Plant Molecular Biology

, Volume 36, Issue 1, pp 55–62 | Cite as

Prosystemin from potato, black nightshade, and bell pepper: primary structure and biological activity of predicted systemin polypeptides

  • C. Peter Constabel
  • Lynn Yip
  • Clarence A. Ryan
Article

Abstract

Prosystemin is the precursor protein of the 18 amino acid wound signal systemin which activates systemic defense in tomato leaves against insect herbivores (McGurl B, Pearce G, Orozco-Cardenas M, Ryan CA, Science 255 (1992) 1570–1573). Here, we report the isolation of cDNA sequences encoding prosystemin from potato (Solanum tuberosum), black nightshade (S. nigrum), and bell pepper (Capsicum annuum), all members of the Solanaceae family, using reverse-transcription polymerase chain reaction (RT-PCR). Pairwise comparisons of the predicted prosystemin proteins from the three species with tomato prosystemin and among each other indicated sequence identities ranging from 73% to 88%. The deduced systemin polypeptides were synthesized and tested for their capacities to induce the synthesis of the defensive proteinase inhibitors in tomato leaves. Potato and pepper systemins were approximately as active as tomato systemin, whereas nightshade systemin was ten-fold less active. The accumulation of proteinase inhibitor mRNA transcripts could be induced in each of these plants by treatment with the homologous systemin. As in the tomato, in potato, black nightshade, and bell pepper plants, prosystemin homologs appear to function as precursors of systemic wound signals.

plant defense proteinase inhibitors systemic wound signaling Solanaceae 

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References

  1. 1.
    Becraft PW, Stinard PS, McCarty DR: CRINKLY4: A TNFRlike receptor kinase involved in maize epidermal differentiation. Science 273: 1406–1409 (1996).PubMedGoogle Scholar
  2. 2.
    Bergey DR, Howe GA, Ryan CA: Polypeptide signaling for plant defensive genes exhibits analogies to defense signaling in animals. Proc Natl Acad Sci USA 93: 12053–12058 (1997).Google Scholar
  3. 3.
    Beuning LL, Spriggs TW, Christeller JT: Evolution of the proteinase inhibitor I family and apparent lack of hypervariability in the proteinase contact loop. J Mol Evol 39: 644–654 (1994).PubMedGoogle Scholar
  4. 4.
    Borson ND, Salo WL, Drewes LR: A lock-docking oligo(dt) primer for 5′ and 3′ RACE PCR. PCR Meth Appl 2: 144–148 (1992)Google Scholar
  5. 5.
    Bradshaw Jr H, Hollick JB, Parsons TJ, Clarke HRG, Gordon MP: Systemically wound-responsive genes in poplar trees encode proteins similar to sweet potato sporamins and legume Kunitz trypsin inhibitors. Plant Mol Biol 14: 51–59 (1989).CrossRefGoogle Scholar
  6. 6.
    Brown WE, Ryan CA: Isolation and characterization of a wound-induced trypsin inhibitor from alfalfa leaves. Biochemistry 23: 3418–3422 (1984).PubMedGoogle Scholar
  7. 7.
    Cleveland TE, Thornburg RW, Ryan CA: Molecular characterization of a wound-inducible inhibitor I gene from potato and the processing of its mRNA and protein. Plant Mol Biol 8: 199–207 (1985).Google Scholar
  8. 8.
    Constabel CP, Bergey DR, Ryan CA: Systemin activates synthesis of wound-inducible tomato leaf polyphenol oxidase via the octadecanoid defense signaling pathway. Proc Natl Acad Sci USA 92: 407–411 (1995).PubMedGoogle Scholar
  9. 9.
    Frohman M: RACE: Rapid amplification of cDNA ends. In: Innis MA (ed) PCR Protocols: AGuide to Methods and Applications, pp. 28–37. Academic Press, San Diego, CA (1990).Google Scholar
  10. 10.
    Graham JS, Hall G, Pearce G, Ryan CA: Regulation of synthesis of proteinase inhibitor I and II mRNAs in leaves of wounded tomato plants. Planta 169: 399–405 (1986).Google Scholar
  11. 11.
    Graham JS, Pearce G, Merryweather J, Titani K, Ericsson L, Ryan CA: Wound-induced proteinase inhibitors from tomato leaves. I. The cDNA-deduced primary structure of preinhibitor I and its post-translational processing. J Biol Chem 260: 6555–6560 (1985).PubMedGoogle Scholar
  12. 12.
    Graham JS, Pearce G, Merryweather J, Titani K, Ericsson LH, Ryan CA: Wound-induced proteinase inhibitors from tomato leaves II: The cDNA-deduced primary structure of preinhibitor. II. J Biol Chem 260: 6561–6564 (1985).PubMedGoogle Scholar
  13. 13.
    Green TR, Ryan CA: Wound-induced proteinase inhibitor in plant leaves: a possible defense mechanism against insects. Science 175: 776–777 (1972).Google Scholar
  14. 14.
    Hummel B: A modified spectrophotometric determination of chymotrypsin, trypsin, and thrombin. Can J Biochem Physiol 37: 1393–1399 (1959).PubMedGoogle Scholar
  15. 15.
    Marchuk D, Dumm M, Saulino A, Collins FS: Construction of T-vectors, a rapid and general system for direct cloning of unmodified PCR products. Nucl Acids Res 19: 1154 (1991).PubMedGoogle Scholar
  16. 16.
    McGurl B, Ryan CA: The organization of the prosystemin gene. Plant Mol Biol 20: 405–409 (1992).PubMedGoogle Scholar
  17. 17.
    McGurl B, Orozco-Cardenas M, Pearce G, Ryan CA: Overexpression of the prosystemin gene in transgenic tomato plants generates a systemic signal that constitutively induces proteinase inhibitor synthesis. Proc Natl Acad Sci USA 91: 9799–9802 (1994).PubMedGoogle Scholar
  18. 18.
    McGurl B, Pearce G, Orozco-Cardenas M, Ryan CA: Structure, expression, and antisense inhibition of the systemin precursor gene. Science 255: 1570–1573 (1992).PubMedGoogle Scholar
  19. 19.
    Pearce G, Johnson S, Ryan CA: Purification and characterization from tobacco (Nicotiana tabacum) leaves of six small, wound-inducible proteinase isoinhibitors of the potato inhibitor II family. Plant Physiol 102: 639–644 (1993).CrossRefPubMedGoogle Scholar
  20. 20.
    Pearce G, Johnson S, Ryan CA: Structure-activity of deleted and substituted systemin, an 18-amino acid polypeptide inducer of plant defensive genes. J Biol Chem 268: 212–216 (1993).PubMedGoogle Scholar
  21. 21.
    Pearce G, Strydom D, Johnson S, Ryan CA: A polypeptide from tomato leaves induces wound-inducible proteinase inhibitor proteins. Science 253: 895–898 (1991).Google Scholar
  22. 22.
    Peña-Cortes H, Sanchez-Serrano J, Rocha-Rosa M, Willmitzer L: Systemic induction of proteinase-inhibitor-II gene expression in potato plants by wounding. Planta 174: 84–89 (1988).Google Scholar
  23. 23.
    Program Manual for theWisconsin Package, Version 8. Genetics Computer Group Inc., Madison, WI (1994).Google Scholar
  24. 24.
    Realini C, Rogers SW, Rechsteiner M: Proposed roles in protein-protein association and presentation of peptides by the MHC class I receptors. FEBS Lett 348: 109–113 (1994).PubMedGoogle Scholar
  25. 25.
    Rohrmeier T, Lehle L: WIP1, a wound-inducible gene from maize with homology to Bowman-Birk proteinase inhibitors. Plant Mol Biol 22: 783–792 (1993).PubMedGoogle Scholar
  26. 26.
    Ryan CA: Protease inhibitors in plants: genes for improving defenses against insects and pathogens. Annu Rev Phytopath 28: 425–429 (1990).CrossRefGoogle Scholar
  27. 27.
    Ryan CA: Quantitative determination of soluble cellular proteins by radial diffusion in agar gel containing antibodies. Anal Biochem 19: 434–440 (1967).PubMedGoogle Scholar
  28. 28.
    Ryan CA: The search for the proteinase inhibitor-inducing factor, PIIF. Plant Mol Biol 19: 123–133 (1992).PubMedGoogle Scholar
  29. 29.
    Saarikoski P, Clapham D, von Arnold S: A wound-inducible gene from Salix viminalis coding for a trypsin inhibitor. Plant Mol Biol 31: 465–478 (1996).PubMedGoogle Scholar
  30. 30.
    Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY (1989).Google Scholar
  31. 31.
    Schaller A, Ryan CA: Systemin: a polypeptide defense signal in plants. BioEssays 18: 27–33 (1995).Google Scholar
  32. 32.
    Seldal T, Andersen K-J, Högstedt G: Grazing-induced proteinase inhibitors: a possible cause for lemming population cycles. Oikos 70: 3–11 (1994).Google Scholar
  33. 33.
    Trautman R, Cowan KM, Wagner GG: Data for processing for radial immunodiffusion. Immunochemistry 8: 901–916 (1971).CrossRefPubMedGoogle Scholar
  34. 34.
    van de Sande K, Pawlowski K, Czaja I, Wieneke U, Schell J, Schmidt J, Walden R, Matvienko M, Wellink J, van Kammen A, Franssen H, Bisseling T: Modification of phytohormone response by a peptide encoded by ENOD40 of legumes and a nonlegume. Science 273: 370–373 (1996).PubMedGoogle Scholar
  35. 35.
    Walker-Simmons M, Ryan CA: Wound-induced accumulation of trypsin inhibitor activities in plant leaves. Plant Physiol 59: 437–439 (1977).Google Scholar
  36. 36.
    Wingate VPM, Broadway RM, Ryan CA: Isolation and characterization of a novel, developmentally regulated proteinase inhibitor I protein and cDNA from the fruit of a wild tomato species. J Biol Chem 264: 17734–17738 (1989).PubMedGoogle Scholar

Copyright information

© Kluwer Academic Publishers 1998

Authors and Affiliations

  • C. Peter Constabel
    • 1
  • Lynn Yip
    • 1
  • Clarence A. Ryan
    • 1
  1. 1.Institute of Biological ChemistryWashington State UniversityPullmanUSA

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