The influence of enriched rhizosphere CO2 on N uptake and metabolism in wild-type and NR-deficient barley plants

Positive influences of high concentrations of dissolved inorganic carbon (DIC) in the growth medium of salinity-stressed plants are associated with carbon assimilation through phosphoenolpyruvate carboxylase (PEPc) activity in roots; and also in salinity-stressed tomato plants, enriched CO₂ in the rhizosphere increases NO^?₃uptake. In the present study, wild-type and nitrate reductase-deficient plants of barley (Hordeum vulgare L. cv. Steptoe) were used to determine whether the influence of enriched CO₂ on NO^?₃ uptake and metabolism is dependent on the activity of nitrate reductase (NR) in the plant. Plants grown in NH₄⁺and aerated with ambient air, were transferred to either NO₃^? or NH₄⁺ solutions and aerated with air containing between 0 and 6 500 μmol mol^?1 CO₂. Nitrogen uptake and tissue concentrations of NO₃^? and NH₄⁺ were measured as well as activities of NR and PEPc. The uptake of NO^?₃ by the wild-type was increased by increasing CO₂. This was associated with increased in vitro NR activity, but increased uptake of NO₃^? was found also in the NR-deficient genotype when exposed to high CO₂ concentrations; so that the influence of CO₂ on NO₃^? uptake was independent of the reduction of NO₃^? and assimilation into amino acids. The increase in uptake of NO₃^? in wild-type plants with enriched CO₂ was the same at pH 7 as at pH 5, indicating that the relative abundance of HCO₃^? or CO₂ in the medium did not influence NO₃^? uptake. Uptake of NH₄⁺ was decreased by enriched CO₂ in a pH (5 or 7) independent fashion. Thus NO₃^? and NH⁺₄ uptakes are influenced by the CO₂ component of DIC independently of anaplerotic carbon provision for amino acid synthesis, and CO₂ may directly affect the uptake of NO₃^? and NH₄⁺ in ways unrelated to the NR activity in the tissue.     

Elevated CO2 plus chronic warming reduce nitrogen uptake and levels or activities of nitrogen‐uptake and ‐assimilatory proteins in tomato roots

Atmospheric CO₂ enrichment is expected to often benefit plant growth, despite causing global warming and nitrogen (N) dilution in plants. Most plants primarily procure N as inorganic nitrate (NO₃^?) or ammonium (NH₄⁺), using membrane‐localized transport proteins in roots, which are key targets for improving N use. Although interactive effects of elevated CO₂, chronic warming and N form on N relations are expected, these have not been studied. In this study, tomato (Solanum lycopersicum) plants were grown at two levels of CO₂ (400 or 700 ppm) and two temperature regimes (30 or 37°C), with NO₃^? or NH₄⁺ as the N source. Elevated CO₂ plus chronic warming severely inhibited plant growth, regardless of N form, while individually they had smaller effects on growth. Although %N in roots was similar among all treatments, elevated CO₂ plus warming decreased (1) N‐uptake rate by roots, (2) total protein concentration in roots, indicating an inhibition of N assimilation and (3) shoot %N, indicating a potential inhibition of N translocation from roots to shoots. Under elevated CO₂ plus warming, reduced NO₃^?‐uptake rate per g root was correlated with a decrease in the concentration of NO₃^?‐uptake proteins per g root, reduced NH₄⁺ uptake was correlated with decreased activity of NH₄⁺‐uptake proteins and reduced N assimilation was correlated with decreased concentration of N‐assimilatory proteins. These results indicate that elevated CO₂ and chronic warming can act synergistically to decrease plant N uptake and assimilation; hence, future global warming may decrease both plant growth and food quality (%N).     

Net fluxes of ammonium and nitrate in association with H+ fluxes in fine roots of Populus popularis

Poplar plants are cultivated as woody crops, which are often fertilized by addition of ammonium (NH₄ ⁺) and/or nitrate (NO₃ ^?) to improve yields. However, little is known about net NH₄ ⁺/NO₃ ^? fluxes and their relation with H⁺ fluxes in poplar roots. In this study, net NH₄ ⁺/NO₃ ^? fluxes in association with H⁺ fluxes were measured non-invasively using scanning ion-selective electrode technique in fine roots of Populus popularis. Spatial variability of NH₄ ⁺ and NO₃ ^? fluxes was found along root tips of P. popularis. The maximal net uptake of NH₄ ⁺ and NO₃ ^? occurred, respectively, at 10 and 15 mm from poplar root tips. Net NH₄ ⁺ uptake was induced by ca. 48 % with provision of NO₃ ^? together, but net NO₃ ^? uptake was inhibited by ca. 39 % with the presence of NH₄ ⁺ in poplar roots. Furthermore, inactivation of plasma membrane (PM) H⁺-ATPases by orthovanadate markedly inhibited net NH₄ ⁺/NO₃ ^? uptake and even led to net NH₄ ⁺ release with NO₃ ^? co-provision. Linear correlations were observed between net NH₄ ⁺/NO₃ ^? and H⁺ fluxes in poplar roots except that no correlation was found between net NH₄ ⁺ and H⁺ fluxes in roots exposed to NH₄Cl and 0 mM vanadate. These results indicate that root tips play a key role in NH₄ ⁺/NO₃ ^? uptake and that net NH₄ ⁺/NO₃ ^? fluxes and the interaction of net fluxes of both ions are tightly associated with H⁺ fluxes in poplar roots.     

Differential responses of root uptake kinetics of NH4+ and NO3− to enriched atmospheric CO2 concentration in field-grown loblolly pine

The nitrogen requirement of plants is predominantly supplied by NH₄⁺ and/or NO₃^? from the soil solution, but the energetic cost of uptake and assimilation is generally higher for NO₃^? than for NH₄⁺. We found that CO₂ enrichment of the atmosphere enhanced the root uptake capacity for NO₃^?, but not for NH₄⁺, in field-grown loblolly pine saplings. Increased preference for NO₃^? at the elevated CO₂ concentration was accompanied by increased carbohydrate levels in roots. The results have important implications for the potential consequences of global climate change on plant-and ecosystem-level processes in many temperate forest ecosystems.     

Nitrogen source and pCO2 synergistically affect carbon allocation,growth and morphology of the coccolithophore Emiliania huxleyi: potential implications of ocean acidification for the carbon cycle

Coccolithophores are unicellular phytoplankton that produce calcium carbonate coccoliths as an exoskeleton. Emiliania huxleyi, the most abundant coccolithophore in the world's ocean, plays a major role in the global carbon cycle by regulating the exchange of CO₂ across the ocean‐atmosphere interface through photosynthesis and calcium carbonate precipitation. As CO₂ concentration is rising in the atmosphere, the ocean is acidifying and ammonium (NH₄⁺) concentration of future ocean water is expected to rise. The latter is attributed to increasing anthropogenic nitrogen (N) deposition, increasing rates of cyanobacterial N₂ fixation due to warmer and more stratified oceans, and decreased rates of nitrification due to ocean acidification. Thus, future global climate change will cause oceanic phytoplankton to experience changes in multiple environmental parameters including CO₂, pH, temperature and nitrogen source. This study reports on the combined effect of elevated pCO₂ and increased NH₄⁺ to nitrate (NO₃^?) ratio (NH₄⁺/NO₃^?) on E. huxleyi, maintained in continuous cultures for more than 200 generations under two pCO₂ levels and two different N sources. Herein, we show that NH₄⁺ assimilation under N‐replete conditions depresses calcification at both low and high pCO₂, alters coccolith morphology, and increases primary production. We observed that N source and pCO₂ synergistically drive growth rates, cell size, and the ratio of inorganic to organic carbon. These responses to N source suggest that, compared to increasing CO₂ alone, a greater disruption of the organic carbon pump could be expected in response to the combined effect of increased NH₄⁺/NO₃^? ratio and CO₂ level in the future acidified ocean. Additional experiments conducted under lower nutrient conditions are needed prior to extrapolating our findings to the global oceans. Nonetheless, our results emphasize the need to assess combined effects of multiple environmental parameters on phytoplankton biology to develop accurate predictions of phytoplankton responses to ocean acidification.     

Photosynthetic energy supply for NO3− assimilation in Scenedesmus

NO₃^?-dependent O₂ in synchronous Scenedesmus obtusiusculus Chod. in the absence of CO₂ is stoichiometric with NH₄⁺ excretion, indicating a close coupling of NO₃^? reduction to non-cyclic electron flow. Also in the presence of CO₂, NO₃^? stimulates O₂ evolution as manifested by an increase in the O₂/CO₂ ratio from 0.96 to 1.11. This quotient was increased to 1.36 by addition of NO₂^?, without competitive interaction with CO₂ fixation, indicating that the capacity for non-cyclic electron transport at saturating light is non-limiting for simultaneous reduction of NO₃^? and CO₂ at high rates. During incubation with NO₃^?+ CO₂, no NH₄⁺ is released to the outer medium, whereas during incubation with NO₂^?+ CO₂, excess NH₄⁺ is formed and excreted. NO₃^? uptake is stimulated by CO₂, and this stimulation is also significant when the cellular energy metabolism is restricted by moderate concentrations of carbonyl cyanide-p-trifluoromethoxyphenylhydrazone, whereas NO₃^? uptake in the absence of CO₂ is severely inhibited by the uncoupler. Also under energy-restricted conditions NO₃^? uptake is not competitive with CO₂ fixation. Antimycin A is inhibitory for NO₃^? uptake in the absence of CO₂, and there is no enhancement of NO₃^? uptake by CO₂ in the presence of antimycin A. It is assumed that the energy demand for NO₃^? uptake is met by energy fixed as triosephosphates in the Calvin cycle. Antimycin A possibly affects the transfer of reduced triose phosphates from the chloroplast to the cytoplasm. Active carbon metabolism also seems to exert a control effect on NO₃^? assimilation, inducing complete incorporation of all NO₃^? taken up into amino acids. This control effect is not functional when NO₂^? is the nitrogen source. Active carbon metabolism thus seems to be essential both for provision of energy for NO₃^? uptake and for regulation of the process.     

AMMONIUM,NITRATE, PHOSPHATE,AND INORGANIC CARBON UPTAKE IN AN OLIGOTROPHIC LAKE: SEASONAL VARIATIONS AMONG LIGHT RESPONSE VARIABLES1

The dependence of substrate saturated uptake of ¹⁵NH₄⁺, ¹⁵NO₃^?, ³²PO₄^3?, and ¹⁴CO₂ on photosynthetic photon flux density (PPFD or photsynthetically active radiation, 400–700 nm) was characterized seasonally in oligotrophic Flathead Lake, Montana. PO₄^3? uptake was not dependent upon PPFD at any time of the year, whereas NH₄⁺, NO₃^?, and CO₂ uptake were consistently dependent on PPFD over all seasons. Maximal rates of NH₄⁺, NO₃^? and CO₂ uptake usually occurred near 40% of surface PPFD, which corresponded to about 5 m in the lake; inhibition was evident at PPFD levels greater than 40%. NH₄⁺, NO₃^? and PO₄^3? were incorporated in the dark at measurable rates most of the year, whereas dark CO₂ uptake was always near 0 relative to light uptake. CO₂ and NO₃^? uptake were more strongly influenced by PPFD than was NH₄^3? uptake. The PPFD dependence of PO₄^3?, NH₄⁺, NO₃^? and CO₂ uptake may affect algal growth and nutrient status by influencing the balance in diel and seasonal C:N:P uptake ratios.     

Responses of soybean to ammonium and nitrate supplied in combination to the whole root system or separately in a split-root system

To address the questions of whether allocation of carbohydrates to roots is influenced by ionic form of nitrogen absorbed and whether allocation of carbohydrates to roots in turn influences proportionality between NH₄⁺ and NO₃^? uptake from mixed sources, NH₄⁺ and NO₃^? were supplied separately to halves of a split-root hydroponic system and were supplied in combination to a whole-root system. Dry matter accumulation in the split-root system was 18% less in the NH₄⁺-fed axis than in the NO₃^?-fed axis. This, however, does not indicate that partitioning of carbohydrate between the two axes was different. Most of the reduction in dry matter accumulation in the NH₄⁺-fed axis can be accounted for by the retransport of CH₂O equivalents from the root back to the shoot with amino acids produced by NH₄⁺ assimilation. Uptake of NH₄⁺ or NO₃^? by the respective halves of the split-root system was proportional to the estimated allocation of carbohydrate to that half. When NH₄⁺ and NO₃^? were supplied to separate halves of the split-root system, the cumulative NH₄⁺ to NO₃^? uptake ratio was 0.81. When supplied in combination to the whole-root system, the cumulative NH₄⁺ to NO₃^? uptake ratio was 1.67. Thus, while the shoot may affect total nitrogen uptake through the export of carbohydrates to roots, the shoot (common for halves of the split-root system) apparently does not exert a direct effect on proportionality of NH₄⁺ and NO₃^? uptake by roots. For whole roots supplied with both NH₄⁺ and NO₃^?, the restriction in uptake of NO₃^? may involve a stimulation of NO₃^? efflux rather than an inhibition of NO₃^? influx. While only the net uptake of NH₄⁺ and NO₃^? was measured by ion chromatography, monitoring at approximately hourly intervals during the first 3 days of treatment revealed irregularly occurring intervals of both depletion (net influx) and enrichment (net efflux) in solutions. In the case of NH₄⁺, numbers of net efflux events were similar (21 to 24 out of 65 sequential sampling intervals) whether NH₄⁺ was supplied with NO₃^? to whole-root systems or separately to an axis of the split-root system. In the case of NO₃^?, however, the number of net efflux events increased from 8 when NO₃^? was supplied to a separate axis of the split-root system to between 19 and 24 when NO₃^? was supplied with NH₄⁺ to whole-root systems.     

Changes in NO3− and K+ uptake by four species in flowing solution culture in response to increased irradiance

Net rates of NO₃^? and K⁺ uptake were compared for oilseed rape (Brassica napus L. cv. Jet neuf), perennial ryegrass (Lolium perenne L. cv. S23), Italian ryegrass (Lolium multiflorum Lam. cv. Augusta) and wheat (Triticum aestivum L. cv. Fen-man) in flowing solution culture during a 4-day sequence of low-low-high-high natural irradiance. Concentrations of NO₃^? (10 μM) and K⁺ (2.5 μM) in solutions were maintained automatically and hourly variation in net uptake of these ions was measured. During the 2 days of low irradiance (<1 MJ m^?2 day^?1) the uptake rates of both ions by all species were low at <1 mmol NO₃^?, m^?2 h^?1 and <0.4 mmol K⁺ m^?2 h^?1. Uptake increased in each species during the first day of high irradiance (7.90 MJ m^?2 day^?1) to >4 mmol NO₃^? m^?2 h^?1 and >1.4 mmol K⁺ m^?1 h^?1. These higher rates were maintained throughout the following night. The lag-time between maximum irradiance and the onset of the highest acceleration in uptake was greater for NO₃^? (5–8 h) than for K⁺ (≤1 h) in rape, wheat and Italian ryegrass. Uptake of NO₃^?, by perennial ryegrass showed an almost constant acceleration for 18 h following maximum irradiance. In all species the measured maximum inflows (uptake rate per unit root length) of both ions were greater than theoretical maximum potential inflows to a non-competing infinite-sink root in soil, by factors of 7 and 36, respectively, for NO₃^? and K⁺, averaged over all species.     

Absorption of nitrate and ammonium ions by Lolium perenne from flowing solution cultures at low root temperatures

Lolium perenne L. cv. 23 (perennial ryegrass) plants were grown in flowing solution culture and acclimatized over 49 d to low root temperature (5°C) prior to treatment at root temperatures of 3, 5, 7 and 9°C for 41 d with common air temperature of 20/15°C day/night and solution pH 5·0. The effects of root temperature on growth, uptake and assimilation of N were compared with N supplied as either NH₄ or NO3 at 10 mmol m^?3. At any given temperature, the relative growth rate (RGR) of roots exceeded that of shoots, thus the root fraction (Rf) increased with time. These effects were found in plants grown with the two N sources. Plants grown at 3 and 5°C had very high dry matter contents as reflected by the fresh weight: freeze-dried weight ratio. This ratio increased sharply, especially in roots at 7 and 9°C. Expressed on a fresh weight basis, there was no major effect of root temperature on the [N] of plants receiving NHJ but at any given temperature, the [N] in plants grown with NHJ was significantly greater than in those grown with NO₃. The specific absorption rate (SAR) of NH⁺₄ was greater at all temperatures than SAR-NO₃. In plants grown with NH⁺, 3–5% of the total N was recovered as NH⁺₄, whereas in those grown with NO^?₃ the unassimilated NO^?₃ rose sharply between 7 and 9°C to become 14 and 28% of the total N in shoots and roots, respectively. The greater assimilation of NH⁺₄ lead to concentrations of insoluble reduced N (= protein) which were 125 and 20% greater, in roots and shoots, respectively, than in NO^?₃-grown plants. Plants grown with NH⁺₄ had very much greater glutamine and asparagine concentrations in both roots and shoots, although other amino acids were more similar in Concentration to those in NO^?₃ grown plants. It is concluded that slow growth at low root temperature is not caused by restriction of the absorption or assimilation of either NH⁺₄ or NO^?₃. The additional residual N (protein) in NH⁺₄ grown plants may serve as a labile store of N which could support growth when external N supply becomes deficient.     

Kinetics of nitrate and ammonium absorption and accompanying H+ fluxes in roots of Lolium perenne L. and N2-fixing Trifolium repens L.

Kinetic parameters for NH₄⁺ and NO₃^? uptake were measured in intact roots of Lolium perenne and actively N₂-fixing Trifolium repens. Simultaneously, net H⁺ fluxes between the roots and the root medium were recorded, as were the net photosynthetic rate and transpiration of the leaves. A Michaelis–Menten-type high-affinity system operated in the concentration range up to about 500 mmol m^?3 NO₃^? or NH₄⁺. In L. perenne, the V_max of this system was 9–11 and 13–14 μmol g^?1 root FW h^?1 for NO₃^? and NH₄⁺, respectively. The corresponding values in T. repens were 5–7 and 2 μmol g^?1 root FW h^?1. The K_m for NH₄⁺ uptake was much lower in L. perenne than in T. repens (c. 40 compared with 170 mmol m^?3), while K_m values for NO₃^? absorption were roughly similar (around 130 mmol m^?3) in the two species. There were no indications of a significant efflux component in the net uptake of the two ions. The translocation rate to the shoots of nitrogen derived from absorbed NO₃^?-N was higher in T. repens than in L. perenne, while the opposite was the case for nitrogen absorbed as NH₄⁺. Trifolium repens had higher rates of transpiration and net photosynthesis than L. perenne. Measurements of net H⁺ fluxes between roots and nutrient solution showed that L. perenne absorbing NO₃^? had a net uptake of H⁺, while L. perenne with access to NH₄⁺ and T. repens, with access to NO₃^? or NH₄⁺, in all cases acidified the nutrient solution. Within the individual combinations of plant species and inorganic N form, the net H⁺ fluxes varied only a little with external N concentration and, hence, with the absorption rate of inorganic N. Based on assessment of the net H⁺ fluxes in T. repens, nitrogen absorption rate via N₂ fixation was similar to that of inorganic N and was not down-regulated by exposure to inorganic N for 2 h. It is concluded that L. perenne will have a competitive advantage over T. repens with respect to inorganic N acquisition.     

Relationships between the kinetics of NH4+ and NO3− absorption and growth in the cultivated tomato (Lycopersicon esculentum Mill, cv T-5)

Tomato growth was examined in solution culture under constant pH and low levels of NH₄⁺ or NO₃^?. There were five nitrogen treatments: 20 mmoles m^?3 NH₄⁺, 50 mmoles m^?3 NO₃^?, 100 mmoles m^?3 NH₄⁺ 200 mmoles m^?3 NO₃^?, and 20 mmoles m^?3 NH₄₊+ 50 mmoles m^?3 NO₃^?. The lower concentrations (20 mmoles m^?3 NH₄⁺ and 50 mmoles m^?3 NO₃^?) were near the apparent K_m for net NH₄⁺ and NO₃^? uptake; the higher concentrations (100 mmoles m^?3 NH₄⁺ and 200 mmoles m^?3 NO₃^?) were near levels at which the net uptake of NH₄⁺ or NO₃^? saturate. Although organic nitrogen contents for the higher NO₃^? and the NH₄⁺+ NO₃^? treatments were 22.2–30.3% greater than those for the lower NO₃^? treatment, relative growth rates were initially only 10–15% faster. After 24 d, relative growth rates were similar among those treatments. These results indicate that growth may be only slightly nitrogen limited when NH₄⁺ or NO₃^? concentrations are held constant over the root surface at near the apparent K_m concentration. Relative growth rates for the two NH₄⁺ treatments were much higher than have been previously reported for tomatoes growing with NH₄⁺ as the sole nitrogen source. Initial growth rates under NH₄⁺ nutrition did not differ significantly (P≥ 0.05) from those under NO₃^? or under combined NH₄⁺+ NO₃^?. Growth rates slowed after 10–15 d for the NH₄⁺ treatments, whereas they remained more constant for the NO₃^? and mixed NH₄⁺+ NO₃^? treatments over the entire observation period of 24–33 d. The decline in growth rate under NH₄⁺ nutrition may have resulted from a reduction in Ca²⁺, K⁺, and/or Mg²⁺ absorption.     

Root growth as a function of ammonium and nitrate in the root zone

We examined the effect of soil NH₄⁺ and NO₃^? content upon the root systems of field-grown tomatoes, and the influence of constant, low concentrations of NH₄⁺ or NO₃^? upon root growth in solution culture. In two field experiments, few roots were present in soil zones with low extractable NH₄⁺ or NO₃^?; they increased to a maximum in zones having 2μg-N NO₃^? g^?1 soil and 6 μg-N NO₃⁼ g^?1 soil, but decreased in zones having higher NH₄⁺ or NO₃^? levels. Root branching was relatively insensitive to available mineral nitrogen. Plants maintained in solution culture at constant levels of NH₄⁺ or NO₃^?, had similar shoot biomass, but all root parameters – biomass, length, branching and area – were greater under NH4 nutrition than under NO₃^?. These results suggest that the size of root system depends on a functional equilibrium between roots and shoots (Brouwer 1967) and on the balance between soil NH₄⁺ and NO₃^?.     

Oxygen consumption during leaf nitrate assimilation in a C3 and C4 plant: the role of mitochondrial respiration

Measurements of net fluxes of CO₂ and O₂ from leaves and chlorophyll a fluorescence were used to determine the role of mitochondrial respiration during nitrate (NO₃^–) assimilation in both a C₃ (wheat) and a C₄ (maize) plant. Changes in the assimilatory quotient (net CO₂ consumed over net O₂ evolved) when the nitrogen source was shifted from NO₃^– to NH₄⁺ (ΔAQ) provided a measure of shoot NO₃^– assimilation. According to this measure, elevated CO₂ inhibited NO₃^– assimilation in wheat but not maize. Net O₂ exchange under ambient CO₂ concentrations increased in wheat plants receiving NO₃^– instead of NH₄⁺, but gross O₂ evolution from the photosynthetic apparatus (J_O2) was insensitive to nitrogen source. Therefore, O₂ consumption within wheat photosynthetic tissue (ΔΟ₂), the difference between J_O2 and net O₂ exchange, decreased during NO₃^– assimilation. In maize, NO₃^– assimilation was insensitive to changes in intercellular CO₂ concentration (C_i); nonetheless, ΔΟ₂ at low C_i values was significantly higher in NO₃^–‐fed than in NH₄⁺‐fed plants. Changes in O₂ consumption during NO₃^– assimilation may involve one or more of the following processes: (a) Mehler ascorbate peroxidase (MAP) reactions; (b) photorespiration; or (c) mitochondrial respiration. The data presented here indicates that in wheat, the last process, mitochondrial respiration, is decreased during NO₃^– assimilation. In maize, NO₃^– assimilation appears to stimulate mitochondrial respiration when photosynthetic rates are limiting.     

Nitrate photo-assimilation in tomato leaves under short-term exposure to elevated carbon dioxide and low oxygen

The role of photorespiration in the foliar assimilation of nitrate (NO₃^–) and carbon dioxide (CO₂) was investigated by measuring net CO₂ assimilation, net oxygen (O₂) evolution, and chlorophyll fluorescence in tomato leaves (Lycopersicon esculentum). The plants were grown under ambient CO₂ with ammonium nitrate (NH₄NO₃) as the nitrogen source, and then exposed to a CO₂ concentration of either 360 or 700 µmol mol^?1, an O₂ concentration of 21 or 2%, and either NO₃^– or NH₄⁺ as the sole nitrogen source. The elevated CO₂ concentration stimulated net CO₂ assimilation under 21% O₂ for both nitrogen treatments, but not under 2% O₂. Under ambient CO₂ and O₂ conditions (i.e. 360 µmol mol^?1 CO₂, 21% O₂), plants that received NO₃^– had 11–13% higher rates of net O₂ evolution and electron transport rate (estimated from chlorophyll fluorescence) than plants that received NH₄⁺. Differences in net O₂ evolution and electron transport rate due to the nitrogen source were not observed at the elevated CO₂ concentration for the 21% O₂ treatment or at either CO₂ level for the 2% O₂ treatment. The assimilatory quotient (AQ) from gas exchange, the ratio of net CO₂ assimilation to net O₂ evolution, indicated more NO₃^– assimilation under ambient CO₂ and O₂ conditions than under the other treatments. When the AQ was derived from gross O₂ evolution rates estimated from chlorophyll fluorescence, no differences could be detected between the nitrogen treatments. The results suggest that short‐term exposure to elevated atmospheric CO₂ decreases NO₃^– assimilation in tomato, and that photorespiration may help to support NO₃^– assimilation.     

Soil nitrogen transformations under Populus tremuloides, Betula papyrifera and Acer saccharum following 3 years exposure to elevated CO2 and O3

Increases in atmospheric CO₂ and tropospheric O₃ may affect forest N cycling by altering plant litter production and the availability of substrates for microbial metabolism. Three years following the establishment of our free‐air CO₂–O₃ enrichment experiment, plant growth has been stimulated by elevated CO₂ resulting in greater substrate input to soil; elevated O₃ has counteracted this effect. We hypothesized that rates of soil N cycling would be enhanced by greater plant productivity under elevated CO₂, and that CO₂ effects would be dampened by O₃. We found that elevated CO₂ did not alter gross N transformation rates. Elevated O₃ significantly reduced gross N mineralization and microbial biomass N, and effects were consistent among species. We also observed significant interactions between CO₂ and O₃: (i) gross N mineralization was greater under elevated CO₂ (1.0 mg N kg^?1 day^?1) than in the presence of both CO₂ and O₃ (0.5 mg N kg^?1 day^?1) and (ii) gross NH₄⁺ immobilization was also greater under elevated CO₂ (0.8 mg N kg^?1 day^?1) than under CO₂ plus O₃ (0.4 mg N kg^?1 day^?1). We used a laboratory ¹⁵N tracer method to quantify transfer of inorganic N to organic pools. Elevated CO₂ led to greater recovery of NH₄⁺‐¹⁵N in microbial biomass and corresponding lower recovery in the extractable NO₃^? pool. Elevated CO₂ resulted in a substantial increase in NO₃^?‐¹⁵N recovery in soil organic matter. We observed no O₃ main effect and no CO₂ by O₃ interaction effect on ¹⁵N recovery in any soil pool. All of the above responses were most pronounced beneath Betula papyrifera and Populus tremuloides, which have grown more rapidly than Acer saccharum. Although elevated CO₂ has increased plant productivity, the resulting increase in plant litter production has yet to overcome the influence of the pre‐existing pool of soil organic matter on soil microbial activity and rates of N cycling. Ozone reduces plant litter inputs and also appears to affect the composition of plant litter in a way that reduces microbial biomass and activity.     

Interaction between atmospheric and pedospheric nitrogen nutrition in spruce (Picea abies L. Karst) seedlings

The effect of NO₂ fumigation on root N uptake and metabolism was investigated in 3-month-old spruce (Picea abics L. Karst) seedlings. In a first experiment, the contribution of NO₂ to the plant N budget was measured during a 48 h fumigation with 100mm³m^?3 NO₂. Plants were pre-treated with various nutrient solutions containing NO₂ and NH₄⁺, NO₃^? only or no nitrogen source for 1 week prior to the beginning of fumigation. Absence of NH₄⁺ in the solution for 6d led to an increased capacity for NO₃^? uptake, whereas the absence of both ions caused a decrease in the plant N concentration, with no change in NO₃^? uptake. In fumigated plants, NO₂ uptake accounted for 20–40% of NO₃^? uptake. Root NO₃^? uptake in plants supplied with NH₄⁺plus NO₃^? solutions was decreased by NO₂ fumigation, whereas it was not significantly altered in the other treatments. In a second experiment, spruce seedlings were grown on a solution containing both NO₂ and NH₄⁺ and were fumigated or not with 100mm³m^?3 NO₂ for 7 weeks. Fumigated plants accumulated less dry matter, especially in the roots. Fluxes of the two N species were estimated from their accumulations in shoots and roots, xylem exudate analysis and ¹⁵N labelling. Root NH₄⁺ uptake was approximately three times higher than NO₃^? uptake. Nitrogen dioxide uptake represented 10–15% of the total N budget of the plants. In control plants, N assimilation occurred mainly in the roots and organic nitrogen was the main form of N transported to the shoot. Phloem transport of organic nitrogen accounted for 17% of its xylem transport. In fumigated plants, neither NO₃^? nor NH₄⁺ accumulated in the shoot, showing that all the absorbed NO₂ was assimilated. Root NO₃^? reduction was reduced whereas organic nitrogen transport in the phloem increased by a factor of 3 in NO₂-fimugated as compared with control plants. The significance of the results for the regulation of whole-plant N utilization is discussed.     

INTERACTIONS BETWEEN LIGHT,NH4+, AND CO2 IN BUOYANCY REGULATION OF ANABAENA FLOS-AQUAE (CYANOPHYCEAE)1

Buoyancy of the gas-vacuolate alga Anabaena flosaquae Brébisson was measured under various levels of light, NH₄⁺, and CO₂. At high irradiance (50 μE · m^?2·^?1) the alga was non-buoyant regardless of the availability of CO₂ and NH₄⁺. At low irradiance (≤10 μE · m ^?2· s^?1) buoyancy was controlled by the availability of NH₄⁺ and CO₂. When NH₄⁺ was abundant, algal buoyancy was high over a wide range of CO₂ concentrations. In the absence of NH₄⁺, algal buoyancy was reduced at high CO₂ concentrations, however as the CO₂ concentration declined below about 5 μmol · L^?1, algal buoyancy increased. These results help explain why gas vacuolate, nitrogen-fixing blue-green algae often form surface blooms in eutrophic lakes.     

Nitrogen Acquisition from Atmospheric NH3 by Lolium perenne

Ammonia (NH₃) is the third most abundant N species in the atmosphere and, due to various natural and anthropogenic sources, can reach high concentrations in some areas. While some plants show effects of toxicity, others are capable of using this N-form and grow well without any utilization of soil-N. Acquisition of atmospheric NH₃ will affect the acid-base balance of the plants as absorption and dissolution causes an alkalinisation (production of OH^?) and assimilation of NH₃ results in an acidification (generation of H⁺). As there is only a limited capacity for biochemical disposal of excess H⁺ in shoots, pH regulation may involve H⁺/OH^? extrusion into the media via roots and transport of (in)organic ions between roots and above-ground parts of the plant. Our aim therefore was to assess NH₃ acquisition by Lolium perenne and to study the effects of gas phase NH₃ on growth, acid-base balance and mineral composition of the plants. The experiments therefore included application of a range of ¹⁴NH₃ to the shoots and of ¹⁵N as NO₃^?, NH₄⁺ or NH₄NO₃ to the roots, from which the amount of gas phase NH₃ acquisition could be quantified. Analysis of the mineral composition provided data for calculation of acid-base balance as well as for water use efficiencies of the plants. The results indicate that over the range of NH₃ supplied, plants from all treatments could utilize gas-phase NH₃ as demonstrated by increases in growth and in N and C use efficiencies. Plants receiving NO₃^? via their roots had a higher capacity to use gaseous NH₃ than those growing with NH₄⁺. NH₃ assimilation in shoots reduced both the acid load with NH₄⁺ nutrition and the alkaline load with NO₃^? supply to the roots. The results of the experiments are discussed in relation to possible acid-base regulation mechanisms of the whole plant.     

Effect of elevated CO2 concentration and nitrate: ammonium ratios on gas exchange and growth of cassava (Manihot esculenta Crantz)

Aims

This study evaluated how different nitrogen forms affect growth and photosynthetic responses of cassava to CO₂ concentration.

Methods

Cassava was grown in 14-L pots in a greenhouse at 390 or 750 ppm of CO₂. Three nitrogen treatments were applied: (a) 12?mM NO₃ ^?, (b) 6?mM NO₃ ^??+?6?mM NH₄ ⁺, and (c) 12?mM NH₄ ⁺.

Results

Thirty-six days after treatments began, plants grown under elevated CO₂ and fertilized only with NO₃ ^? (750_NO₃ ^?) had photosynthetic rates similar to plants grown under 390_NO₃ ^?, indicating significant photosynthetic acclimation to CO₂. In contrast, photosynthetic rates at elevated CO₂ increased as NH₄ ⁺ increased in the nutrient solution, such that photosynthetic acclimation was reduced for plants fertilized with only NH₄ ⁺. However, this positive effect of NH₄ ⁺ on photosynthesis was not observed in more advanced growth stages, and the toxic effects of NH₄ ⁺ severely reduced total dry mass for these plants measured at the end of the experiment.

Conclusions

Our results indicate that cassava will respond with increased biomass accumulation in response to raising atmospheric CO₂ levels, and that N form can have an important impact on the photosynthetic response. However, the positive effect of NH₄ ⁺ fertilization on cassava photosynthetic CO₂ response eventually led to a toxicity problem that reduced biomass production. The challenge is to determine how to manage NH₄ ⁺ fertilization so that the photosynthetic benefit observed in the initial phase may persist throughout the crop cycle.