3142-2_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-3142-2_13

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3141-5

  • Online ISBN: 978-1-4939-3142-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics