Advertisement

Coral Reefs

, Volume 38, Issue 6, pp 1145–1158 | Cite as

N2 fixation, and the relative contribution of fixed N, in corals from Curaçao and Hawaii

  • Michael P. LesserEmail author
  • Kathleen M. Morrow
  • M. Sabrina Pankey
Report

Abstract

Corals from Hawaii (Montipora capitata) and the Caribbean (brown and orange morphs of Montastraea cavernosa) have previously been shown to harbor symbiotic bacteria capable of fixing nitrogen (N2). Using a nitrogen tracer approach, we find that the rates of net photosynthesis and N2 fixation in M. capitata were significantly lower, while steady-state quantum yields of photosystem II (PSII) fluorescence (∆Fv/Fm′) were significantly higher, when compared to both color morphs of M. cavernosa where there was an inverse relationship between rates of photosynthesis and N2 fixation. However, the amount of fixed N contributing to Symbiodiniaceae N demand was consistent with their observed rates of N2 fixation. The lowest values occurred in M. capitata (0.034% ± 0.002 SE), and the brown morph of M. cavernosa had significantly lower values (1.081% ± 0.12 SE) compared to the orange morph (8.141% ± 0.36 SE). Additionally, for both the Symbiodiniaceae and microbial communities there were significant differences between species/color morphs where M. capitata was significantly different from both color morphs of M. cavernosa which were not significantly different from each other. An analysis of predicted metabolic activity, using PICRUSt2, also showed that the corals in this study were not predicted to be differentially enriched in genes involved in carbon metabolism, but genes involved in denitrification were predicted to be significantly enriched in both M. cavernosa orange and brown morphs. Genes involved in N2 fixation were, surprisingly, predicted to be enriched in M. cavernosa brown morphs. We suggest that the inhibition of nitrogenase by hyperoxia is one factor contributing to the low rates of N2 fixation in both M. capitata and M. cavernosa brown morphs.

Keywords

Coral Diazotrophs Microbiome Nitrogen fixation Symbiodiniaceae 

Notes

Acknowledgements

Coral collection in Curaçao was assisted by Elizabeth Kintzing and Abbey Tedford. The coral collections in Curaçao were made under research permit (#2012/48584) issued by the Curaçao Ministry of Health, Environment and Nature (GMN) to the CARMABI foundation. At HIMB, coral collections were permitted under Division of Aquatic Resources Special Activity Permit No. 2015-17. The National Science Foundation (OCE 1437054 to MPL) supported this research.

Authors’ contributions

MPL and KMM designed and conducted the collections and experiments. MPL analyzed all physiological and environmental data. KMM conducted all molecular processing and sequencing. MSP conducted all bioinformatics, statistical and PICRUSt2 analyses on sequence data. MPL, KMM and MSP wrote and approved the content of the manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

338_2019_1863_MOESM1_ESM.docx (12 kb)
Supplementary file1 (DOCX 12 kb)
338_2019_1863_MOESM2_ESM.xlsx (19 kb)
Supplementary file2 (XLSX 18 kb)
338_2019_1863_MOESM3_ESM.xlsx (105 kb)
Supplementary file3 (XLSX 104 kb)
338_2019_1863_MOESM4_ESM.docx (26 kb)
Supplementary file4 (DOCX 26 kb)
338_2019_1863_MOESM5_ESM.xlsx (17 kb)
Supplementary file5 (XLSX 16 kb)

References

  1. Ainsworth TD, Krause L, Bridge T, Torda G, Raina JB, Zakrzewski M, Gates RD, Padilla-Gamiño JL, Spalding HL, Smith C, Woolsey ES, Boourne DG, Bongaerts P, Hoegh-Guldberg O, Leggatt W (2015) The coral core microbiome identifies rare bacterial taxa as ubiquitous endosymbionts. ISME J 9:2261–2274Google Scholar
  2. Apprill A, McNally S, Parsons R, Weber L (2015) Minor revision to V4 region SSU rRNA 806R gene primer greatly increases detection of SAR11 bacterioplankton. Aquat Microb Ecol 75:129–137Google Scholar
  3. Apprill A (2017) Marine animal microbiomes: toward understanding host-microbiome interactions in a changing ocean. Front Mar Sci 4:222Google Scholar
  4. Barbera P, Kozlov AM, Czech L, Morel B, Darriba D, Flouri T, Stamatakis A (2019) EPA-ng: massive parallel evolutionary placement of genetic sequences. Syst Biol 68:365–369PubMedGoogle Scholar
  5. Bednarz VN, Grover R, Maguer J-F, Fine M (2017) The assimilation of diazotroph-derived nitrogen by scleractinian corals depends on their metabolic status. mBio 8:e02058–e2116PubMedPubMedCentralGoogle Scholar
  6. Berman-Frank I, Lundgren P, Falkowski P (2003) Nitrogen fixation and photosynthetic oxygen evolution in cyanobacteria. Res Microbiol 154:157–164PubMedGoogle Scholar
  7. Benavides M, Holbrèque F, Camps M, Lorrain A, Grosso O, Bonnet S (2016) Diazotrophs: a non-negligible source of nitrogen for the tropical coral Stylophora pistillata. J Exp Biol 219:2608–2612PubMedGoogle Scholar
  8. Benavides M, Bednarz VN, Ferrier-Pagès C (2017) Diazotrophs: overlooked key players within coral symbiosis and tropical reef ecosystems? Front Mar Sci 4:10Google Scholar
  9. Bourne DG, Morrow KM, Webster NS (2016) Insights into the coral microbiome: underpinning the health and resilience of reef ecosystems. Ann Rev Microbiol 70:317–340Google Scholar
  10. Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJA, Holmes SP (2016) DADA2: High-resolution sample inference from Illumina amplicon data. Nat Methods 13:581–583PubMedPubMedCentralGoogle Scholar
  11. Cardini U, Bednarz VN, Naumann MS, van Hoytema N, Rix L, Foster RA, Al-Rshaidat MM, Wild C (2015) Functional significance of dinitrogen fixation in sustaining coral production under oligotrophic conditions. Proc R Soc B 282:20152257PubMedGoogle Scholar
  12. Cardini U, van Hoytema N, Bednarz VN, Rix L, Foster RA, Al-Rshaidat MM, Wild C (2016) Microbial dinitrogen fixation in coral holobionts exposed to thermal stress and bleaching. Environ Microbiol 18:2620–2633PubMedGoogle Scholar
  13. Dykens JA, Shick JM (1982) Oxygen production by endosymbiotic algae controls superoxide dismutase activity in their animal host. Nature 297:579–580Google Scholar
  14. Eddy SR (2008) A probabilistic model of local sequence alignment that simplifies statistical significance estimation. PLoS Comput Biol 4:e1000069PubMedPubMedCentralGoogle Scholar
  15. Falkowski PG, Dubinsky Z, Muscatine L, McCloskey L (1993) Population control in symbiotic corals. Bioscience 43:606–611Google Scholar
  16. Fay P (1992) Oxygen relations of nitrogen fixation in cyanobacteria. Microbiol Rev 56:340–373PubMedPubMedCentralGoogle Scholar
  17. Fiore CL, Jarett JK, Olson ND, Lesser MP (2010) Nitrogen fixation and nitrogen transformations in marine symbioses. Trends Microbiol 18:455–463PubMedGoogle Scholar
  18. Gattuso J-P, Jaubert J (1990) Effect of light on oxygen and carbon dioxide fluxes and on metabolic quotients measured in situ in a zooxanthellate coral. Limnol Oceanogr 35:1796–1804Google Scholar
  19. Gloor GB, Macklaim JM, Pawlowsky-Glahn V, Egozcue JJ (2017) Microbiome datasets are compositional: and this is not optional. Front Microbiol 8:2224PubMedPubMedCentralGoogle Scholar
  20. Hernandez-Agreda A, Leggat W, Bongaerts P, Ainsworth TD (2016) The microbial signature provides insight into the mechanistic basis of coral success across reef habitats. mBio 7:e00560–e616PubMedPubMedCentralGoogle Scholar
  21. Hernandez-Agreda A, Gates RD, Ainsworth TD (2017) Defining the core microbiome in corals’ microbial soup. Trends Microbiol 25:125–140PubMedGoogle Scholar
  22. Hernandez-Agreda A, Leggat W, Bongaerts P, Herrera C, Ainsworth TD (2018) Rethinking the coral microbiome: simplicity exists within a diverse microbial biosphere. mBio 9:e00812–e818PubMedPubMedCentralGoogle Scholar
  23. Jarett JK, MacManes MD, Morrow KM, Pankey MS, Lesser MP (2017) Comparative genomics of color morphs in the coral Montastraea cavernosa. Sci Rep 7:16039PubMedPubMedCentralGoogle Scholar
  24. Kühl M, Cohen Y, Dalsgaard T, Jørgensen BB, Revsbech NP (1995) Microenvironment and photosynthesis of zooxanthellae in scleractinian corals studied with microsensors for O2, pH, and light. Mar Ecol Prog Ser 117:159–172Google Scholar
  25. Lahti L, Shetty S, Blake T, Salojarvi J (2017) Tools for microbiome analysis in R. Version 1.9.1. https://microbiome.github.io/tutorials/
  26. Lajeunesse TC, Parkinson JE, Gabrielson PW, Jeong HJ, Reimer JD, Voolstra CR, Santos SR (2018) Systematic revision of Symbiodiniaceae highlights the antiquity and diversity of coral endosymbionts. Curr Biol 28:2570–2580PubMedGoogle Scholar
  27. Lema KA, Willis BL, Bourne DG (2012) Corals form characteristic associations with symbiotic nitrogen-fixing bacteria. Appl Environ Microbiol 78:3136–3144PubMedPubMedCentralGoogle Scholar
  28. Lesser MP, Shick JM (1989) Effects of irradiance and ultraviolet radiation on photoadaptation in the zooxanthellae of Aiptasia pallida: primary production, photoinhibition, and enzymic defenses against oxygen toxicity. Mar Biol 102:243–255Google Scholar
  29. Lesser MP, Mazel CH, Gorbunov MY, Falkowski PG (2004) Discovery of symbiotic nitrogen-fixing cyanobacteria in corals. Science 305:997–1000PubMedPubMedCentralGoogle Scholar
  30. Lesser MP (2006) Oxidative stress in marine environments: biochemistry and physiological ecology. Ann Rev Physiol 68:253–278Google Scholar
  31. Lesser MP, Falcón LI, Rodríguez-Román A, Enríquez S, Hoegh-Guldberg O, Iglesias-Prieto R (2007) Nitrogen fixation by symbiotic cyanobacteria provides a source of nitrogen for the scleractinian coral, Montastraea cavernosa. Mar Ecol Prog Ser 346:143–152Google Scholar
  32. Lesser MP (2011) Coral bleaching: causes and mechanisms. In: Dubinsky Z, Stambler N (eds) Coral reefs: an ecosystem in transition. Springer, Dordrecht, pp 405–420Google Scholar
  33. Lesser MP, Stat M, Gates RD (2013) The endosymbiotic dinoflagellates (Symbiodinium sp.) of corals are parasites and mutualists. Coral Reefs 32:603–611Google Scholar
  34. Lesser MP, Morrow KM, Pankey SM, Noonan SHC (2018) Diazotroph diversity and nitrogen fixation in the coral Stylophora pistillata from the Great Barrier Reef. ISME J 12:813–824PubMedGoogle Scholar
  35. Lesser MP (2019) Phylogenetic signature of light and thermal stress for the symbiotic dinoflagellates of corals (Family Symbiodiniaceae). Limnol Oceanogr 64:1852–1863Google Scholar
  36. Louca S, Doebeli M (2018) Efficient comparative phylogenetics on large trees. Bioinformatics 34:1053–1055PubMedGoogle Scholar
  37. McCarthy MD, Bronk DA (2008) Analytical methods for nitrogen chemical characterization and flux rates. In: Capone DG, Bronk DA, Mulholland M, Carpenter EJ (eds) Nitrogen in the marine environment. Academic Press, New York, pp 1219–1276Google Scholar
  38. McFall-Ngai M, Hadfield MG, Bosch TCG, Carey HV, Domazet-Lošo T, Douglas AE, Dubilier N, Eberl G, Fukami T, Gilbert SF, Hentschel U, King N, Kjelleberg S, Knoll AH, Kremer N, Mazmanian SK, Metcalf JL, Nealson K, Pierce NE, Rawls JF, Reid A, Ruby EG, Rumpho M, Sanders JG, Tautz D, Wernegreen JJ (2013) Animals in a bacterial world, a new imperative for the life sciences. Proc Natl Acad Sci 110:3229–3236PubMedGoogle Scholar
  39. McMurdie PJ, Holmes S (2013) Phyloseq: an R package for reproducible interactive analysis and graphics of microbiome census data. PLoS ONE 8:e61217PubMedPubMedCentralGoogle Scholar
  40. Montoya JP, Voss M, Kähler P, Capone DG (1996) Simple, high-precision, high-sensitivity tracer assay for N2 fixation. Appl Environ Microbiol 62:986–993PubMedPubMedCentralGoogle Scholar
  41. Morán XAG, Gasol JM, Arin L, Estrada M (1999) Comparison between glass fiber and membrane filters for the estimation of phytoplankton POC and DOC production. Mar Ecol Prog Ser 187:31–41Google Scholar
  42. Morrow KM, Muller E, Lesser MP (2018) How does the coral microbiome cause, respond to, or modulate the coral bleaching process? In: van Oppen MJH, Lough JM (eds) Coral bleaching. Ecological Studies, vol 233. Springer, Switzerland, pp 153–188Google Scholar
  43. Oksanen J, Blanchet FG, Friendly M, Kindt R, Legendre P, Minchin OR, O’Hara RB, Simpson GL, Solymos P, Stevens MHH, Wagner H (2019) vegan: Community Ecology Package. R package version 2.4-3. https://CRAN.R-project.org/package=vegan
  44. Olson ND, Ainsworth TD, Gates RD, Takabayashi M (2009) Diazotrophic bacteria associated with Hawaiian Montipora corals: diversity and abundance in correlation with symbiotic dinoflagellates. J Exp Mar Biol Ecol 371:140–146Google Scholar
  45. Olson ND, Lesser MP (2013) Diazotrophic diversity in the Caribbean coral, Montastraea cavernosa. Arch Microbiol 195:853–859PubMedGoogle Scholar
  46. Parada AE, Needham DM, Fuhrman JA (2016) Every base matters: assessing small subunit rRNA primers for marine microbiomes with mock communities, time series and global field samples. Environ Microbiol 18:1403–1414PubMedGoogle Scholar
  47. Piniack GA, Brown EK (2008) Growth and mortality of coral transplants (Pocillopora damicornis) along a range of sediment influences in Maui, Hawai'i. Pac Sci 62:39–55Google Scholar
  48. Pochon X, Pawlowski J, Zainetti L, Rowan R (2001) High genetic diversity and relative specificity among Symbiodinium-like endosymbiotic dinoflagellates in sortid foraminiferans. Mar Biol 139:1069–1078Google Scholar
  49. Pogoreutz C, Rädecker N, Cárdenas A, Gärdes A, Wild C, Voolstra CR (2017) Nitrogen fixation aligns with nifH abundance and expression in two coral trophic groups. Front Microbiol 8:1187PubMedPubMedCentralGoogle Scholar
  50. Rädecker N, Meyer FW, Bednarz VN, Cardini C, Wild C (2014) Ocean acidification rapidly reduces dinitrogen fixation associated with the hermatypic coral Seriatopora hystrix. Mar Ecol Prog Ser 511:297–302Google Scholar
  51. Rädecker N, Pogoreutz C, Voolstra CR, Wiedenmann J, Wild C (2015) Nitrogen cycling in corals: the key to understanding holobiont functioning? Trends Microbiol 23:490–497PubMedGoogle Scholar
  52. Reynolds JM, Bruns BU, Fitt WK, Schmidt GW (2008) Enhanced protection pathways in symbiotic dinoflagellates of shallow-water corals and other cnidarians. Proc Natl Acad Sci 105:13674–13678PubMedGoogle Scholar
  53. Rix L, Bednarz VN, Cardini U, van Hoytema N, Al-Horani A, Wild C, Naumann MS (2015) Seasonality in dinitrogen fixation and primary productivity by coral reef framework substrates from the northern Red Sea. Mar Ecol Prog Ser 533:79–92Google Scholar
  54. Santos HF, Carmo FL, Duarte G, Dini-Andreote F, Castro CB, Rosado AS, van Elsas JD, Peixoto RS (2014) Climate change affects key nitrogen-fixing bacteria populations on coral reefs. ISME J 8:2272–2279PubMedPubMedCentralGoogle Scholar
  55. Seutin G, White BN, Boag PT (1991) Preservation of avian blood and tissue samples for DNA analyses. Can J Zool 69:82–90Google Scholar
  56. Shashar N, Cohen Y, Loya Y (1993) Extreme diel fluctuations of oxygen in diffusive boundary layers surrounding stony corals. Bio Bull 185:455–461Google Scholar
  57. Shore-Maggio A, Runyon CM, Ushijima B, Aeby GS, Callahan SM (2015) Differences in bacterial community structure in two color morphs of the Hawaiian reef coral Montipora capitata. Appl Environ Microbiol 81:7312–7318PubMedPubMedCentralGoogle Scholar
  58. Stat M, Pochon X, Cowie ROM, Gates RD (2009) Specificity in communities of Symbiodinium in corals from Johnston Atoll. Mar Ecol Prog Ser 386:83–96Google Scholar
  59. Suggett DJ, Goyen S, Evenhuis C, Szabó M, Pettay DT, Warner ME, Ralph PJ (2015) Functional diversity of photobiological traits within the genus Symbiodinium appears to be governed by the interaction of cell size with cladal designation. New Phytol 208:370–381Google Scholar
  60. Sunagawa S, Woodley CM, Medina M (2010) Threatened corals provide underexplored microbial habitats. PLoS ONE 5:e9554PubMedPubMedCentralGoogle Scholar
  61. Trinh P, Zaneveld JR, Safranek S, Rabinowitz PM (2018) One health relationships between human, animal, and environmental microbiomes: a mini-review. Front Public Health 6:235PubMedPubMedCentralGoogle Scholar
  62. Warner ME, Lesser MP, Ralph PJ (2010) Chlorophyll fluorescence in reef building corals. In: Suggett D, Borowitzka M, Prásil O (eds) Chlorophyll a fluorescence in aquatic sciences: methods and application. Springer, Dordrecht, pp 209–222Google Scholar
  63. Wilson ST, Böttjer D, Church MJ, Karl DM (2012) Comparative assessment of nitrogen fixation methodologies, conducted in the oligotrophic North Pacific Ocean. Appl Environ Microbiol 78:6516–6523PubMedPubMedCentralGoogle Scholar
  64. Ye Y, Doak TG (2009) A parsimony approach to biological pathway reconstruction/inference for genomes and metagenomes. PLoS Comp Biol 5:e1000465Google Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Michael P. Lesser
    • 1
    • 2
    Email author
  • Kathleen M. Morrow
    • 2
  • M. Sabrina Pankey
    • 2
  1. 1.School of Marine Science and Ocean EngineeringUniversity of New HampshireDurhamUSA
  2. 2.Department of Molecular, Cellular and Biomedical SciencesUniversity of New HampshireDurhamUSA

Personalised recommendations