Skip to main content
PMC home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1987 Jan;84(1):89–93. doi: 10.1073/pnas.84.1.89

Regulation of a plant pathogenesis-related enzyme: Inhibition of chitinase and chitinase mRNA accumulation in cultured tobacco tissues by auxin and cytokinin

Hideaki Shinshi 1,*, Debra Mohnen 1,†, Frederick Meins Jr 1,‡
PMCID: PMC304147  PMID: 16593796

Abstract

Two endochitinases (EC 3.2.1.14) of Mr values of ≈34,000 and ≈32,000 have been purified from cultured tissues of Nicotiana tabacum cv. Havana 425. The chitinase content of cloned tobacco pith tissues subcultured on hormone-free medium increases by ≈5-fold to 8% of the soluble protein over a 7-day period. This induction is inhibited >90% by addition of combinations of the plant hormones auxin and cytokinin to the culture medium. Chitinase is also developmentally regulated in the intact plant. Not detectable in leaves near the top of the plant, it is 1-4% of the soluble protein in roots and lower leaves. A cDNA clone of tobacco chitinase was isolated containing a single, large open reading frame of 310 amino acids that includes the complete amino acid sequence of the mature enzyme. Chitinase and chitinase mRNA measured by RNA blot analysis show similar patterns of regulation indicating that chitinase accumulation is controlled, at least in part, at the mRNA level. The patterns were also similar to those obtained with glucan endo-1,3-β-glucosidase (EC 3.2.1.39) suggesting that the two enzymes are coordinately regulated.

Keywords: Nicotiana tabacum; glucan endo-1,3-β-glucosidase mRNA; β-1,3-glucanase

Full text

PDF
89

Images in this article

90Image
on p.90
92Image
on p.92
92Image
on p.92
92Image
on p.92
92Image
on p.92
92Image
on p.92

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Abeles F. B., Bosshart R. P., Forrence L. E., Habig W. H. Preparation and purification of glucanase and chitinase from bean leaves. Plant Physiol. 1971 Jan;47(1):129–134. doi: 10.1104/pp.47.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bartnicki-Garcia S. Cell wall chemistry, morphogenesis, and taxonomy of fungi. Annu Rev Microbiol. 1968;22:87–108. doi: 10.1146/annurev.mi.22.100168.000511. [DOI] [PubMed] [Google Scholar]
  3. Broglie K. E., Gaynor J. J., Broglie R. M. Ethylene-regulated gene expression: molecular cloning of the genes encoding an endochitinase from Phaseolus vulgaris. Proc Natl Acad Sci U S A. 1986 Sep;83(18):6820–6824. doi: 10.1073/pnas.83.18.6820. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Hewick R. M., Hunkapiller M. W., Hood L. E., Dreyer W. J. A gas-liquid solid phase peptide and protein sequenator. J Biol Chem. 1981 Aug 10;256(15):7990–7997. [PubMed] [Google Scholar]
  5. Hunkapiller M. W., Hood L. E. Analysis of phenylthiohydantoins by ultrasensitive gradient high-performance liquid chromatography. Methods Enzymol. 1983;91:486–493. doi: 10.1016/s0076-6879(83)91045-5. [DOI] [PubMed] [Google Scholar]
  6. Mohnen D., Shinshi H., Felix G., Meins F. Hormonal regulation of beta1,3-glucanase messenger RNA levels in cultured tobacco tissues. EMBO J. 1985 Jul;4(7):1631–1635. doi: 10.1002/j.1460-2075.1985.tb03830.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. POWNING R. F., IRZYKIEWICZ H. STUDIES ON THE CHITINASE SYSTEM IN BEAN AND OTHER SEEDS. Comp Biochem Physiol. 1965 Jan;14:127–133. doi: 10.1016/0010-406x(65)90013-7. [DOI] [PubMed] [Google Scholar]
  8. Pengelly W. L., Meins F., Jr The relationship of indole-3-acetic acid content and growth of crown-gall tumor tissues of tobacco in culture. Differentiation. 1982;21(1):27–31. doi: 10.1111/j.1432-0436.1982.tb01190.x. [DOI] [PubMed] [Google Scholar]
  9. Proudfoot N. J., Brownlee G. G. 3' non-coding region sequences in eukaryotic messenger RNA. Nature. 1976 Sep 16;263(5574):211–214. doi: 10.1038/263211a0. [DOI] [PubMed] [Google Scholar]
  10. Queen C., Korn L. J. A comprehensive sequence analysis program for the IBM personal computer. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):581–599. doi: 10.1093/nar/12.1part2.581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Verma D. P., Maclachlan G. A., Byrne H., Ewings D. Regulation and in vitro translation of messenger ribonucleic acid for cellulase from auxin-treated pea epicotyls. J Biol Chem. 1975 Feb 10;250(3):1019–1026. [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES