Journal of Applied Phycology

, Volume 31, Issue 1, pp 625–635 | Cite as

Nitrogen uptake and assimilation preferences of the main green tide alga Ulva prolifera in the Yellow Sea, China

  • Hongmei Li
  • Yongyu ZhangEmail author
  • Jing Chen
  • Xuan Zheng
  • Feng Liu
  • Nianzhi JiaoEmail author


The successive outbreak of large-scale Ulva prolifera green tides in the Yellow Sea, China, from 2007 to 2017, is a serious regional environmental issue that attracts worldwide attention. The competitive advantage in nitrogen uptake and utilization is an important factor, making U. prolifera the dominant green-tide-forming seaweed. However, the detailed preference characteristics of U. prolifera for nitrogen uptake and assimilation in common inorganic and organic nitrogen sources is poorly understood and is studied using stable nitrogen isotope (15N) analysis. Our results reveal that various nitrogen sources can be simultaneously and directly taken up by U. prolifera. The uptake rates are in the sequence of NO3 (nitrate) > NH4+ (ammonium) > CO(NH2)2 (urea) > C2H5NO2 (glycine) and C3H7NO2 (alanine). In other words, U. prolifera mostly prefers inorganic nitrogen, such as nitrate and ammonium, although it can also utilize different organic nitrogen sources simultaneously. Moreover, the assimilation of NH4+ is inhibited and its uptake cannot be fitted by the Michaelis–Menten equation when the alga is exposed to multiple nitrogen sources. We propose that at the early and middle stages of green tides the rich inorganic nitrogen sources (especially NO3) in seawater are very important to support the fast growth of U. prolifera, while at the later stage of green tides, when inorganic nitrogen sources have been exhausted, organic nitrogen sources may contribute importantly to maintaining the growth of U. prolifera, thus lengthening the duration of green tides.


Harmful macroalgal blooms Nitrogen species Nutrient uptake Stable isotopes Chlorophyta Ulva prolifera 



This study was jointly supported by the National Key Research and Development Program of China (2016YFC1402101-03), Key Laboratory of Marine Ecology and Environmental Science and Engineering SOA (MESE-2016-02), National Natural Science Foundation of China (41606092), National Key Research and Development Programs of China (2016YFA0601402), a Key R&D Project of the Shandong Province (2016GSF115011), and the Key Research Program of Frontier Sciences, Chinese Academy of Sciences (no. QYZDB-SSW-DQC023). This study is a contribution to the international IMBER project.


  1. Abreu MH, Pereira R, Buschmann AH, Sousa-Pinto I, Yarish C (2011) Nitrogen uptake responses of Gracilaria vermiculophylla (Ohmi) Papenfuss under combined and single addition of nitrate and ammonium. J Exp Mar Biol Ecol 407:190–199CrossRefGoogle Scholar
  2. Ale MT, Mikkelsen JD, Meyer AS (2011) Differential growth response of Ulva lactuca to ammonium and nitrate assimilation. J Appl Phycol 23:345–351CrossRefGoogle Scholar
  3. Allanson BR, Human LRD, Claassens L (2016) Observations on the distribution and abundance of a green tide along an intertidal shore, Knysna estuary. S Afr J Bot 107:49–54CrossRefGoogle Scholar
  4. Berges JA (1997) Minireview: algal nitrate reductases. Eur J Phycol 32:3–8CrossRefGoogle Scholar
  5. Berges JA, Franklin DJ, Harrison PJ (2001) Evolution of an artificial seawater medium: improvements in enriched seawater, artificial water over the last two decades. J Phycol 37:1138–1145CrossRefGoogle Scholar
  6. Bracken MES, Stachowicz JJ (2006) Seaweed diversity enhances nitrogen uptake via complementary use of nitrate and ammonium. Ecology 87:2397–2403CrossRefGoogle Scholar
  7. China Fishery Statistical Year Book (CFSYB) (2017) China Agriculture Press, Beijing, pp 155 (in Chinese)Google Scholar
  8. Cohen RA, Fong P (2004) Nitrogen uptake and assimilation in Enteromorpha intestinalis (L.) Link (Chlorophyta): using 15N to determine preference during simultaneous pulses of nitrate and ammonium. J Exp Mar Biol Ecol 309:67–77CrossRefGoogle Scholar
  9. Copertino MDS, Tormena T, Seeliger U (2009) Biofiltering efficiency, uptake and assimilation rates of Ulva clathrata (Roth) J. Agardh (Chlorophyceae) cultivated in shrimp aquaculture waste water. J Appl Phycol 21:31–45CrossRefGoogle Scholar
  10. Du J (2014) The preliminary study on uptake of nutrients by Ulva prolifera and competition of Ulva prolifera with red tide algae. (MsD thesis) Ocean University of China, Qingdao, pp 45 (in Chinese with English abstract)Google Scholar
  11. Fan X, Xu D, Wang YT, Zhang XW, Cao SN, Mou SL, Ye NH (2014) The effect of nutrient concentrations, nutrient ratios and temperature on photosynthesis and nutrient uptake by Ulva prolifera: implications for the explosion in green tides. J Appl Phycol 26:537–544CrossRefGoogle Scholar
  12. Fujita RM, Wheeler PA, Edwards RL (1988) Metabolic regulation of ammonium uptake by Ulva rigida (Chlorophyta): a compartmental analysis of the rate-limiting step for uptake. J Phycol 24:560–566CrossRefGoogle Scholar
  13. Gao KS, Xu JT, Zheng YQ, Ke CY (2012a) Measurement of benthic photosynthesis and calcification in flowing-through seawater with stable carbonate chemistry. Limnol Oceanogr Methods 10:555–559CrossRefGoogle Scholar
  14. Gao S, Shi XY, Wang T (2012b) Variation of nutrient concentrations at the inshore coastal area of northern Jiangsu province and the occurrence of green tide caused by Ulva prolifera. Environ Sci 33:2204–2209 (in Chinese with English abstract)Google Scholar
  15. Gao G, Clare AS, Rose C, Caldwell GS (2017) Eutrophication and warming-driven green tides (Ulva rigida) are predicted to increase under future climate change scenarios. Mar Pollut Bull 114:439–447CrossRefGoogle Scholar
  16. Grasshoff K, Kremling K, Ehrhardt M (1999) Methods of seawater analysis. Verlag Chemie, Weinheim, pp 365–371CrossRefGoogle Scholar
  17. Healey FP (1980) Slope of the Monod equation as an indicator of advantage in nutrient competition. Microb Ecol 5:281–286CrossRefGoogle Scholar
  18. Heisler J, Glibert PM, Burkholder JM, Anderson DM, Cochlan W, Dennison WC, Dortch Q, Gobler CJ, Heil CA, Humphries E, Lewitus A, Magnien R, Marshall HG, Sellner K, Stockwell DA, Stoecker DK, Suddleson M (2008) Eutrophication and harmful algal blooms: a scientific consensus. Harmful Algae 8:3–13CrossRefGoogle Scholar
  19. Huo YZ, Hua L, Wu HL, Zhang JH, Cui JJ, Huang XW, Yu KF, Shi HH, He PM, Ding DW (2014) Abundance and distribution of Ulva microscopic propagules associated with a green tide in the southern coast of the Yellow Sea. Harmful Algae 39:357–364CrossRefGoogle Scholar
  20. Jiang P, Wang JF, Cui YL, Li YX, Lin HZ, Qin S (2008) Molecular phylo-genetic analysis of attached Ulvaceae species and free-floating Enteromorpha from Qingdao coasts in 2007. Chin J Oceanol Limnol 26:276–279CrossRefGoogle Scholar
  21. Kaiser K, Benner R (2005) Hydrolysis-induced racemization of amino acids. Limnol Oceanogr 3:318–325CrossRefGoogle Scholar
  22. Kamer K, Fong P (2001) Nitrogen enrichment ameliorates the negative effects of reduced salinity in the green macroalga Enteromorpha intestinalis. Mar Ecol Prog Ser 218:87–93CrossRefGoogle Scholar
  23. Keesing JK, Liu DY, Shi YJ, Wang Y (2016) Abiotic factors influencing biomass accumulation of green tide causing Ulva spp. on Pyropia culture rafts in the Yellow Sea, China. Mar Pollut Bull 105:88–95CrossRefGoogle Scholar
  24. Lechtenfeld OJ, Herkorn N, Shen Y, Witt M, Benner R (2015) Marine sequestration of carbon in bacterial metabolites. Nat Commun 6:6711CrossRefGoogle Scholar
  25. Li HM, Zhang CS, Han XR, Shi XY (2015) Changes in concentrations of oxygen, dissolved nitrogen, phosphate, and silicate in the southern Yellow Sea, 1980–2012: sources and seaward gradients. Estuar Coast Shelf Sci 163:44–55CrossRefGoogle Scholar
  26. Li HM, Zhang YY, Han XR, Shi XY, Rivkin RB, Legendre L (2016) Growth responses of Ulva prolifera to inorganic and organic nutrients: implications for macroalgal blooms in the southern Yellow Sea, China. Sci Rep 6:26498CrossRefGoogle Scholar
  27. Li HM, Zhang YY, Tang HJ, Shi XY, Rivkin RB, Legendre L (2017) Spatiotemporal variations of inorganic nutrients along the Jiangsu coast, China, and the occurrence of macroalgal blooms (green tides) in the southern Yellow Sea. Harmful Algae 63:164–172CrossRefGoogle Scholar
  28. Liu D, Keesing JK, He PM, Wang ZL, Shi YJ, Wang YJ (2013) The world’s largest macroalgae bloom in the Yellow Sea, China: formation and implications. Estuar Coast Shelf Sci 129:2–10CrossRefGoogle Scholar
  29. Liu XQ, Wang ZL, Zhang XL (2016) A review of the green tides in the Yellow Sea, China. Mar Environ Res 119:189–196CrossRefGoogle Scholar
  30. Liu CC, Xu R, He PM, Zhang ZL, Qin YT, Xiang LY, Deng BP, Liu SH, Ji X (2017) Research on the relations between green tide and Porphyra cultivation in the south Yellow Sea. Mar Sci 41:35–43 (in Chinese with English abstract)Google Scholar
  31. Luo MB, Liu F, Xu ZL (2012) Growth and nutrient uptake capacity of two co-occurring species, Ulva prolifera and Ulva linza. Aquat Bot 100:18–24CrossRefGoogle Scholar
  32. Morand P, Merceron M (2004) Coastal eutrophication and excessive growth of macroalgae. In: Pandalai SG (ed) Recent research developments in environmental biology, vol. 1. Research signpost, Trivandrum, Kerala, pp 395–449Google Scholar
  33. Mulvenna PF, Savidge G (1992) A modified manual method for the determination of urea in seawater using diacetylmonoxime reagent. Estuar Coast Shelf Sci 34:429–438CrossRefGoogle Scholar
  34. Naldi M, Wheeler PA (2002) 15N measurements of ammonium and nitrate uptake by Ulva fenestrata (Chlorophyta) and Gracilaria pacifica (Rhodophyta): comparison of net nutrient disappearance, release of ammonium and nitrate, and 15N accumulation in algal tissue. J Phycol 38:135–144CrossRefGoogle Scholar
  35. National Bureau of Statistics of the People's Republic of China (NBS) (2007–2017)
  36. Ogawa T, Ohkib K, Kamiya M (2015) High heterozygosity and phenotypic variation of zoids in apomictic Ulva prolifera (Ulvophyceae) from brackish environments. Aquat Bot 120 part B:185–195CrossRefGoogle Scholar
  37. Pang SJ, Liu F, Shan TF, Xu N, Zhang ZH, Gao SQ, Chopin T, Sun S (2010) Tracking the algae origin of the Ulva bloom in the Yellow Sea by a combination of molecular, morphological and physiological analyses. Mar Environ Res 69:207–215CrossRefGoogle Scholar
  38. Pedersen MF (1994) Transient ammonium uptake in the macroalga Ulva lactuca (Chlorophyta): nature, regulation, and the consequences for choice of measuring technique. J Phycol 30:980–986CrossRefGoogle Scholar
  39. Rees TAV, Grant CM, Harmens HE, Taylor RB (1998) Measuring rates of ammonium assimilation in marine algae: use of the protonophore carbonyl cyanide m-chlorophenylhydrazone to distinguish between uptake and assimilation. J Phycol 34:264–272CrossRefGoogle Scholar
  40. Rees TAV, Dobson BC, Bijl M, Morelissen B (2007) Kinetics of nitrate uptake by New Zealand marine macroalgae and evidence for two nitrate transporters in Ulva intestinalis L. Hydrobiologia 586:135–141CrossRefGoogle Scholar
  41. Ross ME, Davis K, McColl R, Stanley MS, Day JG, Semião AJC (2018) Nitrogen uptake by the macro-algae Cladophora coelothrix and Cladophora parriaudii: influence on growth, nitrogen preference and biochemical composition. Algal Res 30:1–10CrossRefGoogle Scholar
  42. Runcie JW, Ritchie RJ, Larkum AWD (2003) Uptake kinetics and assimilation of inorganic nitrogen by Catenella nipae and Ulva lactuca. Aquat Bot 76:155–174CrossRefGoogle Scholar
  43. Shi XY, Qi MY, Tang HJ, Han XR (2015) Spatial and temporal nutrient variations in the Yellow Sea and their effects on Ulva prolifera blooms. Estuar Coast Shelf Sci 163:36–43CrossRefGoogle Scholar
  44. Sjöö GL, Mörk E (2009) Tissue nutrient content in Ulva spp. (Chlorophyceae) as bioindicator for nutrient loading along the coast of East Africa. Open J Environ Biol Monit 2:11–17CrossRefGoogle Scholar
  45. State Oceanic Administration People's Republic of China (SOA) (2008–2016) The National Bulletins of marine environment quality status. SOA Publication, BeijingGoogle Scholar
  46. Sun KM, Li RX, Li Y, Xin M, Xiao J, Wang ZL, Tang XX, Pang M (2015) Responses of Ulva prolifera to short-term nutrient enrichment under light and dark conditions. Estuar Coast Shelf Sci 163:56–62CrossRefGoogle Scholar
  47. Tada K, Tada M, Maita Y (1998) Dissolved free amino acids in coastal seawater using a modified fluorometric method. J Oceanogr 54:313–321CrossRefGoogle Scholar
  48. Taylor R, Fletcher RL, Raven JA (2011) Preliminary studies on the growth of selected green tide algae in laboratory culture: effects of irradiance, temperature, salinity and nutrients on growth rate. Bot Mar 44:327–336Google Scholar
  49. Tyler AC, McGlathery KJ, Macko SA (2005) Uptake of urea and amino acids by the macroalgae Ulva lactuca (Chlorophyta) and Gracilaria vermiculophylla (Rhodophyta). Mar Ecol Prog Ser 294:161–172CrossRefGoogle Scholar
  50. Tyrrell T (1999) The relative influences of nitrogen and phosphorus on oceanic primary production. Nature 400:525–531CrossRefGoogle Scholar
  51. Vandermeulen H, Gordin H (1990) Ammonium uptake using Ulva (Chlorophyta) in intensive fishpond systems: mass culture and treatment of effluent. J Appl Phycol 2:363–374CrossRefGoogle Scholar
  52. Vonk JA, Middelburg JJ, Stapel J, Bouma TJ (2008) Dissolved organic nitrogen uptake by seagrasses. Limnol Oceanogr 53:542–548CrossRefGoogle Scholar
  53. Wang XH, Qiao FL, Lu J, Gong F (2011) The turbidity maxima of the northern Jiangsu shoal-water in the Yellow Sea, China. Estuar Coast Shelf Sci 93:202–211CrossRefGoogle Scholar
  54. Wang ZL, Xiao J, Fan SL, Li Y, Liu XQ, Liu DY (2015) Who made the world’s largest green tide in China?—an integrated study on the initiation and early development of the green tide in Yellow Sea. Limnol Oceanogr 60:1105–1117CrossRefGoogle Scholar
  55. Zhou MJ, Liu DY, Anderson DM, Valiela I (2015) Introduction to the special issue on green tides in the Yellow Sea. Estuar Coast Shelf Sci 163:3–8CrossRefGoogle Scholar

Copyright information

© Springer Nature B.V. 2018

Authors and Affiliations

  1. 1.Key Laboratory of Biofuels, Shandong Provincial Key Laboratory of Energy Genetics, Qingdao Institute of Bioenergy and Bioprocess TechnologyChinese Academy of SciencesQingdaoChina
  2. 2.State Key Laboratory for Marine Environmental ScienceXiamen UniversityXiamenChina
  3. 3.CAS Key Laboratory of Experimental Marine Biology, Institute of OceanologyChinese Academy of SciencesQingdaoChina

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