Despite a well-documented rise in nitrate concentration over the past century, Lake Superior has retained an oligotrophic character. In part, this status results from physical attributes of the lake including low temperatures and prolonged isothermy, resulting in deep-mixing and light limitation which constrain primary production. Lake Superior is also phosphorus deficient which limits phytoplankton growth. We conducted large (20 l) volume factorial bioassay experiments to assess the influence of light and nutrients (P, Fe) on nitrate assimilation by a Lake Superior chlorophyte alga. Bioassays seeded with the chlorophyte yielded a strong response to light resulting in the rapid depletion of nitrate. High light resulted in higher activities of the key N-assimilation enzyme nitrate reductase (NR) and increased algal biomass compared to low light treatments. NR activity was highly correlated with rates of nitrate incorporation in bioassays and field surveys suggesting that NR occupies a critical place in nitrate metabolism. In bioassays, the addition of nutrients (P, Fe) only slightly increased the rate at which nitrate became depleted. Parallel trials using a luminescent cyanobacterial bioreporter confirmed the lack of response by added nutrients supporting light as an important factor in constraining nitrate assimilation by phytoplankton in the lake.
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Aleya, L., 1992. The seasonal succession of phytoplankton in an eutrophic lake through the coupling of biochemical-composition of particulates, metabolic parameters and environmental-conditions. Archiv für Hydrobiologie 124: 69–88.
Anagnostou, E. & R. M. Sherrell, 2008. MAGIC method for subnanomolar orthophosphate determination in freshwater. Limnology and Oceanography: Methods 6: 64–74.
Andersen, R. A., J. A. Berges, P. J. Harrison & M. M. Watanabe, 2005. Appendix A. Recipes for freshwater and seawater media. In Andersen, R. A. (ed.), Algal Culturing Techniques. Elsevier, Amsterdam: 429–538.
Barbiero, R. P. & M. L. Tuchman, 2001. Results from U.S. EPA’s biological open water surveillance program of the Laurentian Great Lakes: I. Introduction and phytoplankton results. Journal of Great Lakes Research 27: 134–154.
Barbiero, R. P. & M. L. Tuchman, 2004. The deep chlorophyll maximum in Lake Superior. Journal of Great Lakes Research 30: 256–268.
Beeton, A. M., 1965. Eutrophication of the St. Lawrence Great Lakes. Limnology and Oceanography 10: 240–254.
Bennett, E. B., 1986. The nitrifying of Lake Superior. Ambio 15: 272–275.
Berges, J. A., 1997. Algal nitrate reductases. European Journal of Phycology 32: 3–8.
Berges, J. A. & P. J. Harrison, 1995a. Nitrate reductase activity quantitatively predicts the rate of nitrate incorporation under steady state light limitation: a revised assay and characterization of the enzyme in three species of marine phytoplankton. Limnology and Oceanography 40: 82–93.
Berges, J. A. & P. J. Harrison, 1995b. Relationships between nitrate reductase activity and rates of growth and nitrate incorporation under steady-state light or nitrate limitation in the marine diatom Thalassiosira pseudonana (Bacillariophyceae). Journal of Phycology 31: 85–95.
Berges, J. A., W. P. Cochlan & P. J. Harrison, 1995. Laboratory and field responses of algal nitrate reductase to diel periodicity in irradiance, nitrate exhaustion, and the presence of ammonium. Marine Ecology Progress Series 124: 259–269.
Berges, J. A., C. E. Gibson & B. M. Stewart, 2004. Physiological responses of phytoplankton communities in the Irish Sea to simulated upwelling. Hydrobiologia 517: 121–132.
Boyd, P., J. A. Berges & P. J. Harrison, 1998. In vitro iron enrichment experiments at iron-rich and -poor sites in the NE subarctic Pacific. Journal of Experimental Marine Biology and Ecology 227: 133–151.
Carlton, R. G., G. S. Walker, M. J. Klug & R. G. Wetzel, 1989. Relative values of oxygen, nitrate, and sulfate to terminal microbial processes in the sediments of Lake Superior. Journal of Great Lakes Research 15: 133–140.
Chapra, S. C., A. Dove & G. J. Warren, 2012. Long-term trends of Great Lakes major ion chemistry. Journal of Great Lakes Research 38: 550–560.
Conroy, J. D., D. D. Kane, D. M. Dolan, W. J. Edwards, M. N. Charlton & D. A. Culver, 2005. Temporal trends in Lake Erie plankton biomass: roles of external phosphorus loading and dreissenid mussels. Journal of Great Lakes Research 31: 89–110.
Cullen, J. & E. Renger, 1979. Continuous measurement of the DCMU-induced fluorescence response of natural phytoplankton populations. Marine Biology 53: 13–20.
DeYoe, H. R. & C. A. Suttle, 1994. The inability of the Texas “brown tide” alga to use nitrate and the role of nitrogen in the initiation of a persistent bloom of this organism. Journal of Phycology 30: 800–806.
Dove, A., 2009. Long-term trends in major ions and nutrients in Lake Ontario. Aquatic Ecosystem Health & Management 12: 281–295.
Finlay, J. C., R. W. Sterner & S. Kumar, 2007. Isotopic evidence for in-lake production of accumulating nitrate in Lake Superior. Ecological Applications 17: 2323–2332.
Galloway, J. N., J. D. Aber, J. W. Erisman, S. P. Seitzinger, R. W. Howarth, E. B. Cowling & B. J. Cosby, 2003. The nitrogen cascade. BioScience 53: 341–356.
Gordillo, F. J. L., R. Garcia-Ruiz, A. Corzo, J. Lucena & F. X. Niell, 2001. Nitrate reductase activity in an eutrophic reservoir during the stratification cycle. International Review of Hydrobiology 86: 603–618.
Guildford, S. J. & R. E. Hecky, 2000. Total nitrogen, total phosphorus, and nutrient limitation in lakes and oceans: is there a common relationship? Limnology and Oceanography 45: 1213–1223.
Guildford, S. J., H. A. Bootsma, E. J. Fee, R. E. Hecky & G. Patterson, 2000. Phytoplankton nutrient status and mean water column irradiance in Lakes Malawi and Superior. Aquatic Ecosystem Health & Management 3: 35–45.
Hassler, C. S., S. M. Havens, G. S. Bullerjahn, R. M. L. McKay & M. R. Twiss, 2009. An evaluation of iron bioavailability and speciation in western Lake Superior with the use of combined physical, chemical, and biological assessment. Limnology and Oceanography 54: 987–1001.
Hawley, N., T. H. Johengren, Y. R. Rao, A. Ruberg, D. Beletsky, S. A. Ludsin, B. J. Eadie, D. J. Schwab, T. E. Croley & S. B. Brandt, 2006. Lake Erie hypoxia prompts Canada—U.S. study. Eos, Transactions, American Geophysical Union 87: 313–314.
Healey, F. P. & L. L. Hendzel, 1979. Fluorometric measurement of alkaline phosphatase activity in algae. Freshwater Biology 9: 429–439.
Hecky, R. E. & P. Kilham, 1988. Nutrient limitation of phytoplankton in freshwater and marine environments: a review of recent evidence on the effects of enrichment. Limnology and Oceanography 33: 796–822.
Hochman, A., A. Nissany, D. Wynne, B. Kaplan & T. Berman, 1986. Nitrate reductase: an improved assay method for phytoplankton. Journal of Plankton Research 8: 385–392.
Hung, C. C., G. T. F. Wong, K. K. Liu, F. K. Shiah & G. C. Gong, 2000. The effects of light and nitrate levels on the relationship between nitrate reductase activity and 15NO3 − uptake: field observations in the East China Sea. Limnology and Oceanography 45: 836–848.
Ivanikova, N. V., R. M. L. McKay & G. S. Bullerjahn, 2005. Construction and characterization of a cyanobacterial bioreporter capable of assessing nitrate assimilatory capacity in freshwaters. Limnology and Oceanography: Methods 3: 86–93.
Ivanikova, N. V., R. M. L. McKay, G. S. Bullerjahn & R. W. Sterner, 2007a. Nitrate utilization by phytoplankton in Lake Superior is impaired by low nutrient (P, Fe) availability and seasonal light limitation—a cyanobacterial bioreporter study. Journal of Phycology 43: 475–484.
Ivanikova, N. V., L. C. Popels, R. M. L. McKay & G. S. Bullerjahn, 2007b. Lake Superior supports novel clusters of cyanobacterial picoplankton. Applied and Environmental Microbiology 73: 4055–4065.
Kelly, J. R., P. M. Yurista, S. E. Miller, A. C. Cotter, T. C. Corry, J. V. Scharold, M. E. Sierszen, E. J. Issac & J. D. Stockwell, 2011. Challenges to Lake Superior’s condition, assessment and management: a few observations across a generation of change. Aquatic Ecosystem Health & Management 14: 332–344.
Kumar, S., R. W. Sterner & J. C. Finlay, 2008. Nitrogen and carbon uptake dynamics in Lake Superior. Journal of Geophysical Research 113: G04003.
Li, J., S. A. Crowe, D. Miklesh, M. Kistner, D. E. Canfield & S. Katsev, 2012. Carbon mineralization and oxygen dynamics in sediments with deep oxygen penetration, Lake Superior. Limnology and Oceanography 57: 1634–1650.
Lu, Y., P. A. Meyers, T. H. Johengen, B. J. Eadie, J. A. Robbins & H. Han, 2010. δ15N values in Lake Erie sediments as indicators of nitrogen biogeochemical dynamics during cultural eutrophication. Chemical Geology 273: 1–7.
Mackey, M. D., D. J. Mackey, H. W. Higgens & S. W. Wright, 1996. CHEMTAX—a program for estimating class abundances from chemical markers: application to HPLC measurements of phytoplankton. Marine Ecology Progress Series 144: 265–283.
Mackey, D. J., H. W. Higgins, M. D. Mackey & D. Holdsworth, 1998. Algal class abundances in the western equatorial Pacific: estimation from HPLC measurements of chloroplast pigments using CHEMTAX. Deep Sea Research Part I: Oceanographic Research Papers 45: 1441–1468.
Makarewicz, J. C., P. Bertram & T. W. Lewis, 2000. Chemistry of the offshore surface waters of Lake Erie: pre- and post-Dreissena introduction (1983–1993). Journal of Great Lakes Research 26: 82–93.
Mallet, C., M. Charpin & J. Devaux, 1998. Nitrate reductase activity of phytoplankton populations in eutrophic Lake Aydat and meso-oligotrophic Lake Pavin: a comparison. Hydrobiologia 374: 135–148.
McCarthy, M. J., W. S. Gardner, P. J. Lavrentyev, K. M. Moats, F. J. Jochem & D. M. Klarer, 2007. Effects of hydrological flow regime on sediment–water interface and water column nitrogen dynamics in a Great Lakes coastal watershed (Old Woman Creek, Lake Erie). Journal of Great Lakes Research 33: 219–231.
McDonald, C. P., N. R. Urban & C. M. Casey, 2010. Modeling historical trends in Lake Superior total nitrogen concentrations. Journal of Great Lakes Research 36: 715–721.
McKay, R. M. L., D. Porta, G. S. Bullerjahn, M. M. D. Al-Rshaidat, J. A. Klimowicz, R. W. Sterner, T. A. Smutka, E. T. Brown & R. M. Sherrell, 2005. Bioavailable iron in oligotrophic Lake Superior assessed using biological reporters. Journal of Plankton Research 27: 1033–1044.
Menzel, D. W. & N. Corwin, 1965. The measurement of total phosphorus in seawater based on the liberation of organically bound fractions by persulfate oxidation. Limnology and Oceanography 10: 280–282.
Millie, D. F., G. L. Fahnenstiel, J. Dyble Bressie, R. J. Pigg, R. R. Rediske, D. Klarer, P. A. Tester & R. W. Litaker, 2009. Late-summer phytoplankton in western Lake Erie (Laurentian Great Lakes): bloom distributions, toxicity, and environmental influences. Aquatic Ecology 43: 915–934.
Moore, L. R., A. F. Post, G. Rocap & S. W. Chisholm, 2002. Utilization of different nitrogen sources by the marine cyanobacteria Prochlorococcus and Synechococcus. Limnology and Oceanography 47: 989–996.
Morris, I. & P. J. Syrett, 1963. The development of nitrate reductase in Chlorella and its repression by ammonium. Archiv für Mikrobiologie 47: 32–41.
Nalewajko, C. & D. Voltolina, 1986. Effects of environmental variables on growth rates and physiological characteristics of Lake Superior phytoplankton. Canadian Journal of Fisheries and Aquatic Sciences 43: 1163–1170.
Needoba, J. A. & P. J. Harrison, 2004. Influence of low light and a light: dark cycle on NO3 − uptake, intracellular NO3 −, and nitrogen isotope fractionation by marine phytoplankton. Journal of Phycology 40: 505–516.
Neilson, M. A., D. S. Painter, G. Warren, R. A. Hites, I. Basu, D. V. C. Weseloh, D. M. Whittle, G. Christie, R. Barbiero, M. Tuchman, O. E. Johannsson, T. F. Nalepa, T. A. Edsall, G. Fleischer, C. Bronte, S. B. Smith & P. C. Baumann, 2003. Ecological monitoring for assessing the state of the nearshore and open waters of the Great Lakes. Environmental Monitoring and Assessment 88: 103–117.
Nriagu, J. O., G. Lawson, H. K. T. Wong & V. Cheam, 1996. Dissolved trace metals in Lakes Superior, Erie, and Ontario. Environmental Science and Technology 30: 178–187.
Parsons, T., M. Maita & C. Lalli, 1984. A Manual of Chemical and Biological Methods for Seawater Analysis. Pergammon Press, Oxford.
Rhee, G. Y., 1978. Effects of N:P atomic ratios and nitrate limitation on algal growth, cell composition, and nitrate uptake. Limnology and Oceanography 23: 10–25.
Richards, R. P. & D. B. Baker, 1993. Trends in nutrient and suspended sediment concentrations in Lake Erie tributaries, 1975–1990. Journal of Great Lakes Research 19: 200–211.
Rufty, T. W., C. T. MacKown & D. W. Israel, 1990. Phosphorus stress effects on assimilation of nitrate. Plant Physiology 94: 328–333.
Schindler, D. W., 2006. Recent advances in the understanding and management of eutrophication. Limnology and Oceanography 51: 356–363.
Sinsabaugh, R. L., S. Findlay, P. Franchini & D. Fischer, 1997. Enzymatic analysis of riverine bacterioplankton production. Limnology and Oceanography 42: 29–38.
Small, G. E., G. S. Bullerjahn, R. W. Sterner, B. F. N. Beall, S. Brovold, J. C. Finlay, R. M. L. McKay & M. Mukherjee, 2013a. Rates and controls of nitrification in a large oligotrophic lake. Limnology and Oceanography 58: 276–286.
Small, G. E., J. B. Cotner, J. C. Finlay, R. A. Stark & R. W. Sterner, 2013b. Nitrogen transformations at the sediment–water interface across redox gradients in the Laurentian Great Lakes. Hydrobiologia. doi:10.1007/s10750-013-1569-7.
Sterner, R. W., 2010. In situ measured primary production in Lake Superior. Journal of Great Lakes Research 36: 139–149.
Sterner, R. W., 2011. C:N:P stoichiometry in Lake Superior: freshwater sea as end member. Inland Waters 1: 29–46.
Sterner, R. W., T. M. Smutka, R. M. L. McKay, Q. Xiaoming, E. T. Brown & R. M. Sherrell, 2004. Phosphorus and trace metal limitation of algae and bacteria in Lake Superior. Limnology and Oceanography 49: 495–507.
Sterner, R. W., E. Anagnostou, S. Brovold, G. S. Bullerjahn, J. C. Finlay, S. Kumar, R. M. L. McKay & R. M. Sherrell, 2007. Increasing stoichiometric imbalance in North America’s largest lake: nitrification in Lake Superior. Geophysical Research Letters 34: L10406.
U.S. Environmental Protection Agency, 1971. Algal Assay Procedure: Bottle Test. National Eutrophication Research Program, Corvallis, OR: 82 pp.
Vergara, J. J., J. A. Berges & P. G. Falkowski, 1998. Diel periodicity of nitrate reductase activity and protein levels in the marine diatom Thalassiosira weissflogii (Bacillariophyceae). Journal of Phycology 34: 952–961.
Weiler, R., 1978. Chemistry of Lake Superior. Journal of Great Lakes Research 4: 370–385.
Welschmeyer, N. A., 1994. Fluorometric analysis of chlorophyll a in the presence of chlorophyll b and pheopigments. Limnology and Oceanography 39: 1985–1992.
Wynne, D. & T. Berman, 1990. The influence of environmental factors on nitrate reductase activity in freshwater phytoplankton. I. Field studies. Hydrobiologia 194: 235–245.
This material is based upon work supported by the National Science Foundation under grant numbers OCE-0327738, OCE-0352274 (R.M.L.M. and G.S.B.), and OCE-0352291(R.W.S). The authors are grateful to the Captain and crew of the R/V Blue Heron for their assistance with the collection of samples. We also extend thanks to Michael Twiss, Christel Hassler, Rob Sherrell, Jason Agnich, and Sandra Brovold who provided assistance with shipboard sampling and analysis. HPLC analysis was provided by the Center for Hydro-Optics and Remote Sensing, San Diego State University. We thank Lauren Simmons for assistance with CHEMTAX analyses of HPLC data. Ben Beall provided assistance with figure preparation.
Guest editors: D. Straile, D. Gerdeaux, D. M. Livingstone, P. Nõges, F. Peeters & K.-O. Rothhaupt / European Large Lakes III. Large lakes under changing environmental conditions
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Berges, J.A., Jiang, Y., Sterner, R.W. et al. Identification of factors constraining nitrate assimilation in Lake Superior, Laurentian Great Lakes. Hydrobiologia 731, 81–94 (2014). https://doi.org/10.1007/s10750-013-1637-z
- Lake Superior
- Nitrate reductase