Abstract
The objectives of this work were (i) to use transposon mutagenesis to produce mutants of Pseudomonas fluorescens that were altered in adhesion ability and transport through porous media and (ii) to identify the alterations in surface characteristics that were responsible for the changes in attachment. Mutants of P. fluorescens were generated with TnphoA, which enabled identification of mutants that were altered in surface proteins. Transposon mutants were screened for alterations in adhesion ability by attachment assays on hydrophobic polystyrene and water-wettable polystyrene. Four TnphoA mutants with increased adhesion to the hydrophobic surface and decreased adhesion to the water-wettable surface were obtained. Transport of the strains through porous media was evaluated by passing suspensions of each mutant and the parent through columns containing quartz sand and determining the number of cells retained in the columns. The mutants all demonstrated increased adhesion and retention in the columns. Southern analysis demonstrated two types of mutants with separate transposon insertion sites. Polyacrylamide gel electrophoresis of the strains demonstrated that the O antigen on the lipopolysaccharide was either attenuated or absent. Lack of this polysaccharide, and the consequent increased exposure of the lipid moiety of the lipopolysaccharide, is probably responsible for the increase in adhesion to the hydrophobic substrata and retention in the sand column. This work combined with previous studies of attachment of P. fluorescens demonstrates that more than one type of polymer can mediate the adhesion of this organism to nonbiological surfaces.
Full Text
The Full Text of this article is available as a PDF (243.8 KB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Belas R., Erskine D., Flaherty D. Transposon mutagenesis in Proteus mirabilis. J Bacteriol. 1991 Oct;173(19):6289–6293. doi: 10.1128/jb.173.19.6289-6293.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Bright J. J., Fletcher M. Amino Acid assimilation and electron transport system activity in attached and free-living marine bacteria. Appl Environ Microbiol. 1983 Mar;45(3):818–825. doi: 10.1128/aem.45.3.818-825.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Busscher H. J., Weerkamp A. H., van der Mei H. C., van Pelt A. W., de Jong H. P., Arends J. Measurement of the surface free energy of bacterial cell surfaces and its relevance for adhesion. Appl Environ Microbiol. 1984 Nov;48(5):980–983. doi: 10.1128/aem.48.5.980-983.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Camper A. K., Hayes J. T., Sturman P. J., Jones W. L., Cunningham A. B. Effects of Motility and Adsorption Rate Coefficient on Transport of Bacteria through Saturated Porous Media. Appl Environ Microbiol. 1993 Oct;59(10):3455–3462. doi: 10.1128/aem.59.10.3455-3462.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deflaun M. F., Marshall B. M., Kulle E. P., Levy S. B. Tn5 Insertion Mutants of Pseudomonas fluorescens Defective in Adhesion to Soil and Seeds. Appl Environ Microbiol. 1994 Jul;60(7):2637–2642. doi: 10.1128/aem.60.7.2637-2642.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Deflaun M. F., Tanzer A. S., McAteer A. L., Marshall B., Levy S. B. Development of an Adhesion Assay and Characterization of an Adhesion-Deficient Mutant of Pseudomonas fluorescens. Appl Environ Microbiol. 1990 Jan;56(1):112–119. doi: 10.1128/aem.56.1.112-119.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fletcher M., Marshall K. C. Bubble contact angle method for evaluating substratum interfacial characteristics and its relevance to bacterial attachment. Appl Environ Microbiol. 1982 Jul;44(1):184–192. doi: 10.1128/aem.44.1.184-192.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fontes D. E., Mills A. L., Hornberger G. M., Herman J. S. Physical and chemical factors influencing transport of microorganisms through porous media. Appl Environ Microbiol. 1991 Sep;57(9):2473–2481. doi: 10.1128/aem.57.9.2473-2481.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gannon J. T., Manilal V. B., Alexander M. Relationship between Cell Surface Properties and Transport of Bacteria through Soil. Appl Environ Microbiol. 1991 Jan;57(1):190–193. doi: 10.1128/aem.57.1.190-193.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Gannon J., Tan Y. H., Baveye P., Alexander M. Effect of sodium chloride on transport of bacteria in a saturated aquifer material. Appl Environ Microbiol. 1991 Sep;57(9):2497–2501. doi: 10.1128/aem.57.9.2497-2501.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hitchcock P. J., Brown T. M. Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver-stained polyacrylamide gels. J Bacteriol. 1983 Apr;154(1):269–277. doi: 10.1128/jb.154.1.269-277.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Long S., McCune S., Walker G. C. Symbiotic loci of Rhizobium meliloti identified by random TnphoA mutagenesis. J Bacteriol. 1988 Sep;170(9):4257–4265. doi: 10.1128/jb.170.9.4257-4265.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Manoil C., Mekalanos J. J., Beckwith J. Alkaline phosphatase fusions: sensors of subcellular location. J Bacteriol. 1990 Feb;172(2):515–518. doi: 10.1128/jb.172.2.515-518.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McCarter L. L., Silverman M. Phosphate regulation of gene expression in Vibrio parahaemolyticus. J Bacteriol. 1987 Aug;169(8):3441–3449. doi: 10.1128/jb.169.8.3441-3449.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
- McEldowney S., Fletcher M. Variability of the influence of physicochemical factors affecting bacterial adhesion to polystyrene substrata. Appl Environ Microbiol. 1986 Sep;52(3):460–465. doi: 10.1128/aem.52.3.460-465.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Miller V. L., Mekalanos J. J. A novel suicide vector and its use in construction of insertion mutations: osmoregulation of outer membrane proteins and virulence determinants in Vibrio cholerae requires toxR. J Bacteriol. 1988 Jun;170(6):2575–2583. doi: 10.1128/jb.170.6.2575-2583.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Paul J. H., Jeffrey W. H. Evidence for Separate Adhesion Mechanisms for Hydrophilic and Hydrophobic Surfaces in Vibrio proteolytica. Appl Environ Microbiol. 1985 Aug;50(2):431–437. doi: 10.1128/aem.50.2.431-437.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Sjollema J., van der Mei H. C., Uyen H. M., Busscher H. J. Direct observations of cooperative effects in oral streptococcal adhesion to glass by analysis of the spatial arrangement of adhering bacteria. FEMS Microbiol Lett. 1990 Jun 1;57(3):263–269. doi: 10.1016/0378-1097(90)90078-5. [DOI] [PubMed] [Google Scholar]
- Stenström T. A. Bacterial hydrophobicity, an overall parameter for the measurement of adhesion potential to soil particles. Appl Environ Microbiol. 1989 Jan;55(1):142–147. doi: 10.1128/aem.55.1.142-147.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Vandevivere P., Baveye P. Relationship between Transport of Bacteria and Their Clogging Efficiency in Sand Columns. Appl Environ Microbiol. 1992 Aug;58(8):2523–2530. doi: 10.1128/aem.58.8.2523-2530.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Wiencek K. M., Klapes N. A., Foegeding P. M. Hydrophobicity of Bacillus and Clostridium spores. Appl Environ Microbiol. 1990 Sep;56(9):2600–2605. doi: 10.1128/aem.56.9.2600-2605.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
- de Lorenzo V., Herrero M., Jakubzik U., Timmis K. N. Mini-Tn5 transposon derivatives for insertion mutagenesis, promoter probing, and chromosomal insertion of cloned DNA in gram-negative eubacteria. J Bacteriol. 1990 Nov;172(11):6568–6572. doi: 10.1128/jb.172.11.6568-6572.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]