Skip to main content

Immunolocalization of Proteins in Somatic Embryos

Applications for Studies on the Cytoskeleton

  • Protocol
Plant Embryogenesis

Part of the book series: Methods In Molecular Biology™ ((MIMB,volume 427))

Summary

Plant embryogenesis requires a tight balance between cell proliferation and differentiation. In animals, embryogenesis is dependent on cell migrations, which is in contrast to plant embryogenesis where the rigid cell wall precludes migration. Therefore, plants have to position cells correctly by defining the direction of the division plane during proliferation and control cell shape by local cell expansion. Both these processes are reliant on the organization and dynamics of the cytoskeleton – actin filaments and microtubules. In previous work (7), we have shown that differentiation of the embryo suspensor is accompanied by reorientation of microtubules from random to transverse and reorganization of actin filaments from a fine filamentous network to bundled longitudinal cables. Here, we describe the technique for visualization of cytoskeletal components including actin filaments, microtubules and their associated proteins during the development of plant embryos in whole-mount specimens.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 109.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
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

References

  1. Hyams JS, Lloyd CW, eds. Microtubules. New-York: Wile-Liss, 1994.

    Google Scholar 

  2. Hussey PJ, ed. The Plant Cytoskeleton in Cell Differentiation and Development. Oxford: Blackwell Publishing, 2004.

    Google Scholar 

  3. Goddard RH, Wick SM, Silflow CD, Snustad PD. Microtubule components of the plant cell cytoskeleton. Plant Physiol 1994;104:1–6.

    CAS  PubMed  Google Scholar 

  4. Paredez AR, Somerville CR, Ehrhardt DW. Visualization of cellulose synthase demonstrates functional association with microtubules. Science 2006;312:1491–1495.

    Article  CAS  PubMed  Google Scholar 

  5. Hussey PJ, Ketelaar T, Deeks MJ. Control of the actin cytoskeleton in plant cell growth. Annu Rev Plant Biol 2006;57:109–125.

    Article  CAS  PubMed  Google Scholar 

  6. Ketelaar T, Anthony RG, Hussey PJ. Green fluorescent protein-mTalin causes defects in actin organization and cell expansion in Arabidopsis and inhibits actin depolymerizing factor′s actin depolymerizing activity in vitro. Plant Physiol 2004;136:3990–3998.

    Article  CAS  PubMed  Google Scholar 

  7. Smertenko AP, Bozhkov PV, Filonova LH, von Arnold S, Hussey PJ. Re-organisation of the cytoskeleton during developmental programmed cell death in Picea abies embryos. Plant J 2003;33:813–824.

    Article  CAS  PubMed  Google Scholar 

  8. Bozhkov PV, von Arnold S. Polyethylene glycol promotes maturation but inhibits further development of Picea abies somatic embryos. Physiol Plant 1998;104:211–224.

    Article  CAS  Google Scholar 

  9. von Arnold S, Eriksson T. A revised medium for growth of pea mesophyll protoplasts. Physiol Plant 1977;39:257–260.

    Article  Google Scholar 

  10. Laemmli UK. Cleavage of structural proteins during assembly of head of bacteriophage-T4. Nature 1970;227:680–685.

    Article  CAS  PubMed  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2008 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Smertenko, A.P., Hussey, P.J. (2008). Immunolocalization of Proteins in Somatic Embryos. In: Suárez, M.F., Bozhkov, P.V. (eds) Plant Embryogenesis. Methods In Molecular Biology™, vol 427. Humana Press. https://doi.org/10.1007/978-1-59745-273-1_13

Download citation

  • DOI: https://doi.org/10.1007/978-1-59745-273-1_13

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-931-4

  • Online ISBN: 978-1-59745-273-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics