Skip to main content

Long-Term Cryopreservation of Thylakoid Membranes

  • Protocol
Cryopreservation and Freeze-Drying Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 38))

  • 1438 Accesses

Abstract

Chloroplast thylakoids are the site of photosynthetic light reactions, electron transport, adenosine triphosphate (ATP) synthesis and nicotinamide adenine dinucleotide phosphate (NADP) reduction in plant cells. Isolated thylakoids are therefore the necessary prerequisite for many in vitro studies of photosynthetic mechanisms. They are also used to elucidate aspects of photosynthetic responses to freezing, chilling, heat, and high light stress. For many of these studies it is important to work with membranes of closely similar properties over extended periods of time to facilitate direct comparisons of the results. The pigment, protein, and lipid composition of thylakoid membranes, however, vary with plant growth conditions (13). Even the solute permeability properties of thylakoids are significantly different between warm-and cold-grown spinach plants (4).

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 79.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Webb, M. S. and Green, B R. (1991) Biochemical and biophysical properties of thylakoid acyl lipids. Biochim Biophys. Acta 1060, 133–158

    Article  CAS  Google Scholar 

  2. Demmig-Adams, B. and Adams, W. W., III (1992) Photoprotection and other responses of plants to high light stress. Annu. Rev. Plant Physiol Plant Mol. Biol. 43, 599–626.

    Article  CAS  Google Scholar 

  3. Anderson, J. M. (1986) Photoregulation of the composition, function, and structure of thylakoid membranes. Annu. Rev. Plant Physiol 37, 93–136.

    Article  CAS  Google Scholar 

  4. Hincha, D. K. and Schmitt, J. M. (1988) Mechanical freeze-thaw damage and frost hardening in leaves and isolated thylakoids from spinach. II. Frost hardening reduces solute permeability and increases extensrbihty of thylakoid membranes. Plant Cell Environ. 11, 47–50.

    Article  CAS  Google Scholar 

  5. Hincha, D. K. and Schmitt, J. M. (1992) Freeze-thaw injury and cryoprotection of thylakoid membranes, in Water and Life (Somero, G. N., Osmond, C. B., and Bolis, C. L., eds.), Springer, Berlin, pp. 316–337.

    Chapter  Google Scholar 

  6. Hincha, D. K., Heber, U., and Schmitt, J. M. (1985) Antibodies against individual thylakoid membrane proteins as molecular probes to study chemical and mechanical freezing damage in vitro Biochim. Biophys. Acta 809, 337–344.

    CAS  Google Scholar 

  7. Hincha, D. K (1986) Sucrose influx and mechanical damage by osmotic stress to thylakoid membranes during an in vitro freeze-thaw cycle. Biochim. Biophys. Acta 861, 152–158.

    CAS  Google Scholar 

  8. Bakaltcheva, I., Schmitt, J. M., and Hincha, D. K. (1992) Time and temperature dependent solute loading of isolated thylakoids during freezing. Cryobiology 29, 607–615.

    Article  CAS  Google Scholar 

  9. Lineberger, R. D and Steponkus, P. L. (1980) Cryoprotection by glucose, sucrose, and raffinose to chloroplast thylakoids. Plant Physiol. 65, 298–304.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  10. Santarius, K. A. and Bauer, J. (1983) Cryopreservation of spinach chloroplast membranes by low-molecular-weight carbohydrates. I. Evidence for cryoprotection by a noncolligative-type mechanism. Cryobiology 20, 83–89.

    Article  CAS  PubMed  Google Scholar 

  11. Santarius, K. A. and Giersch, C. (1983) Cryopreservation of spinach chloroplast membranes by low-molecular-weight carbohydrates. II. Discrimination between colligative and noncolligative protection. Cryobiology 20, 90–99.

    Article  CAS  PubMed  Google Scholar 

  12. Steponkus, P. L., Garber, M. P., Myers, S. P., and Lineberger, D. R. (1977) Effects of cold acclimation and freezing on structure and function of chloroplast thylakoids. Cryobiology 14, 303–321

    Article  CAS  PubMed  Google Scholar 

  13. Hincha, D K. (1989) Low concentrations of trehalose protect isolated thylakoids against mechanical freeze-thaw damage. Biochim. Biophys. Acta 987, 231–234.

    Article  CAS  Google Scholar 

  14. Hincha, D. K. (1990) Differential effects of galactose containing saccharides on mechanical freeze-thaw damage to isolated thylakoid membranes. Cryo-Lett. 11, 437–444.

    Google Scholar 

  15. Bakaltcheva, I., Williams, W. P., Schmitt, J. M., and Hincha, D. K. (1994) The solute permeability of thylakoid membranes is reduced by low concentrations of trehalose as a cosolute. Biochim. Biophys. Acta. 1189, 38–44.

    Article  CAS  PubMed  Google Scholar 

  16. Wolter, F. P., Schmitt, J. M., Bohnert, H. J., and Tsugita, A. (1984) Simultaneous isolation of three peripheral proteins/3-a 32 kDa protein, ferredoxin NADP+ reductase and coupling factor/3-from spinach thylakoids and partial characterization of a 32 kDa protein. Plant Sci. Lett. 34, 323–334.

    Article  CAS  Google Scholar 

  17. Mollenhauer, A., Schmitt, J. M., Coughlan, S., and Heber, U. (1983) Loss of membrane proteins from thylakoids during freezing. Biochim. Biophys. Acta 728, 331–338.

    Article  CAS  Google Scholar 

  18. Arnon, D. J. (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol. 24, 1–15.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  19. Hincha, D. K. and Schmitt, J. M. (1985) Mechanical and chemical injury to thylakoid membranes during freezing in vitro. Biochim. Biophys. Acta 812, 173–180.

    Article  CAS  Google Scholar 

  20. Bencini, A. D., Shanley, M. S., Wild, J. R., and O’Donovan, G. A (1983) New assay for enzymatic phosphate release: application to aspartate transcarbamylase and other enzymes. Anal. Biochem. 132, 259–264.

    Article  CAS  PubMed  Google Scholar 

  21. Manson, M. M. (ed.) (1992) Methods in Molecular Biology, vol. 10: Immunochemical Protocols Humana, Totowa, NJ.

    Google Scholar 

  22. Mancini, G., Carbonara, A. O., and Heremans, J. F. (1965) Immunochemical quantitation of antigens by single radial immunodrffusion. Immunochemistry 2, 235–254.

    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

© 1995 Humana Press Inc.

About this protocol

Cite this protocol

K, D., M., J. (1995). Long-Term Cryopreservation of Thylakoid Membranes. In: Day, J.G., Pennington, M.W. (eds) Cryopreservation and Freeze-Drying Protocols. Methods in Molecular Biology™, vol 38. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-296-5:71

Download citation

  • DOI: https://doi.org/10.1385/0-89603-296-5:71

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-296-5

  • Online ISBN: 978-1-59259-525-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics