Factors affecting denitrification in agricultural headwater streams in Northeast Ohio,USA

As a result of increased anthropogenic nitrogen (N) loading in surface waters of agricultural watersheds, there is enhanced interest to understand and quantify N removal mechanisms. Denitrification, an important N removal mechanism in aquatic systems, may contribute to reducing N pollution in agricultural headwater streams. However, the key factors controlling this process in lotic systems remain unclear. The objective of our study was to examine the factors regulating rates of denitrification in the sediments of agricultural headwater streams in the mid-western USA. Denitrification rates were variable among streams and treatments (<0.1–28.0 μg N g AFDM⁻¹ h⁻¹) and on average, were higher than those reported for similar headwater streams. Carbon quantity and quality, and pH had no effect on denitrification, while temperature and nitrate ( ) concentrations had a positive effect on rates of denitrification. Specifically, controlled denitrification following Michaelis-Menten kinetics. We calculated a value of k_m (1.0 mg -N L^-1) that was comparable to other studies in aquatic sediments but was well below the median in-stream concentrations (5.2–17.4 mg -N L⁻¹) observed at the study sites. Despite high rates of denitrification, this removal mechanism is most likely saturated in the agricultural headwater streams we examined, suggesting that these systems are not effective at removing in-stream N. Handling editor: D. Ryder     

Concentration-Dependent Effects on Intracellular and Surface pH of Exposing <Emphasis Type="Italic">Xenopus</Emphasis> oocytes to Solutions Containing NH<Subscript>3</Subscript>/NH<Subscript>4</Subscript><Superscript>+</Superscript>

Others have shown that exposing oocytes to high levels of (10–20 mM) causes a paradoxical fall in intracellular pH (pH_i), whereas low levels (e.g., 0.5 mM) cause little pH_i change. Here we monitored pH_i and extracellular surface pH (pH_S) while exposing oocytes to 5 or 0.5 mM NH₃/NH₄ ⁺. We confirm that 5 mM causes a paradoxical pH_i fall (−ΔpH_i ≅ 0.2), but also observe an abrupt pH_S fall (−ΔpH_S ≅ 0.2)—indicative of NH₃ influx—followed by a slow decay. Reducing [NH₃/NH₄ ⁺] to 0.5 mM minimizes pH_i changes but maintains pH_S changes at a reduced magnitude. Expressing AmtB (bacterial Rh homologue) exaggerates −ΔpH_S at both levels. During removal of 0.5 or 5 mM NH₃/NH₄ ⁺, failure of pH_S to markedly overshoot bulk extracellular pH implies little NH₃ efflux and, thus, little free cytosolic NH₃/NH₄ ⁺. A new analysis of the effects of NH₃ vs. NH₄ ⁺ fluxes on pH_S and pH_i indicates that (a) NH₃ rather than NH₄ ⁺ fluxes dominate pH_i and pH_S changes and (b) oocytes dispose of most incoming NH₃. NMR studies of oocytes exposed to ¹⁵N-labeled show no significant formation of glutamine but substantial accumulation in what is likely an acid intracellular compartment. In conclusion, parallel measurements of pH_i and pH_S demonstrate that NH₃ flows across the plasma membrane and provide new insights into how a protein molecule in the plasma membrane—AmtB—enhances the flux of a gas across a biological membrane.

Leaching losses of inorganic N and DOC following repeated drying and wetting of a spruce forest soil

Forest soils are frequently subjected to dry–wet cycles, but little is known about the effects of repeated drying and wetting and wetting intensity on fluxes of , and DOC. Here, undisturbed soil columns consisting of organic horizons (O columns) and organic horizons plus mineral soil (O + M columns) from a mature Norway spruce stand at the Fichtelgebirge; Germany, were repeatedly desiccated and subsequently wetted by applying different amounts of water (8, 20 and 50 mm day⁻¹) during the initial wetting phase. The constantly moist controls were not desiccated and received 4 mm day⁻¹ during the entire wetting periods. Cumulative inorganic N fluxes of the control were 12.4 g N m⁻² (O columns) and 11.4 g N m⁻² (O + M columns) over 225 days. Repeated drying and wetting reduced cumulative and fluxes of the O columns by 47–60 and 76–85%, respectively. Increasing (0.6–1.1 g N m⁻²) and decreasing fluxes (7.6–9.6 g N m⁻²) indicate a reduction in net nitrification in the O + M columns. The negative effect of dry–wet cycles was attributed to reduced net N mineralisation during both the desiccation and wetting periods. The soils subjected to dry–wet cycles were considerably drier at the final wetting period, suggesting that hydrophobicity of soil organic matter may persist for weeks or even months. Based on results from this study and from the literature we hypothesise that N mineralisation is mostly constrained by hydrophobicity in spruce forests during the growing season. Wetting intensity did mostly not alter N and DOC concentrations and fluxes. Mean DOC concentrations increased by the treatment from 45 mg l⁻¹ to 61–77 mg l⁻¹ in the O tlsbba columns and from 12 mg l⁻¹ to 21–25 mg l⁻¹ in the O + M columns. Spectroscopic properties of DOC from the O columns markedly differed within each wetting period, pointing to enhanced release of rather easily decomposable substrates in the initial wetting phases and the release of more hardly decomposable substrates in the final wetting phases. Our results suggest a small additional DOC input from organic horizons to the mineral soil owing to drying and wetting.     

Effects of common soil anions and pH on the uptake and accumulation of perchlorate in lettuce

A mechanistic understanding of perchlorate () entry into plants is important for establishing the human health risk associated with consumption of contaminated produce and for assessing the effectiveness of phytoremediation. To determine whether common soil anions affect uptake and accumulation in higher plants, a series of competition experiments using lettuce (Lactuca sativa L.) were conducted between (50 nM) and (4–12 mM), (1–10 mM), or Cl⁻ (5–15 mM) in hydroponic solution. The effects of (0–5 mM) and pH (5.5–7.5) on uptake were also examined. Increasing in solution significantly reduced the amount of taken up by green leaf, butter head, and crisphead lettuces. Sulfate and Cl⁻ had no significant effects on uptake in lettuce over the concentrations tested. Increasing pH significantly reduced the amount of taken up by crisphead and green leaf lettuces, whereas increasing significantly reduced uptake in butter head lettuce. The inhibition by across all lettuce genotypes suggests that may share an ion carrier with , and the decrease in uptake with increasing pH or provides macroscopic evidence for cotransport across the plasma membrane.     

(H)NCAHA and (H)CANNH experiments for the determination of vicinal coupling constants related to the ϕ-torsion angle

Summary A set of three-dimensional triple-resonance experiments is described which provide , , and coupling constants. The pulse sequences generate E.COSY-like multiplet patterns and comprise a magnetization transfer from the amide proton to the α-proton or vice versa via the directly bound heteronuclei. For residues with the ¹H^α spin resonating close to the H₂O signal, a modified HNCA experiment can be employed to measure the vicinal ¹H^N,¹H^α couplings. Ambiguities associated with the conversion of values into ϕ-angle constraints for protein structure determination can be resolved with the knowledge of the heteronuclear ³J-couplings. In favourable cases, stereospecific assignments of glycine α-protons can be obtained by employing the experiments described here in combination with NOE data. The methods are applied to flavodoxin from Desulfovibrio vulgaris.     

Soil Respiration in European Grasslands in Relation to Climate and Assimilate Supply

Soil respiration constitutes the second largest flux of carbon (C) between terrestrial ecosystems and the atmosphere. This study provides a synthesis of soil respiration (R _s) in 20 European grasslands across a climatic transect, including ten meadows, eight pastures and two unmanaged grasslands. Maximum rates of R _s ( ), R _s at a reference soil temperature (10°C; ) and annual R _s (estimated for 13 sites) ranged from 1.9 to 15.9 μmol CO₂ m⁻² s⁻¹, 0.3 to 5.5 μmol CO₂ m⁻² s⁻¹ and 58 to 1988 g C m⁻² y⁻¹, respectively. Values obtained for Central European mountain meadows are amongst the highest so far reported for any type of ecosystem. Across all sites was closely related to . Assimilate supply affected R _s at timescales from daily (but not necessarily diurnal) to annual. Reductions of assimilate supply by removal of aboveground biomass through grazing and cutting resulted in a rapid and a significant decrease of R _s. Temperature-independent seasonal fluctuations of R _s of an intensively managed pasture were closely related to changes in leaf area index (LAI). Across sites increased with mean annual soil temperature (MAT), LAI and gross primary productivity (GPP), indicating that assimilate supply overrides potential acclimation to prevailing temperatures. Also annual R _s was closely related to LAI and GPP. Because the latter two parameters were coupled to MAT, temperature was a suitable surrogate for deriving estimates of annual R _s across the grasslands studied. These findings contribute to our understanding of regional patterns of soil C fluxes and highlight the importance of assimilate supply for soil CO₂ emissions at various timescales.     

Nitrogen sink strength of ectomycorrhizal morphotypes of <Emphasis Type="Italic">Quercus douglasii</Emphasis>, <Emphasis Type="Italic">Q. garryana</Emphasis>, and <Emphasis Type="Italic">Q. agrifolia</Emphasis> seedlings grown in a northern California oak woodland

Little information is known on what the magnitude of nitrogen (N) processed by ectomycorrhizal (ECM) fungal species in the field. In a common garden experiment performed in a northern California oak woodland, we investigated transfer of nitrogen applied as ¹⁵NH₄ or ¹⁵NO₃ from leaves to ectomycorrhizal roots of three oak species, Quercus agrifolia, Q. douglasii, and Q. garryana. Oak seedlings formed five common ectomycorrhizal morphotypes on root tips. Mycorrhizal tips were more enriched in ¹⁵N than fine roots. N transfer was greater to the less common morphotypes than to the more common types. ¹⁵N transfer from leaves to roots was greater when , not , was supplied. ¹⁵N transfer to roots was greater in seedlings of Q. agrifolia than in Q. douglasii and Q. garryana. Differential N transfer to ectomycorrhizal root tips suggests that ectomycorrhizal morphotypes can influence flows of N from leaves to roots and that mycorrhizal diversity may influence the total N requirement of plants.     

Energy requirements of Adélie penguin (Pygoscelis adeliae) chicks

Summary The energy requirements of Adélie penguin (Pygoscelis adeliae) chicks were analysed with respect to body mass (W, 0.145–3.35 kg, n=36) and various forms of activity (lying, standing, minor activity, locomotion, walking on a treadmill). Direct respirometry was used to measure O₂ consumption ( ) and CO₂ production. Heart rate (HR, bpm) was recorded from the ECG obtained by both externally attached electrodes and implantable HR-transmitters. The parameters measured were not affected by hand-rearing of the chicks or by implanting transmitters. HR measured in the laboratory and in the field were comparable. Oxygen uptake ranged from in lying chicks to at maximal activity, RQ=0.76. Metabolic rate in small wild chicks (0.14–0.38 kg) was not affected by time of day, nor was their feeding frequency in the colony (Dec 20–21). Regressions of HR on were highly significant (p< 0.0001) in transmitter implanted chicks (n=4), and two relationships are proposed for the pooled data, one for minor activities ( ), and one for walking ( ). Oxygen consumption, mass of the chick (2–3 kg), and duration of walking (T, s) were related as , whereas mass-specific O₂ consumption was related to walking speed (S, m·s^-1) as .Abbreviations bpm beats per minute - D distance walked (m) - ECG electrocardiogram - HR heart rate (bpm) - ns number of steps - RQ respiratory quotient - S walking speed (m·s^-1) - T time walked (s) - W body mass (kg)     

Anaerobic Ammonium Oxidation (Anammox) in Chesapeake Bay Sediments

Anaerobic ammonium oxidation (anammox) has recently been recognized as a pathway for the removal of fixed N from aquatic ecosystems. However, the quantitative significance of anammox in estuarine sediments is variable, and measurements have been limited to a few estuaries. We measured anammox and conventional denitrification activities in sediments along salinity gradients in the Chesapeake Bay and two of its sub-estuaries, the Choptank River and Patuxent River. Homogenized sediments were incubated with ^14/15N amendments of , , and to determine relative activities of anammox and denitrification. The percent of N₂ production due to anammox (ra%) ranged from 0 to 22% in the Chesapeake system, with the highest ra% in the freshwater portion of the main stem of upper Chesapeake Bay, where water column concentrations are consistently high. Intermediate levels of relative anammox (10%) were detected at locations corresponding to tidal freshwater and mesohaline locations in the Choptank River, whereas anammox was not detected in the tidal freshwater location in the Patuxent River. Anammox activity was also not detected in the seaward end of Chesapeake Bay, where water column concentrations are consistently low. The ra% did not correlate with accumulation rate in anoxic sediment incubations, but ra% was related to water column concentrations and salinity. Anammox bacterial communities were also examined by amplifying DNA extracted from the upper Chesapeake Bay sediment with polymerase chain reaction (PCR) primers that are specific for 16S rRNA genes of anammox organisms. A total of 35 anammox-like sequences were detected, and phylogenetic analysis grouped the sequences in two distinct clusters belonging to the Candidatus “Scalindua” genus.     

Carbon membrane-aerated biofilm reactor for synthetic wastewater treatment

A carbon membrane-aerated biofilm reactor (CMABR) was developed to treat synthetic wastewater. Such membrane exhibited a high degree of adhesion and good permeability. Continuous experiments showed that COD and -N removal efficiency were 90 ± 2 and 92 ± 4% at removal rates of 35.6 ± 3.8 g COD/m² per day and 9.3 ± 0.6 g -N/m² per day, respectively. After 108 days, effluent total nitrogen (TN) kept at 35 ± 4 mg/L when influent -N increased to 144–164 mg/L and removal efficiency of TN reached 78 ± 3%. Furthermore, Stoichiometric analysis revealed that 70–90% of oxygen supplied was consumed by nitrifier. Scanning electron microscopic (SEM) images and component analysis of penetrating fluid revealed that extracellular polymeric substance (EPS) adhered to pore and that alkaline washing was an effective method to remove them. The study demonstrated that carbon membrane could be used as effective gas-permeable membrane in MABR for wastewater treatment.     

Soil temperature and biotic factors drive the seasonal variation of soil respiration in a maize (Zea mays L.) agricultural ecosystem

The diurnal and seasonal variation of soil respiration (SR) and their driving environmental factors were studied in a maize ecosystem during the growing season 2005. The diurnal variation of SR showed asymmetric patterns, with the minimum occurring around early morning and the maximum around 13:00 h. SR fluctuated greatly during the growing season. The mean SR rate was 3.16 μmol CO₂ m⁻² s⁻¹, with a maximum of 4.87 μmol CO₂ m⁻² s⁻¹ on July 28 and a minimum of 1.32 μmol CO₂ m⁻² s⁻¹ on May 4. During the diurnal variation of SR, there was a significant exponential relationship between SR and soil temperature (T) at 10 cm depth: . At a seasonal scale, the coefficient α and β fluctuated because the biomass (B) increased α, and the net primary productivity (NPP) of maize markedly increased β of the exponential equation. Based on this, we developed the equation to estimate the magnitude of SR and to simulate its temporal variation during the growth season of maize. Most of the temporal variability (93%) in SR could be explained by the variations in soil temperature, biomass and NPP of maize. This model clearly demonstrated that soil temperature, biomass and NPP of maize combined to drive the seasonal variation of SR during the growing season. However, only taking into account the influence of soil temperature on SR, an exponential equation over- or underestimated the magnitude of SR and resulted in an erroneous representation of the seasonal variation in SR. Our results highlighted the importance of biotic factors for the estimation of SR during the growing season. It is suggested that the models of SR on agricultural sites should not only take into account the influence of soil temperature, but also incorporate biotic factors as they affect SR during the growing season.     

Theoretical study of AlH(2+)n (n=1-7) dications

The structures and stability of 1–7 dications were calculated at the ab initio MP2/aug-cc-pVTZ level of theory. The dications AlH²⁺ 1 and 2 were characterized to be unstable thermodynamically. However, these and the stable dications, 3–7 have considerable kinetic barriers for deprotonation. Each of the structures 3–7 contains one or more two-electron three-center (2e–3c) bonds. Aluminum atoms of these dications carry most of the positive charges, as indicated by NBO charge calculations.Dedicated to Professor Dr. Paul von Ragué Schleyer on the occasion of his 75th birthday     

Natural and constructed littoral zones as nutrient traps in eutrophicated shallow lakes

It is generally known that the water quality of shallow lakes can be influenced significantly by marginal wetlands. In order to study the efficacy of constructed littoral wetlands in the IJsselmeer area (The Netherlands) for water quality improvement, a field survey was carried out in 2003. Vegetation, soil, pore water and surface water characteristics were measured in spring and summer in two types of littoral zones: natural and constructed for 8–16 years. The study showed that constructed wetlands perform well and are suitable to enlarge the vegetated littoral zone in the IJsselmeer area. In both natural and constructed sites vegetation biomass varied between 2,200 g m⁻² for helophyte vegetation and 1,300 g m⁻² for low herbaceous vegetation. Nutrient concentrations in the pore water of constructed sites tended to be higher than in natural sites. and concentrations in pore water were much lower when vegetation was present, probably as a result of plant uptake. The N and P accumulation rate in the soil of constructed wetlands was 20 g N m⁻² y⁻¹ and 3 g P m⁻² y⁻¹ in vegetated plots; without vegetation the rate was much lower (8 g N m⁻² y⁻¹ and 1.8 g P m⁻² y⁻¹). We conclude that concerning their effect on water quality, constructed sites may replace natural sites, at least after 8–16 years. Principal component analysis showed a relationship between vegetation biomass and flooding, and nutrient concentrations in soil and pore water. Biomass was negatively correlated with extractable nutrients and positively with soil total N and P content. Flooding duration was negatively related to pore water salinity and positively to pore water nutrients. Due to their high biomass, helophyte stands retained significantly more nutrients than low pioneer vegetation and are therefore more suitable for improving water quality. Handling editor: S. Declerck     

Soil algal communities inhabiting zinc and lead mine spoils

Algal communities inhabiting four calamine mine spoils differing in time since cessation of exploitation and loaded with high concentrations of zinc (20,284–61,599 μg g⁻¹ soil DW), lead (2,620–3,885 μg g⁻¹ DW) and cadmium (104–232 μg g⁻¹ DW) were studied. In dump soils of slightly alkaline pH (7.28–7.52) and low nutrient (, , ) concentrations, chlorophyll a content ranged from 0.41 to 2.27 μg g⁻¹ soil DW. In total, 23 algal species were recorded. Chlorophyta were the dominant taxonomic group (42–55% of all identified species) followed by Cyanobacteria (28–36%) and Heterokontophyta (13–21%). The highest species richness (18) was observed in the oldest dump (120 years old) with natural succession, while in younger dumps it was lower (11–15). Total algal abundance ranged between 5.5 and 19.1 × 10² ind. g⁻¹ soil DW, and values of Margalef’s diversity indices (1.59–2.25) were low. These results may suggest that both high concentrations of heavy metals and low nutrient content influenced the algal communities in all the dumps studied. The differences in algal microflora observed between tailing dumps may indicate that habitat quality improved with time and that algae isolated from Zn/Pb-loaded soils may be Zn/Pb-resistant ecotypes of ubiquitous species.     

The effect of temperature on post-feeding ammonia excretion and oxygen consumption in the southern catfish

The post-prandial rates of ammonia excretion (TAN) and oxygen consumption in the southern catfish (Silurus meridionalis) were assessed at 2 h intervals post-feeding until the rates returned to those of the fasting rates, at 17.5, 22.5, 27.5, and 32.5°C, respectively. Both fasting TAN and increased with temperature, and were lower than those previously reported for many fish species. The relationship between fasting TAN (mmol NH₃–N kg⁻¹ h⁻¹) and temperature (T, °C) was described as: fasting TAN = 0.144e ^0.0266T (r = 0.526, n = 27, P < 0.05). The magnitude of ammonia excretion and its ratio to total N intake during the specific dynamic action (SDA) tended to increase initially, and then decrease with increasing temperature. The ammonia quotient (AQ), calculated as mol NH₃–N/mol O₂, following feeding decreased as temperature increased. The relationship between AQ during SDA and temperature was described as: AQ_{during SDA} = 0.303e ^−0.0143T (r = 0.739, n = 21, P < 0.05). Our results suggest that ammonia excretion and oxygen consumption post-feeding are operating independently of each other. Furthermore, it appears that the importance of protein as a metabolic substrate in postprandial fish decreases with temperature.     

Hydrochemical fluctuations and crustacean community composition in an ephemeral saline lake (Sua Pan,Makgadikgadi Botswana)

Fluctuating hydrochemistry, as a result of extreme hydrological regimes, imposes major physiological constraints on the biota of ephemeral saline lakes. While the inverse relationship between salinity and zooplankton species richness is well-known across salinity gradients, few studies have documented closely the response of zooplankton to seasonal changes in salinity. Weekly sampling during two flood seasons at Sua Pan, an intermittent saline lake in central Botswana demonstrated the importance of spatial and temporal salinity gradients for crustacean community composition, associated with a decline in species richness, from 11 to three species. Conductivity ranged between 320 and 125,800 μS cm⁻¹ during seasonal flooding; changing from dominance by and , Ca²⁺ and Mg²⁺, at the beginning of the floods, to NaCl dominated waters as the lake dried out and salinities increased. pH estimates generally ranged between 8.6 and 10, with maximum values recorded during initial flooding. Crustaceans comprised mainly Branchinella spinosa, Moina belli, Lovenula africana and Limnocythere tudoranceai, all of which occurred across a wide range of salinities, while halotolerant freshwater species (Metadiaptomus transvaalensis, Leptestheria striatochonca and the ostracods Plesiocypridopsis aldabrae, Cypridopsis newtoni and a newly identified Potamocypris species) disappeared above conductivities of 1,500 μS cm⁻¹. A unique crustacean composition in southern Africa was attributed to Sua Pans’ rare chemical composition among southern African saline lakes; flood waters on Sua Pan contained a higher proportion of Na⁺ and , and less K⁺, Mg²⁺ and than over 80% of records from salt pans elsewhere in southern African. The freshwater species of crustaceans in Sua Pan were similar to those found in other southern Africa lakes, and these similarities decreased in lakes with higher pH and proportions of Na, and less SO₄ and Mg in their chemical composition. The predominant saline tolerant species on Sua Pan, however, showed a greater similarity to those in saline lakes in southern and East Africa with higher proportions of and, particularly, Mg²⁺ in their chemical composition. Handling editor: J. M. Melack     

Determination of the average shear rate in a stirred and aerated tank bioreactor

A method for evaluating the average shear rate () in a stirred and aerated tank bioreactor has been proposed for non-Newtonian fluids. The volumetric oxygen transfer coefficient (k _L a) was chosen as the appropriate characteristic parameter to evaluate the average shear rate (). The correlations for the average shear rate as a function of N and rheological properties of the fluid (K and n) were obtained for two airflow rate conditions (ϕ_air). The shear rate values estimated by the proposed methodology lay within the range of the values calculated by classical correlations. The proposed correlations were utilized to predict the during the Streptomyces clavuligerus cultivations carried out at 0.5 vvm and four different rotational impeller speeds. The results show that the values of the average shear rate () varied from 437 to 2,693 s⁻¹ by increasing with N and flow index (n) and decreasing with the fluid consistency index (K).     

Insect cells respiratory activity in bioreactor

Specific respiration rate ( ) is a key parameter to understand cell metabolism and physiological state, providing useful information for process supervision and control. In this work, we cultivated different insect cells in a very controlled environment, being able to measure . Spodoptera frugiperda (Sf9) cells have been used through virus infection as host for foreign protein expression and bioinsecticide production. Transfected Drosophila melanogaster (S2) cells can be used to produce different proteins. The objective of this work is to investigate respiratory activity and oxygen transfer during the growth of different insect cells lines as Spodoptera frugiperda (Sf9), Drosophila melanogaster (S2) wild and transfected for the expression of GPV and EGFP. All experiments were performed in a well-controlled 1-L bioreactor, with SF900II serum free medium. Spodoptera frugiperda (Sf9) cells reached 10.7 × 10⁶ cells/mL and maximum specific respiration rate () of 7.3 × 10⁻¹⁷ molO₂/cell s. Drosophila melanogaster (S2) cells achieved 51.2 × 10⁶ cells/mL and of 3.1 × 10^–18 molO₂/cell s. S2AcGPV (expressing with rabies virus glycoprotein) reached 24.9 × 10⁶ cells/mL and of 1.7 × 10^–17 molO₂/cell s, while S2MtEGFP (expressing green fluorescent protein) achieved 15.5 × 10⁶ cells/mL and  = 1.9 × 10⁻¹⁷ molO₂/cell s. Relating to the Sf9, S2 cells reached higher maximum cell concentrations and lower specific respiration rate, which can be explained by its smaller size. These results presented useful information for scale-up and process control of insect cells.     

Influences of Lead (II) Chloride on the Nitrogen Metabolism of Spinach

Lead (Pb²⁺) is a well-known highly toxic element. The mechanisms of the Pb²⁺ toxicity are not well understood for nitrogen metabolism of higher plants. In this paper, we studied the effects of various concentrations of PbCl₂ on the nitrogen metabolism of growing spinach. The experimental results showed that Pb²⁺ treatments significantly decreased the nitrate nitrogen absorption and inhibited the activities of nitrate reductase, glutamate dehydrogenase, glutamine synthase, and glutamic–pyruvic transaminase of spinach, and inhibited the synthesis of organic nitrogen compounds such as protein and chlorophyll. However, Pb²⁺ treatments increased the accumulation of ammonium nitrogen in spinach cell. It implied that Pb²⁺ could inhibit inorganic nitrogen to be translated into organic nitrogen in spinach, thus led to the reduction in spinach growth.