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Possible roles of glutamine synthetase in responding to environmental changes in a scleractinian coral

  • Yilu Su
  • Zhi Zhou
  • Xiaopeng Yu
Original Article

Abstract

Glutamine synthetase is an enzyme that plays an essential role in the metabolism of nitrogen by catalyzing the condensation of glutamate and ammonia to form glutamine. In this study, the activity and responses of glutamine synthetase towards environmental changes were investigated in the scleractinian coral Pocillopora damicornis. The identified glutamine synthetase (PdGS) was comprised of 362 amino acids and predicted to contain one Gln-synt_N and one Gln-synt_C domain. Expression of PdGS mRNA increased significantly after 12 h (1.28-fold, p < 0.05) of exposure to elevated ammonium, while glutamine synthetase activity increased significantly from 12 to 24 h, peaking at 12 h (54.80 U mg−1, p < 0.05). The recombinant protein of the mature PdGS (rPdGS) was expressed in E. coli BL21, and its activities were detected under different temperature, pH and glufosinate levels. The highest levels of rPdGS activity were observed at 25 °C and pH 8 respectively, but decreased significantly at lower temperature, and higher or lower pH. Furthermore, the level of rPdGS activities was negatively correlated with the concentration of glufosinate, specifically decreasing at 10−5 mol L−1 glufosinate to be less than 50% (p < 0.05) of that in the blank. These results collectively suggest that PdGS, as a homologue of glutamine synthetase, was involved in the nitrogen assimilation in the scleractinian coral. Further, its physiological functions could be suppressed by high temperature, ocean acidification and residual glufosinate, which might further regulate the coral-zooxanthella symbiosis via the nitrogen metabolism in the scleractinian coral P. damicornis.

Keywords

Glutamine synthetase Scleractinian coral Nitrogen assimilation Environmental factor Glufosinate 

Notes

Acknowledgements

The authors were grateful to all the laboratory members for continuous technical advice and helpful discussion. We thanked Zongxiu Wu for his kind helps with the collection and culturing of the coral samples. This research was supported by a Grant (No. 31772460) from National Science Foundation of China, Natural Science Foundation (No. 20164158) of Hainan Province, and the scientific research foundation (kyqd1554) of Hainan University.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All applicable international, national, and/or institutional guidelines for the care and use of animals were followed. All the experiments were conducted according to the regulations of local and central government, and the study protocol was approved by Hainan University.

References

  1. 1.
    Houlbrèque F, Reynaud S, Godinot C, Oberhänsli F, Rodolfo-Metalpa R, Ferrier-Pagès C (2015) Ocean acidification reduces feeding rates in the scleractinian coral Stylophora pistillata. Limnol Oceanogr 60(1):89–99.  https://doi.org/10.1002/lno.10003 CrossRefGoogle Scholar
  2. 2.
    Leletkin VA (2000) The energy budget of coral polyps. Russ J Mar Biol 26(6):389–398.  https://doi.org/10.1023/a:1009497303413 CrossRefGoogle Scholar
  3. 3.
    Ezzat L, Fine M, Maguer J-F, Grover R, Ferrier-Pagès C (2017) Carbon and nitrogen acquisition in shallow and deep holobionts of the scleractinian coral S. pistillata. Front Mar Sci.  https://doi.org/10.3389/fmars.2017.00102 Google Scholar
  4. 4.
    Barott KL, Venn AA, Perez SO, Tambutte S, Tresguerres M (2015) Coral host cells acidify symbiotic algal microenvironment to promote photosynthesis. Proc Natl Acad Sci USA 112(2):607–612.  https://doi.org/10.1073/pnas.1413483112 CrossRefPubMedGoogle Scholar
  5. 5.
    Ferrier-Pagès C, Godinot C, D’Angelo C, Wiedenmann J, Grover R (2016) Phosphorus metabolism of reef organisms with algal symbionts. Ecol Monogr 86(3):262–277.  https://doi.org/10.1002/ecm.1217 CrossRefGoogle Scholar
  6. 6.
    Tanaka Y, Suzuki A, Sakai K (2018) The stoichiometry of coral-dinoflagellate symbiosis: carbon and nitrogen cycles are balanced in the recycling and double translocation system. ISME J 12(3):860–868.  https://doi.org/10.1038/s41396-017-0019-3 CrossRefPubMedGoogle Scholar
  7. 7.
    Radecker N, Pogoreutz C, Voolstra CR, Wiedenmann J, Wild C (2015) Nitrogen cycling in corals: the key to understanding holobiont functioning? Trends Microbiol 23(8):490–497.  https://doi.org/10.1016/j.tim.2015.03.008 CrossRefPubMedGoogle Scholar
  8. 8.
    Yellowlees D, Rees TA, Leggat W (2008) Metabolic interactions between algal symbionts and invertebrate hosts. Plant Cell Environ 31(5):679–694.  https://doi.org/10.1111/j.1365-3040.2008.01802.x CrossRefPubMedGoogle Scholar
  9. 9.
    Fiore CL, Jarett JK, Olson ND, Lesser MP (2010) Nitrogen fixation and nitrogen transformations in marine symbioses. Trends Microbiol 18(10):455–463.  https://doi.org/10.1016/j.tim.2010.07.001 CrossRefPubMedGoogle Scholar
  10. 10.
    Wang J, Douglas AE (1998) Nitrogen recycling or nitrogen conservation in an alga-invertebrate symbiosis? J Exp Biol 201(Pt 16):2445–2453PubMedGoogle Scholar
  11. 11.
    D’Elia CF, Domotor SL, Webb KL (1983) Nutrient uptake kinetics of freshly isolated zooxanthellae. Mar Biol 75(2–3):157–167.  https://doi.org/10.1007/bf00405998 CrossRefGoogle Scholar
  12. 12.
    Tanaka Y, Miyajima T, Koike I, Hayashibara T, Ogawa H (2006) Translocation and conservation of organic nitrogen within the coral-zooxanthella symbiotic system of Acropora pulchra, as demonstrated by dual isotope-labeling techniques. J Exp Mar Biol Ecol 336(1):110–119.  https://doi.org/10.1016/j.jembe.2006.04.011 CrossRefGoogle Scholar
  13. 13.
    Wang JT, Douglas AE (1999) Essential amino acid synthesis and nitrogen recycling in an alga-invertebrate symbiosis. Mar Biol 135(2):219–222.  https://doi.org/10.1007/s002270050619 CrossRefGoogle Scholar
  14. 14.
    Muller-Parker G, D’Elia CF, Cook CB (2015) Interactions between corals and their symbiotic algae. In: Birkeland C (ed) Coral reefs in the anthropocene. Springer, Dordrecht, pp 99–116.  https://doi.org/10.1007/978-94-017-7249-5_5 CrossRefGoogle Scholar
  15. 15.
    Grover R, Maguer J-F, Reynaud-Vaganay S, Ferrier-Pagès C (2002) Uptake of ammonium by the scleractinian coral Stylophora pistillata: effect of feeding, light, and ammonium concentrations. Limnol Oceanogr 47(3):782–790.  https://doi.org/10.4319/lo.2002.47.3.0782 CrossRefGoogle Scholar
  16. 16.
    Lesser MP, Gorbunov MY (2001) Diurnal and bathymetric changes in chlorophyll fluorescence yields of reef corals measured < em> in situ</em> with a fast repetition rate fluorometer. Mar Ecol Progr Ser 212:69–77CrossRefGoogle Scholar
  17. 17.
    Eisenberg D, Almassy RJ, Janson CA, Chapman MS, Suh SW, Cascio D, Smith WW (1987) Some evolutionary relationships of the primary biological catalysts glutamine synthetase and RuBisCO. Cold Spring Harb Symp Quant Biol 52(0):483–490.  https://doi.org/10.1101/sqb.1987.052.01.055 CrossRefPubMedGoogle Scholar
  18. 18.
    Eisenberg D, Gill HS, Pfluegl GM, Rotstein SH (2000) Structure-function relationships of glutamine synthetases. Biochim Biophys Acta 1477(1–2):122–145CrossRefPubMedGoogle Scholar
  19. 19.
    Liu H-y, Sun W-w, Tan B-p, Chi S-y, Dong X-h, Yang Q-h (2012) Molecular cloning and expression of hepatopancreas glutamine synthetase in the Pacific white shrimp, Litopenaeus vannamei, induced by acute hypo-osmotic stress. Aquaculture 362–363:80–87.  https://doi.org/10.1016/j.aquaculture.2012.07.031 CrossRefGoogle Scholar
  20. 20.
    Meistertzheim AL, Tanguy A, Moraga D, Thebault MT (2007) Identification of differentially expressed genes of the Pacific oyster Crassostrea gigas exposed to prolonged thermal stress. FEBS J 274(24):6392–6402.  https://doi.org/10.1111/j.1742-4658.2007.06156.x CrossRefPubMedGoogle Scholar
  21. 21.
    Niva CC, Lee JM, Myohara M (2008) Glutamine synthetase gene expression during the regeneration of the annelid Enchytraeus japonensis. Dev Genes Evol 218(1):39–46.  https://doi.org/10.1007/s00427-007-0198-4 CrossRefPubMedPubMedCentralGoogle Scholar
  22. 22.
    Grover R, Maguer J-F, Allemand D, Ferrier-Pagés C (2003) Nitrate uptake in the scleractinian coral Stylophora pistillata. Limnol Oceanogr 48(6):2266–2274.  https://doi.org/10.4319/lo.2003.48.6.2266 CrossRefGoogle Scholar
  23. 23.
    Rahav O, Dubinsky Z, Achituv Y, Falkowski PG (1989) Ammonium metabolism in the zooxanthellate coral, Stylophora pistillata. Proc R Soc B 236(1284):325–337.  https://doi.org/10.1098/rspb.1989.0026 CrossRefGoogle Scholar
  24. 24.
    Godinot C, Houlbreque F, Grover R, Ferrier-Pages C (2011) Coral uptake of inorganic phosphorus and nitrogen negatively affected by simultaneous changes in temperature and pH. PLoS ONE 6(9):e25024.  https://doi.org/10.1371/journal.pone.0025024 CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Leggat W, Seneca F, Wasmund K, Ukani L, Yellowlees D, Ainsworth TD (2011) Differential responses of the coral host and their algal symbiont to thermal stress. PLoS ONE 6(10):e26687.  https://doi.org/10.1371/journal.pone.0026687 CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Donner SD, Skirving WJ, Little CM, Oppenheimer M, Hoegh-Guldberg OVE (2005) Global assessment of coral bleaching and required rates of adaptation under climate change. Glob Change Biol 11(12):2251–2265.  https://doi.org/10.1111/j.1365-2486.2005.01073.x CrossRefGoogle Scholar
  27. 27.
    Hoegh-Guldberg O, Mumby PJ, Hooten AJ, Steneck RS, Greenfield P, Gomez E, Harvell CD, Sale PF, Edwards AJ, Caldeira K, Knowlton N, Eakin CM, Iglesias-Prieto R, Muthiga N, Bradbury RH, Dubi A, Hatziolos ME (2007) Coral reefs under rapid climate change and ocean acidification. Science 318(5857):1737–1742.  https://doi.org/10.1126/science.1152509 CrossRefPubMedGoogle Scholar
  28. 28.
    Anthony KR, Kline DI, Diaz-Pulido G, Dove S, Hoegh-Guldberg O (2008) Ocean acidification causes bleaching and productivity loss in coral reef builders. Proc Natl Acad Sci USA 105(45):17442–17446.  https://doi.org/10.1073/pnas.0804478105 CrossRefPubMedGoogle Scholar
  29. 29.
    Loya Y, Sakai K, Yamazato K, Nakano Y, Sambali H, van Woesik R (2001) Coral bleaching: the winners and the losers. Ecol Lett 4(2):122–131.  https://doi.org/10.1046/j.1461-0248.2001.00203.x CrossRefGoogle Scholar
  30. 30.
    Stefanik DJ, Wolenski FS, Friedman LE, Gilmore TD, Finnerty JR (2013) Isolation of DNA, RNA and protein from the starlet sea anemone Nematostella vectensis. Nat Protoc 8(5):892–899.  https://doi.org/10.1038/nprot.2012.151 CrossRefPubMedGoogle Scholar
  31. 31.
    Zhang Y, Zhou Z, Wang L, Huang B (2018) Transcriptome, expression, and activity analyses reveal a vital heat shock protein 70 in the stress response of stony coral Pocillopora damicornis. Cell Stress Chaperones 23(4):711–721.  https://doi.org/10.1007/s12192-018-0883-4 CrossRefPubMedGoogle Scholar
  32. 32.
    Woolfolk CA, Shapiro B, Stadtman ER (1966) Regulation of glutamine synthetase. I: purification and properties of glutamine synthetase from Escherichia coli. Arch Biochem Biophys 116(1):177–192.  https://doi.org/10.1016/0003-9861(66)90026-9 CrossRefPubMedGoogle Scholar
  33. 33.
    Singh J, Joshi MC, Bhatnagar R (2004) Cloning and expression of mycobacterial glutamine synthetase gene in Escherichia coli. Biochem Biophys Res Commun 317(2):634–638.  https://doi.org/10.1016/j.bbrc.2004.03.094 CrossRefPubMedGoogle Scholar
  34. 34.
    Bernard SM, Habash DZ (2009) The importance of cytosolic glutamine synthetase in nitrogen assimilation and recycling. New Phytol 182(3):608–620.  https://doi.org/10.1111/j.1469-8137.2009.02823.x CrossRefPubMedGoogle Scholar
  35. 35.
    Powers SG, Riordan JF (1975) Functional arginyl residues as ATP binding sites of glutamine synthetase and carbamyl phosphate synthetase. Proc Natl Acad Sci 72(7):2616–2620.  https://doi.org/10.1073/pnas.72.7.2616 CrossRefPubMedGoogle Scholar
  36. 36.
    Pandolfi JM, Connolly SR, Marshall DJ, Cohen AL (2011) Projecting coral reef futures under global warming and ocean acidification. Science 333(6041):418–422.  https://doi.org/10.1126/science.1204794 CrossRefPubMedGoogle Scholar
  37. 37.
    Cao D, Cao W, Liang Y, Huang Z (2016) Nutrient variations and isotopic evidences of particulate organic matter provenance in fringing reefs, South China. Sci Total Environ 566–567:378–386.  https://doi.org/10.1016/j.scitotenv.2016.05.112 CrossRefPubMedGoogle Scholar
  38. 38.
    Dayan FE, Owens DK, Duke SO (2012) Rationale for a natural products approach to herbicide discovery. Pest Manag Sci 68(4):519–528.  https://doi.org/10.1002/ps.2332 CrossRefPubMedGoogle Scholar
  39. 39.
    Pornprom T, Prodmatee N, Chatchawankanphanich O (2009) Glutamine synthetase mutation conferring target-site-based resistance to glufosinate in soybean cell selections. Pest Manag Sci 65(2):216–222.  https://doi.org/10.1002/ps.1671 CrossRefPubMedGoogle Scholar
  40. 40.
    Pernice M, Meibom A, Van Den Heuvel A, Kopp C, Domart-Coulon I, Hoegh-Guldberg O, Dove S (2012) A single-cell view of ammonium assimilation in coral-dinoflagellate symbiosis. ISME J 6(7):1314–1324.  https://doi.org/10.1038/ismej.2011.196 CrossRefPubMedPubMedCentralGoogle Scholar
  41. 41.
    Doney SC, Mahowald N, Lima I, Feely RA, Mackenzie FT, Lamarque JF, Rasch PJ (2007) Impact of anthropogenic atmospheric nitrogen and sulfur deposition on ocean acidification and the inorganic carbon system. Proc Natl Acad Sci USA 104(37):14580–14585.  https://doi.org/10.1073/pnas.0702218104 CrossRefPubMedGoogle Scholar
  42. 42.
    Yellowlees D, Rees T, Fitt W (1994) Effect of ammonium-supplemented seawater on glutamine synthetase and glutamate dehydrogenase activities in host tissue and zooxanthellae of Pocillopora damicornis and on ammonium uptake rates of the zooxanthellae. Pac Sci 48(3):291–295Google Scholar
  43. 43.
    Yuan C, Zhou Z, Zhang Y, Chen G, Yu X, Ni X, Tang J, Huang B (2017) Effects of elevated ammonium on the transcriptome of the stony coral Pocillopora damicornis. Mar Pollut Bull 114(1):46–52.  https://doi.org/10.1016/j.marpolbul.2016.08.036 CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Key Laboratory of Tropical Biological Resources of Ministry of EducationHainan UniversityHaikouChina
  2. 2.State Key Laboratory of Marine Resource Utilization in South China SeaHainan UniversityHaikouChina
  3. 3.Hainan Provincial Key Laboratory for Tropical Hydrobiology and Biotechnology, College of Marine ScienceHainan UniversityHaikouChina

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