Effect of transposon-induced motility mutations on colonization of the host light organ by Vibrio fischeri (original) (raw)

Abstract

Vibrio fischeri is found both as a free-living bacterium in seawater and as the specific, mutualistic light organ symbiont of several fish and squid species. To identify those characteristics of symbiosis-competent strains that are required for successful colonization of the nascent light organ of juvenile Euprymna scolopes squids, we generated a mutant pool by using the transposon Mu dI 1681 and screened this pool for strains that were no longer motile. Eighteen independently isolated nonmotile mutants that were either flagellated or nonflagellated were obtained. In contrast to the parent strain, none of these nonmotile mutants was able to colonize the juvenile squid light organ. The flagellated nonmotile mutant strain NM200 possessed a bundle of sheathed polar flagella indistinguishable from that of the wild-type strain, indicating that the presence of flagella alone is not sufficient for colonization and that it is motility itself that is required for successful light organ colonization. This study identifies motility as the first required symbiotic phenotype of V. fischeri.

6986

Images in this article

Selected References

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

  1. Allen R. D., Baumann P. Structure and arrangement of flagella in species of the genus Beneckea and Photobacterium fischeri. J Bacteriol. 1971 Jul;107(1):295–302. doi: 10.1128/jb.107.1.295-302.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Belas R., Mileham A., Simon M., Silverman M. Transposon mutagenesis of marine Vibrio spp. J Bacteriol. 1984 Jun;158(3):890–896. doi: 10.1128/jb.158.3.890-896.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Belas R., Simon M., Silverman M. Regulation of lateral flagella gene transcription in Vibrio parahaemolyticus. J Bacteriol. 1986 Jul;167(1):210–218. doi: 10.1128/jb.167.1.210-218.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boettcher K. J., Ruby E. G. Depressed light emission by symbiotic Vibrio fischeri of the sepiolid squid Euprymna scolopes. J Bacteriol. 1990 Jul;172(7):3701–3706. doi: 10.1128/jb.172.7.3701-3706.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Castilho B. A., Olfson P., Casadaban M. J. Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons. J Bacteriol. 1984 May;158(2):488–495. doi: 10.1128/jb.158.2.488-495.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Csonka L. N., Clark A. J. Construction of an Hfr strain useful for transferring recA mutations between Escherichia coli strains. J Bacteriol. 1980 Jul;143(1):529–530. doi: 10.1128/jb.143.1.529-530.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dunlap P. V., Callahan S. M. Characterization of a periplasmic 3':5'-cyclic nucleotide phosphodiesterase gene, cpdP, from the marine symbiotic bacterium Vibrio fischeri. J Bacteriol. 1993 Aug;175(15):4615–4624. doi: 10.1128/jb.175.15.4615-4624.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Dunlap P. V., Greenberg E. P. Control of Vibrio fischeri luminescence gene expression in Escherichia coli by cyclic AMP and cyclic AMP receptor protein. J Bacteriol. 1985 Oct;164(1):45–50. doi: 10.1128/jb.164.1.45-50.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Dunlap P. V., Kuo A. Cell density-dependent modulation of the Vibrio fischeri luminescence system in the absence of autoinducer and LuxR protein. J Bacteriol. 1992 Apr;174(8):2440–2448. doi: 10.1128/jb.174.8.2440-2448.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dunlap P. V. Regulation of luminescence by cyclic AMP in cya-like and crp-like mutants of Vibrio fischeri. J Bacteriol. 1989 Feb;171(2):1199–1202. doi: 10.1128/jb.171.2.1199-1202.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gillen K. L., Hughes K. T. Negative regulatory loci coupling flagellin synthesis to flagellar assembly in Salmonella typhimurium. J Bacteriol. 1991 Apr;173(7):2301–2310. doi: 10.1128/jb.173.7.2301-2310.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Groisman E. A. In vivo genetic engineering with bacteriophage Mu. Methods Enzymol. 1991;204:180–212. doi: 10.1016/0076-6879(91)04010-l. [DOI] [PubMed] [Google Scholar]
  13. Hanahan D. Studies on transformation of Escherichia coli with plasmids. J Mol Biol. 1983 Jun 5;166(4):557–580. doi: 10.1016/s0022-2836(83)80284-8. [DOI] [PubMed] [Google Scholar]
  14. Martin M., Showalter R., Silverman M. Identification of a locus controlling expression of luminescence genes in Vibrio harveyi. J Bacteriol. 1989 May;171(5):2406–2414. doi: 10.1128/jb.171.5.2406-2414.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. 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]
  16. McFall-Ngai M. J., Ruby E. G. Symbiont recognition and subsequent morphogenesis as early events in an animal-bacterial mutualism. Science. 1991 Dec 6;254(5037):1491–1494. doi: 10.1126/science.1962208. [DOI] [PubMed] [Google Scholar]
  17. Nachamkin I., Yang X. H., Stern N. J. Role of Campylobacter jejuni flagella as colonization factors for three-day-old chicks: analysis with flagellar mutants. Appl Environ Microbiol. 1993 May;59(5):1269–1273. doi: 10.1128/aem.59.5.1269-1273.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Norqvist A., Wolf-Watz H. Characterization of a novel chromosomal virulence locus involved in expression of a major surface flagellar sheath antigen of the fish pathogen Vibrio anguillarum. Infect Immun. 1993 Jun;61(6):2434–2444. doi: 10.1128/iai.61.6.2434-2444.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Richardson K., Nixon L., Mostow P., Kaper J. B., Michalski J. Transposon-induced non-motile mutants of Vibrio cholerae. J Gen Microbiol. 1990 Apr;136(4):717–725. doi: 10.1099/00221287-136-4-717. [DOI] [PubMed] [Google Scholar]
  20. Richardson K. Roles of motility and flagellar structure in pathogenicity of Vibrio cholerae: analysis of motility mutants in three animal models. Infect Immun. 1991 Aug;59(8):2727–2736. doi: 10.1128/iai.59.8.2727-2736.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ruby E. G., Asato L. M. Growth and flagellation of Vibrio fischeri during initiation of the sepiolid squid light organ symbiosis. Arch Microbiol. 1993;159(2):160–167. doi: 10.1007/BF00250277. [DOI] [PubMed] [Google Scholar]
  22. Ruby E. G., McFall-Ngai M. J. A squid that glows in the night: development of an animal-bacterial mutualism. J Bacteriol. 1992 Aug;174(15):4865–4870. doi: 10.1128/jb.174.15.4865-4870.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Schmitt C. K., Darnell S. C., Tesh V. L., Stocker B. A., O'Brien A. D. Mutation of flgM attenuates virulence of Salmonella typhimurium, and mutation of fliA represses the attenuated phenotype. J Bacteriol. 1994 Jan;176(2):368–377. doi: 10.1128/jb.176.2.368-377.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Silverman M., Showalter R., McCarter L. Genetic analysis in vibrio. Methods Enzymol. 1991;204:515–536. doi: 10.1016/0076-6879(91)04026-k. [DOI] [PubMed] [Google Scholar]
  25. Simon R., O'Connell M., Labes M., Pühler A. Plasmid vectors for the genetic analysis and manipulation of rhizobia and other gram-negative bacteria. Methods Enzymol. 1986;118:640–659. doi: 10.1016/0076-6879(86)18106-7. [DOI] [PubMed] [Google Scholar]
  26. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]