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Increased biofilm formation due to high-temperature adaptation in marine Roseobacter


Ocean temperatures will increase significantly over the next 100 years due to global climate change1. As temperatures increase beyond current ranges, it is unclear how adaptation will impact the distribution and ecological role of marine microorganisms2. To address this major unknown, we imposed a stressful high-temperature regime for 500 generations on a strain from the abundant marine Roseobacter clade. High-temperature-adapted isolates significantly improved their fitness but also increased biofilm formation at the air–liquid interface. Furthermore, this altered lifestyle was coupled with genomic changes linked to biofilm formation in individual isolates, and was also dominant in evolved populations. We hypothesize that the increasing biofilm formation was driven by lower oxygen availability at elevated temperature, and we observe a relative fitness increase at lower oxygen. The response is uniquely different from that of Escherichia coli adapted to high temperature3 (only 3% of mutated genes were shared in both studies). Thus, future increased temperatures could have a direct effect on organismal physiology and an indirect effect via a decrease in ocean oxygen solubility, leading to an alteration in microbial lifestyle.

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Fig. 1: Increased fitness of HTLs competed against control low temperature line 10.
Fig. 2: Changes in phenotypes due to adaptation.
Fig. 3: Mutation distribution across the genome and compared to E. coli.
Fig. 4: Genotypic associations with phenotypic variation.


  1. IPCC Climate Change 2007: Impacts, Adaptation and Vulnerability (eds Parry, M. L. et al.) 976 (Cambridge Univ. Press, 2007).

  2. Hutchins, D. A. & Fu, F. Microorganisms and ocean global change. Nat. Microbiol. 2, 17058 (2017).

    Article  CAS  PubMed  Google Scholar 

  3. Tenaillon, O. et al. The molecular diversity of adaptive convergence. Science 335, 457–461 (2012).

    Article  CAS  PubMed  Google Scholar 

  4. Franks, S. J. & Hoffmann, A. A. Genetics of climate change adaptation. Annu. Rev. Genet. 46, 185–208 (2012).

    Article  CAS  PubMed  Google Scholar 

  5. Bergmann, N. et al. Population-specificity of heat stress gene induction in northern and southern eelgrass Zostera marina populations under simulated global warming. Mol. Ecol. 19, 2870–2883 (2010).

    Article  PubMed  Google Scholar 

  6. Thomas, M. K., Kremer, C. T., Klausmeier, C. A. & Litchman, E. A global pattern of thermal adaptation in marine phytoplankton. Science 338, 1085–1088 (2012).

    Article  CAS  PubMed  Google Scholar 

  7. Collins, S. Many possible worlds: expanding the ecological scenarios in experimental evolution. Evol. Biol. 38, 3–14 (2010).

    Article  Google Scholar 

  8. Hug, S. M. & Gaut, B. S. The phenotypic signature of adaptation to thermal stress in Escherichia coli. BMC Evol. Biol. 15, 177 (2015).

    Article  PubMed  PubMed Central  Google Scholar 

  9. Nielsen, E. S. & Jørgensen, E. G. The adaptation of plankton algae. Physiol. Plant. 21, 401–413 (1968).

    Article  Google Scholar 

  10. Toseland, A. et al. The impact of temperature on marine phytoplankton resource allocation and metabolism. Nat. Clim. Change 3, 979–984 (2013).

    Article  CAS  Google Scholar 

  11. Linzner, K. A., Kent, A. G. & Martiny, A. C. Evolutionary pathway determines the stoichiometric response of Escherichia coli adapted to high temperature. Front. Ecol. Evol. 5, 173 (2018).

    Article  Google Scholar 

  12. Wagner-Döbler, I. & Biebl, H. Environmental biology of the marine Roseobacter lineage. Annu. Rev. Microbiol. 60, 255–280 (2006).

    Article  PubMed  Google Scholar 

  13. Moran, M. A. et al. Genome sequence of Silicibacter pomeroyi reveals adaptations to the marine environment. Nature 432, 910–913 (2004).

    Article  CAS  PubMed  Google Scholar 

  14. Giebel, H. A., Brinkhoff, T., Zwisler, W., Selje, N. & Simon, M. Distribution of Roseobacter RCA and SAR11 lineages and distinct bacterial communities from the subtropics to the Southern Ocean. Environ. Microbiol. 11, 2164–2178 (2009).

    Article  CAS  PubMed  Google Scholar 

  15. Buchan, A., González, J. M. & Moran, M. A. Overview of the marine Roseobacter lineage. Appl. Environ. Microbiol. 71, 5665–5677 (2005).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Benson, B. B. & Krause, D. The concentration and isotopic fractionation of oxygen dissolved in freshwater and seawater in equilibrium with the atmosphere. Limnol. Oceanogr. 29, 620–632 (1984).

    Article  CAS  Google Scholar 

  17. Zan, J. et al. A complex LuxR-LuxI type quorum sensing network in a roseobacterial marine sponge symbiont activates flagellar motility and inhibits biofilm formation. Mol. Microbiol. 85, 916–933 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  18. Moller, T. et al. Hfq: A Bacterial Sm-like protein that mediates RNA–RNA interaction. Mol. Cell 9, 23–30 (2002).

    Article  CAS  PubMed  Google Scholar 

  19. Lenz, D. H. et al. The small RNA chaperone Hfq and multiple small RNAs control quorum sensing in Vibrio harveyi and Vibrio cholerae. Cell 118, 69–82 (2004).

    Article  CAS  PubMed  Google Scholar 

  20. Zeng, Q., Mcnally, R. R. & Sundin, G. W. Global small RNA chaperone Hfq and regulatory small RNAs are important virulence regulators in Erwinia amylovora. J. Bacteriol. 195, 1706–1717 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  21. Rempe, K. A., Hinz, A. K. & Vadyvaloo, V. Hfq regulates biofilm gut blockage that facilitates flea-borne transmission of Yersinia pestis. J. Bacteriol. 194, 2036–2040 (2012).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Geng, H. & Belas, R. Molecular mechanisms underlying roseobacter-phytoplankton symbioses. Curr. Opin. Biotechnol. 21, 332–338 (2010).

    Article  CAS  PubMed  Google Scholar 

  23. Rainey, P. B. & Travisano, M. Adaptive radiation in a heterogeneous environment. Nature 394, 69–72 (1998).

    Article  CAS  PubMed  Google Scholar 

  24. Spiers, A. J., Kahn, S. G., Bohannon, J., Travisano, M. & Rainey, P. B. Adaptive divergence in experimental populations of Pseudomonas fluorescens. I. Genetic and phenotypic bases of wrinkly spreader fitness. Genetics 161, 33–46 (2002).

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Hansen, S. K., Rainey, P. B., Haagensen, J. A. J. & Molin, S. Evolution of species interactions in a biofilm community. Nature 445, 533–536 (2007).

    Article  CAS  PubMed  Google Scholar 

  26. Moore, L., Rocap, G. & Chisholm, S. Physiology and molecular phylogeny of coexisting Prochlorococcus ecotypes. Nature 576, 220–223 (1998).

    Google Scholar 

  27. Shiah, F. K. & Ducklow, H. W. Temperature regulation of heterotrophic bacterioplankton production, and specific growth rate in Chesapeake Bay. Limnol. Oceanogr. 39, 1243–1258 (1994).

    Article  Google Scholar 

  28. Bianchi, D., Weber, T. S., Kiko, R. & Deutsch, C. Global niche of marine anaerobic metabolisms expanded by particle microenvironments. Nat. Geosci. 11, 263–268 (2018).

    Article  CAS  Google Scholar 

  29. Keeling, R. E., Körtzinger, A. & Gruber, N. Ocean deoxygenation in a warming world. Ann. Rev. Mar. Sci. 2, 199–229 (2010).

    Article  PubMed  Google Scholar 

  30. Miller, T. R. & Belas, R. Dimethylsulfoniopropionate metabolism by Pfiesteria-associated Roseobacter spp. Appl. Environ. Microbiol. 70, 3383–3391 (2004).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. Alavi, M., Miller, T., Erlandson, K., Schneider, R. & Belas, R. Bacterial community associated with Pfesteria-like dinoflagellate cultures. Environ. Microbiol. 3, 380–396 (2001).

    Article  CAS  PubMed  Google Scholar 

  32. Lenski, R. E. & Bennett, A. F. Evolutionary response of Escherichia coli to thermal stress. Am. Nat. 142(Suppl. 1), S47–S64 (1993).

  33. Rodriguez-Verdugo, A., Carrillo-Cisneros, D., Gonzalez-Gonzalez, A., Gaut, B. S. & Bennett, A. F. Different tradeoffs result from alternate genetic adaptations to a common environment. Proc. Natl Acad. Sci. USA 111, 12121–12126 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Lenski, R. E., Rose, M. R., Simpson, S. C. & Tadler, S. C. Long-term experimental evolution in Escherichia coli. I. Adaptation and divergence during 2,000 generations. Am. Nat. 138, 1315–1341 (1991).

    Article  Google Scholar 

  35. Applebee, M. K., Herrgård, M. J. & Palsson, B. Ø. Impact of individual mutations on increased fitness in adaptively evolved strains of Escherichia coli. J. Bacteriol. 190, 5087–5094 (2008).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. O’Toole, G. A. Microtiter dish biofilm formation assay. J. Vis. Exp. 2011, 2437 (2011).

  37. Somerville, G. A. & Proctor, R. A. Cultivation conditions and the diffusion of oxygen into culture media: the rationale for the flask-to-medium ratio in microbiology. BMC Microbiol. 13, 9 (2013).

    Article  PubMed  PubMed Central  Google Scholar 

  38. Sharp, J. H. Improved analysis for ‘particulate’ organic carbon and nitrogen from seawater. Limnol. Ocean. 19, 984–989 (1974).

    Article  CAS  Google Scholar 

  39. Lomas, M. W. et al. Sargasso Sea phosphorus biogeochemistry: an important role for dissolved organic phosphorus (DOP). Biogeosciences 7, 695–710 (2010).

    Article  CAS  Google Scholar 

  40. Hunt, M. et al. REAPR: a universal tool for genome assembly evaluation. Genome Biol. 14, R47 (2013).

    Article  PubMed  PubMed Central  Google Scholar 

  41. Aziz, R. K. et al. The RAST Server: rapid annotations using subsystems technology. BMC Genomics 9, 75 (2008).

    Article  PubMed  PubMed Central  Google Scholar 

  42. Deatherage, D. E. & Barrick, J. E. Identification of mutations in laboratory-evolved microbes from next-generation sequencing data using breseq. Methods Mol. Biol. 1151, 165–188 (2014).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Li, L., Stoeckert, C. J. J. & Roos, D. S. OrthoMCL: identification of ortholog groups for eukaryotic genomes. Genome Res. 13, 2178–2189 (2003).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. R Development Core Team. R: A language and environment for statistical computing. (R Foundation for Statistical Computing, 2016);

  45. Riahi, K. et al. RCP 8.5—a scenario of comparatively high greenhouse gas emissions. Clim. Change 109, 33–57 (2011).

    Article  CAS  Google Scholar 

  46. Bates, D., Mächler, M., Bolker, B. & Walker, S. Fitting linear mixed-effects models using lme4. J. Stat. Softw. 67, 1–48 (2014).

    Google Scholar 

  47. Best, D. J. & Roberts, D. E. Algorithm AS 89: the upper tail probabilities of Spearman’s rho. Appl. Stat. 24, 377–379 (1975).

    Article  Google Scholar 

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The authors thank J. Martiny and B. Gaut for helpful comments, C. Mouginot, T. Kooner and K. Linzner for laboratory assistance, and R. Belas for permission to use this strain. The authors also acknowledge the World Climate Research Programme’s Working Group on Coupled Modelling and thank the climate modelling groups listed in the Methods for creating and making available their CMIP5 model output. For CMIP, the US Department of Energy’s Program for Climate Model Diagnosis and Intercomparison leads research in partnership with the Global Organization for Earth System Science Portals. A.G.K. was supported by the National Science Foundation Graduate Research Fellowship Program (DGE-1321846) and the National Institute of Biomedical Imaging and Bioengineering, National Research Service Award EB009418 from the University of California, Irvine, Center for Complex Biological Systems. C.A.G. was supported by NASA Headquarters under the NASA Earth and Space Science Fellowship (15-EARTH15F-0335). A.C.M. was supported by the National Science Foundation (OCE-1559002 and OCE-1046297).

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Authors and Affiliations



A.G.K. and C.A.G. performed the experiments and analysed the data. A.G.K. and A.C.M. conceived and designed the experiments. A.G.K., C.A.G. and A.C.M. wrote the paper.

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Correspondence to Adam C. Martiny.

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Supplementary Figures 1–18, Supplementary Tables 1–6, 8, 9, 11–14, and Supplementary References.

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Kent, A.G., Garcia, C.A. & Martiny, A.C. Increased biofilm formation due to high-temperature adaptation in marine Roseobacter. Nat Microbiol 3, 989–995 (2018).

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