Protection of Thylakoid Membranes from Freeze-Thaw Damage by Proteins

  • Dirk K. Hincha
  • Frank Sieg
  • Jürgen M. Schmitt


The degree of frost hardiness differs vastly between different plant species, from around −1.5°C in tender plants such as tobacco (Hincha et al., 1996b) to the temperature of liquid nitrogen (−196°C) in some extremely hardy trees and shrubs. In addition, most plants from temperate climates follow an annual cycle of hardening and dehardening, with the maximum frost hardiness in the winter and the minimum during summer. In herbaceous plants, hardening/dehardening is triggered by growth temperature. Hardening occurs under low, non-freezing temperatures, usually in the range between 10 and 0°C (cold acclimation) over several days to weeks.


Thylakoid Membrane Cold Acclimation Frost Hardiness Frost Hardening Unfrozen Control 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Bakaltcheva I, Schmitt JM, Hincha DK (1992) Time and temperature dependent solute loading of isolated thy-lakoids during freezing. Cryobiology 29: 607–615CrossRefGoogle Scholar
  2. Block MA, Dorne A-J, Joyard J, Douce R (1983) Preparation and characterization of membrane fractions enriched in outer and inner envelope membranes from spinach chloroplasts. II. Biochemical characterization. J. Biol. Chem. 258: 13281–13286PubMedGoogle Scholar
  3. Dwek RA, Edge CJ, Harvey DJ, Wormald MR (1993) Analysis of glycoprotein-associated oligosaccharides. Annu. Rev. Biochem. 62: 65–100PubMedCrossRefGoogle Scholar
  4. Etzler ME (1985) Plant lectins: molecular and biological aspects. Annu. Rev. Plant Physiol. 36: 209–234CrossRefGoogle Scholar
  5. Grafflage S, Krause GH (1986) Simulation of in situ freezing damage of the photosynthetic apparatus by freezing in vitro of thylakoids suspended in complex media. Planta 168: 67–76CrossRefGoogle Scholar
  6. Grant CWM, Peters MW (1984) Lectin-membrane interactions-information from model systems. Biochim. Biophys. Acta 779: 403–422PubMedCrossRefGoogle Scholar
  7. Griffith OH, Jost PC (1976) Lipid spin labeles in biological membranes. In: Berliner L J (ed) Spin labelling: Theory and applications. Academic Press, New York, pp 454–519Google Scholar
  8. Guy CL (1990) Cold acclimation and freezing stress tolerance: role of protein metabolism. Annu. Rev. Plant Physiol. Plant Mol. Biol. 41: 187–223CrossRefGoogle Scholar
  9. Guy CL, Niemi KJ, Brambl R (1985) Altered gene expression during cold acclimation of spinach. Proc. Natl. Acad. Sci. USA 82: 3673–3677PubMedCrossRefGoogle Scholar
  10. Haehnel W (1986) Plastocyanin. In: Staehelin L A, Arntzen C J (ed) Encyclopedia of plant physiology, New Series Vol 19. Springer, Berlin, pp 547–559Google Scholar
  11. Haschke H-P, Kaiser G, Martinoia E, Hammer U, Teucher T, Dorne AJ, Heinz E (1990) Lipid profiles of leaf tonoplasts from plants with different CO2-fixation mechanisms. Bot. Acta 103: 32–38Google Scholar
  12. Heber U, Kempfle M (1970) Proteine als Schutzstoffe gegenüber dem Gefriertod der Zelle. Z. Naturforsch. 25b: 834–842Google Scholar
  13. Hincha DK (1986) Sucrose influx and mechanical damage by osmotic stress to thylakoid membranes during an in vitro freeze-thaw cycle. Biochim. Biophys. Acta 861: 152–158Google Scholar
  14. Hincha DK, Bakaltcheva I, Schmitt JM (1993) Galactose-specific lectins protect isolated thylakoids against freeze-thaw damage. Plant Physiol. 103: 59–65PubMedGoogle Scholar
  15. Hincha DK, Bratt PJ, Williams WP (1997) A cryoprotective lectin reduces the solute permeability and lipid fluidity of thylakoid membranes, submittedGoogle Scholar
  16. Hincha DK, Heber U, Schmitt JM (1985) Antibodies against individual thylakoid membrane proteins as molecular probes to study chemical and mechanical freezing damage in vitro. Biochim. Biophys. Acta 809: 337–344CrossRefGoogle Scholar
  17. Hincha DK, Heber U, Schmitt JM (1989b) Freezing ruptures thylakoid membranes in leaves, and rupture can be prevented in vitro by cryoprotective proteins. Plant Physiol. Biochem. 27: 795–801Google Scholar
  18. Hincha DK, Heber U, Schmitt JM (1990) Proteins from frost-hardy leaves protect thylakoids against mechanical freeze-thaw damage in vitro. Planta 180: 416–419CrossRefGoogle Scholar
  19. Hincha DK, Höfner R, Schwab KB, Heber U, Schmitt JM (1987) Membrane rupture is the common cause of damage to chloroplast membranes in leaves injured by freezing or excessive wilting. Plant Physiol. 83: 251–253PubMedCrossRefGoogle Scholar
  20. Hincha DK, Müller M, Hillmann T, Schmitt JM (1989a) Osmotic stress causes mechanical freeze-thaw damage to thylakoids in vitro and in vivo. In: Cherry J H (ed) Environmental stress in plants. Springer, Berlin, pp 303–315CrossRefGoogle Scholar
  21. Hincha DK, Schmidt JE, Heber U, Schmitt JM (1984) Colligative and non-colligative freezing damage to thylakoid membranes. Biochim. Biophys. Acta 769: 8–14CrossRefGoogle Scholar
  22. Hincha DK, Schmitt JM (1988a) Mechanical freeze-thaw damage and frost hardening in leaves and isolated thylakoids from spinach. I. Mechanical freeze-thaw damage in an artificial stroma medium. Plant Cell Environ. 11:41–46CrossRefGoogle Scholar
  23. Hincha DK, Schmitt JM (1988b) Mechanical freeze-thaw damage and frost hardening in leaves and isolated thy-lakoids from spinach. II. Frost hardening reduces solute permeability and increases extensibility of thylak-oid membranes. Plant Cell Environ. 11: 47–50CrossRefGoogle Scholar
  24. Hincha DK, Schmitt JM (1992a) Freeze-thaw injury and cryoprotection of thylakoid membranes. In: Somero G N, Osmond C B, Bolis C L (ed) Water and life. Springer, Berlin, pp 316–337CrossRefGoogle Scholar
  25. Hincha DK, Schmitt JM (1992b) Cryoprotective leaf proteins: assay methods and heat stability. J. Plant Physiol. 140:236–240CrossRefGoogle Scholar
  26. Hincha DK, Sieg F, Bakaltcheva I, Köth H, Schmitt JM (1996a) Freeze-thaw damage to thylakoid membranes: specific protection by sugars and proteins. In: Steponkus P L (ed) Advances in low-temperature biology. JAI Press, London, pp 141–183CrossRefGoogle Scholar
  27. Hincha DK, Sonnewald U, Whlmitzer L, Schmitt JM (1996b) The role of sugar accumulation in leaf frost hardiness-investigations with transgenic tobacco expressing a bacterial pyrophosphatase or a yeast invertase gene. J. Plant Physiol. 147: 604–610CrossRefGoogle Scholar
  28. Kates M (1990) Handbook of lipid research. Vol. 6, Plenum Press, New York.Google Scholar
  29. Lelkes PI, Miller IR (1980) Perturbations of membrane structure by optical probes: I. Location and structural sensitivity of merocyanine 540 bound to phospholipid membranes. J. Membrane Biol. 52: 1–15CrossRefGoogle Scholar
  30. Lentz BR (1993) Use of fluorescent probes to monitor molecular order and motions within liposome bilayers. Chem. Phys. Lipids 64: 99–116PubMedCrossRefGoogle Scholar
  31. Levitt J (1980) Responses of plants to environmental stresses Vol. 1: Chilling, freezing, and high temperature stresses. Academic Press, OrlandoGoogle Scholar
  32. Lis H, Sharon N (1986) Lectins as molecules and as tools. Annu. Rev. Biochem. 55: 35–67PubMedCrossRefGoogle Scholar
  33. Loganathan D, Osborne SE, Glick GD, Goldstein IJ (1992) Synthesis of high-affinity, hydrophobic monosac-charide derivatives and study of their interaction with Concanavalin A, the pea, the lentil, and fava bean lectins. Arch. Biochem. Biophys. 299: 268–274PubMedCrossRefGoogle Scholar
  34. Lynch DV, Steponkus PL (1987) Plasma membrane lipid alterations associated with cold acclimation of winter rye seedlings (Secale cereale L. cv Puma). Plant Physiol. 83: 761–767PubMedCrossRefGoogle Scholar
  35. Quinn PJ (1982) The molecular biology of cell membranes. Macmillan Press, LondonGoogle Scholar
  36. Ramalingam TS, Das PK, Podder SK (1994) Ricin-membrane interaction: membrane penetration depth by fluorescence quenching and resonance energy transfer. Biochemistry 33: 12247–12254PubMedCrossRefGoogle Scholar
  37. Roberts DD, Goldstein IJ (1982) Hydrophobic binding properties of the lectin from lima beans (Phaseolus lu-natus). J. Biol. Chem. 257: 11274–11277PubMedGoogle Scholar
  38. Roberts DD, Goldstein IJ (1983) Binding of hydrophobic ligands to plant lectins: titration with arylaminonaph-talenesulfonates. Arch. Biochem. Biophys. 224: 479–484PubMedCrossRefGoogle Scholar
  39. Rosas A, Alberdi M, Delseny M, Meza-Basso L (1986) A cryoprotective polypeptide isolated from Nothofagus dombeyi seedlings. Phytochemistry 25: 2497–2500CrossRefGoogle Scholar
  40. Santarius KA (1986) Freezing of isolated thylakoid membranes in complex media II. Simulation of the conditions in the chloroplast stroma. Cryo-Lett. 7: 31–40Google Scholar
  41. Schmidt JE, Schmitt JM, Kaiser WM, Hincha DK (1986) Salt treatment induces frost hardiness in leaves and isolated thylakoids from spinach. Planta 168: 50–55CrossRefGoogle Scholar
  42. Sharon N (1993) Lectin-carbohydrate complexes of plants and animals: an atomic view. TIBS 18: 221–226PubMedGoogle Scholar
  43. Sieg F, Schröder W, Schmitt JM, Hincha DK (1996) Purification and characterization of a cryoprotective protein (cryoprotectin) from the leaves of cold-acclimated cabbage. Plant Physiol. 111: 215–221PubMedGoogle Scholar
  44. Thomashow MF (1990) Molecular genetics of cold acclimation in higher plants. In: Scandalios J G (ed) Genomic responses to environmental stress. Academic Press, San Diego, pp 99–131CrossRefGoogle Scholar
  45. Thomashow MF (1993) Genes induced during cold acclimation in higher plants. In: Steponkus P L (ed) Advances in low-temperature biology. JAI Press, London, England, pp 183–210Google Scholar
  46. Volger HG, Heber U (1975) Cryoprotective leaf proteins. Biochim. Biophys. Acta 412: 335–349PubMedCrossRefGoogle Scholar
  47. Webb MS, Green BR (1991) Biochemical and biophysical properties of thylakoid acyl lipids. Biochim. Biophys. Acta 1060: 133–158CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1997

Authors and Affiliations

  • Dirk K. Hincha
    • 1
  • Frank Sieg
    • 1
  • Jürgen M. Schmitt
    • 1
  1. 1.Institut für Pflanzenphysiologie und MikrobiologieFreie UniversitätBerlinGermany

Personalised recommendations