Uptake and inhibition kinetics of nitrogen in Microcystis aeruginosa: Results from cultures and field assemblages collected in the San Francisco Bay Delta,CA

The nitrogen (N) uptake kinetic parameters for Microcystis field assemblages collected from the San Francisco Bay Delta (Delta) in 2012 and non-toxic and toxic laboratory culture strains of M. aeruginosa were assessed. The ¹⁵N tracer technique was used to investigate uptake of ammonium (NH₄⁺), nitrate (NO₃⁻), urea and glutamic acid over short-term incubations (0.5–1 h), and to study inhibition of NO₃⁻, NH₄⁺ and urea uptake by NH₄⁺, NO₃⁻ and NH₄⁺, respectively. This study demonstrates that Delta Microcystis can utilize different forms of inorganic and organic N, with the greatest capacity for NH₄⁺ uptake and the least for glutamic acid uptake, although N uptake did not always follow the classic Michaelis–Menten hyperbolic relationship at substrate concentrations up to 67 μmol N L⁻¹. Current ambient N concentrations in the Delta may be at sub-saturating levels for N uptake, indicating that if N loading (especially NH₄⁺) were to increase, Delta Microcystis assemblages have the potential for increased N uptake rates. Delta Microcystis had the highest specific affinity, α, for NH₄⁺ and the lowest for NO₃⁻. In culture, N uptake by non-toxic and toxic M. aeruginosa strains was much higher than from the field, but followed similar N utilization trends to those in the field. Neither strain showed severe inhibition of NO₃⁻ uptake by NH₄⁺ or inhibition of NH₄⁺ uptake on NO₃⁻, but both strains showed some inhibition of urea uptake by NH₄⁺.     

Organic and inorganic nitrogen utilization by nitrogen-stressed cyanobacteria during bloom conditions

Cyanobacterial blooms often occur in lakes that have high phosphorus (P) and low nitrogen (N) concentrations, and the growth rate of the blooms is often constrained by N. For these reasons, many researchers have suggested that regulation of both P and N is required to control eutrophication. However, because N occurs in many bioavailable forms, regulation of a particular form may be beneficial rather than regulation of all N forms. To address how N-stressed cyanobacteria respond to various N inputs, N enrichment experiments (nitrate, ammonium, urea, and alanine) were performed during N-limited cyanobacterial blooms in Maumee and Sandusky Bays of Lake Erie and in Grand Lake St. Marys (GLSM). Bioavailable N (nitrate, urea, and ammonium) concentrations were also determined. Microcystis aeruginosa dominated the Maumee Bay bloom, where the highest growth rates were in response to ammonium additions, and lowest growth rates were in response to nitrate. Urea and the amino acid alanine resulted in intermediate growth rates. Planktothrix agardhii dominated the Sandusky Bay and GLSM blooms, where nitrate, ammonium, and urea addition resulted in similar growth rates. Additions of alanine did not stimulate growth of the Planktothrix blooms. Incubations using stable isotope ¹⁵N showed the cyanobacteria had a preference for ammonium, but the other forms were also assimilated in the presence of ammonium. These results show that cyanobacterial blooms will assimilate multiple forms of N to support growth. Thus, if lake managers do decide that N abatement is necessary, then all forms of bioavailable N need to be constrained.     

Nutrient limitation of Microcystis aeruginosa in northern California Klamath River reservoirs

Nutrient limitations were investigated in Copco and Iron Gate Reservoirs, on the Klamath River in California, where blooms of the toxin-producing cyanobacterium Microcystis aeruginosa were first reported in 2005. Nutrient enrichment experiments conducted in situ in June and August, 2007 and 2008, determined responses in phytoplankton biomass, Microcystis abundance and microcystin concentration to additions of phosphorus and different forms of nitrogen (NH₄⁺, NO₃, and urea). Microcystis abundance was determined using quantitative PCR targeting the phycocyanin intergenic spacer cpcBA.Total phytoplankton biomass increased with additions of N both before and during Microcystis blooms, with no primary effects from P, suggesting overall N limitation for phytoplankton growth during the summer season. NH₄⁺ generally produced the greatest response in phytoplankton growth, while Microcystis abundance increased in response to all forms of N. Microcystis doubling time in the in situ experiments was 1.24–1.39 days when N was not limiting growth. The results from this study suggest availability of N during the summer is a key growth-limiting factor for the initiation and maintenance of toxic Microcystis blooms in Copco and Iron Gate Reservoirs in the Klamath River.     

Nitrogen dynamics at the sediment–water interface in shallow, sub-tropical Florida Bay: why denitrification efficiency may decrease with increased eutrophication

Nitrogen (N) dynamics at the sediment–water interface were examined in four regions of Florida Bay to provide mechanistic information on the fate and effects of increased N inputs to shallow, subtropical, coastal environments. Dissimilatory nitrate (NO₃ ^?) reduction to ammonium (DNRA) was hypothesized to be a significant mechanism retaining bioreactive N in this warm, saline coastal ecosystem. Nitrogen dynamics, phosphorus (P) fluxes, and sediment oxygen demand (SOD) were measured in north-central (Rankin Key; eutrophic), north-eastern (Duck Key; high N to P seston ratios), north-western (Murray Key; low N to P ratios), and central (Rabbit Key; typical central site) Florida Bay in August 2004, January 2005, and November 2006. Site water was passed over intact sediment cores, and changes in oxygen (O₂), phosphate (o-PO₄ ^3?), ammonium (NH₄ ⁺), NO₃ ^?, nitrite (NO₂ ^?), and N₂ concentrations were measured, without and with addition of excess ¹⁵NO₃ ^? or ¹⁵NH₄ ⁺ to inflow water. These incubations provided estimates of SOD, nutrient fluxes, N₂ production, and potential DNRA rates. Denitrification rates were lowest in summer, when SOD was highest. DNRA rates and NH₄ ⁺ fluxes were high in summer at the eutrophic Rankin site, when denitrification rates were low and almost no N₂ came from added ¹⁵NO₃ ^?. Highest ¹⁵NH₄ ⁺ accumulation, resulting from DNRA, occurred at Rabbit Key during a picocyanobacteria bloom in November. ¹⁵NH₄ ⁺ accumulation rates among the stations correlated with SOD in August and January, but not in November during the algal bloom. These mechanistic results help explain why bioreactive N supply rates are sometimes high in Florida Bay and why denitrification efficiency may decrease with increased NO₃ ^? inputs in sub-tropical coastal environments.     

Inorganic carbon uptake by phytoplankton in Vineyard Sound,Massachusetts. II. Comparative primary productivity and nutritional status of winter and summer assemblages

Using time-course, natural-light incubations, we assessed the rate of carbon uptake at a range of light intensities, the effect of supplemental additions of nitrogen (as NH₄⁺ or urea) on light and dark carbon uptake, and the rates of uptake of NH₄⁺ and urea by phytoplankton from Vineyard Sound, Massachusetts from February through August 1982. During the winter, photoinhibition was severe, becoming manifested shortly after the start of an incubation, whereas during the summer, there was little to no evidence of photoinhibition during the first several hours after the start of an incubation. At light levels which were neither photoinhibiting nor light limiting, rates of carbon uptake normalized per liter were high and approximately equal during winter and summer (22–23 μg C·l^?1 · h^?1), and low during spring (<10 μgC·l^?1· h^?1). In contrast, on a chlorophyll a basis, rates of carbon fixation were as high during spring (15–20μg C·μg Chl a^?1·h^?1), when concentrations of chlorophyll a were at the yearly minimum (<0.5 μg · l^?1) as during the summer, when chlorophyll a concentrations were substantially higher (0.8–1.3 μg · l^?1). Highest rates of NH₄⁺ and urea uptake were observed during summer, and at no time of the year was there evidence for severe nitrogen deficiency, although moderate nitrogen nutritional stress was apparent during the summer months.     

Nutrient limitation of Microcystis aeruginosa in northern California Klamath River reservoirs

Nutrient limitations were investigated in Copco and Iron Gate Reservoirs, on the Klamath River in California, where blooms of the toxin-producing cyanobacterium Microcystis aeruginosa were first reported in 2005. Nutrient enrichment experiments conducted in situ in June and August, 2007 and 2008, determined responses in phytoplankton biomass, Microcystis abundance and microcystin concentration to additions of phosphorus and different forms of nitrogen (NH₄⁺, NO₃, and urea). Microcystis abundance was determined using quantitative PCR targeting the phycocyanin intergenic spacer cpcBA.Total phytoplankton biomass increased with additions of N both before and during Microcystis blooms, with no primary effects from P, suggesting overall N limitation for phytoplankton growth during the summer season. NH₄⁺ generally produced the greatest response in phytoplankton growth, while Microcystis abundance increased in response to all forms of N. Microcystis doubling time in the in situ experiments was 1.24–1.39 days when N was not limiting growth. The results from this study suggest availability of N during the summer is a key growth-limiting factor for the initiation and maintenance of toxic Microcystis blooms in Copco and Iron Gate Reservoirs in the Klamath River.     

Urea dynamics during Lake Taihu cyanobacterial blooms in China

Lake Taihu, the third largest freshwater lake in China, suffers from harmful cyanobacteria blooms caused by Microcystis spp., which do not fix nitrogen (N). Reduced N (i.e., NH₄⁺, urea and other labile organic N compounds) is an important factor affecting the growth of Microcystis. As the world use of urea as fertilizer has escalated during the past decades, an understanding of how urea cycling relates to blooms of Microcystis is critical to predicting, controlling and alleviating the problem. In this study, the cycling rates of urea-N in Lake Taihu ranged from non-detectable to 1.37 μmol N L⁻¹ h⁻¹ for regeneration, and from 0.042 μmol N L⁻¹ h⁻¹ to 2.27 μmol N L⁻¹ h⁻¹ for potential urea-N removal. The fate of urea-N differed between light and dark incubations. Increased ¹⁵NH₄⁺ accumulated and higher quantities of the removed urea-¹⁵N remained in the ¹⁵NH₄⁺ form were detected in the dark than in the light. A follow-up incubation experiment with ¹⁵N-urea confirmed that Microcystis can grow on urea but its effects on urea dynamics were minor, indicating that Microcystis was not the major factor causing the observed fates of urea under different light conditions in Lake Taihu. Bacterial community composition and predicted functional gene data suggested that heterotrophic bacteria metabolized urea, even though Microcystis spp. was the dominant bloom organism.     

SHORT TERM UPTAKE OF NUTRIENTS BY ENTEROMORPHA PROLIFERA (CHLOROPHYCEAE)1

Rates of NH₄⁺ and NO₃^? uptake were determined by accumulation of ¹⁵N in plant tissue and by disappearance of nutrient from the medium. Agreement between rates calculated by the two methods was good, averaging 82.7% (SD = 15.8%) and 91.2% (SD = 13.7%) for NH₄⁺ and NO₃^? uptake, respectively. An average of 93.4 and 96.0% of added ¹⁵NH₄⁺ and ¹⁵NO₃^? was recovered from the medium and /or plant tissue at the end of the incubations. Both bacterial uptake and regeneration of NH₄⁺ may contribute to discrepancies between NH₄⁺ uptake rates calculated by ¹⁵N accumulation and disappearance of NH₄⁺ from the medium. The influence of tissue composition on uptake of NH₄⁺, NO₃^? and PO₄^3- by Enteromorpha prolifera (Müller) J. Agardh was examined. For NH₄⁺ uptake, V_max was 188 μmol NH₄^+. g dry wt^?1. h^?1 and K_s ranged from 9.3 to 13.4 μM, but there was no correlation between kinetic parameters and tissue nitrogen content. For NO₃^?, both kinetic parameters were higher for plants with low tissue nitrogen than for plants with high tissue nitrogen. Maximum rates were 169 and 75.4 μmol NO₃^?. g dry wt^?1. h^?1, and K_s was 13.3 and 2.31 μM for low and high tissue nitrogen plants, respectively. Estimates of uptake in the field suggested that NH₄⁺ accounted for 65% and NO₃^? for up to 35% of total nitrogen uptake during the summer. Nutrient uptake rates of field-collected plants also indicated that E. prolifera in Yaquina Bay, Oregon was not likely to have been nitrogen-limited, but may have been phosphorus-limited.     

Longitudinal and seasonal variation of stream N uptake in an urbanizing watershed: effect of organic matter, stream size, transient storage and debris dams

We examined seasonal and spatial linkages between N cycling and organic matter for a suburban stream in Maryland and addressed the question: How do longitudinal NH₄ ⁺ uptake patterns vary seasonally and what is the effect of organic matter, stream size, transient storage and debris dams? We applied a longitudinal (stream channel corridor) approach in a forested stream section and conducted short-term nutrient addition experiments (adapted to account for the effect of nutrient saturation) covering 14–16 reaches, and compared two distinct seasons (late fall 2003 and late summer 2004). Longitudinal NH₄ ⁺ uptake rate patterns had a distinct seasonal reversal; fall had the highest uptake rates in the upper reaches, while summer had the highest uptake rates in the lower reaches. This seasonal reversal was attributed to organic matter and evidenced by DON patterns. Transient storage did not have an expected effect on uptake rates in fall because it was confounded by leaf litter; litter produced higher uptakes, but also may have reduced transient storage. In summer however, uptake rates had a positive correlation with transient storage. Debris dams had no distinct effect on uptake in fall because of their recent formation. In summer however, the debris dam effect was significant; although the debris dams were hydraulically inactive then, the upstream reaches had 2–5 fold higher uptake rates. The seasonal and longitudinal differences in NH₄ ⁺ uptake reflect interactions between flow conditions and the role of organic matter. Urbanization can alter both of these characteristics, hence affect stream N processing.     

CHARACTERIZATION OF NITROGEN UPTAKE BY NATURAL POPULATIONS OF AUREOCOCCUS ANOPHAGEFFERENS (CHRYSOPHYCEAE) AS A FUNCTION OF INCUBATION DURATION,SUBSTRATE CONCENTRATION,LIGHT, AND TEMPERATURE1

Nitrogen uptake studies were conducted during an aestival “brown tide” bloom in Shinnecock Bay, Long Island, New York. The same station was sampled in late July and mid-August 1995 when Aureococcus anophagefferens composed >90% and 30–40% of the total cell density, respectively. Experiments were designed to examine the effect of incubation duration on the uptake kinetics, and the effect of light and temperature dependencies of NH₄⁺, urea, and NO₃^? uptake. Maximum specific uptake rates (V'_max) decreased in the order NH₄⁺, urea, NO₃^? and were nonlinear with time for NH₄⁺ and urea, both of which exhibited an exponential decline between 1 and 10 min and then did nut significantly change for 60 min. Nitrogen uptake kinetic experiments exhibited a typical hyperbolic response for urea and NO₃^?. Half-saturation constants. (K_s) were calculated to he 0.03 and 0.12 μmol · L^?1 for urea and NO₃^?; respectively, but could not be calculated for NH₄⁺ under these experimental conditions. Nutrient uptake rate versus, irradiance (NI) experiments showed that maximum uptake rates occurred at ≤% of incident irradiance on both sampling dates and that values of V′_max-cell (NH₄⁺) were on average 30% greater than V′_max-cell (urea). A7°–9°C temperature decrease in incubation temperature between the two NI experiments in August resulted in a 30% decrease in V′_max-cell(NH₄⁺), no change in V′_max-cell(urea), and a 3–4-fold decrease in calculated K_lt values for both NH₄⁺ and urea. The results from these experiments demonstrate that A. anophagefferens has a higher affinity for NH₄⁺ and urea than for NO₃^? and that this particular species is adapted to use these substrates at low irradiances and concentrations. The data presented in this study are also consistent with the hypothesis that A. anophagefferens may be an oceanic clone that was displaced by an anomalous oceanographic event.     

Uptake of dissolved inorganic and organic nitrogen by the benthic toxic dinoflagellate Ostreopsis cf. ovata

Environmental factors that shape dynamics of benthic toxic blooms are largely unknown. In particular, for the toxic dinoflagellate Ostreopsis cf. ovata, the importance of the availability of nutrients and the contribution of the inorganic and organic pools to growth need to be quantified in marine coastal environments. The present study aimed at characterizing N-uptake of dissolved inorganic and organic sources by O. cf. ovata cells, using the ¹⁵N-labelling technique. Experiments were conducted taking into account potential interactions between nutrient uptake systems as well as variations with the diel cycle. Uptake abilities of O. cf. ovata were parameterized for ammonium (NH₄⁺), nitrate (NO₃⁻) and N-urea, from the estimation of kinetic and inhibition parameters. In the range of 0 to 10 μmol N L⁻¹, kinetic curves showed a clear preference pattern following the ranking NH₄⁺ > NO₃⁻ > N-urea, where the preferential uptake of NH₄⁺ relative to NO₃⁻ was accentuated by an inhibitory effect of NH₄⁺ concentration on NO₃⁻ uptake capabilities. Conversely, under high nutrient concentrations, the preference for NH₄⁺ relative to NO₃⁻ was largely reduced, probably because of the existence of a low-affinity high capacity inducible NO₃⁻ uptake system. Ability to take up nutrients in darkness could not be defined as a competitive advantage for O. cf. ovata. Species competitiveness can also be defined from nutrient uptake kinetic parameters. A strong affinity for NH₄⁺ was observed for O. cf. ovata cells that may partly explain the success of this toxic species during the summer season in the Bay of Villefranche-sur-mer (France).     

Effects of NH4+ concentration on growth,morphology and NH4+ uptake kinetics of Salvinia natans

Many plants develop toxicity symptoms and have reduced growth rates when supplied with ammonium (NH₄⁺) as the only source of inorganic nitrogen. In the present study, the growth, morphology, NH₄⁺ uptake kinetics and mineral concentrations in the tissues of the free-floating aquatic plant Salvinia natans (water fern) supplied exclusively with NH₄⁺–N at concentrations of 0.25–15 mM were investigated. S. natans grew well, with relative growth rates of c. 0.25 g g^?1 d^?1 at external NH₄⁺ concentrations up to 5 mM, but at higher levels growth was suppressed and the plants had small leaves and short roots with stunted growth. The high-affinity transport system (HATS) that mediate NH₄⁺ uptake at dilute NH₄⁺ levels was downregulated at high NH₄⁺ concentrations with lower velocities of maximum uptake (V_max) and higher half-saturation constants (K_1/2). High NH₄⁺ levels also barely affected the concentrations of mineral cations and anions in the plant tissue. It is concluded that S. natans can be characterized as NH₄⁺-tolerant in line with a number of other species of wetland plants as growth was unaffected at NH₄⁺ concentrations as high as 5 mM and as symptoms of toxicity at higher concentrations were relatively mild. Depolarization of the plasma membrane to the equilibrium potential for NH₄⁺ at high external concentrations may be a mechanism used by the plant to avoid excessive futile transmembrane cycling. S. natans is tolerant to the high NH₄⁺ levels that prevail in domestic and agricultural wastewaters, and the inherent high growth rate and the ease of biomass harvesting make S. natans a primary candidate for use in constructed wetland systems for the treatment of various types of nitrogen-rich wastewaters.     

Dominant genera of cyanobacteria in Lake Taihu and their relationships with environmental factors

Cyanobacterial blooms in freshwaters have become one of the most widespread of environmental problems and threaten water resources worldwide. Previous studies on cyanobacteria in Lake Taihu often collected samples from one site (like Meiliang Bay or Zhushan Bay) and focused on the variation in patterns or abundance of Microcystis during the blooming season. However, the distribution of cyanobacteria in Lake Taihu shows differing pattern in various seasons. In this study, water samples were collected monthly for one year at five sites in Lake Taihu with different trophic status and a physicochemical analysis and denaturing gradient gel electrophoresis (DGGE) were conducted. DGGE fingerprint analysis showed that Microcystis (7/35 bands) and Synechococcus (12/35 bands) were the two most dominant genera present during the study period at all five sites. Cyanobium (3/35 bands) was the third most common genus which has seldom been previously reported in Lake Taihu. Redundancy analysis (RDA) indicated that the cyanobacterial community structure was significantly correlated with NO₃^--N, COD_Mn, and NH₄⁺-N in the winter and spring, whereas it was correlated with water temperature in the summer and autumn. Limiting the nutrient input (especially of N and C loading) in Lake Taihu would be a key factor in controlling the growth of different genera of cyanobacteria.     

THE RELATIVE IMPORTANCE OF WATER MOTION ON NITROGEN UPTAKE BY THE SUBTIDAL MACROALGA ADAMSIELLA CHAUVINII (RHODOPHYTA) IN WINTER AND SUMMER1

The influence of seawater velocity (1.5–12 cm · s^?1) on inorganic nitrogen (N) uptake by the soft‐sediment perennial macroalga Adamsiella chauvinii (Harv.) L. E. Phillips et W. A. Nelson (Rhodophyta) was determined seasonally by measuring uptake rate in a laboratory flume. Regardless of N tissue content, water velocity had no influence on NO₃^? uptake in either winter or summer, indicating that NO₃^?‐uptake rate was biologically limited. However, when thalli were N limited, increasing water velocity increased NH₄⁺ uptake, suggesting that mass‐transfer limitation of NH₄⁺ is likely during summer for natural populations. Uptake kinetics (V_max, K_s) were similar among three populations of A. chauvinii at sites with different mean flow speeds; however, uptake rates of NO₃^? and NH₄⁺ were lower in summer (when N status was generally low) than in winter. Our results highlight how N uptake can be affected by seasonal changes in the physiology of a macroalga and that further investigation of N uptake of different macroalgae (red, brown, and green) during different seasons is important in determining the relative influence of water velocity on nutrient uptake.     

Microcystin mcyA and mcyE Gene Abundances Are Not Appropriate Indicators of Microcystin Concentrations in Lakes

Cyanobacterial harmful algal blooms (cyanoHABs) are a primary source of water quality degradation in eutrophic lakes. The occurrence of cyanoHABs is ubiquitous and expected to increase with current climate and land use change scenarios. However, it is currently unknown what environmental parameters are important for indicating the presence of cyanoHAB toxins making them difficult to predict or even monitor on time-scales relevant to protecting public health. Using qPCR, we aimed to quantify genes within the microcystin operon (mcy) to determine which cyanobacterial taxa, and what percentage of the total cyanobacterial community, were responsible for microcystin production in four eutrophic lakes. We targeted Microcystis-16S, mcyA, and Microcystis, Planktothrix, and Anabaena-specific mcyE genes. We also measured microcystins and several biological, chemical, and physical parameters—such as temperature, lake stability, nutrients, pigments and cyanobacterial community composition (CCC)—to search for possible correlations to gene copy abundance and MC production. All four lakes contained Microcystis-mcyE genes and high percentages of toxic Microcystis, suggesting Microcystis was the dominant microcystin producer. However, all genes were highly variable temporally, and in few cases, correlated with increased temperature and nutrients as the summer progressed. Interestingly, toxin gene abundances (and biomass indicators) were anti-correlated with microcystin in all lakes except the largest lake, Lake Mendota. Similarly, gene abundance and microcystins differentially correlated to CCC in all lakes. Thus, we conclude that the presence of microcystin genes are not a useful tool for eliciting an ecological role for toxins in the environment, nor are microcystin genes (e.g. DNA) a good indicator of toxins in the environment.     

Ammonium uptake in Lemna gibba G 1, related membrane potential changes, and inhibition of anion uptake

In N-starved (?N) fronds of Lemna gibba L. G 1, NH₄⁺ uptake rates were several-fold those of NO₃^?-supplied (+N) fronds. NO₃^?, uptake in +N-plants was slow and not inhibited by addition of NH₄⁺. However, in ?N-plants with higher NO₃^? and still higher NH₄⁺ uptake rates, addition of NH₄⁺ immediately reduced the NO₃^? uptake rates to about one third until the NH₄⁺ was consumed. The membrane potential (E_m) decreased immediately upon addition of NH₄⁺ in all fronds, but whereas depolarisation was moderate and transient in +N-plants, it was strong, up to 150 mV, in N-starved plants, where E_m remained at the level of the K⁺ diffusion potential (E_D) until NH₄⁺ was removed. In N-starved plants NH₄⁺ uptake and membrane depolarisation showed the same concentration dependence, except for an apparent linear component for uptake. Phosphate uptake was inhibited by NH₄⁺ similarly to NO₃^? uptake, but only in P- and N-starved plants, not after mere P starvation. Influx of NO₃^? and H₂PO ₄^? into the negatively charged cells of Lemna is mediated by anion/H⁺ cotransport, but NH₄⁺ influx can follow the electrochemical gradient. Its saturating component may reflect a carrier-mediated NH₄⁺ uniport, the linear component diffusion of NH₄⁺ or NH₃. Inhibition of anion/H⁺ cotransport by high NH₄⁺ influx rates may be due to loss of the proton-driving force, Δμ?H⁺, across the plasmalemma. Reversible inhibition by NH₄⁺ of the H⁺ extrusion pump may contribute to the finding that Δμ?H⁺ cannot be reconstituted in the presence of higher NH₄⁺ concentrations.     

Nitrogen uptake and regeneration (ammonium regeneration,nitrification and photoproduction) in waters of the West Florida Shelf prone to blooms of Karenia brevis

The West Florida Shelf (WFS) encompasses a range of environments from inshore estuarine to offshore oligotrophic waters, which are frequently the site of large and persistent blooms of the toxic dinoflagellate, Karenia brevis. The goals of this study were to characterize the nitrogen (N) nutrition of plankton across the range of environmental conditions on the WFS, to quantify the percentage of the plankton N demand met through in situ N regeneration, and to determine whether planktonic N nutrition changes when high concentrations of Karenia are present. In the fall of 2007, 2008, and 2009 we measured ambient nutrient concentrations and used stable isotope techniques to measure rates of primary production and uptake rates of inorganic N (ammonium, NH₄⁺, and nitrate, NO₃⁻), and organic N and carbon (C; urea and amino acids, AA) in estuarine, coastal, and offshore waters, as well as coastal sites with Karenia blooms present. In parallel, we also measured rates of in situ N regeneration – NH₄⁺ regeneration, nitrification, and photoproduction of NH₄⁺, nitrite and AA. Based on microscope observations, ancillary measurements, and previous monitoring history, Karenia blooms sampled represented three bloom stages – initiation in 2008, maintenance in 2007, and late maintenance/stationary phase in 2009. Nutrient concentrations were highest at estuarine sampling sites and lowest at offshore sites. Uptake of NH₄⁺ and NO₃⁻ provided the largest contribution to N nutrition at all sites. At the non-Karenia sites, in situ rates of NH₄⁺ regeneration and nitrification were generally sufficient to supply these substrates equal to the rates at which they were taken up. At Karenia sites, NO₃⁻ was the most important N substrate during the initiation phase, while NH₄⁺ was the most important N form used during bloom maintenance and stationary phases. Rates of NH₄⁺ regeneration were high but insufficient (85 ± 36% of uptake) to support the measured NH₄⁺ uptake at all the Karenia sites although nitrification rates far exceeded uptake rates of NO₃⁻. Taken together our results support the “no smoking gun” nutrient hypothesis that there is no single nutrient source or strategy that can explain Karenia's frequent dominance in the waters where it occurs. Consistent with other papers in this volume, our results indicate that Karenia can utilize an array of inorganic and organic N forms from a number of N sources.     

The effect of nutrient concentrations,nutrient ratios and temperature on photosynthesis and nutrient uptake by Ulva prolifera: implications for the explosion in green tides

The green-tide macroalga, Ulva prolifera, was tested in the laboratory to determine its nutrient uptake and photosynthesis under different conditions. In the nutrient concentration experiments U. prolifera showed a saturated uptake for nitrate but an escalating uptake in the tested range for phosphorus. Both N/P and NO₃ ^?/NH₄ ⁺ ratios influenced nutrient uptake significantly (p?<?0.05) while the PSII quantum yield [Y(II)] (p?>?0.05) remained unaffected. The maximum N uptake rate (33.9?±?0.8 μmol g^?1 DW h^?1) and P uptake rate (11.1?±?4.7) was detected at N/P ratios of 7.5 and 2.2, respectively. U. prolifera preferred NH₄ ⁺-N to NO₃ ^?-N when the NO₃ ^?-N/NH₄ ⁺-N ratio was less than 2.2 (p?<?0.05). But between ratios of 2.2 and 12.9, the uptake of NO₃ ^?-N surpassed that of NH₄ ⁺-N. In the temperature experiments, the highest N uptake rate and [Y(II)] were observed at 20 °C, while the lowest rates were detected at 5 °C. P uptake rates were correlated with increasing temperature.     

High NH4+ efflux from roots of the common alpine grass,Festuca nigrescens,at field-relevant concentrations restricts net uptake

Alpine meadows of high ecological value could be severely endangered by anthropogenic N enrichment, modifying the relationships between species and the environment. While a constraint exerted by N availability on alpine plant development has been demonstrated by some fertilization experiments, in others no effect was observed. Basically, the problem is that mineral N absorption has not been characterized in alpine plants. In growth chamber experiments, we investigated the component fluxes of ¹⁵NO₃^? and ¹⁵NH₄⁺ uptake in a tussock grass (Festuca nigrescens) very common and representative of the dominant plant growth form in European alpine meadows. Rates of influx supported data already published for low elevation herbaceous species. These rates were up to ten times higher for NH₄⁺ than for NO₃^? but rates of net uptake were similar for both ions demonstrating the occurrence of elevated NH₄⁺ efflux (80% of primary influx). An increase in external N in the range of field-relevant concentrations did not substantially enhance net uptake. Thus, the alpine plant which is assumed to be adapted to relatively high soil NH₄⁺ responded like an NH₄⁺-sensitive species: as if it was unable to use the incoming nitrogen. It is suggested that the ability of this typical alpine grass to respond to increasing N availability due to global changes is limited.     

Regulation of Nitrate Uptake in Penicillium chrysogenum by Ammonium Ion

A nitrate uptake system is induced (along with nitrate reductase) when NH₄⁺-grown Penicillium chrysogenum is incubated with inorganic nitrate in synthetic medium in the absence of NH₄⁺. Nitrate uptake and nitrate reduction are probably in steady state in fully induced mycelium, but the ratios of the two activities are not constant during the induction period. Substrate concentrations of ammonium cause a rapid decay of nitrate uptake and nitrate reductase activity. The two activities are differentially inactivated (the uptake activity being more sensitive). Glutamine and asparagine are as effective as NH₄⁺ in suppressing nitrate uptake activity. Glutamate and alanine were about half as effective as NH₄⁺. Cycloheximide interferes with the NH₄⁺-induced decay of nitrate uptake activity. The ammonium transport system is almost maximally deinhibited (or derepressed) in nitrate-grown mycelium.