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).     

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.     

Effects of elevated atmospheric carbon dioxide on amino acid and NH4+-N cycling in a temperate pine ecosystem

Rising atmospheric carbon dioxide (CO₂) is expected to increase forest productivity, resulting in greater carbon (C) storage in forest ecosystems. Because elevated atmospheric CO₂ does not increase nitrogen (N) use efficiency in many forest tree species, additional N inputs will be required to sustain increased net primary productivity (NPP) under elevated atmospheric CO₂. We investigated the importance of free amino acids (AAs) as a source for forest N uptake at the Duke Forest Free Air CO₂ Enrichment (FACE) site, comparing its importance with that of better‐studied inorganic N sources. Potential proteolytic enzyme activity was monitored seasonally, and individual AA concentrations were measured in organic horizon extracts. Potential free AA production in soils ranged from 190 to 690 nmol N g⁻¹ h⁻¹ and was greater than potential rates of soil NH₄⁺ production. Because of this high potential rate of organic N production, we determined (1) whether intact AA uptake occurs by Pinus taeda L., the dominant tree species at the FACE site, (2) if the rate of cycling of AAs is comparable with that of ammonium (NH₄⁺), and (3) if atmospheric CO₂ concentration alters the aforementioned N cycling processes. A field experiment using universally labeled ammonium (¹⁵NH₄⁺) and alanine (¹³C₃H₇¹⁵NO₂) demonstrated that ¹⁵N is more readily taken up by plants and heterotrophic microorganisms as NH₄⁺. Pine roots and microbes take up on average 2.4 and two times as much NH₄^{+ 15}N compared with alanine ¹⁵N 1 week after tracer application. N cycling through soil pools was similar for alanine and NH₄⁺, with the greatest ¹⁵N tracer recovery in soil organic matter, followed by microbial biomass, dissolved organic N, extractable NH₄⁺, and fine roots. Stoichiometric analyses of ¹³C and ¹⁵N uptake demonstrated that both plants and soil microorganisms take up alanine directly, with a ¹³C : ¹⁵N ratio of 3.3 : 1 in fine roots and 1.5 : 1 in microbial biomass. Our results suggest that intact AA (alanine) uptake contributes substantially to plant N uptake in loblolly pine forests. However, we found no evidence supporting increased recovery of free AAs in fine roots under elevated CO₂, suggesting plants will need to acquire additional N via other mechanisms, such as increased root exploration or increased N use efficiency.     

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.     

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.     

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.     

Ammonium, nitrate, and proton fluxes along the maize root

Ion-selective microelectrodes were used to measure NH₄⁺, NO₃^– and H⁺ fluxes along the primary root of maize seedlings. Plants were exposed to nutrient solutions containing NH₄⁺, NO₃^– or both ions. Nitrogen fluxes along the root varied substantially among the different treatments. Net NH₄⁺ and NO₃^– uptake and H⁺ extrusion were low at the very apex of the root and generally increased in the more basal regions. In the absence of nitrogen or in the presence of NO₃^– alone, net H⁺ uptake (and root surface alkalinization) occurred at the root tip (0–1 mm), whereas net H⁺ extrusion occurred in all other regions. In the presence of NH₄⁺ alone, a dramatic increase in net H⁺ extrusion was detected in all regions except for the region 6–11 mm from the apex. In contrast, when NO₃^– alone was supplied, net H⁺ extrusion was depressed at all locations except for the tip (0–1 mm). When both NH₄⁺ and NO₃^– were supplied, NO₃^– uptake was suppressed at all locations while net H⁺ extrusion was increased relative to NO₃^– alone. The capacities to absorb NH₄⁺ and NO₃^– at the tip were similar, as indicated by flux rates when NH₄⁺ or NO₃^– were supplied as sole sources, but when supplied together, net NO₃^– uptake was half that of net NH₄⁺ uptake, indicating that NH₄⁺ may satisfy the nitrogen requirements of the poorly vascularized apical tissue in the most energy-efficient way. The high spatial resolution of the measurements enabled us to establish that acidification in the root expansion zone is maintained regardless of nitrogen source.     

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.     

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.     

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.     

Regenerating temperate forest mesocosms in elevated CO2: belowground growth and nitrogen cycling

The response of temperate forest ecosystems to elevated atmospheric CO₂ concentrations is important because these ecosystems represent a significant component of the global carbon cycle. Two important but not well understood processes which elevated CO₂ may substantially alter in these systems are regeneration and nitrogen cycling. If elevated CO₂ leads to changes in species composition in regenerating forest communities then the structure and function of these ecosystems may be affected. In most temperate forests, nitrogen appears to be a limiting nutrient. If elevated CO₂ leads to reductions in nitrogen cycling through increased sequestration of nitrogen in plant biomass or reductions in mineralization rates, long-term forest productivity may be constrained. To study these processes, we established mesocosms of regenerating forest communities in controlled environments maintained at either ambient (375 ppm) or elevated (700 ppm) CO₂ concentrations. Mesocosms were constructed from intact monoliths of organic forest soil. We maintained these mesocosms for 2 years without any external inputs of nitrogen and allowed the plants naturally present as seeds and rhizomes to regenerate. We used ¹⁵N pool dilution techniques to quantify nitrogen fluxes within the mesocosms at the end of the 2 years. Elevated atmospheric CO₂ concentration significantly affected a number of plant and soil processes in the experimental regenerating forest mesocosms. These changes included increases in total plant biomass production, plant C/N ratios, ectomycorrhizal colonization of tree fine roots, changes in tree fine root architecture, and decreases in plant NH₄ ⁺ uptake rates, gross NH₄ ⁺ mineralization rates, and gross NH₄ ⁺ consumption rates. In addition, there was a shift in the relative biomass contribution of the two dominant regenerating tree species; the proportion of total biomass contributed by white birch (Betula papyrifera) decreased and the proportion of total biomass contributed by yellow birch (B. alleghaniensis) increased. However, elevated CO₂ had no significant effect on the total amount of nitrogen in plant and soil microbial biomass. In this study we observed a suite of effects due to elevated CO₂, some of which could lead to increases in potential long term growth responses to elevated CO₂, other to decreases. The reduced plant NH₄ ⁺ uptake rates we observed are consistent with reduced NH₄ ⁺ availability due to reduced gross mineralization rates. Reduced NH₄ ⁺ mineralization rates are consistent with the increases in C/N ratios we observed for leaf and fine root material. Together, these data suggest the positive increases in plant root architectural parameters and mycorrhizal colonization may not be as important as the potential negative effects of reduced nitrogen availability through decreased decomposition rates in a future atmosphere with elevated CO₂. Received: 10 January 1997 / Accepted: 25 July 1997     

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.     

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.     

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.     

A field method of determining NH 4 + and NO 3 - uptake kinetics in intact roots: Effects of CO2 enrichment on trees and crop species

Models describing plant and ecosystem N cycles require an accurate assessment of root physiological uptake capacity for NH ₄ ⁺ and NO ₃ ^- under field conditions. Traditionally, rates of ion uptake in field-grown plants are determined by using excised root segments incubated for a short period in an assay solution containing N either as a radioactive or stable isotope tracer (e.g., ³⁶ClO₃ as a NH ₄ ⁺ analogue, ¹⁴CH₃NH₃ as an NO ₃ ^- analogue or ¹⁵NH ₄ ⁺ and ¹⁵NO ₃ ^- ). Although reliable, this method has several drawbacks. For example, in addition to radioactive safety issues, purchase and analysis of radioactive and stable isotopes is relatively expensive and can be a major limitation. More importantly, because excision effectively interrupts exchange of compounds between root and shoot (e.g., carbohydrate supply to root and N transport to shoot), the assay must be conducted quickly to avoid such complications. Here we present a novel field method for simultaneous measurements of NH ₄ ⁺ and NO ₃ ^- uptake kinetics in intact root systems. The application of this method is demonstrated using two tree species; red maple (Acer rubrum) and sugar maple (Acer saccharum) and two crop species soybean (Glycine max) and sorghum (Sorghum bicolor). Plants were grown in open-top chambers at either ambient or elevated levels of atmospheric CO₂ at two separate US national sites involved in CO₂ research. Absolute values of net uptake rates and the kinetic parameters determined by our method were found to be in agreement with the literature reports. Roots of the crop species exhibited a greater uptake capacity for both N forms relative to tree species. Elevated CO₂ did not significantly affect kinetics of N uptake in species tested except in red maple where it increased root uptake capacity, V, for NH ₄ ⁺ . The application, reliability, advantages and disadvantages of the method are discussed in detail. This revised version was published online in June 2006 with corrections to the Cover Date.     

Influence of elevated CO2 and nitrogen nutrition on photosynthesis and nitrate photo-assimilation in maize (Zea mays L.)

Measurements of CO₂ and O₂ gas exchange and chlorophyll a fluorescence were used to test the hypothesis that elevated atmospheric CO₂ inhibits nitrate (NO₃ ^– ) photo‐assimilation in the C₄ plant, maize (Zea mays L.). The assimilatory quotient (AQ), the ratio of net CO₂ assimilation to net O₂ evolution, decreases as NO₃ ^– photo‐assimilation increases so that the difference in AQ between the ammonium‐ and nitrate‐fed plants (ΔAQ) provided an in planta estimate of NO₃ ^– photo‐assimilation. In fully expanded maize leaves, NO₃ ^– photo‐assimilation was detectable only under high light and was not affected by CO₂ treatments. Furthermore, CO₂ assimilation and O₂ evolution were higher under NO₃ ^– than ammonia (NH₄⁺) regardless of CO₂ levels. In conclusion, NO₃ ^– photo‐assimilation in maize primarily occurred at high light when reducing equivalents were presumably not limiting. Nitrate photo‐assimilation enhanced C₄ photosynthesis, and in contrast to C₃ plants, elevated CO₂ did not inhibit foliar NO₃^– photo‐assimilation.     

Altered cell wall properties are responsible for ammonium‐reduced aluminium accumulation in rice roots

The phytotoxicity of aluminium (Al) ions can be alleviated by ammonium (NH₄⁺) in rice and this effect has been attributed to the decreased Al accumulation in the roots. Here, the effects of different nitrogen forms on cell wall properties were compared in two rice cultivars differing in Al tolerance. An in vitro Al‐binding assay revealed that neither NH₄⁺ nor NO₃^? altered the Al‐binding capacity of cell walls, which were extracted from plants not previously exposed to N sources. However, cell walls extracted from NH₄⁺‐supplied roots displayed lower Al‐binding capacity than those from NO₃^?‐supplied roots when grown in non‐buffered solutions. Fourier‐transform infrared microspectroscopy analysis revealed that, compared with NO₃^?‐supplied roots, NH₄⁺‐supplied roots possessed fewer Al‐binding groups (‐OH and COO‐) and lower contents of pectin and hemicellulose. However, when grown in pH‐buffered solutions, these differences in the cell wall properties were not observed. Further analysis showed that the Al‐binding capacity and properties of cell walls were also altered by pHs alone. Taken together, our results indicate that the NH₄⁺‐reduced Al accumulation was attributed to the altered cell wall properties triggered by pH decrease due to NH₄⁺ uptake rather than direct competition for the cell wall binding sites between Al³⁺ and NH₄⁺.     

Complementarity in the use of nitrogen forms in a temperate broad-leaved mixed forest

Background: The complementary use of different forms of soil nitrogen (N) might lead to a higher productivity of mixed forests than monocultures, but convincing evidence for temperate mixed forests is scarce.

Aims: We searched for species differences in N uptake rates and the preference for NH₄⁺, NO₃^? or glycine among five temperate broad?leaved tree species (Acer pseudoplatanus, Carpinus betulus, Fagus sylvatica, Fraxinus excelsior, Tilia cordata) in a mature mixed stand.

Methods: ¹⁵N tracer was added to the soil and its accumulation in fine root biomass was analysed after 10 min, 1 h and 1 d.

Results: The estimated root uptake rates of the species were in the range of 5–46 µg N g^?1 root h^?1 for NH₄⁺, 6–86 µg N g^?1 h^?1 for NO₃^? and 4–29 µg N g^?1 h^?1 for glycine during the first hour after tracer application. Carpinus, Tilia and Acer tended to prefer NH₄⁺ over NO₃^?, while Fraxinus showed equal preference for both N forms and Fagus seemed to prefer NO₃^?.

Conclusions: The five co-existing tree species differed in uptake rates and partly in their N form preference, but complementarity in the use of different N forms seems to be of minor importance in this forest because tree species appear to be rather flexible in their N form use.     

The role of N-remobilisation and the uptake of NH4+ and NO3- by Lolium perenne L. in laminae growth following defoliation under field conditions

Several studies have previously shown that shoot removal of forage species, either by cutting or herbivore grazing, results in a large decline in N uptake (60%) and/or N₂ fixation (80%). The source of N used for initial shoot growth following defoliation relies mainly on mobilisation of N reserves from tissues remaining after defoliation. To date, most studies investigating N-mobilisation have been conducted, with isolated plants grown in controlled conditions. The objectives of this study were for Lolium perenne L., grown in a dense canopy in field conditions, to determine: 1) the contribution of N-mobilisation, NH₄ ⁺ uptake and NO₃ ^- uptake to growing shoots after defoliation, and 2) the contribution of the high (HATS) and low (LATS) affinity transport systems to the total plant uptake of NH₄ ⁺ and NO₃ ^-. During the first seven days following defoliation, decreases in biomass and N-content of roots (34% and 47%, respectively) and to a lesser extent stubble (18% and 43%, respectively) were observed, concomitant with mobilisation of N to shoots. The proportion and origin of N used by shoots (derived from reserves or uptake) was similar to data reported for isolated plants. Both HATS and LATS contributed to the total root uptake of NH₄ ⁺ and NO₃ ^-. The V_max of both the NH₄ ⁺ and NO₃ ^- HATS increased as a function of time after defoliation, and both HATS systems were saturated by substrate concentrations in the soil at all times. The capacity of the LATS was reduced as soil NO₃ ^- and NH₄ ⁺ concentrations decreased following defoliation. Data from ¹⁵N uptake by field-grown plants, and uptake rates of NH₄ ⁺ and NO₃ ^- estimated by excised root bioassays, were significantly correlated, though uptake was over-estimated by the later method. The results are discussed in terms of putative mechanisms for regulating N uptake following severe defoliation. This revised version was published online in June 2006 with corrections to the Cover Date.     

Redefining the dose of the entomopathogenic fungus Metarhizium brunneum (Ascomycota,Hypocreales) to increase Fe bioavailability and promote plant growth in calcareous and sandy soils

Background and aims

Plants differ in their ability to use different nitrogen (N) chemical forms, these differences can be related to their ecology and drive community structure. The capacity to uptake intact organic N has been observed in plants of several ecosystems. However, soil organic N uptake by Mediterranean plants is unknown despite organic N being abundant in Mediterranean ecosystems. We compare the uptake of different N forms in two widespread coexisting Mediterranean forest trees with contrasting ecophysiological characteristics: Quercus ilex and Pinus halepensis.

Methods

To estimate root uptake rate of each N form we used equimolar solutions (1 mM N) of ¹⁵NO₃ ^?, ¹⁵NH₄ ⁺ and ¹⁵N-¹³C glycine.

Results

NH₄ ⁺ and glycine were taken up at a similar rate, but faster than NO₃ ^? in both species. Intact dual labeled glycine was found in both species, demonstrating that both species can absorb intact organic N.

Conclusions

Despite their ecological differences, both species had similar preference for N forms suggesting no fundamental niche complementarity for N uptake. The higher preference for NH₄ ⁺ and glycine over NO₃ ^? possibly reflects adaptation to the differing proportions of N forms in Mediterranean soils.