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Plant Molecular Biology

, Volume 33, Issue 6, pp 979–987 | Cite as

Molecular cloning, structure and expression of an elicitor-inducible chitinase gene from pine trees

  • Haiguo Wu
  • Craig S. Echt
  • Michael P. Popp
  • John M. Davis*
Article

Abstract

We have cloned, sequenced, and examined the expression of genes from pine trees that appear to encode extracellular class II chitinase. Nucleotide sequence analysis indicates a coding sequence composed of three exons interrupted by two introns at locations identical to those found in other chitinase genes that possess introns. One of the genes, Pschi4, potentially encodes a protein that shares 62% amino acid sequence identity through the catalytic domain with class II chitinase from tobacco. In contrast, Pschi1 contains a stop codon in the first exon and may be a pseudogene. Pschi4 genes are conserved in several species of pine, and appear to comprise a small multigene family. Treatment of pine cell suspension cultures with the general elicitor chitosan induced Pschi4 expression. The regulatory sequences associated with the Pschi4 gene were sufficient to direct chitosan-inducible expression of Pschi4 in transgenic tobacco plants, which indicates that Pschi4 is an actively expressed member of the multigene family. The observation that the Pschi4 gene from pine (a gymnosperm) was appropriately regulated by chitosan in tobacco (an angiosperm) suggests that the signaling pathways that mediate chitosan-induced transcription are highly conserved in the plant kingdom.

chitosan class II chitinase defense response gymnosperm Pinus transgenic plants 

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References

  1. 1.
    Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ: Basic local alignment search tool. J Mol Biol 215: 403–410 (1990).Google Scholar
  2. 2.
    An G, Ebert PR, Mitra A, Ha SB: Binary vectors. In: Gelvin SB, Schilperoort RA, Verma DPS (eds) Plant Molecular Biology Manual, pp. A3/1–A3/19. Kluwer Academic Publishers, Dordrecht (1988).Google Scholar
  3. 3.
    Balandin T, van der Does C, Albert JMB, Bol JF, Linthorst HJM: Structure and induction pattern of a novel proteinase inhibitor class II gene of tobacco. Plant Mol Biol 27: 1197–1204 (1995).Google Scholar
  4. 4.
    Campbell MM, Ellis BE: Fungal elicitor-mediated responses in pine cell cultures. Plant Physiol 98: 62–70 (1992).Google Scholar
  5. 5.
    Church GM, Gilbert W: Genomic sequencing. Proc Natl Acad Sci USA 81: 1991–1995 (1984).Google Scholar
  6. 6.
    Collinge DB, Kragh KM, Mikkelsen JD, Nielsen KK, Rasmussen U, Vad K: Plant chitinases. Plant J 3: 31–40 (1993).Google Scholar
  7. 7.
    Davis JM, Clarke HRG, Bradshaw HD Jr, Gordon MP: Populus chitinase genes: structure, organization, and similarity of translated sequences to herbaceous plant chitinases. Plant Mol Biol 17: 631–639 (1991).Google Scholar
  8. 8.
    Feinberg AP, Vogelstein B: Addendum, A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 137: 266–267 (1984).Google Scholar
  9. 9.
    Harlow WM, Harrar ES, White FM: Textbook of Dendrology. McGraw-Hill, New York (1979).Google Scholar
  10. 10.
    Jaeck E, Dumas B, Geoffrey P, Favet N, Inze D, Van Montagu M, Fritig B, Legrand M: Regulation of enzymes involved in lignin biosynthesis: induction of Omethyltransferase mRNAs during the hypersensitive reaction of tobacco to tobacco mosaic virus. Mol Plant-Microbe Interact 5: 294–300 (1992).Google Scholar
  11. 11.
    Keen NT, Dawson WO: Pathogen avirulence genes and elicitors of plant defense. In: Boller T, Meins F (eds) Genes Involved in Plant Defense, pp. 85–114, Springer-Verlag, New York (1992).Google Scholar
  12. 12.
    Kojima K, Sasaki S, Yamamoto N: Light-independent and tissue-specific expression of a reporter gene mediated by the pine Cab-6 promoter in transgenic tobacco. Plant J 6: 591–596 (1994).Google Scholar
  13. 13.
    Kozak M: Structural features in eukaryotic mRNAs that modulate the initiation of translation. J BiolChem 266: 19867–19870 (1991).Google Scholar
  14. 14.
    Lesney MS: Growth responses and lignin production in cell suspensions of ‘elicited’ by chitin, chitosan or mycelium of Cronartium quercuum f. sp. fusiforme. Plant Cell Tiss Org Cult 19: 23–31 (1989).Google Scholar
  15. 15.
    Martinussen I, Bate N, Weterings K, Junttila O, Twell D: Analysis of gene-regulation in growing pollen tubes of angiosperm and gymnosperm species using microprojectile bombardment. Physiol Plant 93: 445–450 (1995).Google Scholar
  16. 16.
    Mason ME: Identification of elicitor-responsive cDNAs in pine cells using differential display. M.S. thesis, University of Florida (1995).Google Scholar
  17. 17.
    Mauch F, Staehelin LA: Functional implications of the subcellular localization of ethylene-induced chitinase and β-1,3-glucanase in bean leaves. Plant Cell 1: 447–457 (1989).Google Scholar
  18. 18.
    Mauch F, Mauch-Mani B, Boller T: Antifungal hydrolases in pea tissue II. Inhibition of fungal growth by combinations of chitinase and β-1,3-glucanase. Plant Physiol 88: 936–942 (1988).Google Scholar
  19. 19.
    Neuhaus JM, Sticher L, Meins F, Boller T: A short C-terminal sequence is necessary and sufficient for the targeting of chitinases to the plant vacuole. Proc Natl Acad Sci USA 88: 10362–10366 (1991).Google Scholar
  20. 20.
    Palm CJ, Costa MA, An G, Ryan CA: Wound-inducible nuclear protein binds DNA fragments that regulate a proteinase inhibitor II gene from potato. Proc Natl Acad Sci USA 87: 603–607 (1990).Google Scholar
  21. 21.
    Popp MP, Lesney MS, Davis JM: Defense responses elicited in pine cell suspension cultures. Plant Cell Tiss Org Cult (in press).Google Scholar
  22. 22.
    Popp MP: Induction of putative defense responses of slash and loblolly pine to bark beetle vectored fungi. Ph. D. dissertation, University of Florida (1993).Google Scholar
  23. 23.
    Rogers SG, Horsch RB, Fraley RT: Gene transfer in plants: production of transformed plants using Ti plasmid vectors. Meth Enzymol 118: 627–640 (1986).Google Scholar
  24. 24.
    Rothstein SJ, Lahners KN, Lotstein RJ, Carozzi NB, Jayne SM, Rice DA: Promoter cassettes, antibiotic-resistance genes, and vectors for plant transformation. Gene 53: 153–161 (1987).Google Scholar
  25. 25.
    Sambrook J, Fritsch EF, Maniatis T: Molecular Cloning: A Laboratory Manual, 2nd ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1989).Google Scholar
  26. 26.
    Sanger F, Nicklen S, Coulson AR: DNAsequencingwith chainterminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).Google Scholar
  27. 27.
    Schneiderbauer A, Heinrich S Jr, Ernst D: Isolation of functional RNA from plant tissues rich in phenolic compounds. Anal Biochem 197: 91–95 (1991).Google Scholar
  28. 28.
    Sela-Buurlage MB, Ponstein AS, Bres-Vloemans SA, Melchers LS, van den Elzen PJM, Cornelissen BJC: Only specific tobacco (Nicotiana tabacum) chitinases and β1,3-glucanases exhibit antifungal activity. Plant Physiol 101: 857–863 (1993).Google Scholar
  29. 29.
    Stintzi A, Heitz T, Prasad V, Wiedemann-Merdinoglu S, Kauffmann S, Geoffroy P, Legrand M, Fritig B: Plant ‘pathogenesisrelated’ proteins and their role in defense against pathogens. Biochimie 75: 687–706 (1993).Google Scholar
  30. 30.
    Wagner DB, Furnier GR, Saghai-Maroof MA, Williams SM, Dancik BD, Allard RW: Chloroplast DNA polymorphisms in lodgepole and jack pines and their hybrids. Proc Natl Acad Sci USA 84: 2097–2100 (1987).Google Scholar
  31. 31.
    Yamamoto N, Tada Y, Fujimura T: The promoter of a pine photosynthetic gene allows expression of a beta-glucuronidase reporter gene in transgenic rice plants in a light-independent but tissue-specific manner. Plant Cell Physiol 35: 773–778 (1994).Google Scholar

Copyright information

© Kluwer Academic Publishers 1997

Authors and Affiliations

  • Haiguo Wu
    • 1
  • Craig S. Echt
    • 3
  • Michael P. Popp
    • 2
  • John M. Davis*
    • 1
    • 2
  1. 1.Plant Molecular and Cellular Biology ProgramUSA
  2. 2.School of Forest Resources and ConservationUniversity of FloridaGainesvilleUSA
  3. 3.Forestry Sciences LaboratoryUSDA-FSRhinelanderUSA

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