Skip to main content
Applied and Environmental Microbiology logoLink to Applied and Environmental Microbiology
. 1977 Oct;34(4):424–432. doi: 10.1128/aem.34.4.424-432.1977

Bacterial lipopolysaccharides as inducers of disease resistance in tobacco.

T L Graham, L Sequeira, T S Huang
PMCID: PMC242674  PMID: 21613

Abstract

The cell wall component of Pseudomonas solanacearum that induces disease resistance in tobacco was highly heat stable at neutral or alkaline pH but highly labile at acid pH. Activity was unaffected by nucleases and proteases but destroyed by a mixture of beta-glycosidases. Washing of bacterial cell walls released a lipopolysaccharide (LPS) fraction with high inducer activity. Purified LPS, extracted by a variety of procedures from whole cells, isolated cell walls, and culture filtrates of both smooth and rough forms of P. solanacearum, induced disease resistance in tobacco at concentrations as low as 50 microgram/ml. The LPS from the non-plant pathogens Escherichia coli B, E. coli K, and Serratia marcescens was also active. Cell wall protein, free phospholipid, and nucleic acids were not necessary for activity. Moreover, since LPS from rough forms was active, the O-specific polysaccharide of the LPS was not required for activity. Hydrolysis of the remaining core-lipid A linkage or deacylation of lipid A destroyed inducer activity. When injected into tobacco leaves, purified LPS attached to tobacco mesophyll cell walls and induced ultrastructural changes in the host cell similar to those induced by attachment of whole heat-killed bacteria.

Full text

PDF
424

Images in this article

Selected References

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

  1. BRAGDON J. H. Colorimetric determination of blood lipides. J Biol Chem. 1951 Jun;190(2):513–517. [PubMed] [Google Scholar]
  2. Corpe W. A., Salton M. R. Properties of surface materials released from cells of Chromobacterium violaceum. Biochim Biophys Acta. 1966 Jul 27;124(1):125–135. doi: 10.1016/0304-4165(66)90320-5. [DOI] [PubMed] [Google Scholar]
  3. DAVIES D. A. The specific polysaccharides of some gram-negative bacteria. Biochem J. 1955 Apr;59(4):696–704. doi: 10.1042/bj0590696. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. DISCHE Z. Qualitative and quantitative colorimetric determination of heptoses. J Biol Chem. 1953 Oct;204(2):983–997. [PubMed] [Google Scholar]
  5. Galanos C., Lüderitz O., Westphal O. A new method for the extraction of R lipopolysaccharides. Eur J Biochem. 1969 Jun;9(2):245–249. doi: 10.1111/j.1432-1033.1969.tb00601.x. [DOI] [PubMed] [Google Scholar]
  6. Gray G. W., Wilkinson S. G. The effect of ethylenediaminetetra-acetic acid on the cell walls of some gram-negative bacteria. J Gen Microbiol. 1965 Jun;39(3):385–399. doi: 10.1099/00221287-39-3-385. [DOI] [PubMed] [Google Scholar]
  7. Homma J. Y., Suzuki N. The protein moiety of the endotoxin of Pseudomonas aeruginosa. Ann N Y Acad Sci. 1966 Jun 30;133(2):508–526. doi: 10.1111/j.1749-6632.1966.tb52386.x. [DOI] [PubMed] [Google Scholar]
  8. Jones G. W., Rutter J. M. Role of the K88 antigen in the pathogenesis of neonatal diarrhea caused by Escherichia coli in piglets. Infect Immun. 1972 Dec;6(6):918–927. doi: 10.1128/iai.6.6.918-927.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Knox K. W., Cullen J., Work E. An extracellular lipopolysaccharide-phospholipid-protein complex produced by Escherichia coli grown under lysine-limiting conditions. Biochem J. 1967 Apr;103(1):192–201. doi: 10.1042/bj1030192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. LOWRY O. H., ROBERTS N. R., LEINER K. Y., WU M. L., FARR A. L. The quantitative histochemistry of brain. I. Chemical methods. J Biol Chem. 1954 Mar;207(1):1–17. [PubMed] [Google Scholar]
  11. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  12. Leive L., Shovlin V. K., Mergenhagen S. E. Physical, chemical, and immunological properties of lipopolysaccharide released from Escherichia coli by ethylenediaminetetraacetate. J Biol Chem. 1968 Dec 25;243(24):6384–6391. [PubMed] [Google Scholar]
  13. Loebenstein G., Lovrekovich L. Interference with tobacco mosaic virus local lesion formation in tobacco by injection heat-killed cells of Pseudomonas syringae. Virology. 1966 Dec;30(4):587–591. doi: 10.1016/0042-6822(66)90162-0. [DOI] [PubMed] [Google Scholar]
  14. Marsh D. G., Crutchley M. J. Purification and physico-chemical analysis of fractions from the culture supernatant of Escherichia coli O78K80: free endotoxin and a non-toxic fraction. J Gen Microbiol. 1967 Jun;47(3):405–420. doi: 10.1099/00221287-47-3-405. [DOI] [PubMed] [Google Scholar]
  15. NETER E., WESTPHAL O., LUDERITZ O., GORZYNSKI E. A., EICHENBERGER E. Studies of enterobacterial lipopolysaccharides; effects of heat and chemicals on erythrocyte-modifying, antigenic, toxic and pyrogenic properties. J Immunol. 1956 May;76(5):377–385. [PubMed] [Google Scholar]
  16. Ng A. K., Butler R. C., Chen C. L., Nowotny A. Relationship of structure to function in bacterial endotoxins. IX. Differences in the lipid moiety of endotoxic glycolipids. J Bacteriol. 1976 Apr;126(1):511–515. doi: 10.1128/jb.126.1.511-515.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Nowotny A. Molecular aspects of endotoxic reactions. Bacteriol Rev. 1969 Mar;33(1):72–98. doi: 10.1128/br.33.1.72-98.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Roberts N. A., Gray G. W., Wilkinson S. G. Release of lipopolysaccharide during the preparation of cell walls of Pseudomonas aeruginosa. Biochim Biophys Acta. 1967;135(5):1068–1071. doi: 10.1016/0005-2736(67)90079-x. [DOI] [PubMed] [Google Scholar]
  19. Rogers S. W., Gilleland H. E., Jr, Eagon R. G. Characterization of a protein-lipopolysaccharide complex released from cell walls of Pseudomonas aeruginosa by ethylenediaminetetraacetic acid. Can J Microbiol. 1969 Jul;15(7):743–748. doi: 10.1139/m69-130. [DOI] [PubMed] [Google Scholar]
  20. SALTON M. R. J., HORNE R. W. Studies of the bacterial cell wall. II. Methods of preparation and some properties of cell walls. Biochim Biophys Acta. 1951 Jul;7(2):177–197. doi: 10.1016/0006-3002(51)90017-0. [DOI] [PubMed] [Google Scholar]
  21. STROMINGER J. L., PARK J. T., THOMPSON R. E. Composition of the cell wall of Staphylococcus aureus: its relation to the mechanism of action of penicillin. J Biol Chem. 1959 Dec;234:3263–3268. [PubMed] [Google Scholar]
  22. Smith H. W., Linggood M. A. Observations on the pathogenic properties of the K88, Hly and Ent plasmids of Escherichia coli with particular reference to porcine diarrhoea. J Med Microbiol. 1971 Nov;4(4):467–485. doi: 10.1099/00222615-4-4-467. [DOI] [PubMed] [Google Scholar]
  23. TREVELYAN W. E., PROCTER D. P., HARRISON J. S. Detection of sugars on paper chromatograms. Nature. 1950 Sep 9;166(4219):444–445. doi: 10.1038/166444b0. [DOI] [PubMed] [Google Scholar]
  24. Tsang J. C., Wang C. S., Alaupovic P. Degradative effect of phenol on endotoxin and lipopolysaccharide preparations from Serratia marcescens. J Bacteriol. 1974 Feb;117(2):786–795. doi: 10.1128/jb.117.2.786-795.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. WARREN L. Thiobarbituric acid spray reaction for deoxy sugars and sialic acids. Nature. 1960 Apr 16;186:237–237. doi: 10.1038/186237a0. [DOI] [PubMed] [Google Scholar]
  26. WEISSBACH A., HURWITZ J. The formation of 2-keto-3-deoxyheptonic acid in extracts of Escherichia coli B. I. Identification. J Biol Chem. 1959 Apr;234(4):705–709. [PubMed] [Google Scholar]
  27. Whatley M. H., Bodwin J. S., Lippincott B. B., Lippincott J. A. Role of Agrobacterium cell envelope lipopolysaccharide in infection site attachment. Infect Immun. 1976 Apr;13(4):1080–1083. doi: 10.1128/iai.13.4.1080-1083.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wolpert J. S., Albersheim P. Host-symbiont interactions. I. The lectins of legumes interact with the o-antigen-containing lipopolysaccharides of their symbiont Rhizobia. Biochem Biophys Res Commun. 1976 Jun 7;70(3):729–737. doi: 10.1016/0006-291x(76)90653-7. [DOI] [PubMed] [Google Scholar]
  29. Wright B. G., Rebers P. A. Procedure for determining heptose and hexose in lipopolysaccharides. Modification of the cysteine-sulfuric acid method. Anal Biochem. 1972 Oct;49(2):307–319. doi: 10.1016/0003-2697(72)90433-2. [DOI] [PubMed] [Google Scholar]

Articles from Applied and Environmental Microbiology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES