Logo of plntphysLink to Publisher's site
Plant Physiol. 1996 Aug; 111(4): 1219–1225.
PMCID: PMC160999

Novel osmotically induced antifungal chitinases and bacterial expression of an active recombinant isoform.


NaCl (428 mM)-adapted tobacco (Nicotiana tabacum L. var Wisconsin 38) cells accumulate and secrete several antifungal chitinases. The predominant protein secreted to the culture medium was a 29-kD peptide that, based on internal amino acid sequence, was determined to be a class II acidic chitinase with similarity to PR-Q. The four predominant chitinases (T1, T2, T3, and T4) that accumulated intracellularly in 428 mM NaCl-adapted cells were purified. Based on N-terminal sequence analyses, two of these were identified as class I chitinase isoforms, one similar to the N. tomentosiformis (H. Shinshi, J.M. Neuhaus, J. Ryals, F. Meins [1990] Plant Mol Biol 14:357-368) protein (T1) and the other homologous to the N. sylvestris (Y. Fukuda, M. Ohme, H. Shinshi [1991] Plant Mol Biol 16:1-10) protein (T2). The other two proteins (T3 and T4) were determined to be novel chitinases that have sequence similarity with class I chitinases, but each lacks a chitin-binding domain. All four chitinases inhibited Fusarium oxysporum f. sp. lycopersici and Trichoderma longibrachiatum hyphal growth in vitro, although the isoforms containing a chitin-binding domain were somewhat more active. Conditions were established for the successful expression of soluble and active bacterial recombinant T2. Expression of soluble recombinant T2 was achieved when isopropyl beta-D-thiogalactopyranoside induction occurred at 18 degrees C but not at 25 or 37 degrees C. The purified recombinant protein exhibited antifungal activity comparable to a class I chitinase purified from NaCl-adapted tobacco cells.

Full Text

The Full Text of this article is available as a PDF (2.2M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Chrispeels MJ, Raikhel NV. Lectins, lectin genes, and their role in plant defense. Plant Cell. 1991 Jan;3(1):1–9. [PMC free article] [PubMed]
  • Fukuda Y, Ohme M, Shinshi H. Gene structure and expression of a tobacco endochitinase gene in suspension-cultured tobacco cells. Plant Mol Biol. 1991 Jan;16(1):1–10. [PubMed]
  • Guan KL, Dixon JE. Eukaryotic proteins expressed in Escherichia coli: an improved thrombin cleavage and purification procedure of fusion proteins with glutathione S-transferase. Anal Biochem. 1991 Feb 1;192(2):262–267. [PubMed]
  • Huynh QK, Hironaka CM, Levine EB, Smith CE, Borgmeyer JR, Shah DM. Antifungal proteins from plants. Purification, molecular cloning, and antifungal properties of chitinases from maize seed. J Biol Chem. 1992 Apr 5;267(10):6635–6640. [PubMed]
  • Jach G, Görnhardt B, Mundy J, Logemann J, Pinsdorf E, Leah R, Schell J, Maas C. Enhanced quantitative resistance against fungal disease by combinatorial expression of different barley antifungal proteins in transgenic tobacco. Plant J. 1995 Jul;8(1):97–109. [PubMed]
  • Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. [PubMed]
  • Larosa PC, Chen Z, Nelson DE, Singh NK, Hasegawa PM, Bressan RA. Osmotin gene expression is posttranscriptionally regulated. Plant Physiol. 1992 Sep;100(1):409–415. [PMC free article] [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]
  • Mauch F, Mauch-Mani B, Boller T. Antifungal Hydrolases in Pea Tissue : II. Inhibition of Fungal Growth by Combinations of Chitinase and beta-1,3-Glucanase. Plant Physiol. 1988 Nov;88(3):936–942. [PMC free article] [PubMed]
  • Melchers LS, Apotheker-de Groot M, van der Knaap JA, Ponstein AS, Sela-Buurlage MB, Bol JF, Cornelissen BJ, van den Elzen PJ, Linthorst HJ. A new class of tobacco chitinases homologous to bacterial exo-chitinases displays antifungal activity. Plant J. 1994 Apr;5(4):469–480. [PubMed]
  • Nielsen KK, Bojsen K, Roepstorff P, Mikkelsen JD. A hydroxyproline-containing class IV chitinase of sugar beet is glycosylated with xylose. Plant Mol Biol. 1994 May;25(2):241–257. [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]
  • Raghothama KG, Liu D, Nelson DE, Hasegawa PM, Bressan RA. Analysis of an osmotically regulated pathogenesis-related osmotin gene promoter. Plant Mol Biol. 1993 Dec;23(6):1117–1128. [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]
  • 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]
  • Song CS, Yu JH, Bai DH, Hester PY, Kim KH. Antibodies to the alpha-subunit of insulin receptor from eggs of immunized hens. J Immunol. 1985 Nov;135(5):3354–3359. [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]
  • Stintzi A, Heitz T, Prasad V, Wiedemann-Merdinoglu S, Kauffmann S, Geoffroy P, Legrand M, Fritig B. Plant 'pathogenesis-related' proteins and their role in defense against pathogens. Biochimie. 1993;75(8):687–706. [PubMed]
  • Takeda S, Sato F, Ida K, Yamada Y. Nucleotide sequence of a cDNA for osmotin-like protein from cultured tobacco cells. Plant Physiol. 1991 Oct;97(2):844–846. [PMC free article] [PubMed]
  • van Loon LC, van Kammen A. Polyacrylamide disc electrophoresis of the soluble leaf proteins from Nicotiana tabacum var. "Samsun" and "Samsun NN". II. Changes in protein constitution after infection with tobacco mosaic virus. Virology. 1970 Feb;40(2):190–211. [PubMed]

Articles from Plant Physiology are provided here courtesy of American Society of Plant Biologists


Save items

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • Compound
    PubChem chemical compound records that cite the current articles. These references are taken from those provided on submitted PubChem chemical substance records. Multiple substance records may contribute to the PubChem compound record.
  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles
  • Substance
    PubChem chemical substance records that cite the current articles. These references are taken from those provided on submitted PubChem chemical substance records.

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...