Skip to main content
Log in

Characterization of a pine multigene family containing elicitor- responsive stilbene synthase genes

  • Published:
Plant Molecular Biology Aims and scope Submit manuscript

Abstract

Young pine seedlings respond to environmental stress by induced synthesis of pinosylvin, a stilbene phytoalexin. Heartwood of pine trees is characterized by a high content of pinosylvin. The formation of pinosylvin from cinnamoyl-CoA and three molecules malonyl-CoA catalysed by pinosylvin synthase is typical of the genus Pinus. Its enzyme activity not detectable in unstressed seedlings is substantially increased upon application of stimuli like UV-light or infection with the phytopathogenic fungus Botrytis cinerea. A genomic DNA library was screened with pinosylvin synthase cDNA pSP-54 as a probe. Ten clones were isolated and grouped into five subclasses according to the size of their introns. After subcloning into plasmid T7T3, four different members of the five gene subclasses were characterized by sequencing. Emphasis was put on isolating various promoters and analyzing and comparing their responsiveness. The amino acid sequences deduced from genes PST-1, PST-2, PST-3 and PST-5 shared an overall identity of more than 95%. In gene PST-5, the putative translation start site ATG was replaced by CTG. While promoter regions near the TATAA box were almost identical PST-1, PST-2 and PST-3, further upstream sequences differed substantially. Differences in promoter strength were analysed both in transgenic tobacco plants and by transient expression in tobacco protoplasts. Constructs used contained the bacterial β-glucuronidase under the control of the promoters of pine genes PST-1, PST-2 and PST-3. Upon treatment with UV light or fungal elicitor, the promoter of PST-1 showed highest responsiveness and led to tissue-specific expression in vascular bundles. The data suggest that in pine the gene product of PST-1 is responsible for both the stress response in seedlings and pinosylvin formation in the heartwood.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  1. Bradley DJ, Kjellbom P, Lamb CJ: Elicitor-induced and wound-induced oxidative cross-linking of a proline-rich plant cell wall protein: a novel, rapid defense response. Cell 70: 21–30 (1992).

    Google Scholar 

  2. Chomczynski P: One-hour downward alkaline capillary transfer for blotting of DNA and RNA. Anal Biochem 201: 134–139 (1992).

    Google Scholar 

  3. Ding X, Shintkau MH, Carter SA, Nelson RS: Invasion of minor veins of tobacco leaves inoculated with tobacco mosaic virus mutants defective in phloem-dependent movement. Proc Nat Acad Sci USA 93: 11155–11160 (1996).

    Google Scholar 

  4. Estabrock EM, Segupta-Gopalan: Differential expression of phenylalanine ammonia-lyase and chalcone synthase during soybean nodule development. Plant Cell 3: 299–308 (1991).

    Google Scholar 

  5. Fischer R, Hain R: Plant disease resistance resulting from the expression of foreign phytoalexins. Curr Opin Biotechnol 5: 125–130 (1994).

    Google Scholar 

  6. Fliegmann J, Schröder G, Schanz S, Britsch L, Schröder J: Molecular analysis of chalcone and dihydropinosylvin synthase from scots pine (Pinus sylvestris), and differential regulation of these and related enzyme activities in stressed plants. Plant Mol Biol 18: 489–503 (1992).

    Google Scholar 

  7. Gehlert R, Schöppner A, Kindl H: Stilbene synthase from seedlings of Pinus sylvestris. Purification and induction in response to fungal infection. Mol Plant-Microbe Interact 3: 444–449 (1990).

    Google Scholar 

  8. Hahlbrock K, Scheel D: Physiology and molecular biology of phenylpropanoid metabolism. Annu Rev Plant Physiol Plant Mol Biol 40: 347–369 (1989).

    Google Scholar 

  9. Hain R, Reif HJ, Krause E, Langebartels R, Kindl H, Vornam B,WieseW, Schmelzer E, Schreier PH, Stöcker RH, Thomzik JE, Stenzel K: Disease resistance results from foreign phytoalexin expression in a novel plant. Nature 361: 153–156 (1993).

    Google Scholar 

  10. Hain R, Stabel P, Czernilofsky AP, Steinbiss H-H, Herrera-Estrella L, Schell J: Uptake, integration, expression and genetic transmission of a selectable chimaeric gene by plant protoplasts. Mol Gen Genet 199: 161–168 (1985).

    Google Scholar 

  11. Herget T, Schell J, Schreier PH: Elicitor-specific induction of one member of the chitinase gene family in Arachis hypogaea. Mol Gen Genet 224: 469–476 (1990).

    Google Scholar 

  12. Horsch RB, Fry JE, Hoffmann NL, Eichholtz D, Rogers SG, Fraley RT: A simple and general method for transferring genes into plants. Science 227: 1229–1231 (1985).

    Google Scholar 

  13. Jefferson RA: Assaying chimeric genes in plants: the GUS gene fusion system. Plant Mol Biol Rep 5: 387–405 (1987).

    Google Scholar 

  14. Jefferson RA, Kavanagh TA, Bevan MW: GUS-fusion: β-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6: 3901–3907 (1987).

    Google Scholar 

  15. Kindl H: Biosynthesis of stilbenes. In: Higuchi T (ed) Biosynthesis and Biodegradation ofWood Components, pp. 349–377. Academic Press, New York (1985).

    Google Scholar 

  16. Klee H, Horsch R, Rogers S: Agrobacterium-mediated plant transformation and its further applications to plant biology. Annu Rev Plant Physiol 38: 467–486 (1987).

    Google Scholar 

  17. Langcake P, McCarthy WV: The relationship of resveratrol production to infection of grapevine leaves by Botrytis cinerea. Vitis 18: 244–253 (1979).

    Google Scholar 

  18. Liswidowati, Melchior F, Hohmann F, Schwer B, Kindl H: Induction of stilbene synthase by Botrytis cinerea in cultured grapevine cells. Planta 183: 307–314 (1991).

    Google Scholar 

  19. Melchior F, Kindl H: Coordinate and elicitor dependent expression of stilbene synthase and phenylalanine ammonialyase genes in Vitis cv. Optima. Arch Biochem Biophys 288: 552–557 (1991).

    Google Scholar 

  20. Preisig-Müller R, Gnau P, Kindl H: The inducible 9,10-dihydrophenanthrene pathway: characterization and expression of bibenzyl synthase and S-adenosylhomocysteine hydrolase. Arch Biochem Biophys 317: 201–207 (1995).

    Google Scholar 

  21. Rodgers SO, Bendich AJ: Extraction of DNA from plant tissue. In: Gelvin SB, Schilperoort RA (eds) Plant Molecular Biology Manual, pp. A6: 1–10. Kluwer Academic Publishers, Dordrecht, Netherlands (1988)

    Google Scholar 

  22. Schanz S, Schröder G, Schröder J: Stilbene synthase from scots pine (Pinus sylvestris). FEBS Lett 313: 71–74 (1992).

    Google Scholar 

  23. Schöppner A, Kindl H: Stilbene synthase (pinosylvin synthase) and its induction by UV light. FEBS Lett 108: 349–352 (1979).

    Google Scholar 

  24. Schöppner A, Kindl H: Purification and properties of a stilbene synthase from induced cell suspension cultures of peanut. J Biol Chem 259: 6806–6811 (1984).

    Google Scholar 

  25. Schröder G, Brown JWS, Schröder J: Molecular analysis of resveratrol synthase: cDNA clones and relationship with chalcone synthase. Eur J Biochem172: 161–169 (1988).

    Google Scholar 

  26. Schwekendiek A, Pfeffer G, Kindl H: Pine stilbene synthase cDNA, a tool for probing environmental stress. FEBS Lett 301: 41–44 (1992).

    Google Scholar 

  27. Vornam B, Schön H, Kindl H: Control of gene expression during induction of cultured peanut cells: mRNA levels, protein synthesis and enzyme activity of stilbene synthase. Plant Mol Biol 10: 235–243 (1988).

    Google Scholar 

  28. Wakamiya I, Newton RJ, Johnston JS, Price HJ: Genome size and environmental factors in the genus Pinus. Am J Bot 80: 1235–1241 (1993).

    Google Scholar 

  29. Wiese W, Vornam B, Krause E, Kindl H: Structural organization and differential expression of three stilbene synthase genes located on a 13 kb grapevine DNA fragment. Plant Mol Biol 26: 667–677 (1994).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Preisig-müller, R., Schwekendiek, A., Brehm, I. et al. Characterization of a pine multigene family containing elicitor- responsive stilbene synthase genes. Plant Mol Biol 39, 221–229 (1999). https://doi.org/10.1023/A:1006163030646

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1006163030646

Navigation