Plant Molecular Biology

, Volume 19, Issue 2, pp 193–204 | Cite as

The identification of leaf thionin as one of the main jasmonate-induced proteins of barley (Hordeum vulgare)

  • Ingrid Andresen
  • Walter Becker
  • Kirsten Schlüter
  • Jan Burges
  • Benno Parthier
  • Klaus Apel
Research Articles


Jasmonic acid (JA) and its methyl ester (JA-Me) are able to introduce the accumulation of several specific polypeptides in cut leaf segments of barley. Two of the most prominent JA-induced proteins of Mr 15 000 and 23 000 have been characterized by isolating and sequencing complete cDNA sequences. While the sequence of the Mr 23 000 polypeptide shows no similarity to published sequences, the sequence of the Mr 15 000 polypeptide corresponds to the higher-molecular-weight precursor of a leaf thionin previously characterized.

Transcripts for the Mr 23 000 and Mr 15 000 polypeptides accumulate in leaf segments shortly after the beginning of JA treatment. JA and JA-Me induce the appearance of the two proteins not only in leaf segments but also in intact barley seedlings. However, in seedlings the accumulation of JA-induced proteins occurs much more slowly and requires high concentrations of volatile JA-Me. Thus, in barley it seems unlikely that volatile JA-Me is involved in the interaction between different members of this species, as has been proposed recently for tomato seedlings.

Key words

jasmonate-induced proteins (JIPs) leaf thionin stress response barley 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Anderson JM: Jasmonic acid-dependent increase in the level of specific polypeptides in soybean suspension cultures and seedlings. J Plant Growth Regul 7: 203–211 (1988).Google Scholar
  2. 2.
    Anderson JM: Jasmonic acid-dependent increase in vegetative storage protein in soybean tissue cultures. J Plant Growth Regul 10: 5–10 (1991).Google Scholar
  3. 3.
    Anderson JM, Spilatro SR, Klauer SF, Franceschi VR: Jasmonic acid-dependent increase in the level of vegetative storage proteins in soybean. Plant Sci 62: 45–52 (1989).CrossRefGoogle Scholar
  4. 4.
    Apel K, Bohlmann H, Reimann-Philipp U: Leaf thionins, a novel class of putative defence factors. Physiol Plant 80: 315–321 (1990).CrossRefGoogle Scholar
  5. 5.
    Aviv H, Leder P: Purification of biologically active globin messenger RNA by chromatography on oligothymedilic acid-cellulose. Proc Natl Acad Sci USA 69: 1408–1412 (1972).PubMedGoogle Scholar
  6. 6.
    Batschauer A, Mösinger E, Kreuz K, Dörr I, Apel K: The implication of a plastid-derived factor in the transcriptional control of nuclear genes encoding the light-harvesting chlorophyll a/b protein. Eur J Biochem 154: 625–634 (1986).PubMedGoogle Scholar
  7. 7.
    Bohlmann H, Apel K: Isolation and characterization of cDNAs coding for leaf-specific thionins closely related to the endosperm-specific hordothionin of barley (Hordeum vulgare L). Mol Gen Genet 207: 446–454 (1987).CrossRefGoogle Scholar
  8. 8.
    Bohlmann H, Clausen S, Behnke S, Giese H, Hiller C, Reimann-Philipp U, Schrader G, Barkholt V, Apel K: Leaf-specific thionins—a novel class of cell wall proteins toxic to plant-pathogenic fungi and and possibly involved in the defence mechanism of plants. EMBO J 7: 1559–1565 (1988).Google Scholar
  9. 9.
    Bohlmann H, Apel K: Thionins. Annu Rev Plant Physiol Plant Mol Biol 42: 227–240 (1991).CrossRefGoogle Scholar
  10. 10.
    Bonner WM, Laskey RA: A film detection method for tritium labelled proteins and nucleic acids in polyacrylamide gels. Eur J Biochem 46: 83–88 (1974).PubMedGoogle Scholar
  11. 11.
    Chirgwin JN, Przybyla AE, MacDonald RJ, Rutter WJ: Isolation of biologically active ribonucleic acid from sources enriched in ribonuclease. Biochemistry 18: 5294–5299 (1979).PubMedGoogle Scholar
  12. 12.
    Dehesh K, Klaas M, Häuser I, Apel K: Light-induced changes in the distribution of the 36 000-M r polypeptide of NADPH-protochlorophyllide oxidoreductase within different cellular compartements of barley (Hordeum vulgare L). Planta 169: 162–171 (1986).Google Scholar
  13. 13.
    Denhardt DT: A membrane filter technique for the detection of complementary DNA. Biochem Biophys Res Comm 23: 641–646 (1966).PubMedGoogle Scholar
  14. 14.
    Dure L, Crouch M, Harada J, Ho TH, Mundy J, Quatrano R, Thomas T, Sung ZR: Common amino acid sequence domains among the LEA proteins of higher plants. Plant Mol Biol 12: 475–486 (1989).Google Scholar
  15. 15.
    Farmer EE, Ryan CA: Interplant communication: Airborne methyl jasmonate induces synthesis of proteinase inhibitors in plant leaves. Proc Natl Acad Sci USA 87: 7713–7716 (1990).PubMedGoogle Scholar
  16. 16.
    Feinberg AP, Vogelstein B: A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 132: 6–13 (1983).PubMedGoogle Scholar
  17. 17.
    Forreiter C, van Cleve B, Schmidt A, Apel K: Evidence for a general light-dependent negative control of NADPH-protochlorophyllide oxidoreductase in angiosperms. Planta 183: 126–132 (1990).Google Scholar
  18. 18.
    Gollmer I, Apel K: The phytochrome-controlled accumulation of mRNA sequences encoding the light-harvesting chlorophyll a/b protein of barley (Hordeum vulgare L.). Eur J Biochem 133: 309–313 (1983).PubMedGoogle Scholar
  19. 19.
    Herrmann G, Lehmann J, Peterson A, Sembdner G, Weidhase RA, Parthier B: Species and tissue specificity of jasmonate-induced abundant proteins. J Plant Physiol 134: 703–709 (1989).Google Scholar
  20. 20.
    Huyn TV, Young RA, Davis RW: Construction and screening of cDNA libraries in lambda gt10 and lambda gt11. In: Glover DM (ed) DNA Cloning: A Practical Approach, vol. 1, pp. 49–78, JRL Press, Oxford (1985).Google Scholar
  21. 21.
    Laemmli UK: Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680–685 (1970).PubMedGoogle Scholar
  22. 22.
    Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).Google Scholar
  23. 23.
    Mason HS, Mullet JE: Expression of two soybean vegetative storage protein genes during development and in response to water deficit, wounding, and jasmonic acid. Plant Cell 2: 569–579 (1990).CrossRefPubMedGoogle Scholar
  24. 24.
    Meyer A, Miersch O, Büttner C, Dathe W, Sembdner G: Occurrence of the plant growth regulator jasmonic acid in plants. J Plant Growth Regul 3: 1–8 (1984).Google Scholar
  25. 25.
    Mueller-Uri F, Parthier B, Nover L: Jasmonate-induced alteration of gene expression in barley leaf segments analyzed by in vivo and in vitro protein synthesis. Planta 176: 241–247 (1988).Google Scholar
  26. 26.
    Murray HG, Thompson WF: Rapid isolation of high molecular weight plant DNA. Nucl Acids Res 8: 4321–4325 (1980).PubMedGoogle Scholar
  27. 27.
    Parthier B: Jasmonates: Hormonal regulators or stress factors in leaf senescence? J Plant Growth Regul 9: 57–63 (1990).Google Scholar
  28. 28.
    Parthier B: Jasmonates, new regulators of plant growth and development: many facts and few hypotheses on their action. Bot Acta, in press (1991).Google Scholar
  29. 29.
    Parthier B, Brückner C, Dathe W, Hause B, Herrmann G, Knöfel HD, Kramell HM, Kramell R, Lehmann J, Miersch O, Reinbothe St, Sembdner G, Wasternack C, zur Nieden U: Jasmonates: Metabolism, biological activities, and modes of action in senescence and stress responses. In: Karssen CM (ed) Plant Growth Substances. Kluwer Academic Publishers, Dordrecht, Netherlands (1991).Google Scholar
  30. 30.
    Pelham HRB, Jackson RJ: An efficient mRNA-dependent translation system from reticulocyte lysates. Eur J Biochem 67: 247–256 (1976).PubMedGoogle Scholar
  31. 31.
    Reimann-Philipp U, Schrader G, Martincia E, Barkholt V, Apel K: Intracellular thionins of barley. A second group of leaf thionins closely related to but distinct from cell wall-bound thionins. J Biol Chem 264: 8978–8984 (1989).PubMedGoogle Scholar
  32. 32.
    Reinbothe S, Machmudova C, Wasternack C, Reinbothe C, Parthier B: Jasmonate-induced proteins in cotton: immunological relationship to the respective barley proteins and homology of transcripts to late embryogenesis abundant (LEA) mRNA. J Plant Growth Regul, in press (1991).Google Scholar
  33. 33.
    Ryan CA: Proteinase inhibitors in plant leaves: A biochemical model for pest-induced natural plant protection. Trends Biochem Sci 3: 148–150 (1978).CrossRefGoogle Scholar
  34. 34.
    Sanger F, Nicklen S, Coulson AR: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).PubMedGoogle Scholar
  35. 35.
    Satler SO, Thimann KV: Le jasmonate de méthyle: nouveau et puissant promoteur de la senescence de feuilles. C R Acad Sci Paris, Ser III 293: 735–740 (1981).Google Scholar
  36. 36.
    Staswick PE: Novel regulation of vegetative storage protein genes. Plant Cell 2: 1–6 (1990).CrossRefPubMedGoogle Scholar
  37. 37.
    Staswick PE, Huang JF, Rhee Y: Nitrogen and methyl-jasmonate induction of soybean vegetative storage protein genes. Plant Physiol 96: 130–136 (1991).Google Scholar
  38. 38.
    Towbin H, Staehelin T, Gordon J: Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76: 4350–4354 (1979).PubMedGoogle Scholar
  39. 39.
    Ueda J, Kato J: Isolation and identification of a senescence-promoting substance from wormwood (Artemisia absinthium L.). Plant Physiol 66: 246–249 (1980).Google Scholar
  40. 40.
    Weidhase RA, Lehmann J, Kramell H, Sembdner G, Parthier B: Degradation of ribulose-1,5-bisphosphate carboxylase and chlorophyll in senescing barley leaf segments triggered by jasmonic acid methylester, and counteraction by cytokinin. Physiol Plant 69: 161–166 (1987).Google Scholar
  41. 41.
    Weidhase RA, Kramell HM, Lehmann J, Liebisch HW, Lerbs W, Parthier B: Methyljasmonate-induced changes in the polypeptide pattern of senescing barley leaf segments. Plant Sci 51: 177–186 (1987).CrossRefGoogle Scholar
  42. 42.
    Wilen RW, van Rooijen GJH, Pearce DW, Pharis P, Holbrook LA, Moloney MM: Effects of jasmonic acid on embryo-specific processes in Brassica and Linum oilseeds. Plant Physiol 95: 399–405 (1991).Google Scholar
  43. 43.
    Yamane H, Takagi H, Abe T, Yokota T, Takahashi N: Identification of jasmonic acid in three species of higher plants and its biological activities. Plant Cell Physiol 22: 689–697 (1981).Google Scholar

Copyright information

© Kluwer Academic Publishers 1992

Authors and Affiliations

  • Ingrid Andresen
    • 1
  • Walter Becker
    • 1
  • Kirsten Schlüter
    • 2
  • Jan Burges
    • 3
  • Benno Parthier
    • 3
  • Klaus Apel
    • 1
    • 2
  1. 1.Botanisches Institut der Universität KielKielGermany
  2. 2.Institut für Pflanzenwissenschaften der ETH ZürichZürichSwitzerland
  3. 3.Institut für Biochemie der PflanzenHalle/SaaleGermany

Personalised recommendations