Skip to main content
Plant Physiology logoLink to Plant Physiology
. 1997 Jul;114(3):1077–1083. doi: 10.1104/pp.114.3.1077

[beta]-1,3-Glucanase Is Cryoprotective in Vitro and Is Accumulated in Leaves during Cold Acclimation.

D K Hincha 1, F Meins Jr 1, J M Schmitt 1
PMCID: PMC158397  PMID: 12223761

Abstract

We have used isolated spinach (Spinacea oleracea L.) thylakoid membranes to investigate the possible cryoprotective properties of class I [beta]-1,3-glucanase (1,3-[beta]-D-glucan 3-glucanohydrolase; EC 3.2.1.39) and chitinase. Class I [beta]-1,3-glucanase that was purified from tobacco (Nicotiana tabacum L.) protected thylakoids against freeze-thaw injury in our in vitro assays, whereas class I chitinase from tobacco had no effect under the same conditions. The [beta]-1,3-glucanase acted by reducing the influx of solutes into the membrane vesicles during freezing and thereby reduced osmotic stress and vesicle rupture during thawing. Western blots probed with antibodies directed against tobacco class I [beta]-1,3-glucanase showed that in spinach and cabbage (Brassica oleracea L.) leaves an isoform of 41 kD was accumulated during frost hardening under natural conditions.

Full Text

The Full Text of this article is available as a PDF (1.9 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Arnon D. I. COPPER ENZYMES IN ISOLATED CHLOROPLASTS. POLYPHENOLOXIDASE IN BETA VULGARIS. Plant Physiol. 1949 Jan;24(1):1–15. doi: 10.1104/pp.24.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beffa R. S., Hofer R. M., Thomas M., Meins F., Jr Decreased Susceptibility to Viral Disease of [beta]-1,3-Glucanase-Deficient Plants Generated by Antisense Transformation. Plant Cell. 1996 Jun;8(6):1001–1011. doi: 10.1105/tpc.8.6.1001. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Beffa R. S., Neuhaus J. M., Meins F., Jr Physiological compensation in antisense transformants: specific induction of an "ersatz" glucan endo-1,3-beta-glucosidase in plants infected with necrotizing viruses. Proc Natl Acad Sci U S A. 1993 Oct 1;90(19):8792–8796. doi: 10.1073/pnas.90.19.8792. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Duman J. G. Purification and characterization of a thermal hysteresis protein from a plant, the bittersweet nightshade Solanum dulcamara. Biochim Biophys Acta. 1994 May 18;1206(1):129–135. doi: 10.1016/0167-4838(94)90081-7. [DOI] [PubMed] [Google Scholar]
  5. Griffith M., Ala P., Yang D. S., Hon W. C., Moffatt B. A. Antifreeze protein produced endogenously in winter rye leaves. Plant Physiol. 1992 Oct;100(2):593–596. doi: 10.1104/pp.100.2.593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hincha D. K., Bakaltcheva I., Schmitt J. M. Galactose-Specific Lectins Protect Isolated Thylakoids against Freeze-Thaw Damage. Plant Physiol. 1993 Sep;103(1):59–65. doi: 10.1104/pp.103.1.59. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hincha D. K., DeVries A. L., Schmitt J. M. Cryotoxicity of antifreeze proteins and glycoproteins to spinach thylakoid membranes--comparison with cryotoxic sugar acids. Biochim Biophys Acta. 1993 Mar 14;1146(2):258–264. doi: 10.1016/0005-2736(93)90364-6. [DOI] [PubMed] [Google Scholar]
  8. Hincha D. K., Schmitt J. M. Long-term cryopreservation of thylakoid membranes. Methods Mol Biol. 1995;38:71–80. doi: 10.1385/0-89603-296-5:71. [DOI] [PubMed] [Google Scholar]
  9. Hon W. C., Griffith M., Chong P., Yang DSC. Extraction and Isolation of Antifreeze Proteins from Winter Rye (Secale cereale L.) Leaves. Plant Physiol. 1994 Mar;104(3):971–980. doi: 10.1104/pp.104.3.971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hon W. C., Griffith M., Mlynarz A., Kwok Y. C., Yang D. S. Antifreeze proteins in winter rye are similar to pathogenesis-related proteins. Plant Physiol. 1995 Nov;109(3):879–889. doi: 10.1104/pp.109.3.879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kauffmann S., Legrand M., Geoffroy P., Fritig B. Biological function of ;pathogenesis-related' proteins: four PR proteins of tobacco have 1,3-beta-glucanase activity. EMBO J. 1987 Nov;6(11):3209–3212. doi: 10.1002/j.1460-2075.1987.tb02637.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  13. Lotan T., Ori N., Fluhr R. Pathogenesis-related proteins are developmentally regulated in tobacco flowers. Plant Cell. 1989 Sep;1(9):881–887. doi: 10.1105/tpc.1.9.881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Mauch F., Mauch-Mani B., Boller T. Antifungal Hydrolases in Pea Tissue : II. Inhibition of Fungal Growth by Combinations of Chitinase and beta-1,3-Glucanase. Plant Physiol. 1988 Nov;88(3):936–942. doi: 10.1104/pp.88.3.936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mauch F., Staehelin L. A. Functional Implications of the Subcellular Localization of Ethylene-Induced Chitinase and [beta]-1,3-Glucanase in Bean Leaves. Plant Cell. 1989 Apr;1(4):447–457. doi: 10.1105/tpc.1.4.447. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Moore A. E., Stone B. A. A -I,3-glucan hydrolase from Nicotiana glutinosa. I. Extraction, purification and physical properties. Biochim Biophys Acta. 1972 Jan 20;258(1):238–247. doi: 10.1016/0005-2744(72)90982-5. [DOI] [PubMed] [Google Scholar]
  17. Ori N., Sessa G., Lotan T., Himmelhoch S., Fluhr R. A major stylar matrix polypeptide (sp41) is a member of the pathogenesis-related proteins superclass. EMBO J. 1990 Nov;9(11):3429–3436. doi: 10.1002/j.1460-2075.1990.tb07550.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Payne G., Ward E., Gaffney T., Goy P. A., Moyer M., Harper A., Meins F., Jr, Ryals J. Evidence for a third structural class of beta-1,3-glucanase in tobacco. Plant Mol Biol. 1990 Dec;15(6):797–808. doi: 10.1007/BF00039420. [DOI] [PubMed] [Google Scholar]
  19. Rubinsky B., Arav A., Fletcher G. L. Hypothermic protection--a fundamental property of "antifreeze" proteins. Biochem Biophys Res Commun. 1991 Oct 31;180(2):566–571. doi: 10.1016/s0006-291x(05)81102-7. [DOI] [PubMed] [Google Scholar]
  20. Schaffner W., Weissmann C. A rapid, sensitive, and specific method for the determination of protein in dilute solution. Anal Biochem. 1973 Dec;56(2):502–514. doi: 10.1016/0003-2697(73)90217-0. [DOI] [PubMed] [Google Scholar]
  21. Schraudner M., Ernst D., Langebartels C., Sandermann H. Biochemical Plant Responses to Ozone : III. Activation of the Defense-Related Proteins beta-1,3-Glucanase and Chitinase in Tobacco Leaves. Plant Physiol. 1992 Aug;99(4):1321–1328. doi: 10.1104/pp.99.4.1321. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Sheffield J. B., Graff D., Li H. P. A solid-phase method for the quantitation of protein in the presence of sodium dodecyl sulfate and other interfering substances. Anal Biochem. 1987 Oct;166(1):49–54. doi: 10.1016/0003-2697(87)90544-6. [DOI] [PubMed] [Google Scholar]
  23. Shinshi H., Mohnen D., Meins F. Regulation of a plant pathogenesis-related enzyme: Inhibition of chitinase and chitinase mRNA accumulation in cultured tobacco tissues by auxin and cytokinin. Proc Natl Acad Sci U S A. 1987 Jan;84(1):89–93. doi: 10.1073/pnas.84.1.89. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Shinshi H., Wenzler H., Neuhaus J. M., Felix G., Hofsteenge J., Meins F. Evidence for N- and C-terminal processing of a plant defense-related enzyme: Primary structure of tobacco prepro-beta-1,3-glucanase. Proc Natl Acad Sci U S A. 1988 Aug;85(15):5541–5545. doi: 10.1073/pnas.85.15.5541. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sieg F., Schroder W., Schmitt J. M., Hincha D. K. Purification and Characterization of a Cryoprotective Protein (Cryoprotectin) from the Leaves of Cold-Acclimated Cabbage. Plant Physiol. 1996 May;111(1):215–221. doi: 10.1104/pp.111.1.215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Towbin H., Staehelin T., Gordon J. Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad Sci U S A. 1979 Sep;76(9):4350–4354. doi: 10.1073/pnas.76.9.4350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Turner D. M., Kassell N. F., Sasaki T., Comair Y. G., Beck D. O., Klein S. L. Cerebral and systemic vascular effects of naloxone in pentobarbital-anesthetized normal dogs. Neurosurgery. 1984 Mar;14(3):276–282. doi: 10.1227/00006123-198403000-00003. [DOI] [PubMed] [Google Scholar]
  28. Urrutia M. E., Duman J. G., Knight C. A. Plant thermal hysteresis proteins. Biochim Biophys Acta. 1992 May 22;1121(1-2):199–206. doi: 10.1016/0167-4838(92)90355-h. [DOI] [PubMed] [Google Scholar]
  29. Wong Y. S., Maclachlan G. A. 1,3-beta-D-glucanases from Pisum sativum seedlings. I. Isolation and purification. Biochim Biophys Acta. 1979 Dec 7;571(2):244–255. doi: 10.1016/0005-2744(79)90095-0. [DOI] [PubMed] [Google Scholar]
  30. Worrall D., Hird D. L., Hodge R., Paul W., Draper J., Scott R. Premature dissolution of the microsporocyte callose wall causes male sterility in transgenic tobacco. Plant Cell. 1992 Jul;4(7):759–771. doi: 10.1105/tpc.4.7.759. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Yeh Yin, Feeney Robert E. Antifreeze Proteins: Structures and Mechanisms of Function. Chem Rev. 1996 Mar 28;96(2):601–618. doi: 10.1021/cr950260c. [DOI] [PubMed] [Google Scholar]

Articles from Plant Physiology are provided here courtesy of Oxford University Press

RESOURCES