The kinetics of NH4 + and NO3 − uptake by Douglas fir from single N-solutions and from solutions containing both NH4 + and NO3 −

The kinetics of NH₄ ⁺ and NO₃ ⁻ uptake in young Douglas fir trees (Pseudotsuga menziesii [Mirb.] Franco) were studied in solutions, containing either one or both N species. Using solutions containing a single N species, the V_max of NH₄ ⁺ uptake was higher than that of NO₃ ⁻ uptake. The K_m of NH₄ ⁺ uptake and K_m of NO₃ ⁻ uptake differed not significantly. When both NH₄ ⁺ and NO₃ ⁻ were present, the V_max for NH₄ ⁺ uptake became slightly higher, and the K_m for NH₄ ⁺ uptake remained in the same order. Under these conditions the NO₃ ⁻ uptake was almost totally inhibited over the whole range of concentrations used (10–1000 μM total N). This inhibition by NH₄ ⁺ occurred during the first two hours after addition. ei]{gnA C}{fnBorstlap}     

Antioxidative system in maize roots as affected by osmotic stress and different nitrogen sources

The activities of antioxidative enzymes and contents of proline and total phenolics were assayed in roots of two maize (Zea mays L.) genotypes grown in a medium containing nitrate (NO₃ ⁻) or both nitrogen forms, nitrate and ammonium (NH₄ ⁺/NO₃ ⁻). An increase in the activities of class III peroxidases (POD), superoxide dismutase (SOD), ascorbate peroxidase (APX), ascorbate oxidase (AO) and proline content, and decrease in phenolic content were observed in NH₄ ⁺/NO₃ ⁻ in comparison with NO₃ ⁻ grown plants. When polyethylene glycol (PEG) was added to both nitrogen treatments, the content of total phenolics and proline was increased, especially in NH₄ ⁺/NO₃ ⁻ treatment. The PEG treatment decreased enzyme activities in NH₄ ⁺/NO₃ ⁻ grown plants, but in NO₃ ⁻ grown plants activities of POD and SOD were increased, opposite to decreased APX and AO. Isoelectric focusing demonstrated increased activities of acidic POD isoforms in PEG treated NO₃ ⁻ grown plants, and lower activities of both, acidic and basic isoforms in NH₄ ⁺/NO₃ ⁻grown plants.     

Functional role of DNRA and nitrite reduction in a pristine south Chilean <Emphasis Type="Italic">Nothofagus</Emphasis> forest

Nitrite (NO₂ ⁻) is an intermediate in a variety of soil N cycling processes. However, NO₂ ⁻ dynamics are often not included in studies that explore the N cycle in soil. Within the presented study, nitrite dynamics were investigated in a Nothofagus betuloides forest on an Andisol in southern Chile. We carried out a ¹⁵N tracing study with six ¹⁵N labeling treatments, including combinations of NO₃ ⁻, NH₄ ⁺ and NO₂ ⁻. Gross N transformation rates were quantified with a ¹⁵N tracing model in combination with a Markov chain Monte Carlo optimization routine. Our results indicate the occurrence of functional links between (1) NH₄ ⁺ oxidation, the main process for NO₂ ⁻ production (nitritation), and NO₂ ⁻ reduction, and (2) oxidation of soil organic N, the dominant NO₃ ⁻ production process in this soil, and dissimilatory NO₃ ⁻ reduction to NH₄ ⁺ (DNRA). The production of NH₄ ⁺ via DNRA was approximately ten times higher than direct mineralization from recalcitrant soil organic matter. Moreover, the rate of DNRA was several magnitudes higher than the rate of other NO₃ ⁻ reducing processes, indicating that DNRA is able to outcompete denitrification, which is most likely not an important process in this ecosystem. These functional links are most likely adaptations of the microbial community to the prevailing pedo-climatic conditions of this Nothofagus ecosystem.     

Role of cytokinin in enhanced productivity of maize supplied with NH4 + and NO3 −

Supplying both N forms (NH₄ ⁺+NO₃ ⁻) to the maize (Zea mays L.) plant can optimize productivity by enhancing reproductive development. However, the physiological factors responsible for this enhancement have not been elucidated, and may include the supply of cytokinin, a growth-regulating substance. Therefore, field and gravel hydroponic studies were conducted to examine the effect of N form (NH₄ ⁺+NO₃ ⁻ versus predominantly NO₃ ⁻) and exogenous cytokinin treatment (six foliar applications of 22 μM 6-benzylaminopurine (BAP) during vegetative growth versus untreated) on productivity and yield of maize. For untreated plants, NH₄ ⁺+NO₃ ⁻ nutrition increased grain yield by 11% and whole shoot N content by 6% compared with predominantly NO₃ ⁻. Cytokinin application to NO₃ ⁻-grown field plants increased grain yield to that of NH₄ ⁺+NO₃ ⁻-grown plants, which was the result of enhanced dry matter partitioning to the grain and decreased kernel abortion. Likewise, hydroponically grown maize supplied with NH₄ ⁺+NO₃ ⁻ doubled anthesis earshoot weight, and enhanced the partitioning of dry matter to the shoot. NH₄ ⁺+NO₃ ⁻ nutrition also increased earshoot N content by 200%, and whole shoot N accumulation by 25%. During vegetative growth, NH₄ ⁺+NO₃ ⁻ plants had higher concentrations of endogenous cytokinins zeatin and zeatin riboside in root tips than NO₃ ⁻-grown plants. Based on these data, we suggest that the enhanced earshoot and grain production of plants supplied with NH₄ ⁺+NO₃ ⁻ may be partly associated with an increased endogenous cytokinin supply.     

Variability of leaf photosynthetic characteristics in rice and its relationship with resistance to water stress under different nitrogen nutrition regimes

The negative effects of water stress on rice can be alleviated by NH₄⁺ nutrition. However, the effects of mixed nitrogen (N) nutrition (NO₃^? + NH₄⁺) on resistance to water stress are still not well known. To investigate the response of rice growth to water stress and its relationship with photosynthetic characteristics, a hydroponic experiment supplying different N forms was conducted. Compared with NO₃^? nutrition, mixed‐N and NH₄⁺ nutrition greatly alleviated the reduction of leaf area, chlorophyll content, and photosynthesis under water stress, whilst subsequently maintaining higher biomass. In contrast, water stress inhibited the root‐shoot ratios in NH₄⁺‐ and mixed‐N‐supplied plants, indicating reduced root growth and higher photosynthate availability to shoots. The following key observations were made: (1) a similar stomatal limitation and low proportion of activated Rubisco were observed among the three different N nutrition regimes; (2) increased mesophyll conductance in NH₄⁺‐ and mixed‐N‐supplied plants simultaneously stimulated leaf photosynthesis and improved the water use efficiency and (3), the maximum carboxylation rate and actual photochemical efficiency of photosystem II in NH₄⁺‐ and mixed‐N‐supplied plants were significantly higher than that in NO₃^?‐supplied plants, thus resulting in higher photochemical efficiency under water stress. In conclusion, mixed‐N and NH₄⁺ nutrition may be used to develop strategies for improved water stress resistance and stimulated biomass production under conditions of osmotic stress and possibly drought.     

Nitrate reductase activity and nitrogen compounds in xylem exudate of Juglans nigra seedlings: relation to nitrogen source and supply

Nitrogen (N) limits plant productivity and its uptake and assimilation may be regulated by N source, N availability, and nitrate reductase activity (NRA). Knowledge of how these factors interact to affect N uptake and assimilation processes in woody angiosperms is limited. We fertilized 1-year-old, half-sib black walnut (Juglans nigra L.) seedlings with ammonium (NH₄ ⁺) [as (NH₄)₂SO₄], nitrate (NO₃ ⁻) (as NaNO₃), or a mixed N source (NH₄NO₃) at 0, 800, or 1,600 mg N plant⁻¹ season⁻¹. Two months following final fertilization, growth, in vivo NRA, plant N status, and xylem exudate N composition were assessed. Specific leaf NRA was higher in NO₃ ⁻-fed and NH₄NO₃-fed plants compared to observed responses in NH₄ ⁺-fed seedlings. Regardless of N source, N addition increased the proportion of amino acids (AA) in xylem exudate, inferring greater NRA in roots, which suggests higher energy cost to plants. Root total NRA was 37% higher in NO₃ ⁻-fed than in NH₄ ⁺-fed plants. Exogenous NO₃ ⁻ was assimilated in roots or stored, so no difference was observed in NO₃ ⁻ levels transported in xylem. Black walnut seedling growth and physiology were generally favored by the mixed N source over NO₃ ⁻ or NH₄ ⁺ alone, suggesting NH₄NO₃ is required to maximize productivity in black walnut. Our findings indicate that black walnut seedling responses to N source and level contrast markedly with results noted for woody gymnosperms or herbaceous angiosperms.     

The kinetics of ammonium and nitrate uptake by young rice plants

Summary An important process which affects the fate of fertilizer nitrogen (N) applied to a rice crop is crop N uptake. This uptake rate is controlled by many factors including the N-ion species and its concentration. In this study the relation between N concentration at the root surface and N uptake was characterized using Michaelis-Menten kinetics. The equation considers two parameters, Vmax and Km, which are measures of the maximum rate of uptake and the affinity of the uptake sites for the nutrient, respectively. Uptake rates of intact rice plants growing in a continuously flowing nutrient solution system were fitted to the Michaelis-Menten model using a weighted regression analysis. For NH₄−N the Km values for 4- and 9-week-old rice plants indicated a high affinity for the ammonium ions relative to concentrations reported for rice soils after fertilization. The Vmax values expressed on a unit-root-mass basis decreased with plant age, indicating a reduction in the average density of uptake sites on the root surface. The kinetics of NO₃−N uptake was similar to that of NH₄−N when NO₃−N was the only N source. However, if NH₄−N and NO₃−N were present simultaneously in the solution the Vmax for the uptake of NO₃−N was severely reduced, while the Km was affected very little. This inhibition appears to be noncompetitive. Fertilization of young rice plants leading to concentration of N at the root surface above approximately 900 μM will not increase crop uptake and may contribute to inefficient N recovery by the crop. The existence of NH₄−N and NO₃−N simultaneously at the root surface may also lead to inefficient N recovery because of reduced uptake of NO₃−N.     

Nitrate reductase activity of two leafy vegetables as affected by nickel and different nitrogen forms

The author studied the effect of different nickel concentrations (0, 0.4, 40 and 80 μM Ni) on the nitrate reductase (NR) activity of New Zealand spinach (Tetragonia expansa Murr.) and lettuce (Lactuca sativa L. cv. Justyna) plants supplied with different nitrogen forms (NO₃ ⁻–N, NH₄ ⁺–N, NH₄NO₃). A low concentration of Ni (0.4 μM) did not cause statistically significant changes of the nitrate reductase activity in lettuce plants supplied with nitrate nitrogen (NO₃ ⁻–N) or mixed (NH₄NO₃) nitrogen form, but in New Zealand spinach leaves the enzyme activity decreased and increased, respectively. The introduction of 0.4 μM Ni in the medium containing ammonium ions as a sole source of nitrogen resulted in significantly increased NR activity in lettuce roots, and did not cause statistically significant changes of the enzyme activity in New Zealand spinach plants. At a high nickel level (Ni 40 or 80 μM), a significant decrease in the NR activity was observed in New Zealand spinach plants treated with nitrate or mixed nitrogen form, but it was much more marked in leaves than in roots. An exception was lack of significant changes of the enzyme activity in spinach leaves when plants were treated with 40 μM Ni and supplied with mixed nitrogen form, which resulted in the stronger reduction of the enzyme activity in roots than in leaves. The statistically significant drop in the NR activity was recorded in the aboveground parts of nickel-stressed lettuce plants supplied with NO₃ ⁻–N or NH₄NO₃. At the same time, there were no statistically significant changes recorded in lettuce roots, except for the drop of the enzyme activity in the roots of NO₃ ⁻-fed plants grown in the nutrient solution containing 80 μM Ni. An addition of high nickel doses to the nutrient solution contained ammonium nitrogen (NH₄ ⁺–N) did not affect the NR activity in New Zealand spinach plants and caused a high increase of this enzyme in lettuce organs, especially in roots. It should be stressed that, independently of nickel dose in New Zealand spinach plants supplied with ammonium form, NR activity in roots was dramatically higher than that in leaves. Moreover, in New Zealand spinach plants treated with NH₄ ⁺–N the enzyme activity in roots was even higher than in those supplied with NO₃ ⁻–N.     

Chronic Atmospheric NO<Stack><Subscript>3</Subscript><Superscript>−</Superscript></Stack> Deposition Does Not Induce NO<Stack><Subscript>3</Subscript><Superscript>−</Superscript></Stack> Use by <Emphasis Type="Italic">Acer saccharum</Emphasis> Marsh.

The ability of an ecosystem to retain anthropogenic nitrogen (N) deposition is dependent upon plant and soil sinks for N, the strengths of which may be altered by chronic atmospheric N deposition. Sugar maple (Acer saccharum Marsh.), the dominant overstory tree in northern hardwood forests of the Lake States region, has a limited capacity to take up and assimilate NO₃⁻. However, it is uncertain whether long-term exposure to NO₃⁻ deposition might induce NO₃⁻ uptake by this ecologically important overstory tree. Here, we investigate whether 10 years of experimental NO₃⁻ deposition (30 kg N ha⁻¹ y⁻¹) could induce NO₃⁻ uptake and assimilation in overstory sugar maple (approximately 90 years old), which would enable this species to function as a direct sink for atmospheric NO₃⁻ deposition. Kinetic parameters for NH₄⁺ and NO₃⁻ uptake in fine roots, as well as leaf and root NO₃⁻ reductase activity, were measured under conditions of ambient and experimental NO₃⁻ deposition in four sugar maple-dominated stands spanning the geographic distribution of northern hardwood forests in the Upper Lake States. Chronic NO₃⁻ deposition did not alter the V _max or K _m for NO₃⁻ and NH₄⁺ uptake nor did it influence NO₃⁻ reductase activity in leaves and fine roots. Moreover, the mean V _max for NH₄⁺ uptake (5.15 μmol ¹⁵N g⁻¹ h⁻¹) was eight times greater than the V _max for NO₃⁻ uptake (0.63 μmol ¹⁵N g⁻¹ h⁻¹), indicating a much greater physiological capacity for NH₄⁺ uptake in this species. Additionally, NO₃⁻ reductase activity was lower than most values for woody plants previously reported in the literature, further indicating a low physiological potential for NO₃⁻ assimilation in sugar maple. Our results demonstrate that chronic NO₃⁻ deposition has not induced the physiological capacity for NO₃⁻ uptake and assimilation by sugar maple, making this dominant species an unlikely direct sink for anthropogenic NO₃⁻ deposition.     

Optimization and elucidation of interactions between ammonium, nitrate and phosphate inCentella asiatica cell culture using response surface methodology

The effects of macronutrients (NO₃ ⁻, NH₄ ⁺ and PO₄ ³⁻) on cell growth and triterpenoids production inCentella asiatica cell suspension cultures were analyzed using the Box-Behnken response surface model experimental design. In screening and optimization experiments, PO₄ ³⁻ as a single factor significantly influenced cell growth where increasing the phosphate level from 0.1 to 2.4 or 2.6 mM, elevated cell growth from 3.9 to 14–16 g/L. The optimum values predicted from the response surface model are 5.05 mM NH₄ ⁺, 15.0 mM NO₃ ⁻ and 2.6 mM PO₄ ³⁻, yielding 16.0 g/L cell dry weight with 99% fitness to the experimental data. While the NH₄ ⁺-NO₃ ⁻ interaction influenced cell growth positively in the optimization experiment, NH₄ ⁺ and NO₃ ⁻ as single factors; and interactions of NO₃ ⁻-PO₄ ³⁻, NH₄ ⁺-PO₄ ³⁻ and NH₄ ⁺-NO₃ ⁻ were all negative in the screening experiment. Cell growth and the final pH level were positively affected by PO₄ ³⁻, but negatively affected by NH₄ ⁺ and NH₄ ⁺-PO₄ ³⁻ interactions. The different effects of factors and their interactions on cell growth and final pH are influenced by a broad or narrow range of macronutrient concentrations. The productions of triterpenoids however were lower than 4 mg/g cell dry weight.     

Organic matter quality and management effects on enrichment of soil organic matter fractions in contrasting soils in Zimbabwe

Spatial variability of soil total nitrogen (N), available N (KCl extractable NH₄⁺ and NO₃⁻), and spatial patterns of N mineralization and nitrification at a stand scale were characterized with geostatistical and univariate analysis. Two extensive soil spatial samplings were conducted in an evergreen broadleaf forest in Sichuan province, southwestern China in June and August 2000. In a study area of 90 × 105 m², three soil samples were collected from each 5 × 5 m² plot (n = 378) in June and August, and were analyzed for total N and available N contents. Net N mineralization and nitrification were measured by in situ core incubation and the rates were estimated based on the difference of NH₄⁺ and NO₃⁻ contents between the two sampling dates. Total N, NH₄⁺, and NO₃⁻ were all spatially structured with different semivariogram ranges (from high to low: NH₄⁺, NO₃⁻, and total N). The semivariograms of mineralization and nitrification were not as spatially structured as available N. NH₄⁺ was the dominant soil inorganic N form in the system. Both NH₄⁺ and NO₃⁻ affected spatial patterns of soil available N, but their relative importance switched in August, probably due to high nitrification as indicated by greatly increased soil NO₃⁻ content. High spatial auto-correlations (>0.7) were found between available N and NH4⁺, available N and NO₃⁻ on both sampling dates, as well as total N measurements between both sampling dates. Although significant, the spatial auto-correlation between NH₄⁺ and NO₃⁻ were generally low. Topography had significant but low correlations with mineralization (r = −0.16) and nitrification (r = −0.14), while soil moisture did not. The large nugget values of the calculated semivariograms and high-semivariance values, particularly for mineralization and nitrification, indicate that some fine scale (<5 m) variability may lie below the threshold for detection in this study.     

Effect of nitrogen forms on growth,cell composition and N<Subscript>2</Subscript> fixation of <Emphasis Type="Italic">Cylindrospermopsis raciborskii</Emphasis> in phosphorus-limited chemostat cultures

The aim of this research was to test whether NH₄ ⁺ and NO₃ ⁻ affect the growth, P demand, cell composition and N₂ fixation of Cylindrospermopsis raciborskii under P limitation. Experiments were carried out in P-limited (200 μg l⁻¹ PO₄-P) chemostat cultures of C. raciborskii using an inflowing medium containing either 4,000 μg l⁻¹ NH₄-N, 4,000 μg l⁻¹ NO₃-N or no combined N. The results showed the cellular N:P and C:P ratios of C. raciborskii decreased towards the Redfield ratio with increasing dilution rate (D) due to the alleviation of P limitation. The cellular C:N and carotenoids:chlorophyll-a ratios also decreased with D, predominantly as a result of an increase in the chlorophyll-a and N content. The NH₄ ⁺ and NO₃ ⁻ supply reduced the P maintenance cell quota of C. raciborskii. Consequently, the biomass yield of the N₂-grown culture was significantly lower. The maximum specific growth rate of N₂-grown culture was also the lowest observed. It is suggested that these differences in growth parameters were caused by the P and energy requirement for heterocyte formation, nitrogenase synthesis and N₂ fixation. N₂ fixation was partially inhibited by NO₃ ⁻ and completely inhibited by NH₄ ⁺. It was probably repressed through the high N content of cells at high dissolved N concentrations. These results indicate that C. raciborskii is able to grow faster and maintain a higher biomass under P limitation where a sufficient supply of NH₄ ⁺ or NO₃ ⁻ is maintained. Information gained about the species-specific nutrient and pigment stoichiometry of C. raciborskii could help to access the degree of nutrient limitation in water bodies. Handling editor: Luigi Naselli-Flores     

Ammonium nutrition increases water absorption in rice seedlings (Oryza sativa L.) under water stress

Water stress is a primary limitation on plant growth. In previous studies, it has been found that ammonium enhances the tolerance of rice plants to water stress, but how water is related to nitrogen form and water stress remains unknown. To study the effects of nitrogen form (NH ₄ ⁺ , NO ₃ ^? , and a mixture of NH ₄ ⁺ and NO ₃ ^? ) on the growth and water absorption of rice (Oryza sativa L.) seedlings, a hydroponic experiment with water stress, simulated by the addition of polyethylene glycol (PEG, 10% w/v, MW 6000), was conducted in a greenhouse. The results showed that, compared with non-water stress, under water stress, the fresh weight of rice seedlings increased by 14% with NH ₄ ⁺ nutrition, whereas it had decreased by about 20% with either NO ₃ ^? or mixed nitrogen nutrition. No significant difference was found in the transpiration rate of excised shoots or in xylem exudation of excised roots in NH ₄ ⁺ supply between the two water situations, whereas xylem flow decreased by 57% and 24% under water stress in NO ₃ ^? and mixed nutrition, and root hydraulic conductivity decreased by 29% and 54% in plants in NH ₄ ⁺ and NO ₃ ^? nutrition conditions, respectively. Although water absorption ability decreased in both NH ₄ ⁺ and NO ₃ ^? nutrition, aquaporin activity was higher in NH ₄ ⁺ than in NO ₃ ^? nutrition, regardless of water stress. We conclude that NH ₄ ⁺ nutrition can improve water handling in rice seedlings and subsequently enhance their resistance to drought.     

Seasonal variation in nutrient limitation of microbial biofilms colonizing organic and inorganic substrata in streams

Humans have increased the availability of nutrients including nitrogen and phosphorus worldwide; therefore, understanding how microbes process nutrients is critical for environmental conservation. We examined nutrient limitation of biofilms colonizing inorganic (fritted glass) and organic (cellulose sponge) substrata in spring, summer, and autumn in three streams in Michigan, USA. Biofilms were enriched with nitrate (NO₃ ⁻), phosphate (PO₄ ³⁻), ammonium (NH₄ ⁺), NO₃ ⁻ + PO₄ ³⁻, NH₄ ⁺ + PO₄ ³⁻, or none (control). We quantified biofilm structure and function as chlorophyll a (i.e., primary producer biomass) and community respiration on all substrata. In one stream, we characterized bacterial and fungal communities on cellulose in autumn using clone library sequencing and denaturing gradient gel electrophoresis to determine if community structure was linked to nutrient limitation status. Despite oligotrophic conditions, primary producer biomass was infrequently nutrient limited. In contrast, respiration on organic substrata was frequently limited by N + P combinations. We found no difference between biofilm response to NH₄ ⁺ versus NO₃ ⁻ enrichment, although the response to both N-species was positively related to water column PO₄ ³⁻ concentrations and temperature. Molecular analysis for fungal community composition suggested no relationship to nutrient limitation, but the dominant members of the bacterial community on cellulose were different on NO₃ ⁻, PO₄³, and NO₃ ⁻ + PO₄ ³⁻ treatments relative to control, NH₄ ⁺, and NH₄ ⁺ + PO₄ ³⁻ treatments, which matched patterns for biofilm respiration rates from each treatment. Our results show discrete patterns of nutrient limitation dependent upon substratum type and season, and imply changes in bacterial community structure and function may be linked following nutrient enrichment in streams.     

Epidemiology of wheat take-all as influenced by soil pH and temporal changes in inorganic soil N

Summary Soil pH, NH ₄ ⁺ and NO ₃ ⁻ concentrations in soil, and take-all root rot of winter wheat grown in the field were measured concurrently from sowing to anthesis in order to relate disease development to liming and N fertilization practices. Experimental variables included soil pH (5.5 and 6.0) and three N sources (NH₄NO₃, (NH₄)₂SO₄, NH₄Cl) banded with the seed at sowing in factorial combination with the same three N sources topdressed in the spring. Take-all severity was increased by increasing soil pH and by fertilization with NO ₃ ⁻ . Disease severity on crown roots increased exponentially following spring N fertilization and was affected more by soil pH and N-form than was severity on seminal roots. Grain yield ranged from 4.70 Mgha⁻¹ with spring NH₄NO₃ at soil pH 6.0 to 7.65 Mgha⁻¹ with spring NH₄Cl at soil pH 5.5. Sixty-six percent of the variability in grain yield was explained by the number of take-all infected crown roots per tiller at anthesis. Oregon Agric. Exp. Stn. technical paper no. 7707.     

Effect of activated charcoal and 6-benzyladenine on in vitro nitrogen uptake by Lagerstroemia indica

A sterile hydroponic culture system suitable for studying nitrogen (N) uptake ofLagerstroemia indica L.in vitro was developed. Four different treatments were assayed: with and without activated charcoal (AC and NAC, respectively), with and without 50 μM of 6-benzyladenine (+BA and −BA, respectively). Medium pH, electrical conductivity (EC), NO₃ ⁻ and NH₄ ⁺ concentrations were measured weekly. At the end of the culture, propagules were sampled and SPAD indices, and shoot and root fresh weights were determined. Explants grown in media with activated charcoal were able to take up both NO₃ ⁻ and NH₄ ⁺, although NH₄ ⁺ uptake was lower. Subsequently the pH of the media was maintained between 5.5–6.0. In treatments with no addition of activated charcoal, NH₄ ⁺ uptake was preferential and the pH dropped to 3.1. Explants in these conditions were unable to raise the pH by taking up NO₃ ⁻, especially when root morphogenesis was inhibited by addition of BA. Supply of this PGR produced root growth inhibition, which was almost complete in the treatment without activated charcoal. This component significantly reduced the inhibitory effect of 50 μM BA on root growth. This revised version was published online in June 2006 with corrections to the Cover Date.     

NO<Subscript>3</Subscript><Superscript>−</Superscript>-Driven SO<Subscript>4</Subscript><Superscript>2−</Superscript> Production in Freshwater Ecosystems: Implications for N and S Cycling

Massive anthropogenic acceleration of the global nitrogen (N) cycle has stimulated interest in understanding the fate of excess N loading to aquatic ecosystems. Nitrate (NO₃ ⁻) is traditionally thought to be removed mainly by microbial respiratory denitrification coupled to carbon (C) oxidation, or through biomass assimilation. Alternatively, chemolithoautotrophic bacterial metabolism may remove NO₃ ⁻ by coupling its reduction with the oxidation of sulfide to sulfate (SO₄ ²⁻). The NO₃ ⁻ may be reduced to N₂ or to NH₄ ⁺, a form of dissimilatory nitrate reduction to ammonium (DNRA). The objectives of this study were to investigate the importance of S oxidation as a NO₃ ⁻ removal process across diverse freshwater streams, lakes, and wetlands in southwestern Michigan (USA). Simultaneous NO₃ ⁻ removal and SO₄ ²⁻ production were observed in situ using modified “push-pull” methods in nine streams, nine wetlands, and three lakes. The measured SO₄ ²⁻ production can account for a significant fraction (25–40%) of the overall NO₃ ⁻ removal. Addition of ¹⁵NO₃ ⁻ and measurement of ¹⁵NH₄ ⁺ production using the push–pull method revealed that DNRA was a potentially important process of NO₃ ⁻ removal, particularly in wetland sediments. Enrichment cultures suggest that Thiomicrospira denitrificans may be one of the organisms responsible for this metabolism. These results indicate that NO₃ ⁻-driven SO₄ ²⁻ production could be widespread and biogeochemically important in freshwater sediments. Removal of NO₃ ⁻ by DNRA may not ameliorate problems such as eutrophication because the N remains bio-available. Additionally, if sulfur (S) pollution enhances NO₃ ⁻ removal in freshwaters, then controls on N processing in landscapes subject to S and N pollution are more complex than previously appreciated. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Distribution of nitrogen-15 tracers applied to the canopy of a mature spruce-hemlock stand,Howland, Maine,USA

In N-limited ecosystems, fertilization by N deposition may enhance plant growth and thus impact C sequestration. In many N deposition–C sequestration experiments, N is added directly to the soil, bypassing canopy processes and potentially favoring N immobilization by the soil. To understand the impact of enhanced N deposition on a low fertility unmanaged forest and better emulate natural N deposition processes, we added 18 kg N ha⁻¹ year⁻¹ as dissolved NH₄NO₃ directly to the canopy of 21 ha of spruce-hemlock forest. In two 0.3-ha subplots, the added N was isotopically labeled as ¹⁵NH₄ ⁺ or ¹⁵NO₃ ⁻ (1% final enrichment). Among ecosystem pools, we recovered 38 and 67% of the ¹⁵N added as ¹⁵NH₄ ⁺ and ¹⁵NO₃ ⁻, respectively. Of ¹⁵N recoverable in plant biomass, only 3–6% was recovered in live foliage and bole wood. Tree twigs, branches, and bark constituted the most important plant sinks for both NO₃ ⁻ and NH₄ ⁺, together accounting for 25–50% of ¹⁵N recovery for these ions, respectively. Forest floor and soil ¹⁵N retention was small compared to previous studies; the litter layer and well-humified O horizon were important sinks for NH₄ ⁺ (9%) and NO₃ ⁻ (7%). Retention by canopy elements (surfaces of branches and boles) provided a substantial sink for N that may have been through physico-chemical processes rather than by N assimilation as indicated by poor recoveries in wood tissues. Canopy retention of precipitation-borne N added in this particular manner may thus not become plant-available N for several years. Despite a large canopy N retention potential in this forest, C sequestration into new wood growth as a result of the N addition was only ~16 g C m⁻² year⁻¹ or about 10% above the current net annual C sequestration for this site.     

Changes in Canopy Processes Following Whole-Forest Canopy Nitrogen Fertilization of a Mature Spruce-Hemlock Forest

Abstract Most experimental additions of nitrogen to forest ecosystems apply the N to the forest floor, bypassing important processes taking place in the canopy, including canopy retention of N and/or conversion of N from one form to another. To quantify these processes, we carried out a large-scale experiment and determined the fate of nitrogen applied directly to a mature coniferous forest canopy in central Maine (18–20 kg N ha⁻¹ y⁻¹ as NH₄NO₃ applied as a mist using a helicopter). In 2003 and 2004 we measured NO₃ ⁻, NH₄ ⁺, and total dissolved N (TDN) in canopy throughfall (TF) and stemflow (SF) events after each of two growing season applications. Dissolved organic N (DON) was greater than 80% of the TDN under ambient inputs; however NO₃ ⁻ accounted for more than 50% of TF N in the treated plots, followed by NH₄ ⁺ (35%) and DON (15%). Although NO₃ ⁻ was slightly more efficiently retained by the canopy under ambient inputs, canopy retention of NH₄ ⁺as a percent of inputs increased markedly under fertilization. Recovery of less than 30% of the fertilizer N in TF suggested that the forest canopy retained more than 70% of the applied N (>80% when corrected for N which bypassed tree surfaces at the time of fertilizer addition). Results from plots receiving ¹⁵N enriched NO₃ ⁻ and NH₄ ⁺ confirmed bulk N estimations that more NO₃ ⁻ than NH₄ ⁺ was washed from the canopy by wet deposition. The isotope data did not show evidence of canopy nitrification, as has been reported in other spruce forests receiving much higher N inputs. Conversions of fertilizer-N to DON were observed in TF for both ¹⁵NH₄ ⁺ and ¹⁵NO₃ ⁻ additions, and occurred within days of the application. Subsequent rain events were not significantly enriched in ¹⁵N, suggesting that canopy DON formation was a rapid process related to recent N inputs to the canopy. We speculate that DON may arise from lichen and/or microbial N cycling rather than assimilation and re-release by tree tissues in this forest. Canopy retention of experimentally added N may meet and exceed calculated annual forest tree demand, although we do not know what fraction of retained N was actually physiologically assimilated by the plants. The observed retention and transformation of DIN within the canopy demonstrate that the fate and ecosystem consequences of N inputs from atmospheric deposition are likely influenced by forest canopy processes, which should be considered in N addition studies. Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users.