Skip to main content
SpringerLink
Log in

Comparative analysis of the lipopolysaccharides of a rough and a smooth strain of Pseudomonas syringae pv. phaseolicola

  • Original Papers
  • Published:
Archives of Microbiology Aims and scope Submit manuscript

Abstract

The lipopolysaccharides (LPS) of a rough (R) and a smooth (S) strain of Pseudomonas syringae pv. phaseolicola were analysed. The S-LPS revealed markedly more rhamnose and fucose, but less glucose, than the R-LPS. The presence of 3-O-methyl-rhamnose (acofriose) in the S-LPS was confirmed by cochromatography with authentic acofriose. SDS polyacrylamide gel electrophoresis of the S-LPS demonstrated a cluster of regularly spaced high molecular weight fractions, which was almost lacking in the R-LPS. The main fatty acids of the lipid A of both LPS species were 3-OH-10:0,3-OH-12:0,2-OH-12:0, and 12:0. Two N-linked diesters were demonstrated: 3-O(12:0)-12:0 and 3-O(2-OH-12:0)-12:0. S-LPS was subjected to mild hydrolysis and the “degraded polysaccharide” separated into three fractions by gel permeation chromatography on a Fractogel TSK HW-50 column. Fraction I, representing nearly only the O-specific side chain, consisted of rhamnose and fucose in a molar ratio of 4:1, with 4% of the rhamnose being 3-O-methylated (acofriose). Fraction II, representing mostly core material, was composed of glucose, rhamnose, heptose, glucosamine, galactosamine, alanine, and a still unidentified amino compound, in an approximate molar ratio of 3:1:1:1:1:1:1, and KDO. Fraction III consisted of released monomers and salts. The LPS was highly phosphorylated (3.28% phosphorus in the “core fraction”). The thus characterized composition of the LPS O-chain seems to be unique for the pathovar phaseolicola of P. syringae, although many similarities exist to other pathovars as well as to other bacterial species.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

LPS:

lipopolysacchairdes

GC/MS:

combined gas liquid chromatography-mass spectrometry

HVE:

high voltage electrophoresis

KDO:

2-keto-3-deoxyoctonic acid

PAGE:

polyacrylamide gel electrophoresis

SDS:

sodium dodecylsulfate

References

  • Adam DB, Pugsley AT (1934) “Smooth-rough” variation in Phytomonas medicaginis phaseolicola Burk. Austr J Exp Biol Med Sci 12:193–201

    Google Scholar 

  • Anderson AJ (1984) Differences between lipopolysaccharide compositions of plant pathogenic and saprophytic Pseudomonas species. Appl Env Microbiol 48:31–35

    Google Scholar 

  • Anderson PJ (1966) A sensitive reagent for detecting 2-deoxysugars and 3-deoxypolyols. J Chromatogr 21:163–164

    Google Scholar 

  • Baker CJ, Neilson MJ, Sequeira L, Keegstra KG (1984) Chemical characterization of the lipopolysaccharide of Pseudomonas solanacearum. Appl Env Microbiol 47:1096–1100

    Google Scholar 

  • Bartlett GR (1959) Phosphorus assay in column chromatography. J Biol Chem 234:466–468

    Google Scholar 

  • Barton-Willis PA, Wang MC, Holliday MJ, Long MR, Keen NT (1984) Purification and composition of lipopolysaccharides from Pseudomonas syringae pv. glycinea. Physiol Plant Pathol 25:387–398

    Google Scholar 

  • Barton-Willis PA, Wang MC, Staskawicz B, Keen NT (1987) Structural studies on the O-chain polysaccharides of lipopolysaccharides from Pseudomonas syringae pv. glycinea. Physiol Mol Plant Pathol 30:187–197

    Google Scholar 

  • Basu S, Radziejewska-Lebrecht J, Mayer H (1986) Lipopolysaccharides of Providencia rettgeri. Arch Microbiol 144:213–218

    Google Scholar 

  • DeVos P, DeLey J (1983) Intra- and intergenic similarities of Pseudomonas and Xanthomonas ribosomal ribonucleic acid cistrons. Int J Syst Bacteriol 33:487–509

    Google Scholar 

  • Drewry DT, Symes KC, Gray GW, Wilkinson SG (1975) Studies of polysaccharide fractions from the lipopolysaccharide of Pseudomonas aeruginosa NCTC 1999. Biochem J 149:93–106

    Google Scholar 

  • Evans LR, Linker A (1973) Production and characterization of the slime polysaccharide of Pseudomonas aeruginosa. J Bacteriol 116:915–924

    Google Scholar 

  • Fett WF, Osman SF, Fishman ML, Siebles TS (1986) Alginate production by plant-pathogenic pseudomonads. Appl Env Microbiol 52:466–473

    Google Scholar 

  • Galanos C, Lüderitz O, Westphal O (1969) A new method for the extraction of R-lipopolysaccharides. Eur J Biochem 9:245–249

    Google Scholar 

  • Gerwe PG, Rudolph K, Köhn S (1987) Vergleich eines “glatten” und eines “rauhen” Stammes von Pseudomonas syringae pv. phaseolicola. Phytopath 118:326–334

    Google Scholar 

  • Gross M, Rudolph K (1987) Demonstration of levan and alginate in bean plants (Phaseolus vulgaris) infected by Pseudomonas syringae pv. phaseolicola. J Phytopathol 120:9–19

    Google Scholar 

  • Hendrick CA, Sequeira L (1984) Lipopolysaccharide-defective mutants of the wilt pathogen Pseudomonas solanacearum. Appl Env Microbiol 48:94–101

    Google Scholar 

  • Jansson PE, Lindberg AA, Lindberg B, Wollin R (1981) Structural studies on the hexose region of the core in lipopolysaccharides from Enterobacteriaceae. Eur J Biochem 115:571–577

    Google Scholar 

  • Karkhanis YD, Zetner J, Jackson JJ, Carlo DJ (1978) A new and improved micro-assay to determine 2-keto-3-deoxyoctonate in lipopolysaccharide of Gram-negative bacteria. Anal Biochem 85:595–601

    Google Scholar 

  • Keen NT, Williams PH (1971) Chemical and biological properties of a lipomucopolysaccharide from Pseudomonas lachrymans. Physiol Plant Pathol 1:247–264

    Google Scholar 

  • King EO, Ward MK, Raney DE (1954) Two simple media for the demonstration of pyocyanin and fluorescin. J Lab Clin Med 44:301–307

    Google Scholar 

  • Klement Z, Rudolph K, Ebrahim-Nesbat F (1984) Comparison of rough and smooth strains of Pseudomonas phaseolicola: Symptom development on a macroscopic and microscopic scale and multiplication rates in the leaves. Proc. 2nd Working Group on Pseudomonas syringae Pathovars. Sounion, Greece, pp 76–77

  • Kotelko K (1986) Proteus mirabilis: Taxonomic position, pecularities of growth, components of the cell envelope. Curr Top Microbiol Immunol 129:181–215

    Google Scholar 

  • Lowry OH, Roberts NR, Leiner KY, Mu ML, Farr AL (1954) The quantitative histochemistry of brain. I. Chemical methods. J Biol Chem 207:1–17

    Google Scholar 

  • Palleroni NJ (1984) Family 1. Pseudomonaceae. In: Krieg NR, Holt JG (eds) Bergey's manual of systematic bacteriology, vol 1. Williams and Wilkens, Baltimore, pp 141–219

    Google Scholar 

  • Romanowska E, Gamian A, Dabrowski J (1986) Core region of Citrobacter lipopolysaccharide from strain PCM 1487. Structure elucidation by two-dimensional 1H-NMR spectroscopy at 500 MHz and methylation analysis/mass spectrometry. Eur J Biochem 161:557–564

    Google Scholar 

  • Rowe PSN, Meadow PM (1983) Structure of the core oligosaccharide from the lipopolysaccharide of Pseudomonas aeruginosa PAC 1 R and its defective mutants. Eur J Biochem 132:329–337

    Google Scholar 

  • Sawardeker JS, Sloneker JH, Jeanes A (1965) Quantitative determination of monosaccharides as their alditol acetates by gas liquid chromatography. Anal Chem 37:1602–1604

    Google Scholar 

  • Sequeira L, Graham TL (1977) Agglutination of avirulent strains of Pseudomonas phaseolicola by potato lectin. Physiol Plant Pathol 11:43–54

    Google Scholar 

  • Smith ARW, Zamze SE, Hignett RC (1985) Composition of lipopolysaccharide from Pseudomonas syringae pv. morsprunorum and its digestion by bacteriophage A7 J Gen Microbiol 131:963–974

    Google Scholar 

  • Tharanathan RN, Weckesser J, Mayer H (1978) Location of O-methyl sugars in antigenic (lipo-)polysaccharides of photosynthetic bacteria and cyanobacteria. Biochem J 171:403–408

    Google Scholar 

  • Tsai CM, Frash CE (1982) A sensitive silver stain for detecting lipopolysaccharides in polyacrylamide gels. Anal Biochem 119:115–119

    Google Scholar 

  • Westphal O, Lüderitz O, Bister F (1952) Über die Extraktion von Bakterien mit Phenol/Wasser. Z Naturforsch 7b:148–155

    Google Scholar 

  • Wilkinson SG (1977) Composition and structure of bacterial lipopolysaccharides. In: Sutherland I (ed) Surface carbohydrates of the procaryotic cell. Academic Press, London, pp 97–175

    Google Scholar 

  • Whatley MH, Hunter, N, Cantrell MA, Hendrick C, Keegstra K, Sequeira L (1980) Lipopolysaccharide composition of the wilt pathogen, Pseudomonas solanacearum. Plant Physiol 65:557–559

    Google Scholar 

  • Wollenweber HW, Broady KW, Lüderitz O, Rietschel ET (1982) The chemical structure of lipid A. Demonstration of amide-linked 3-acyloxyacyl residues in Salmonella minnesota Re lipopolysaccharide. Eur J Biochem 124:191–198

    Google Scholar 

  • Wollenweber H-W, Seydel U, Lindner B, Lüderitz O, Rietschel ET (1984) Nature and location of amide-bound (R)-3-acyloxyacyl groups in lipid A of lipopolysaccharides from various Gramnegative bacteria. Eur J Biochem 145:265–272

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Institut für Pflanzenpathologie und Pflanzenschutz, Grisebachstrasse 6, D-3400, Göttingen, Federal Republic of Germany

    M. Gross & K. Rudolph

  2. Max Planck-Institut für Immunbiologie, Stübeweg 51, D-7800, Freiburg, Federal Republic of Germany

    H. Mayer & C. Widemann

Authors
  1. M. Gross
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. H. Mayer
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. C. Widemann
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. K. Rudolph
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

P.s. pv. phaseolicola is termed P. phaseolicola in the text

Rights and permissions

Reprints and permissions

About this article

Cite this article

Gross, M., Mayer, H., Widemann, C. et al. Comparative analysis of the lipopolysaccharides of a rough and a smooth strain of Pseudomonas syringae pv. phaseolicola . Arch Microbiol 149, 372–376 (1988). https://doi.org/10.1007/BF00411658

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00411658

Key words

Search

Navigation