DETECTION OF LIMITING OR SURPLUS NITROGEN IN ALGAE AND AQUATIC WEEDS1

The rate of NH₄+-N absorption by algae and aquatic weeds in the dark has been shown to be 4-5 times greater for plants which are N-limited as compared to plants with sufficient available N. Eight species of green algae, 2 blue-green algae, 2 diatoms, and 3 aquatic weeds were used to demonstrate the usefulness of the test in determining if available N was in surplus or limited supply in a particular environment. The test was shown not to differentiate between blue-green algae capable of fixing N (4 species) from media with NO₃-N or without combined nitrogen. The factors influencing the results of NH4⁺-N absorption tests have been investigated. In order to differentiate between plants with sufficient available N and those which are N limited, the rate of NH₄⁺-N absorption (0.1 mg N) over 1-hr incubation in the dark by 10-20 mg of algae or aquatic weed tissues is measured. The relatively simple analysis for NH₄⁺-N in the samples makes it very easy to follow the changing N nutrition of plants in cultures with a limited N supply or in the presence of possible N sources.     

AMMONIUM STIMULATION OF DARK CARBON FIXATION AS AN INDICATOR OF NITROGEN DEFICIENCY IN PHYTOPLANKTON: POTENTIAL ERRORS CAUSED BY AMMONIUM-OXIDIZING BACTERIA1

Stimulation of dark fixation of carbon by NH₄⁺ is often used as an indicator of phytoplankton N deficiency. This assay is based on the influence of available NH₄⁺ on anaplerotic CO₂ fixation by algae. However, carbon fixation by chemoautotrophic NH₄⁺-oxidizing bacteria may also be stimulated by NH₄⁺ enrichment, a process that can mask the algal response in natural communities. NH₄⁺ addition enhanced dark carbon fixation up to 300%, relative to unamended controls, in organisms collected on a 0.7-μm retention filter in oligotrophic Flathead Lake, Montana, but the effect was not detectable in the presence of nitrapyrin, an inhibitor of NH₄⁺-oxidizing bacteria. Dark carbon fixation was enhanced with addition of NH₄⁺ in organisms retained on 2-μm filters (which should allow passage of most bacteria). NH₄⁺ stimulated dark carbon fixation in N-deficient axenic cultures of Chlamydomonas reinhardtii Dang but not in N-replete cultures in both the presence and absence of nitrapyrin. Application of nitrapyrin or size fractionation treatments, to separate the processes of dark carbon fixation by nitrifiers and phytoplankton, may improve the efficacy of assays using NH₄⁺ stimulation of dark carbon fixation to specifically indicate N deficiency in natural algal communities.     

INFLUENCE OF UV-B RADIATION ON NITROGEN UTILIZATION BY A NATURAL ASSEMBLAGE OF PHYTOPLANKTON

A 7‐day mesocosm experiment was conducted in July 1996 to investigate the effects of ambient UV‐B radiation (UVBR) exclusion and two UVBR enhancements above ambient levels on NO₃^?, NH₄⁺ and urea utilization in a natural plankton community (<240 μm) from the Lower St. Lawrence Estuary. The phytoplankton community was dominated by diatoms during the first 3 days and, afterward, by flagellates and dinoflagellates. The results of 4‐h incubations just below the water surface show that, compared with ambient UVBR conditions, UVBR exclusion generally increased NO₃^?, NH₄⁺, and urea uptakes. During the last 4 days of the experiment, the percent increase in the specific uptake rate of urea under excluded UVBR conditions varied between 17% and 130% and was a linear function of the ambient UVBR dose removed. During the first 3 days, the phytoplankton community dominated by diatoms was able to withstand UVBR enhancements without any perceptible effect on nitrogen uptake. However, during the post‐diatom bloom period, UVBR enhancements resulted in decreases in NO₃^?, NH₄⁺, and urea uptake compared with ambient UVBR conditions. The reduction of urea uptake under UVBR enhancements during the last 3 days varied between 23% and 64% and was linearly related to the enhanced UVBR dose. However, the different UVBR treatments did not affect the internal organic nitrogen composition (internal urea, free amino acids, and proteins) of the phytoplankton community experiencing vertical mixing in the mesocosms. The discrepancy between short‐term uptake measurements at the surface and long‐term effects in the mesocosms emphasizes the importance of vertical mixing on UVBR effects in natural ecosystems. This suggests that an increase in ambient UVBR would have a minimal effect on nitrogen utilization by natural phytoplankton assemblages if these are vertically mixed.     

Is cell composition related to the phylogenesis of microalgae? An investigation using Hierarchical Cluster Analysis of Fourier Transform Infrared spectra of whole cells

Fourier Transform Infra-Red (FTIR) spectroscopy was used to investigate whether algae belonging to the green evolutionary lineage and algae belonging to the red evolutionary lineage consistently differ for their biomass composition and for the way their composition responds to changes in NO₃⁻ availability. For this purpose, the FTIR spectra of eleven algal species cultured in media containing different amount of NO₃⁻ were subjected to Hierarchical Cluster Analysis (HCA). The same analysis was conducted on the differential FTIR spectra of cells grown at different NO₃⁻ concentrations. Although the organic composition of cells was affected by the availability of NO₃⁻, the analysis of its change did not elicit a clear relationship with the phylogenetic position of the species used. However, when the two evolutionary lineages, and not the individual species, are compared, some differences emerge: the species belonging to the green lineage reorganize their biomass composition similarly in response to variations in N availability, whereas the species of the red lineage modulate their allocation patterns more heterogeneously. This difference may contribute to the ecological fitness of microalgae and may have had an influence on the evolutionary trajectories of phytoplankton groups.     

A comparative kinetic analysis of nitrate and ammonium influx in two early-successional tree species of temperate and boreal forest ecosystems

Root NO₃ ^? and NH₄ ⁺ influx systems of two early‐successional species of temperate (trembling aspen: Populus tremuloides Michx.) and boreal (lodgepole pine: Pinus contorta Dougl. ex Loud. var. latifolia Engelm.) forest ecosystems were characterized. NO₃ ^? and NH₄ ⁺ influxes were biphasic, consisting of saturable high‐affinity (HATS) and constitutive non‐saturable low‐affinity transport systems (LATS) that were evident at low and relatively high N concentrations, respectively. NO₃ ^? influx via HATS was inducible (IHATS); nitrate pre‐treatment resulted in 8–10‐fold increases in the V_max for influx in both species. By contrast, HATS for NH₄ ⁺ were entirely constitutive. In both species, V_max values for NH₄ ⁺ influx were higher than those for NO₃ ^? uptake; the differences were larger in pine (6‐fold) than aspen (1·8‐fold). In aspen, the K_m for NH₄ ⁺ influx by HATS was approximately 3‐fold higher than for IHATS NO₃ ^? influx, while in pine the K_m for IHATS NO₃ ^? influx was approximately 3‐fold higher than for NH₄ ⁺ influx. The aspen IHATS for NO₃ ^? influx appeared to be more efficient than that of pine (V_max values for aspen being approximately 10‐fold higher and K_m values being approximately 13‐fold lower than for pine). By contrast, only small differences in values for the NH₄ ⁺ HATS were evident between the two species. The kinetic parameters observed here probably result from adaptations to the N availabilities in their respective natural habitats; these may contribute to the distribution and niche separation of these species.     

Variation in growth rate,carbon assimilation,and photosynthetic efficiency in response to nitrogen source and concentration in phytoplankton isolated from upper San Francisco Bay

Six species of phytoplankton recently isolated from upper San Francisco Bay were tested for their sensitivity to growth inhibition by ammonium (NH₄⁺), and for differences in growth rates according to inorganic nitrogen (N) growth source. The quantum yield of photosystem II (F_v/F_m) was a sensitive indicator of NH₄⁺ toxicity, manifested by a suppression of F_v/F_m in a dose‐dependent manner. Two chlorophytes were the least sensitive to NH₄⁺ inhibition, at concentrations of >3,000 μmoles NH₄⁺ · L^?1, followed by two estuarine diatoms that were sensitive at concentrations >1,000 μmoles NH₄⁺ · L^?1, followed lastly by two freshwater diatoms that were sensitive at concentrations between 200 and 500 μmoles NH₄⁺ · L^?1. At non‐inhibiting concentrations of NH₄⁺, the freshwater diatom species grew fastest, followed by the estuarine diatoms, while the chlorophytes grew slowest. Variations in growth rates with N source did not follow taxonomic divisions. Of the two chlorophytes, one grew significantly faster on nitrate (NO₃^?), whereas the other grew significantly faster on NH₄⁺. All four diatoms tested grew faster on NH₄⁺ compared with NO₃^?. We showed that in cases where growth rates were faster on NH₄⁺ than they were on NO₃^?, the difference was not larger for chlorophytes compared with diatoms. This holds true for comparisons across a number of culture investigations suggesting that diatoms as a group will not be at a competitive disadvantage under natural conditions when NH₄⁺ dominates the total N pool and they will also not have a growth advantage when NO₃^? is dominant, as long as N concentrations are sufficient.     

Different Nutritional Histories Affect the Susceptibility of Algae to Grazing

We hypothesize that algae with different cell compositions are differently perceived by their predators and consequently subjected to selective grazing. Five populations of the diatom Phaeodactylum tricornutum that differed in organic and elemental composition, but were otherwise identical, were generated by acclimation to distinct growth regimes. The different populations were then mixed in pairs and subjected to predation by either the rotifer Brachionus plicatilis or the copepod Acartia tonsa. The presence of rotifers had no impact on the ratio between any two algal populations. The presence of copepods, however, affected the ratio between algae previously acclimated to a medium containing 1 mM NH₄⁺ and algae acclimated to 0.5 mM NO₃^?, and to either a lower irradiance or a higher CO₂ concentration. We discuss the possible reason for the influence of different nutritional histories on the vulnerability of algae to predators. The differential impact of grazers on the growth of algae with different nutritional histories may result from direct selective grazing (i.e., grazers can detect algae with the most palatable cell composition), alone or combined to an asymmetric utilization of the nutrients regenerated after predation by co‐existing algal populations. Our results strongly suggest that the nutritional history of algae can influence the relationships between phytoplankton and grazers and hint at the possibility that algal cell composition is potentially subject to natural selection, because it influences the probability that algae survive predation.     

15N MEASUREMENTS OF AMMONIUM AND NITRATE UPTAKE BY ULVA FENESTRATA (CHLOROPHYTA) AND GRACILARIA PACIFICA (RHODOPHYTA): COMPARISON OF NET NUTRIENT DISAPPEARANCE,RELEASE OF AMMONIUM AND NITRATE,AND 15N ACCUMULATION IN ALGAL TISSUE 1

Ammonium and nitrate uptake rates in the macroalgae Ulva fenestrata (Postels and Ruprecht) (Chlorophyta) and Gracilaria pacifica (Abbott) (Rhodophyta) were determined by ¹⁵N accumulation in algal tissue and by disappearance of nutrient from the medium in long‐term (4–13 days) incubations. Nitrogen‐rich algae (total nitrogen> 4% dry weight [dw]) were used to detect isotope dilution by release of inorganic unlabeled N from algal thalli. Uptake of NH₄ ⁺ was similar for the two macroalgae, and the highest rates were observed on the first day of incubation (45 μmol N·g dw ^{? 1}·h ^{? 1} in U. fenestrata and 32 μmol N·g dw ^{? 1}·h ^{? 1} in G. pacifica). A significant isotope dilution (from 10 to 7.9 atom % enrichment) occurred in U. fenestrata cultures during the first day, corresponding to a NH₄ ⁺ release rate of 11 μmol N·g dw ^{? 1}·h ^{? 1}. Little isotope dilution occurred in the other algal cultures. Concurrently to net NH₄ ⁺ uptake, we observed a transient free amino acid (FAA) release on the first day in both macroalgal cultures. The uptake rates estimated by NH₄ ⁺ disappearance and ¹⁵N incorporation in algal tissue compare well (82% agreement, defined as the percentage ratio of the lower to the higher rate) at high NH₄ ⁺ concentrations, provided that isotope dilution is taken into account. On average, 96% of added ¹⁵NH₄ ⁺ was recovered from the medium and algal tissue at the end of the incubation. Negligible uptake of NO₃ ^? was observed during the first 2–3 days in both macroalgae. The lag of uptake may have resulted from the need for either some N deprivation (use of NO₃ ^? pools) or physiological/metabolic changes required before the uptake of NO₃ ^? . During the subsequent days, NO₃ ^? uptake rates were similar for the two macroalgae but much lower than NH₄ ⁺ uptake rates (1.97–3.19 μmol N·g dw ^{? 1}·h ^{? 1}). Very little isotope dilution and FAA release were observed. The agreement between rates calculated with the two different methods averaged 91% in U. fenestrata and 95% in G. pacifica. Recovery of added ¹⁵NO₃ ^? was virtually complete (99%). These tracer incubations show that isotope dilution can be significant in NH₄ ⁺ uptake experiments conducted with N‐rich macroalgae and that determination of ¹⁵N atom % enrichment of the dissolved NH₄ ⁺ is recommended to avoid poor isotope recovery and underestimation of uptake rates.     

Segregation of nitrogen use between ammonium and nitrate of ectomycorrhizas and beech trees

Here, we characterized nitrogen (N) uptake of beech (Fagus sylvatica) and their associated ectomycorrhizal (EM) communities from NH₄⁺ and NO₃^?. We hypothesized that a proportional fraction of ectomycorrhizal N uptake is transferred to the host, thereby resulting in the same uptake patterns of plants and their associated mycorrhizal communities. ¹⁵N uptake was studied under various field conditions after short‐term and long‐term exposure to a pulse of equimolar NH₄⁺ and NO₃^? concentrations, where one compound was replaced by ¹⁵N. In native EM assemblages, long‐term and short‐term ¹⁵N uptake from NH₄⁺ was higher than that from NO₃^?, regardless of season, water availability and site exposure, whereas in beech long‐term ¹⁵N uptake from NO₃^? was higher than that from NH₄⁺. The transfer rates from the EM to beech were lower for ¹⁵N from NH₄⁺ than from NO₃^?. ¹⁵N content in EM was correlated with ¹⁵N uptake of the host for ¹⁵NH₄⁺, but not for ¹⁵NO₃^?‐derived N. These findings suggest stronger control of the EM assemblage on N provision to the host from NH₄⁺ than from NO₃^?. Different host and EM accumulation patterns for inorganic N will result in complementary resource use, which might be advantageous in forest ecosystems with limited N availability.     

Ion complexation in nonactin,monactin, and dinactin: A Raman spectroscopic study

The ability of the macrotetrolide nactins to complex selectivity with a wide variety of cations makes these ionophorous antibiotics important model systems for the study of biologic ionic transport. We report a Raman spectroscopic investigation of the Na⁺, K⁺, Rb⁺, Cs⁺, Tl⁺, NH₄⁺, NH₃OH⁺, C(NH₂)₃⁺, and Ba⁺⁺ complexes of nonactin, monactin, and dinactin in 4:1 (v/v) CH₃OH/CHCl₃ and in the solid state. The nactins display characteristic spectral changes upon complexation, some of which are specific for a given cation. In the K⁺, Rb⁺, Cs⁺, NH₃OH⁺, and C(NH₂)₃⁺ complexes, which are apparently isosteric, the ester carbonyl stretch frequency is found to be linearly proportional to the cation–carbonyl electrostatic interaction energy, as calculated from a simplified model. Deviations for the Na⁺, NH₄⁺, Tl⁺, and Ba⁺⁺ complexes are interpreted as arising from additional nonelectrostatic interactions. Additional information is obtained from other spectral regions and from measurements of depolarization ratios. Spectra of the nactin complexes differ from each other more in the solid state than in solution, reflecting the effects of crystalline contact forces.     

Optimization of ammonium acquisition and metabolism by potassium in rice (Oryza sativa L. cv. IR-72)

We present the first characterization of K⁺ optimization of N uptake and metabolism in an NH₄⁺‐tolerant species, tropical lowland rice (cv. IR‐72). ¹³N radiotracing showed that increased K⁺ supply reduces futile NH₄⁺ cycling at the plasma membrane, diminishing the excessive rates of both unidirectional influx and efflux. Pharmacological testing showed that low‐affinity NH₄⁺ influx may be mediated by both K⁺ and non‐selective cation channels. Suppression of NH₄⁺ influx by K⁺ occurred within minutes of increasing K⁺ supply. Increased K⁺ reduced free [NH₄⁺] in roots and shoots by 50–75%. Plant biomass was maximized on 10 mm NH₄⁺ and 5 mm K⁺, with growth 160% higher than 10 mm NO₃^‐‐grown plants, and 220% higher than plants grown at 10 mm NH₄⁺ and 0.1 mm K⁺. Unlike in NH₄⁺‐sensitive barley, growth optimization was not attributed to a reduced energy cost of futile NH₄⁺ cycling at the plasma membrane. Activities of the key enzymes glutamine synthetase and phosphoenolpyruvate carboxylase (PEPC) were strongly stimulated by elevated K⁺, mirroring plant growth and protein content. Improved plant performance through optimization of K⁺ and NH₄⁺ is likely to be of substantial agronomic significance in the world's foremost crop species.     

POTASSIUM ALLEVIATES THE INHIBITORY ACTION OF AMMONIUM UPON THE PHOTOSYNTHETIC RECOVERY OF NOSTOC FLAGELLIFORME (CYANOPHYCEAE) DURING REHYDRATION1

Effects of ammonium on the photosynthetic recovery of Nostoc flagelliforme Berk. et M. A. Curtis were assayed when being rehydrated in low‐K⁺ or high‐K⁺ medium. Its photosynthetic recovery was K⁺ limited after 3 years of dry storage. The potassium absorption of N. flagelliforme reached the maximum after 3 h rehydration in low‐K⁺ medium but at 5 min in high‐K⁺ medium. The K⁺ content of N. flagelliforme rehydrated in high‐K⁺ medium was much higher than that in low‐K⁺ medium. The maximal PSII quantum yield (F_v/F_m) value of N. flagelliforme decreased significantly when samples were rehydrated in low‐K⁺ medium treated with 5 mM NH₄Cl. However, the treatment of 20 mM NH₄Cl had little effect on its F_v/F_m value in high‐K⁺ medium. The relative F_v/F_m 24 h EC₅₀ (concentration at which 50% inhibition occurred) value of NH₄⁺ in high‐K⁺ medium (64.35 mM) was much higher than that in low‐K⁺ medium (22.17 mM). This finding indicated that high K⁺ could alleviate the inhibitory action of NH₄⁺ upon the photosynthetic recovery of N. flagelliforme during rehydration. In the presence of 10 mM tetraethylammonium chloride (TEACl), the relative F_v/F_m 24 h EC₅₀ value of NH₄⁺ was increased to 46.34 and 70.78 mM, respectively, in low‐K⁺ and high‐K⁺ media. This observation suggested that NH₄⁺ entered into N. flagelliforme cells via the K⁺ channel. Furthermore, NH₄⁺ could decrease K⁺ absorption in high‐K⁺ medium.     

The effect of land use on dissolved organic carbon and nitrogen uptake in streams

1. Agricultural and urban land use may increase dissolved inorganic nitrogen (DIN) concentrations in streams and saturate biotic nutrient demand, but less is known about their impacts on the cycling of organic nutrients. To assess these impacts we compared the uptake of DIN (as ammonium, NH₄⁺), dissolved organic carbon (DOC, as acetate), and dissolved organic nitrogen (DON, as glycine) in 18 low‐gradient headwater streams in southwest Michigan draining forested, agricultural, or urban land‐use types. Over 3 years, we quantified uptake in two streams in each of the three land‐use types during three seasons (spring, summer and autumn). 2. We found significantly higher NH₄⁺ demand (expressed as uptake velocity, V_f) in urban compared to forested streams and NH₄⁺V_f was greater in spring compared to summer and autumn. Acetate V_f was significantly higher than NH₄⁺ and glycine V_f, but neither acetate nor glycine V_f were influenced by land‐use type or season. 3. We examined the interaction between NH₄⁺ and acetate demand by comparing simultaneous short‐term releases of both solutes to releases of each solute individually. Acetate V_f did not change during the simultaneous release with NH₄⁺, but NH₄⁺V_f was significantly higher with increased acetate. Thus, labile DOC V_f was not limited by the availability of NH₄⁺, but NH₄⁺V_f was limited by the availability of labile DOC. In contrast, neither glycine nor NH₄⁺V_f changed when released simultaneously indicating either that overall N‐uptake was saturated or that glycine and NH₄⁺ uptake were controlled by different factors. 4. Our results suggest that labile DOC and DON uptake can be equivalent to, or even higher than NH₄⁺ uptake, a solute known to be highly bioreactive, but unlike NH₄⁺ uptake, may not differ among land‐use types and seasons. Moreover, downstream export of nitrogen may be exacerbated by limitation of NH₄⁺ uptake by the availability of labile DOC in headwater streams from the agricultural Midwestern United States. Further research is needed to identify the factors that influence cycling of DOC and DON in streams.     

Evidence that exogenous urea acts as a potent cue to alleviate ammonium‐inhibition of root system growth of cotton plant (Gossypium hirsutum)

Many plants grown with low‐millimolar concentration of NH₄⁺ as a sole nitrogen source develop NH₄⁺‐toxicity symptoms. To date, crucial molecular identities and a practical approach involved in the improvement of plant NH₄⁺‐tolerance remain largely unknown. By phenotyping of upland cotton grown on varied nitrogen forms, we came across a phenomenon that caused sub‐millimolar concentrations of urea (e.g., up 50 μM) to repress the growth inhibition of roots and whole plant cultivated in a NH₄⁺‐containing nutrient solution. A growth‐recovery assay revealed that the relief in NH₄⁺‐inhibited growth required only a short‐term exposure (≧12 h) of the roots to urea, implying that urea could elicit an internal signaling and be involved in antagonizing NH₄⁺‐sensitivity. Intriguingly, split‐root experiments demonstrated that low urea occurrence in one root‐half could efficaciously stimulate not only supplied root but also the root‐half grown in NH₄⁺‐solution without urea, indicating the existence of urea‐triggered local and systemic long‐distance signaling. In the split‐root experiment we also observed high arginase activity, strong arginine reduction and remarkable upregulation of polyamine biosynthesis‐related genes (ADC1/2, SPDS and SPMS). Therefore, we suggest that external urea might serve as an effective cue (signal molecule) in an arginine‐/polyamine‐related process for ameliorating NH₄⁺‐suppressed root growth, providing a novel aspect for deeper exploring and understanding plant NH₄⁺‐tolerance.     

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.     

Interspecies variation in nitrogen uptake kinetic responses of temperate forest species to elevated CO2: potential causes and consequences

Despite the recognition that the capacity to acquire N is critical in plant response to CO₂ enrichment, there is little information on how elevated CO₂ affects root N uptake kinetics. The few available data indicate a highly variable pattern of response to elevated CO₂, but it is presently unclear if the observed inconsistencies are caused by differences in experimental protocols or by true species differences. Furthermore, if there are interspecific variations in N uptake responses to elevated CO₂, it is not clear whether these are associated with different functional groups. Accordingly, we examined intact root‐system NH₄⁺ and NO₃^– uptake kinetic responses to elevated CO₂ in seedlings of six temperate forest tree species, representing (i) fast‐ vs. slow‐growers and (ii) broad‐leaves vs. conifers, that were cultured and assayed in otherwise similar conditions. In general, the species tested had a higher uptake capacity (V_max) for NH₄⁺ than for NO₃^–. Species substantially differed in their NO₃^– and NH₄⁺ uptake capacities, but the interspecific differences were markedly greater for NO₃^– than NH₄⁺ uptake. Elevated CO₂ had a species‐dependent effect on root uptake capacity for NH₄⁺ ranging from an increase of 215% in Acer negundo L. to a decrease of about 40% in Quercus macrocarpa Michx. In contrast, NO₃^– uptake capacity responded little to CO₂ in all the species except A. negundo in which it was significantly down‐regulated at elevated CO₂. Across species, the capacity for NH₄⁺ uptake was positively correlated with the relative growth rate (RGR) of species; however, the CO₂ effect on NH₄⁺ uptake capacity could not be explained by changes in RGR. The observed variation in NH₄⁺ uptake response to elevated CO₂ was also inconsistent with life‐form differences. Other possible mechanisms that may explain why elevated CO₂ elicits a species‐specific response in root N uptake kinetics are discussed. Despite the fact that the exact mechanism(s) for such interspecific variation remains unresolved, these differences may have a significant implication for competitive interactions and community responses to elevated CO₂ environment. We suggest that differential species responses in nutrient uptake capacity could be one potential mechanism for the CO₂‐induced shifts in net primary productivity and species composition that have been observed in experimental communities exposed to elevated levels of CO₂.     

Impaired electron transfer accounts for the photosynthesis inhibition in wheat seedlings (Triticum aestivum L.) subjected to ammonium stress

No single mechanism can provide an adequate explanation for the inhibition of photosynthesis when plants are supplied with ammonium (NH₄⁺) as the sole nitrogen (N) source. We performed a hydroponic experiment using two N sources [5 mM NH₄⁺ and 5 mM nitrate (NO₃^?)] to investigate the effects of NH₄⁺ stress on the photosynthetic capacities of two wheat cultivars (NH₄⁺‐sensitive AK58 and NH₄⁺‐tolerant XM25). NH₄⁺ significantly inhibited the growth and light‐saturated photosynthesis (A_sat) of both cultivars, but the extent of such inhibition was greater in the NH₄⁺‐sensitive AK58. The CO₂ concentration did not limit CO₂ assimilation under NH₄⁺ nutrition; though both stomatal and mesophyll conductance were significantly suppressed. Carboxylation efficiency (CE), light‐saturated potential rate of electron transport (J_max), the quantum efficiency of PSII (Φ_PSII), electron transport rate through PSII [Je(PSII)], and F_v/F_m were significantly reduced by NH₄⁺. As a result, NH₄⁺ nutrition resulted in a significant increase in the production of hydrogen peroxide (H₂O₂) and superoxide anion radicals (O₂^??), but these symptoms were less severe in the NH₄⁺‐tolerant XM25, which had a higher capacity of removing elevated reactive oxygen species (ROS). Thus, NH₄⁺ N sources might decreased electron transport efficiency and increased the production of ROS, exacerbating damage to the electron transport chain, leading to a reduced plant photosynthetic capacity.     

Novel Na+ doped Alq3 hybrid materials for organic light‐emitting diode (OLED) devices and flat panel displays

Pure and Na⁺‐doped Alq₃ complexes were synthesized by a simple precipitation method at room temperature, maintaining a stoichiometric ratio. These complexes were characterized by X‐ray diffraction, Fourier transform infrared (FTIR), UV/Vis absorption and photoluminescence (PL) spectra. The X‐ray diffractogram exhibits well‐resolved peaks, revealing the crystalline nature of the synthesized complexes, FTIR confirms the molecular structure and the completion of quinoline ring formation in the metal complex. UV/Vis absorption and PL spectra of sodium‐doped Alq₃ complexes exhibit high emission intensity in comparison with Alq₃ phosphor, proving that when doped in Alq₃, Na⁺ enhances PL emission intensity. The excitation spectra of the synthesized complexes lie in the range 242–457 nm when weak shoulders are also considered. Because the sharp excitation peak falls in the blue region of visible radiation, the complexes can be employed for blue chip excitation. The emission wavelength of all the synthesized complexes lies in the bluish green/green region ranging between 485 and 531 nm. The intensity of the emission wavelength was found to be elevated when Na⁺ is doped into Alq₃. Because both the excitation and emission wavelengths fall in the visible region of electromagnetic radiation, these phosphors can also be employed to improve the power conversion efficiency of photovoltaic cells by using the solar spectral conversion principle. Thus, the synthesized phosphors can be used as bluish green/green light‐emitting phosphors for organic light‐emitting diodes, flat panel displays, solid‐state lighting technology – a step towards the desire to reduce energy consumption and generate pollution free light. Copyright © 2014 John Wiley & Sons, Ltd.     

A critical experimental evaluation of methods for determination of NH4+ in plant tissue, xylem sap and apoplastic fluid

Ammonium (NH₄⁺) is a central intermediate in the N metabolism of plants, but the quantitative importance of NH₄⁺ in transporting N from root to shoot and the capability of plants to store NH₄⁺ in leaves are still matters of substantial controversy. This paper shows that some of these controversies have to be related to the use of inadequate analytical procedures used for extraction and quantification of NH₄⁺ in plants. The most frequently used methods for determination of NH₄⁺, viz. colorimetric methods based on the classical Berthelot reaction, suffered severely from interference caused by amino acids, amines, amides and proteins. For some of these metabolites the interference was positive, while for others it was negative, making correction impossible. Consequently, colorimetric analysis is inapplicable for determination of NH₄⁺ in plants. Results obtained by ion chromatography may overestimate the NH₄⁺ concentration due to co‐elution of NH₄⁺ with amines like methylamine, ethylamine, ethanolamine and the non‐protein amino acid Γ‐aminobutyric acid. Derivatization of NH₄⁺ with o‐phthalaldehyde at alkaline pH and subsequent quantification of NH₄⁺ by fluorescence spectroscopy was also associated with interference. However, when pH was lowered to 6.8 during derivatization and 2‐mercaptoethanol was used as reductant, NH₄⁺ could be determined with a high selectivity and sensitivity down to a detection limit of 3.3 μM in a 10‐μl sample volume. Derivatization was performed on‐line using a column‐less HPLC system, enabling rapid quantification of NH₄⁺ in a few minutes. Flow injection analysis with on‐line gas dialysis was, likewise, free from interference, except when applied on highly senescent plant material containing volatile amines. Labile N metabolites in leaf tissue extract, xylem sap and apoplastic fluid were degraded to NH₄⁺ during extraction and subsequent instrumental analysis if the samples were not stabilised. A simple and efficient stabilisation could be obtained by addition of 10 mM ice‐cold HCOOH to the plant extraction medium or to the samples of apoplastic fluid or xylem sap. We conclude that significant concentrations of NH₄⁺, exceeding 1 mM, may occur in xylem sap, leaf apoplastic fluid and leaf tissue water of nitrate‐grown tomato and oilseed rape plants. The measured NH₄⁺ concentrations were not a result of excessive N supplies, as even plants grown under mildly N‐deficient conditions contained NH₄⁺.     

Algal biomass production,N uptake and N2 fixation in a synthetic medium

An experiment was conducted in the growth chamber to quantify the biomass production, N removal and N₂ fixation from a synthetic medium by Chlamydomonas reinhardtii and Anabaena flos-aquae. Nitrogen was supplied at a concentration of 100 mg liter⁻¹ of NO₃⁻−¹⁵N and NH₄⁺−¹⁵ (3·5 atom %), respectively. After 21 days Chlamydomonas reinhardtii removed an average of 83·8 and 78·7 mg N liter⁻¹ as NO₃⁻ and NH₄⁺, respectively. Averages of 0·89 and 0·71 g liter⁻¹ (first batch), 1·63 and 0·95 g liter⁻ (second batch) algal biomass were collected from NO₃⁻ and NH₄⁺ media, respectively. Uptake rates of 0·11 mg ¹⁵N g⁻¹ algae day⁻¹ from NO₃⁻ medium and 0·10 mg ¹⁵N g⁻¹ algae day⁻¹ from NH₄⁺ medium were calculated. Algal cells grown in NO₃⁻ and NH₄⁺ medium contained 71 and 65 g N kg⁻¹ (first batch), 39 and 58 g N kg⁻¹ (second batch), respectively. Anabaena flos-aquae produced averages of 0·58 and 0·46 g liter⁻¹ (first batch), 0·55 and 0·48 g liter⁻¹ (second batch) after 14 days of growth from NO₃⁻ and NH₄⁺ media, respectively. Blue-green algal biomass contained higher N (81–98 g kg⁻¹) than green algae. Isotope dilution method for determining N₂ fixation indicated that 55% and 77% of total N of blue-green algae grown in NO₃⁻ and NH₄⁺ media, respectively, was derived from the atmosphere.