The Marine Bacterium Shewanella woodyi Produces C8-HSL to Regulate Bioluminescence
Quorum sensing (QS), a cell-to-cell communication system involved in the synchronization of bacterial behavior in a cell-density-dependent manner has been shown to control phenotypes such as luminescence, virulence, and biofilm formation. The marine strain, Shewanella woodyi MS32 has been identified as a luminous bacterium. Very little information is known on this bacterium, in particular if its luminescence and biofilm formation are controlled by QS. In this study, we have demonstrated that S. woodyi MS32 emits luminescence in planktonic and sessile conditions. The putative QS regulatory genes homologous to luxI and luxR identified in the S. woodyi MS32 genome, named swoI and swoR, are divergently transcribed and are not genetically linked to the lux operon in contrast with its closest parent Shewanella hanedai and with Aliivibrio fischeri. Interestingly, the phylogenetic analysis based on the SwoI and SwoR sequences shows that a separate horizontal gene transfer (HGT) occurred for the regulatory genes and for the lux operon. Functional analyses demonstrate that the swoI and swoR mutants were non-luminescent. Expression of lux genes was impaired in the QS regulatory mutants. N-octanoyl-L-homoserine lactone (C8-HSL) identified using liquid chromatography mass spectrometry in the wild-type strain (but not in ΔswoI) can induce S. woodyi luminescence. No significant difference has been detected between the wild-type and mutants on adhesion and biofilm formation in the conditions tested. Therefore, we have demonstrated that the luxCDABEG genes of S. woodyi MS32 are involved in luminescence emission and that the swoR/swoI genes, originated from a separate HGT, regulate luminescence through C8-HSL production.
KeywordsMarine bacteria Shewanella woodyi Quorum sensing Luminescence HGT Biofilm
We thank J.A. Gralnick from the BioTechnology Institute and Department of Plant and Microbial Biology, University of Minnesota-Twin Cities for kindly providing the Shewanella strain and the pSMV3 and pBBR1MCS-2 plasmids. We thank Yoan Ferandin for the advisory help in the qPCR. We thank P. Poupin from the Laboratory of Microbial Biodiversity and Biotechnology of the Sorbonne Universities and CNRS for his help in the construction of the complementation plasmids. We also thank anonymous reviewers for their commentaries and suggestions.
Mahmoud Hayek is the beneficiary of the Lebanese grants from the Association of Specialization and Scientific Guidance (ASSG). This research was also partly funded by the EMBRC network.
Compliance with Ethical Standards
Conflict of Interest
The authors declare that they have no conflict of interest.
This article does not contain any studies with human participants or animals performed by any of the authors.
Informed consent was obtained from all individual participants included in the study.
- 1.Urbanczyk H, Ast JC, Higgins MJ, Carson J, Dunlap PV (2007) Reclassification of Vibrio fischeri, Vibrio logei, Vibrio salmonicida and Vibrio wodanis as Aliivibrio fischeri gen. nov., comb. nov., Aliivibrio logei comb. nov., Aliivibrio salmonicida comb. nov. and Aliivibrio wodanis comb. nov. Int J Syst Evol Microbiol lin 57:2823–2829. https://doi.org/10.1099/ijs.0.65081-0 CrossRefGoogle Scholar
- 2.Hmelo LR (2017) Quorum sensing in marine microbial environments. Annu Rev Mar Sci 9:257–281. https://doi.org/10.1146/annurev-marine-010816-060656 CrossRefGoogle Scholar
- 4.Khajanchi BK, Sha J, Kozlova EV, Erova TE, Suarez G, Sierra JC, Popov VL, Horneman AJ, Chopra AK (2009) N-Acylhomoserine lactones involved in quorum sensing control the type VI secretion system, biofilm formation, protease production, and in vivo virulence in a clinical isolate of Aeromonas hydrophila. Microbiology 155:3518–3531. https://doi.org/10.1099/mic.0.031575-0 CrossRefPubMedPubMedCentralGoogle Scholar
- 5.Waters CM, Bassler BL (2005) Quorum sensing: cell-to-cell communication in bacteria. Annu Rev Cell Dev Biol 21:319–346. https://doi.org/10.1146/annurev.cellbio.21.012704.131001 CrossRefPubMedGoogle Scholar
- 8.Kita-Tsukamoto K, Yao K, Kamiya A, Yoshizawa S, Uchiyama N, Kogure K, Wada M (2006) Rapid identification of marine bioluminescent bacteria by amplified 16S ribosomal RNA gene restriction analysis. FEMS Microbiol Lett 256:298–303. https://doi.org/10.1111/j.1574-6968.2006.00129.x CrossRefPubMedGoogle Scholar
- 10.Lin B, Wang Z, Malanoski AP, O'Grady EA, Wimpee CF, Vuddhakul V, Alves Jr N, Thompson FL, Gomez-Gil B, Vora GJ (2010) Comparative genomic analyses identify the Vibrio harveyi genome sequenced strains BAA-1116 and HY01 as Vibrio campbellii. Environ Microbiol Rep 2:81–89. https://doi.org/10.1111/j.1758-2229.2009.00100.x CrossRefPubMedPubMedCentralGoogle Scholar
- 18.Bassler BL, Wright M, Silverman MR (1994) Multiple signalling systems controlling expression of luminescence in Vibrio harveyi: sequence and function of genes encoding a second sensory pathway. Mol Microbiol 13:273–286. https://doi.org/10.1111/j.1365-2958.1994.tb00422.x CrossRefPubMedGoogle Scholar
- 19.McClean KH, Winson MK, Fish L, Taylor A, Chhabra SR, Camara M, Daykin M, Lamb JH, Swift S, Bycroft BW, Stewart GS, Williams P (1997) Quorum sensing and Chromobacterium violaceum: exploitation of violacein production and inhibition for the detection of N-acylhomoserine lactones. Microbiology 143(Pt 12):3703–3711. https://doi.org/10.1099/00221287-143-12-3703 CrossRefPubMedGoogle Scholar
- 23.Aye M, Bonnin-Jusserand M, Brian-Jaisson F, Ortalo-Magne A, Culioli G, Koffi Nevry R, Rabah N, Blache Y, Molmeret M (2015) Modulation of violacein production and phenotypes associated with biofilm by exogenous quorum sensing N-acylhomoserine lactones in the marine bacterium Pseudoalteromonas ulvae TC14. Microbiology 161:2039–2051. https://doi.org/10.1099/mic.0.000147 CrossRefGoogle Scholar
- 28.Parsek MR, Schaefer AL, Greenberg EP (1997) Analysis of random and site-directed mutations in rhlI, a Pseudomonas aeruginosa gene encoding an acylhomoserine lactone synthase. Mol Microbiol 26:301–310. https://doi.org/10.1046/j.1365-2958.1997.5741935.x CrossRefPubMedGoogle Scholar
- 32.Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425. https://doi.org/10.1093/oxfordjournals.molbev.a040454 CrossRefGoogle Scholar
- 33.Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–791. https://doi.org/10.1111/j.1558-5646.1985.tb00420.x CrossRefPubMedGoogle Scholar
- 35.Liu J, Fu K, Wang Y, Wu C, Li F, Shi L, Ge Y, Zhou L (2017) Detection of diverse N-Acyl-Homoserine Lactones in Vibrio alginolyticus and regulation of biofilm formation by N-(3-Oxodecanoyl) Homoserine Lactone In vitro. Front Microbiol 8:1097. https://doi.org/10.3389/fmicb.2017.01097 CrossRefPubMedPubMedCentralGoogle Scholar
- 45.Guillonneau R, Baraquet C, Bazire A, Molmeret M (2018) Multispecies biofilm development of marine bacteria implies complex relationships through competition and synergy and modification of matrix components. Front Microbiol 9:1960. https://doi.org/10.3389/fmicb.2018.01960 CrossRefPubMedPubMedCentralGoogle Scholar