Letter | Published:

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.

Access optionsAccess options

Rent or Buy article

Get time limited or full article access on ReadCube.


All prices are NET prices.

Additional information

Publisher’s note: Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.


  1. 1.

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

  2. 2.

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

  3. 3.

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

  4. 4.

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

  5. 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).

  6. 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).

  7. 7.

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

  8. 8.

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

  9. 9.

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

  10. 10.

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

  11. 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).

  12. 12.

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

  13. 13.

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

  14. 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).

  15. 15.

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

  16. 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).

  17. 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).

  18. 18.

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

  19. 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).

  20. 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).

  21. 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).

  22. 22.

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

  23. 23.

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

  24. 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).

  25. 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).

  26. 26.

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

  27. 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).

  28. 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).

  29. 29.

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

  30. 30.

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

  31. 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).

  32. 32.

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

  33. 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).

  34. 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).

  35. 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).

  36. 36.

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

  37. 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).

  38. 38.

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

  39. 39.

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

  40. 40.

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

  41. 41.

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

  42. 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).

  43. 43.

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

  44. 44.

    R Development Core Team. R: A language and environment for statistical computing. (R Foundation for Statistical Computing, 2016); http://www.R-project.org

  45. 45.

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

  46. 46.

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

  47. 47.

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

Download references


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).

Author information

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.

Competing interests

The authors declare no competing interests.

Correspondence to Adam C. Martiny.

Supplementary information

  1. Supplementary Information

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

  2. Reporting Summary

  3. Supplementary Table 7

    Supplementary Table 7.

  4. Supplementary Table 10

    Supplementary Table 10.

Rights and permissions

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Further reading

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.