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Plant Physiol. 1994 Jan; 104(1): 109–118.
PMCID: PMC159168

A novel pathogen- and wound-inducible tobacco (Nicotiana tabacum) protein with antifungal activity.


A novel pathogen- and wound-inducible antifungal protein of 20 kD was purified from tobacco (Nicotiana tabacum) Samsun NN leaves inoculated with tobacco mosaic virus (TMV). The protein, designated CBP20, was purified by chitin-affinity chromatography and gel filtration. In vitro assays demonstrated that CBP20 exhibits antifungal activity toward Trichoderma viride and Fusarium solani by causing lysis of the germ tubes and/or growth inhibition. In addition it was shown that CBP20 acts synergistically with a tobacco class I chitinase against F. solani and with a tobacco class I beta-1,3-glucanase against F. solani and Alternaria radicina. Analysis of the protein and corresponding cDNAs revealed that CBP20 contains an N-terminal chitin-binding domain that is present also in the class I chitinases of tobacco, the putative wound-induced (WIN) proteins of potato, WIN1 and WIN2, and several plant lectins. The C-terminal domain of CBP20 showed high identity with tobacco pathogenesis-related (PR) proteins, PR-4a and PR-4b, tomato PR-P2, and potato WIN1 and WIN2. CBP20 is synthesized as a preproprotein, which is processed into the mature protein by the removal of an N-terminal signal peptide and a C-terminal propeptide, most likely involved in the vacuolar targeting of the protein. The intracellular localization of CBP20 and its induction upon TMV infection and wounding indicate that CBP20 is the first class I PR-4 type protein purified.

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Selected References

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  • Boller T, Vögeli U. Vacuolar localization of ethylene-induced chitinase in bean leaves. Plant Physiol. 1984 Feb;74(2):442–444. [PMC free article] [PubMed]
  • Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. [PubMed]
  • Brederode FT, Linthorst HJ, Bol JF. Differential induction of acquired resistance and PR gene expression in tobacco by virus infection, ethephon treatment, UV light and wounding. Plant Mol Biol. 1991 Dec;17(6):1117–1125. [PubMed]
  • Broekaert I, Lee HI, Kush A, Chua NH, Raikhel N. Wound-induced accumulation of mRNA containing a hevein sequence in laticifers of rubber tree (Hevea brasiliensis). Proc Natl Acad Sci U S A. 1990 Oct;87(19):7633–7637. [PMC free article] [PubMed]
  • Broekaert WF, VAN Parijs J, Leyns F, Joos H, Peumans WJ. A chitin-binding lectin from stinging nettle rhizomes with antifungal properties. Science. 1989 Sep 8;245(4922):1100–1102. [PubMed]
  • Chen EY, Seeburg PH. Supercoil sequencing: a fast and simple method for sequencing plasmid DNA. DNA. 1985 Apr;4(2):165–170. [PubMed]
  • Cleveland DW, Fischer SG, Kirschner MW, Laemmli UK. Peptide mapping by limited proteolysis in sodium dodecyl sulfate and analysis by gel electrophoresis. J Biol Chem. 1977 Feb 10;252(3):1102–1106. [PubMed]
  • Collinge DB, Kragh KM, Mikkelsen JD, Nielsen KK, Rasmussen U, Vad K. Plant chitinases. Plant J. 1993 Jan;3(1):31–40. [PubMed]
  • Cornelissen BJ, Horowitz J, van Kan JA, Goldberg RB, Bol JF. Structure of tobacco genes encoding pathogenesis-related proteins from the PR-1 group. Nucleic Acids Res. 1987 Sep 11;15(17):6799–6811. [PMC free article] [PubMed]
  • Hejgaard J, Jacobsen S, Bjørn SE, Kragh KM. Antifungal activity of chitin-binding PR-4 type proteins from barley grain and stressed leaf. FEBS Lett. 1992 Aug 3;307(3):389–392. [PubMed]
  • Ishii S, Yokosawa H, Kumazaki T, Nakamura I. Immobilized anhydrotrypsin as a specific affinity adsorbent for tryptic peptides. Methods Enzymol. 1983;91:378–383. [PubMed]
  • Joosten MH, Bergmans CJ, Meulenhoff EJ, Cornelissen BJ, De Wit PJ. Purification and serological characterization of three basic 15-kilodalton pathogenesis-related proteins from tomato. Plant Physiol. 1990 Oct;94(2):585–591. [PMC free article] [PubMed]
  • 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. [PMC free article] [PubMed]
  • Kumazaki T, Nakako T, Arisaka F, Ishii S. A novel method for selective isolation of C-terminal peptides from tryptic digests of proteins by immobilized anhydrotrypsin: application to structural analyses of the tail sheath and tube proteins from bacteriophage T4. Proteins. 1986 Sep;1(1):100–107. [PubMed]
  • Lawton K, Ward E, Payne G, Moyer M, Ryals J. Acidic and basic class III chitinase mRNA accumulation in response to TMV infection of tobacco. Plant Mol Biol. 1992 Aug;19(5):735–743. [PubMed]
  • Lee HI, Broekaert WF, Raikhel NV, Lee H. Co- and post-translational processing of the hevein preproprotein of latex of the rubber tree (Hevea brasiliensis) J Biol Chem. 1991 Aug 25;266(24):15944–15948. [PubMed]
  • Lerner DR, Raikhel NV. The gene for stinging nettle lectin (Urtica dioica agglutinin) encodes both a lectin and a chitinase. J Biol Chem. 1992 Jun 5;267(16):11085–11091. [PubMed]
  • Linthorst HJ, Danhash N, Brederode FT, Van Kan JA, De Wit PJ, Bol JF. Tobacco and tomato PR proteins homologous to win and pro-hevein lack the "hevein" domain. Mol Plant Microbe Interact. 1991 Nov-Dec;4(6):586–592. [PubMed]
  • Linthorst HJ, van Loon LC, van Rossum CM, Mayer A, Bol JF, van Roekel JS, Meulenhoff EJ, Cornelissen BJ. Analysis of acidic and basic chitinases from tobacco and petunia and their constitutive expression in transgenic tobacco. Mol Plant Microbe Interact. 1990 Jul-Aug;3(4):252–258. [PubMed]
  • Lischwe MA, Ochs D. A new method for partial peptide mapping using N-chlorosuccinimide/urea and peptide silver staining in sodium dodecyl sulfate-polyacrylamide gels. Anal Biochem. 1982 Dec;127(2):453–457. [PubMed]
  • Melchers LS, Sela-Buurlage MB, Vloemans SA, Woloshuk CP, Van Roekel JS, Pen J, van den Elzen PJ, Cornelissen BJ. Extracellular targeting of the vacuolar tobacco proteins AP24, chitinase and beta-1,3-glucanase in transgenic plants. Plant Mol Biol. 1993 Feb;21(4):583–593. [PubMed]
  • Molano J, Durán A, Cabib E. A rapid and sensitive assay for chitinase using tritiated chitin. Anal Biochem. 1977 Dec;83(2):648–656. [PubMed]
  • Moos M, Jr, Nguyen NY, Liu TY. Reproducible high yield sequencing of proteins electrophoretically separated and transferred to an inert support. J Biol Chem. 1988 May 5;263(13):6005–6008. [PubMed]
  • Payne G, Ahl P, Moyer M, Harper A, Beck J, Meins F, Jr, Ryals J. Isolation of complementary DNA clones encoding pathogenesis-related proteins P and Q, two acidic chitinases from tobacco. Proc Natl Acad Sci U S A. 1990 Jan;87(1):98–102. [PMC free article] [PubMed]
  • Sela-Buurlage MB, Ponstein AS, Bres-Vloemans SA, Melchers LS, Van Den Elzen PJM, Cornelissen BJC. Only Specific Tobacco (Nicotiana tabacum) Chitinases and [beta]-1,3-Glucanases Exhibit Antifungal Activity. Plant Physiol. 1993 Mar;101(3):857–863. [PMC free article] [PubMed]
  • Selsted ME, Martinez RJ. A simple and ultrasensitive enzymatic assay for the quantitative determination of lysozyme in the picogram range. Anal Biochem. 1980 Nov 15;109(1):67–70. [PubMed]
  • Shinshi H, Neuhas JM, Ryals J, Meins F., Jr Structure of a tobacco endochitinase gene: evidence that different chitinase genes can arise by transposition of sequences encoding a cysteine-rich domain. Plant Mol Biol. 1990 Mar;14(3):357–368. [PubMed]
  • Shinshi H, Wenzler H, Neuhaus JM, 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. [PMC free article] [PubMed]
  • Singh NK, Bracker CA, Hasegawa PM, Handa AK, Buckel S, Hermodson MA, Pfankoch E, Regnier FE, Bressan RA. Characterization of osmotin : a thaumatin-like protein associated with osmotic adaptation in plant cells. Plant Physiol. 1987 Oct;85(2):529–536. [PMC free article] [PubMed]
  • Stanford A, Bevan M, Northcote D. Differential expression within a family of novel wound-induced genes in potato. Mol Gen Genet. 1989 Jan;215(2):200–208. [PubMed]
  • Verwoerd TC, Dekker BM, Hoekema A. A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res. 1989 Mar 25;17(6):2362–2362. [PMC free article] [PubMed]
  • Vogelsang R, Barz W. Purification, characterization and differential hormonal regulation of a beta-1,3-glucanase and two chitinases from chickpea (Cicer arietinum L.). Planta. 1993 Jan;189(1):60–69. [PubMed]
  • Ward ER, Uknes SJ, Williams SC, Dincher SS, Wiederhold DL, Alexander DC, Ahl-Goy P, Metraux JP, Ryals JA. Coordinate Gene Activity in Response to Agents That Induce Systemic Acquired Resistance. Plant Cell. 1991 Oct;3(10):1085–1094. [PMC free article] [PubMed]
  • Yokosawa H, Ishii S. Immobilized anhydrotrypsin as a biospecific affinity adsorbent for the peptides produced by trypsin-like proteases. Anal Biochem. 1979 Sep 15;98(1):198–203. [PubMed]

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