Response of coastal phytoplankton to ammonium and nitrate pulses: seasonal variations of nitrogen uptake and regeneration

Seasonal variation in uptake and regeneration of ammonium and nitrate in a coastal lagoon was studied using ¹⁵N incorporation in particulate matter and by measuring changes in particulate nitrogen. Uptake and regeneration rates were two orders of magnitude lower in winter than in summer. Summer uptake values were 2.8 and 2.2 mol N.l^–1.d^–1 for ammonium and nitrate, respectively. Regeneration rates were 2.9 and 2.1 mol N.l^–1.d^–1 for ammonium and nitrate respectively. Regeneration/uptake ratios were often below one, indicating that water column processes were not sufficient to satisfy the phytoplankton nitrogen demand. This implies a role of other sources of nitrogen, such as macrofauna (oysters and epibionts) and sediment. Phytoplankton was well adapted to the seasonal variations in resources, with mixotrophic dinoflagellates dominant in winter, and fast growing diatoms in summer. In winter and spring, ammonium was clearly preferred to nitrate as a nitrogen source, but nitrate was an important nitrogen source in summer because of high nitrification rates. Despite low nutrient levels, the high rates of nitrogen regeneration in summer as well as the simultaneous uptake of nitrate and ammonium allow high phytoplankton growth rates which in turn enable high oyster production.     

The fate of labelled15N urea and ammonium nitrate applied to a winter wheat crop

Labelled urea or ammonium nitrate was applied to winter wheat growing on a loamy soil in Northern France. Two applications of fertilizer were given: 50 kg N ha^–1 at tillering (early March) and 110 kg N ha^–1 at the beginning of stem elongation (mid-April). The kinetics of urea hydrolysis, nitrification of ammonium and the disappearance of inorganic nitrogen were followed at frequent intervals. Inorganic nitrogen soon disappeared, mainly immobilized by soil microflora and absorbed by the crop. Net immobilization of fertilizer N occured at a very similar rate for urea and ammonium nitrate. Maximum immobilization (16 kg N ha¹) was found at harvest for the first dressing and at anthesis for the second dressing (23 kg N ha¹). During the nitrification period, the labelled ammonium pool was immobilized two to three times faster than the labelled nitrate pool. No significant net¹⁵N remineralization was found during the growth cycle.The actual denitrification and volatilization losses were probably more important than indicated from calculations made by extrapolation of fluxes measured over short intervals. However microbial immobilization was the most important of the processes which compete with plant uptake for nitrogen.     

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.     

Inorganic and organic nitrogen uptake by the toxigenic diatom Pseudo-nitzschia australis (Bacillariophyceae)

The nitrogen uptake capabilities of the toxigenic diatom Pseudo-nitzschia australis (Frenguelli), freshly isolated from Monterey Bay California, were examined in unialgal laboratory cultures at saturating photosynthetic photon flux densities (100 μmol photons m⁻² s⁻¹) and 15 °C. The kinetics of nitrogen (nitrate, ammonium, urea and glutamine) uptake as a function of substrate concentration were estimated from short (20.5 min) incubations using the ¹⁵N-tracer technique. Based on the estimated maximum specific uptake rates and measures of N affinity (the initial slope of the uptake versus nutrient concentration curve), nitrate is the preferred nitrogen substrate, followed by glutamine and ammonium, which are equivalent. Rates of urea uptake by P. australis did not saturate at concentrations as high as 36 μg-at N L⁻¹, and urea uptake as a function of concentration could not be described by Michaelis–Menten kinetics over the concentration gradient tested. Although there is a clear preference for nitrate at equivalent concentrations (compared to ammonium, urea, and glutamine), these laboratory results demonstrate the capability of this pennate diatom to utilize both inorganic and organic forms of nitrogen, supporting field observations that P. australis blooms during both upwelling and non-upwelling conditions off the west coast of North America. Substantial differences in the nitrogenous nutrition of P. australis can be expected in these environments, and anthropogenic inputs of N substrates such as ammonium and urea can support its growth, and may contribute significantly to both harmful diatom blooms and the maintenance of seed populations at non-bloom abundances, particularly during periods of reduced or absent upwelling.     

Phytoplankton nitrogen demand and the significance of internal and external nitrogen sources in a large shallow lake (Lake Balaton,Hungary)

Since the middle of 1990s the trend of Lake Balaton towards an increasingly trophic status has been reversed, but N₂-fixing cyanobacteria are occasionally dominant, endangering water quality in summer. The sources of nitrogen and its uptake by growing phytoplankton were therefore studied. Experiments were carried out on samples collected from the middle of the Eastern (Siófok) and Western (Keszthely) basins between February and October 2001. Ammonium, urea and nitrate uptake and ammonium regeneration were measured in the upper 5-cm layer of sediment using the ¹⁵N-technique. Ammonium was determined by an improved microdiffusion assay. N₂ fixation rates were measured by the acetylene-reduction method. Ammonium regeneration rates in the sediment were similar in the two basins. They were relatively low in winter (0.13 and 0.16 μg N cm^?3 day^?1 in the Eastern and Western basin, respectively), increased slowly in the spring (0.38 and 0.45 μg N cm^?3 day^?1) and peaked in late summer (0.82 and 1.29 μg N cm^?3 day^?1, respectively). Ammonium uptake was predominant in spring in the Eastern basin and in summer in the Western basin, coincident with the cyanobacterial bloom. The amount of N₂ fixed was less than one third of the internal load during summer when external N loading was insignificant. Potentially, the phytoplankton N demand could be supported entirely by the internal N load via ammonium regeneration in the water column and sediment. However, the quantity of N from ammonium regeneration in the upper layer of sediment combined with that from the water column would limit the standing phytoplankton crop in spring in both basins and in late summer in the Western basin, especially when the algal biomass increases suddenly.     

Growth, uptake, and assimilation of ammonium, nitrate, and urea, by three strains of Karenia brevis grown under low light

Observations of near-bottom populations of Karenia brevis suggest that these cells may derive nutrients from the sediment–water interface. Cells undergoing a metabolic-mediated migration may be in close proximity to enhanced concentrations of nutrients associated with the sediment during at least a fraction of their diel cycle. In this study, the growth, uptake and assimilation rates of ammonium, nitrate, and urea by K. brevis were examined on a diel basis to better understand the potential role of these nutrients in the near-bottom ecology of this species. Three strains of K. brevis, C6, C3, and CCMP 2229, were grown under 12:12 light dark cycle under 30 μmol photons m⁻² s⁻¹ delivered to the surface plain of batch cultures. Nitrogen uptake was evaluated using ¹⁵N tracer techniques and trichloroacetic acid extraction was used to evaluate the quantity of nitrogen (N) assimilated into cell protein. Growth rates ranged from a low of 0.12 divisions day⁻¹ for C6 and C3 grown on nitrate to a high of 0.18 divisions day⁻¹ for C3 grown on urea. Diurnal maximum uptake rates, ρ_max, varied from 0.41 pmol-N cell⁻¹ h⁻¹ for CCMP 2229 grown on nitrate, to 1.29 pmol-N cell⁻¹ h⁻¹ for CCMP 2229 grown on urea. Average nocturnal uptake rates were 29% of diurnal rates for nitrate, 103% of diurnal uptake rates for ammonium and 56% of diurnal uptake rates for urea. Uptake kinetic parameters varied between substrates, between strains and between day and night measurements. Highest maximum uptake rates were found for urea for strains CCMP2229 and C3 and for ammonium for strain C6. Rates of asmilation into protein also varied day and night, but overall were highest for urea. The comparison of maximal uptake rates as well as assimilation efficiencies indicate that ammonium and urea are utilized (taken up and assimilated) more than twice was fast as nitrate on a diel basis.     

Nutrient uptake and growth responses of three intertidal macroalgae with perennial, opportunistic and summer-annual strategies

An important life history trait of macroalgae species is the physiological ability to cope with nutrient limiting conditions, which seasonally occur in temperate coasts while other environmental factors are adequate (e.g., sufficient light). Nitrogen (N) and Phosphorus (P) uptake kinetics and field growth limitation were investigated in the perennial Bifucaria bifurcata, the opportunistic Ulva intestinalis, and the summer-annual Nemalion helminthoides from Asturian coasts (N Spain). We performed 4 nutrient uptake experiments (ammonium, nitrate, nitrate + ammonium, and phosphate) and monitored the growth and N content of field individuals in the presence/absence of artificial nutrient supply to assess potential growth limitations. B. bifurcata was not actively growing during summer thus low nutrient demands probably relied on stored pools and/or the low background nutrient levels in seawater, as generally observed for perennials. Corresponding N content and uptake rates in this species were the lowest. The opportunistic U. intestinalis showed kinetics suitable for assimilating N quickly at high external concentrations in order to fulfill the high nutrient demands that support its fast-growing strategy. This response is well adapted to seasons and sites of high nutrient loading but signs of nutrient starvation during summer (decreasing growth and N content) were found in the pristine studied area. N. helminthoides showed an intermediate response in terms of thallus N content and uptake affinity, together with an inducible activation of nitrate uptake. This response assured the uptake of transient nutrient pulses without the nutrient storage response of perennials, or the costly enzymatic machinery of opportunistics. This allows N. helminthoides to effectively exploit low background nutrient conditions interrupted by transient peaks during spring-summer, when most ephemerals found difficulties to survive and perennials suspend their active growth. P uptake did not differ greatly between species suggesting its secondary importance compared to N in the tested algae.     

Urea uptake by the scleractinian coral Stylophora pistillata

Urea can be one of the major sources of nitrogen for phytoplankton, but little is known about its importance for corals. Experiments were therefore designed to assess the uptake rates of urea by the scleractinian coral Stylophora pistillata; ¹⁵N-urea was used to follow the incorporation of nitrogen into the zooxanthellae and animal tissue. The uptake kinetics of urea in the tissue of S. pistillata showed that there is a concentration-dependent uptake of urea. The transport of urea was composed of a linear component (diffusion) at concentrations higher than 6 μmol N-urea l^− 1 and an active carrier-mediated component, at lower concentrations. The value of the carrier affinity (K_m = 1.05 μmol urea l^− 1) indicates a good adaptation of the corals to low levels of urea in seawater. At the in situ concentration of ca. 0.2 μmol N-urea l^− 1, the uptake rate was equal to 0.1 nmol N h^− 1 cm^− 2. Urea uptake was at least four times higher in the animal than in the algal fraction, and five times higher when corals were incubated in the light than in the dark. These results could be explained by the involvement of urea in the calcification process, which is also enhanced by light. Comparison of urea uptake rates with nitrate or ammonium uptake rates for the same S. pistillata species, at in situ concentrations, showed that urea is preferred to nitrate and may therefore be an important source of nitrogen for scleractinian corals.     

Seasonal nitrate physiology of Macrocystis integrifolia Bory

Ambient sea-water nitrate and tissue nitrogen (ethanol soluble nitrate and amino acids, as well as total nitrogen) of Macrocystis integrifolia Bory were monitored over a 2-yr period in Bamfield, Vancouver Island, British Columbia. Sea-water nitrate varied from a high of 12 μmol · 1^?1 (individual values as high as 23 μmol · 1^?1 were recorded) in late winter to below detection limits for most of the summer. Tissue nitrate and total nitrogen paralleled the ambient nitrate levels and showed summer minima and winter maxima (from 0 to 70 μmol · g fresh wt^?1 for nitrate and from 0.8 to 2.9% of dry wt for total N). The nitrate uptake capacity was inversely proportional to tissue nitrate concentration and, furthermore, was much higher for subapical surface blades (60–70 nmol · cm^?2 · h^?1) than for older, deeper blades (5–10 nmol · cm^?2 · h^?1). Nitrate uptake by subapical blade disks in summer is apparently higher in dark (1.0–1.7 μmol · g fresh wt^?1 · h^?1) than in light (0.6–1.3 μmol · g fresh wt^?1 · h^?1) and the data obtained in 36–108 h experiments indicate nitrate pool sizes of 30–90 μmol · g fresh wt^?1. These pools are $\text{2}\text{3}$ to nearly full in winter. Ammonium does not inhibit nitrate uptake. It is taken up and apparently utilized much faster than nitrate and it may well be an important source of nitrogen for marine macrophytes.     

Use of Inorganic and Organic Nitrogen by Synechococcus spp. and Diatoms on the West Florida Shelf as Measured Using Stable Isotope Probing

The marine nitrogen (N) cycle is a complex network of biological transformations in different N pools. The linkages among these different reservoirs are often poorly understood. Traditional methods for measuring N uptake rely on bulk community properties and cannot provide taxonomic information. ¹⁵N-based stable isotope probing (SIP), however, is a technique that allows detection of uptake of individual N sources by specific microorganisms. In this study we used ¹⁵N SIP methodology to assess the use of different nitrogen substrates by Synechococcus spp. and diatoms on the west Florida shelf. Seawater was incubated in the presence of ¹⁵N-labeled ammonium, nitrate, urea, glutamic acid, and a mixture of 16 amino acids. DNA was extracted and fractionated using CsCl density gradient centrifugation. Quantitative PCR was used to quantify the amounts of Synechococcus and diatom DNA as a function of density, and ¹⁵N tracer techniques were used to measure rates of N uptake by the microbial community. The ammonium, nitrate, urea, and dissolved primary amine uptake rates were 0.077, 0.065, 0.013, and 0.055 μmol N liter⁻¹ h⁻¹, respectively. SIP data indicated that diatoms and Synechococcus spp. actively incorporated N from [¹⁵N]nitrate, [¹⁵N]ammonium, and [¹⁵N]urea. Synechococcus also incorporated nitrogen from [¹⁵N]glutamate and ¹⁵N-amino acids, but no evidence indicating uptake of labeled amino acids by diatoms was detected. These data suggest that N flow in communities containing Synechococcus spp. and diatoms has more plasticity than the new-versus-recycled production paradigm suggests and that these phytoplankters should not be viewed strictly as recycled and new producers, respectively.The marine nitrogen (N) cycle is a complex network of biological transformations in different inorganic and organic N reservoirs (58). Processes related to the N cycle can at times limit productivity in marine systems (47) and influence the rate at which carbon (C) is exported from the euphotic zone to the deep ocean and marine sediments, where it can be sequestered (21). The historical paradigm with respect to the marine N and C cycles is deeply interwoven with the concepts of new and regenerated primary production in the euphotic zone (17, 20). New and export production have traditionally been equated with large nutrient influxes, particularly influxes of nitrate, which lead to diatom productivity. When high levels of nitrate are present, diatoms often dominate and exhibit high sinking rates due to aggregation and/or packaging into fecal pellets (18, 48). By contrast, the subtropical and tropical oligotrophic surface oceans have been viewed primarily as areas where recycled productivity dominates.In recent years, however, our view of the linkages between the marine N and C cycles has become increasingly complex (58). For example, geochemical rate estimates have suggested that N fixation rates in surface waters of the tropical and subtropical oceans may be many times greater than previously thought (13, 34). The divergence between in situ observations and data obtained using the geochemical mass balance approach is attributed, among other things, to meso-scale physical forcing (38) and to diazotrophic activity of planktonic cyanobacteria (14, 57). Furthermore, it is now appreciated that the ability to use nitrate is more widely distributed among the marine bacteria than previously thought and that bacteria are capable of competing with phytoplankton for both ammonium and nitrate (29, 31). Despite these advances, direct measurements of uptake of specific forms of N by individual populations of phyto- and bacterioplankton are scarce. This is primarily due to the fact that most measurements of N uptake are made using glass fiber filters that collect autotrophs and some variable fraction of heterotrophic bacteria (3). Uptake rates thus represent bulk uptake by hundreds of different phytoplankton and bacterial species. Methods that could be used to investigate uptake of N by specific species (e.g., Synechococcus spp.) or groups of species (e.g., diatoms) would therefore greatly improve our ability to elucidate N fluxes in marine systems.DNA stable isotope probing (SIP) is a technique that is based on the observation that DNA molecules with different densities can be separated by ultracentrifugation in a concentrated solution of cesium chloride (CsCl). CsCl density gradient centrifugation has a long history in biological research and was first used to demonstrate the semiconservative nature of DNA replication (39). In their experiments, Meselson and Stahl grew Escherichia coli in medium in which all available forms of N contained the heavy, stable isotope ¹⁵N. Fully labeled with ¹⁵N, DNA has an average density of 1.722 g cm⁻³, whereas ¹⁴N-containing DNA has an average density of 1.700 g cm⁻³ (10, 11). This small, yet significant difference in density is enough to allow separation of ¹⁴N-containing DNA from ¹⁵N-containing DNA. DNA SIP has been used to study the dynamics of microbial communities (46). Radajewski et al. (46) used DNA SIP to identify the microbial species involved in the biotransformation of specific ¹³C-labeled substrates among the large pool of bacterial species that typically are present in environmental communities. These authors demonstrated that ¹³C-labeled DNA could be recovered from microbial populations after incubation. ¹³C-labeled DNA was then taxonomically characterized using routine molecular ecology methods to identify active community members, demonstrating that SIP can be a powerful technique for taxonomic identification of microbes performing specific metabolic processes under in situ conditions. A series of studies have since been performed using the ¹³C-based technique to examine microbial communities in different environments (for reviews, see references 19 and 40).More recently, ¹⁵N-based SIP techniques have been developed to facilitate identification of the free-living diazotrophs responsible for in situ N fixation in soil (10). This work demonstrated that ¹⁵N-based SIP techniques could be used to study N flow in environmental communities. In the present study, ¹⁵N-based SIP techniques (10, 11) were employed to assess the use of a suite of inorganic and organic nitrogen substrates by Synechococcus spp. and diatoms in a coastal marine system. Our goal was to investigate the traditional characterization of Synechococcus spp. as recycled producers (mainly ammonium uptake) and diatoms as new producers (nitrate uptake). To do this, seawater was incubated with a series of ¹⁵N-labeled N substrates. DNA was then extracted at the end of the incubation period, and quantitative PCR (qPCR) was used to determine the amounts of Synechococcus and diatom DNA as a function of density in fractionated gradients. Shifts in the densities of Synechococcus and diatom DNA as the result of incubation with ¹⁵N-labeled N substrates were interpreted as evidence of uptake. Our data indicate that Synechococcus spp. and diatoms both actively incorporated [¹⁵N]ammonium, [¹⁵N]nitrate, and [¹⁵N]urea. Synechococcus spp. appeared to also incorporate N from [¹⁵N]glutamate and ¹⁵N-amino acids. These data suggest that N flow in communities containing Synechococcus spp. and diatoms has more plasticity than the new-versus-regenerated production paradigm suggests and that these two types of phytoplankton should not be viewed strictly as recycled and new producers, respectively.     

不同作物对旱地农田残留硝态氮的利用差异

在黄土高原南部娄土上,通过2a田间试验研究了小麦和苜蓿对土壤中不同累积量的残留硝态氮的利用差异。研究包括0—3 m土壤残留硝态氮累积量(设N1、N2、N3、N4、N5和N6共6个水平,残留硝态氮量依次增加)和作物种类(冬小麦和苜蓿)2个因素,分别采用冬小麦-夏休闲-冬小麦和苜蓿连作种植方式。结果表明,不施用氮肥条件下,冬小麦-休闲-冬小麦轮作周期与苜蓿连作2a内,土壤残留硝态氮的消长有明显差异。在第1季小麦生长期间,小麦的氮素携出量(63.9—130.3 kg/hm2)、氮素携出量占播前残留硝态氮量的比例(18%—27%)及氮素携出量占该生长季硝态氮减少量的比例(29%—62%)均显著高于同期的苜蓿处理。在第2个生长季内,苜蓿的氮素携出量是小麦当季氮素携出量的近6倍,但由于苜蓿固氮作用强烈,至第2生长季结束后,0—3 m土壤硝态氮量与苜蓿播前相比平均只减少了72.4 kg/hm2,而麦田0—3 m土壤硝态氮量与小麦播前相比减少了158.3 kg/hm2。在短期内如果通过种植作物消耗土壤剖面的残留硝态氮,冬小麦比苜蓿更有优势。第1季小麦氮素携出量与小麦播前0—2 m(r=0.920**)和0—3 m(r=0.857*)土层残留硝态氮量呈显著或极显著正相关,与0—1 m土层残留硝态氮量没有显著相关性;第1生长季苜蓿氮素携出量与播前0—1 m土壤硝态氮累积量呈显著正相关关系(r=0.846*),而与0—2 m和0—3 m土壤硝态氮累积量的相关性并不显著。小麦比苜蓿能利用更深土层中的硝态氮。随着播前0—3 m土壤残留硝态氮的增加,小麦和苜蓿地上部氮素携出量呈增加的趋势,硝态氮表观损失也显著增加。     

NITROGENOUS NUTRITION OF ALEXANDRIUM CATENELLA (DINOPHYCEAE) IN CULTURES AND IN THAU LAGOON,SOUTHERN FRANCE1

Alexandrium catenella (Whedon et Kofoid) Balech was isolated from Thau lagoon (northern Mediterranean) and its growth and uptake characteristics measured for nitrate, ammonium, and urea. Although affinity constants did not indicate a preference for ammonium over nitrate, there was a strong inhibition of nitrate uptake by ammonium when both nitrogen (N) sources were present. Nitrogen budgets during growth in cultures revealed major imbalances between decreases in dissolved N and increases in particulate N, indicating excretion of dissolved organic N during the early part of the growth phase and uptake during the later part. A quasi‐unialgal bloom in November 2001 (4×10⁶ cells·L^?1) allowed measurements of uptake of nitrate, nitrite, ammonium, and urea; net and gross growth rate of A. catenella; and grazing rates on this organism. The affinity constants indicate that it is not a strong competitor for the N nutrients tested when these are in low concentrations (<10 μgat N·L^?1), compared with other members of the phytoplankton community. Indirect evidence from cultures indicate that dissolved organic N compounds could be important in triggering those blooms. Finally, the strongly unbalanced growth observed in the field indicates that A. catenella exhibits a storage rather than a growth response to a nutrient pulse and is adapted to low frequency events such as the passage of frontal disturbances. The disappearance of A. catenella was due to grazing that balanced growth at the peak of the bloom.     

Contribution of several nitrogen sources to growth of Alexandrium catenella during blooms in Thau lagoon,southern France

A monitoring program with a weekly sampling frequency over a 15-month period indicates that urea concentrations above a certain threshold level may trigger the blooms of Alexandrium catenella in Thau lagoon. However, urea concentrations were also sometimes related to ammonium and dissolved organic nitrogen concentrations, indicating that the role of urea may not be a direct one. An original approach is used to assess the relative contribution of several nitrogen sources (nitrate, nitrite, ammonium, urea) to growth of A. catenella by comparing nitrogen uptake rates to nitrogen-based growth rates estimated from dilution experiments during four blooms over a 4-year period (2001–2004) in Thau lagoon. Nitrate and nitrite contributed 0.1–14% and 0.1–5% respectively of growth requirements. Ammonium and urea were the main N sources fueling growth of A. catenella (30–100% and 2–59%, respectively). Indirect estimates indicated that an unidentified N source could also contribute significantly to growth at specific times. Concerning ammonium and urea uptake kinetics, half-saturation constants varied between 0.2 and 20 μgat N L⁻¹ for ammonium and between 0.1 and 44 μgat N L⁻¹ over the 4-year period, indicating that A. catenella can have a competitive advantage over other members of the phytoplankton even under low concentrations of ammonium and urea. However, the observed large changes in ammonium and urea uptake kinetics on a short time scale (days) during blooms preclude more precise estimates of those contributions to growth and require further investigation.     

δ<Superscript>15</Superscript>N dynamics of ammonium and particulate nitrogen in a temperate eutrophic estuary

We monitored the stable nitrogen isotopic composition (δ¹⁵N) of suspended matter and ammonium in the freshwater stretch of the Scheldt estuary (Belgium) over a full year to investigate for seasonal evolution and possible co-variation between isotopic signatures. The δ¹⁵N value of ammonium remained rather constant during winter (average = +11.4‰) but increased significantly with the spring and summer bloom, reaching values as high as +70‰. This enrichment of the ammonium pool in ¹⁵N coincided with significant ammonium depletion during summer period, suggesting a close causal relationship. Based on a semi-closed system approach we deduced an apparent fractionation factor associated with NH₄⁺ utilization (i.e. combining effects of uptake and nitrification) of 18.4‰ (SE = 2.0‰), which is similar to values reported in literature. Observed variations of ammonium δ¹⁵N could account for about 69% of δ¹⁵N variation in suspended matter.     

Biochar application to rice with 15N-labelled fertilizers,enhanced leaf nitrogen concentration and assimilation by improving morpho-physiological traits and soil quality

Leaf nitrogen (N) concentration plays an important role in biochemical and physiological functions, and N availability directly influences rice yield. However, excessive N fertilization is considered to be a root cause of environmental issues and low nitrogen use efficiency. Therefore, the selection of appropriate nutrient management practices and organic amendments is key to maximizing nitrogen uptake and maintaining high and sustainable rice production. Here, we evaluated the effects of different ¹⁵N-labelled nitrogen sources (urea, ammonium nitrate, and ammonium sulfate at 315 kg ha⁻¹) with or without biochar (30 t ha⁻¹) on paddy soil properties, root growth, leaf gas exchange, N metabolism enzymes, and N uptake in the early and late seasons of 2019. We found significant differences among N fertilizer sources applied with or without biochar (P < 0.05). Across the seasons, the combination of biochar with N fertilizers significantly increased soil organic carbon by 51.21% and nitrogen availability by 27.51% compared with N fertilizers alone. Correlation analysis showed that rice root morphological traits were strongly related to soil chemical properties, and higher root growth was measured in the biochar treatments. Similarly, net leaf photosynthetic rate averaged 9.34% higher, chlorophyll (Chl) a concentration 12.91% higher, and Chl b concentration 10.05% higher in the biochar treatments than in the biochar-free treatments across the seasons. Notably, leaf ¹⁵N concentration was 23.19% higher in the biochar treatments in both seasons. These results illustrated higher activities of N metabolism enzymes such as NR, GS, and GOGAT by an average 23.44%, 11.26% and 18.16% in the biochar treatments across the seasons, respectively. The addition of biochar with synthetic N fertilizers is an ecological nutrient management strategy that can increase N uptake and assimilation by ameliorating soil properties and improving the morpho-physiological factors of rice.     

Diversity, abundance, and activity of ammonia-oxidizing bacteria and archaea in Chongming eastern intertidal sediments

Ammonia oxidation plays a pivotal role in the cycling and removal of nitrogen in aquatic sediments. Certain bacterial groups and a novel group of archaea, which is affiliated with the novel phylum Thaumarchaeota, can perform this initial nitrification step. We examined the diversity and abundance of ammonia-oxidizing β-Proteobacteria (β-AOB) and ammonia-oxidizing archaea (AOA) in the sediments of Chongming eastern tidal flat using the ammonia monooxygenase-α subunit (amoA) gene as functional markers. Clone library analysis showed that AOA had a higher diversity of amoA gene than β-AOB. The β-Proteobacterial amoA community composition correlated significantly with water soluble salts in the sediments, whereas the archaeal amoA community composition was correlated more with nitrate concentrations. Quantitative PCR (qPCR) results indicated that the abundance of β-AOB amoA gene (9.11?×?10⁴–6.47?×?10⁵?copies?g^?1 sediment) was always greater than that of AOA amoA gene (7.98?×?10³–3.51?×?10⁵?copies?g^?1 sediment) in all the samples analyzed in this study. The β-Proteobacterial amoA gene abundance was closely related to organic carbon, while no significant correlations were observed between archaeal amoA gene abundance and the environmental factors. Potential nitrification rates were significantly greater in summer than in winter and correlated strongly with the abundance of amoA genes. Additionally, a greater contribution of single amoA gene to potential nitrification occurred in summer (1.03–5.39 pmol?N?copy^?1?day^?1) compared with winter (0.16–0.38 pmol?N?copy^?1?day^?1), suggesting a higher activity of ammonia-oxidizing prokaryotes in warm seasons.     

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.     

Seasonal Patterns of Soil Respiration and Related Soil Biochemical Properties under Nitrogen Addition in Winter Wheat Field

Understanding the changes of soil respiration under increasing N fertilizer in cropland ecosystems is crucial to accurately predicting global warming. This study explored seasonal variations of soil respiration and its controlling biochemical properties under a gradient of Nitrogen addition during two consecutive winter wheat growing seasons (2013–2015). N was applied at four different levels: 0, 120, 180 and 240 kg N ha^-1 year^-1 (denoted as N0, N12, N18 and N24, respectively). Soil respiration exhibited significant seasonal variation and was significantly affected by soil temperature with Q₁₀ ranging from 2.04 to 2.46 and from 1.49 to 1.53 during 2013–2014 and 2014–2015 winter wheat growing season, respectively. Soil moisture had no significant effect on soil respiration during 2013–2014 winter wheat growing season but showed a significant and negative correlation with soil respiration during 2014–2015 winter wheat growing season. Soil respiration under N24 treatment was significantly higher than N0 treatment. Averaged over the two growing seasons, N12, N18 and N24 significantly increased soil respiration by 13.4, 16.4 and 25.4% compared with N0, respectively. N addition also significantly increased easily extractable glomalin-related soil protein (EEG), soil organic carbon (SOC), total N, ammonium N and nitrate N contents. In addition, soil respiration was significantly and positively correlated with β-glucosidase activity, EEG, SOC, total N, ammonium N and nitrate N contents. The results indicated that high N fertilization improved soil chemical properties, but significantly increased soil respiration.     

Rapid and slow modulation of nitrate reductase activity in the red macroalga <Emphasis Type="Italic">Gracilaria chilensis</Emphasis> (Gracilariales,Rhodophyta): influence of different nitrogen sources

Nitrate is one of the most important stimuli in nitrate reductase (NR) induction, while ammonium is usually an inhibitor. We evaluated the influence of nitrate, ammonium or urea as nitrogen sources on NR activity of the agarophyte Gracilaria chilensis. The addition of nitrate rapidly (2 min) induced NR activity, suggesting a fast post-translational regulation. In contrast, nitrate addition to starved algae stimulated rapid nitrate uptake without a concomitant induction of NR activity. These results show that in the absence of nitrate, NR activity is negatively affected, while the nitrate uptake system is active and ready to operate as soon as nitrate is available in the external medium, indicating that nitrate uptake and assimilation are differentially regulated. The addition of ammonium or urea as nitrogen sources stimulated NR activity after 24 h, different from that observed for other algae. However, a decrease in NR activity was observed after the third day under ammonium or urea. During the dark phase, G. chilensis NR activity was low when compared to the light phase. A light pulse of 15 min during the dark phase induced NR activity 1.5-fold suggesting also fast post-translational regulation. Nitrate reductase regulation by phosphorylation and dephosphorylation, and by protein synthesis and degradation, were evaluated using inhibitors. The results obtained for G. chilensis show a post-translational regulation as a rapid response mechanism by phosphorylation and dephosphorylation, and a slower mechanism by regulation of RNA synthesis coupled to de novo NR protein synthesis.     

Distribution and recovery of15N after fertilization of Douglas-fir saplings with different nitrogen sources

Summary Four-year-old Douglas-fir (Pseudotsuga menziesii (Mirb.) Franco) saplings planted in pots with a sand and peat mix (11) were fertilized at the rate of 200 kg N/ha of (¹⁵NH₂)₂CO (U-15),¹⁵NH₄NO₃ (A-15) and NH₄ ¹⁵NO₃(An-15). They were placed in a shadehouse and watered regularly to maintain soil moisture at field capacity over periods of one and two years. Quantity of¹⁵N in foliage generally increased from old to current growth, irrespective of the nitrogen source. Utilization of¹⁵N fertilizers by saplings after the first and second growing seasons following fertilization was greatest with nitrate labelled ammonium nitrate AN-15, and nearly equal for urea U-15 and ammonium labelled ammonium nitrate A-15. The soil immobilized more fertilizer nitrogen-15 from U-15 and A-15 than from AN-15. Data from the present study, in which leaching losses of fertilizer were minimized, demonstrated that in terms of nitrogen uptake by the saplings the nitrate fertilizer was superior to ammonium fertilizer.