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.     

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.     

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.     

Effect of nitrogen form and phosphorus source on the growth,nutrient uptake and rhizosphere soil property of Camellia sinensis L.

The effects of nitrogen form and phosphorus source on the growth, nutrient uptake and rhizosphere soil property of tea (Camellia sinensis L.) were investigated in a pot experiment. The experiment was performed with a compartmental cropping device, which enables the collection of rhizosphere soil at defined distances from the root of tea plant. Nitrogen was supplied as nitrate or ammonium in combination with soluble phosphorus as Ca(H₂PO₄)₂ or insoluble P as rock phosphate. The leaf dry matter production of tea was significantly greater in the treatments with NH₄ ⁺ than NO₃ ^-, whereas dry matter production of root and stem was not significantly affected. Addition of phosphorus as either source did not influence the dry matter production. The concentrations of K in root, Mg and Ca in both the shoot and root supplied with NO₃ ^- were significantly higher than in NH₄ ⁺ and influence of P sources was minor. On the contrary, Al and Mn concentrations were significantly larger in NH₄ ^--fed plants which could be attributed to remarkably increased availability of Al and Mn caused by acidification of the rhizosphere soil (the first 1-mm soil section from the root surface) with NH₄–N nutrition. The concentration of N in shoot was also significantly higher in NH₄- than in NO₃-fed plants, indicating higher use efficiency of NH₄–N. Whatever the phosphate source, rhizosphere pH declined in ammonium compared to in nitrate treatment. The pH decrease was much larger when no P or soluble P were applied and reached 0.85–1.30 units which extended to 3–5 mm away from the root surface. Exchangeable acidity, content of exchangeable Al and Mn were also considerably higher in the rhizosphere soils of NH₄ ⁺ fed tea plants. Significant amounts of P dissolved from rock phosphate accumulated in rhizosphere of NH₄ ⁺, not NO₃ ^-, suggesting that the dissolution of rock phosphate was induced by the proton excreted by tea root fed with ammonium. With soluble P addition, shoot and root P concentrations were greater in NH₄ ⁺ than in NO₃ ^- treatment and it appeared that this difference could not be sufficiently explained by the available P content in soil which was only slightly higher in NH₄ ⁺ treatment. With rock phosphate addition, the shoot and root P concentrations were hardly affected by nitrogen form, although the available P content was much higher and accumulated in the rhizosphere soil supplied with ammonium. The reason for this was discussed with regard to the inter-relationship of Al with P uptake. This revised version was published online in June 2006 with corrections to the Cover Date.     

Simultaneous measurement of ammonium,nitrate and proton fluxes along the length of eucalypt roots

Knowledge of the preferred source of N for Eucalyptus nitens will lead to improved fertiliser management practices in plantations. Ion selective microelectrodes were used non-invasively to measure simultaneously net fluxes of NH₄ ⁺, NO₃ ^– and H⁺ along the tap root of solution-cultured E. nitens. Measurements were conducted in solutions containing 100 m NH₄NO₃. The pattern of fluxes was such that there was a large influx of NH₄ ⁺, a smaller influx of NO₃ ^– and large H⁺ efflux. The ratio of these fluxes was constant, according to the ratio 3:1:–6 (NH₄ ⁺:NO₃ ^–:H⁺). Within the region 20–60 mm from the root apex of E. nitens seedlings there was spatial and temporal variation in fluxes but flux patterns remained constant. Root hair density did not affect fluxes nor did proximity to lateral roots. Variation was less than that found in previous studies of localised root fluxes using similar high-resolution measurement techniques. It was concluded that small-scale spatial variation in fluxes may have confounded previous studies. There were associations between fluxes of all three ions, the strongest associations being between NH₄ ⁺ and H⁺, and NH₄ ⁺ and NO₃ ^–. Overall, these results are consistent with NH₄ ⁺ being the preferred source N for E. nitens.     

Influence of Nitrate and Ammonium Nutrition on the Uptake, Assimilation, and Distribution of Nutrients in Ricinus communis

Ricinus communis L. plants were grown in nutrient solutions in which N was supplied as NO₃⁻ or NH₄⁺, the solutions being maintained at pH 5.5. In NO₃⁻-fed plants excess nutrient anion over cation uptake was equivalent to net OH⁻ efflux, and the total charge from NO₃⁻ and SO₄²⁻ reduction equated to the sum of organic anion accumulation plus net OH⁻ efflux. In NH₄⁺-fed plants a large H⁺ efflux was recorded in close agreement with excess cation over anion uptake. This H⁺ efflux equated to the sum of net cation (NH₄⁺ minus SO₄²⁻) assimilation plus organic anion accumulation. In vivo nitrate reductase assays revealed that the roots may have the capacity to reduce just under half of the total NO₃⁻ that is taken up and reduced in NO₃⁻-fed plants. Organic anion concentration in these plants was much higher in the shoots than in the roots. In NH₄⁺-fed plants absorbed NH₄⁺ was almost exclusively assimilated in the roots. These plants were considerably lower in organic anions than NO₃⁻-fed plants, but had equal concentrations in shoots and roots. Xylem and phloem saps were collected from plants exposed to both N sources and analyzed for all major contributing ionic and nitrogenous compounds. The results obtained were used to assist in interpreting the ion uptake, assimilation, and accumulation data in terms of shoot/root pH regulation and cycling of nutrients.     

Nitrate and ammonium as nitrogen sources for lupins prior to nodulation

Two cultivars of Lupinus angustifolius L. were grown in a glasshouse in solutions containing NO₃ ^-, NH₄ ⁺ or NH₄NO₃ with a total nitrogen concentration of 2.8 M m^-3 in each treatment. One cultivar chosen (75A-258) was relatively tolerant to alkaline soils whereas the other (Yandee) was intolerant to alkalinity. Controlled experiments were used to assess the impact of cationic vs. anionic forms of nitrogen on the relative performance of these cultivars. Relative growth rates (dry weight basis) were not significantly different between the two cultivars when grown in the presence of NO₃ ^-, NH₄ ⁺ or NH₄NO₃. However, when NO₃ ^- was supplied, there was a modest decline in relative growth rates in both cultivars over time. When plants grown on the three sources of nitrogen for 9 days were subsequently supplied with ¹⁵NH₄NO₃ or NH₄ ¹⁵NO₃ for 30 h, NH₄ ⁺ uptake was generally twice as fast as NO₃ ^- uptake, even for plants grown in the presence of NO₃ ^-. Low rates of NO₃ ^- uptake accounted for the decrease in growth rates over time when plants were grown in the presence of NO₃ ^-. It is concluded that the more rapid growth of 75A-258 than Yandee in alkaline conditions was not due to preferential uptake of NH₄ ⁺ and acidification of the external medium. In support of this view, acidification of the root medium was not significantly different between cultivars when NH₄ ⁺ was the sole nitrogen source.     

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.     

Ammonium uptake in Lemna gibba G 1, related membrane potential changes, and inhibition of anion uptake

In N-starved (?N) fronds of Lemna gibba L. G 1, NH₄⁺ uptake rates were several-fold those of NO₃^?-supplied (+N) fronds. NO₃^?, uptake in +N-plants was slow and not inhibited by addition of NH₄⁺. However, in ?N-plants with higher NO₃^? and still higher NH₄⁺ uptake rates, addition of NH₄⁺ immediately reduced the NO₃^? uptake rates to about one third until the NH₄⁺ was consumed. The membrane potential (E_m) decreased immediately upon addition of NH₄⁺ in all fronds, but whereas depolarisation was moderate and transient in +N-plants, it was strong, up to 150 mV, in N-starved plants, where E_m remained at the level of the K⁺ diffusion potential (E_D) until NH₄⁺ was removed. In N-starved plants NH₄⁺ uptake and membrane depolarisation showed the same concentration dependence, except for an apparent linear component for uptake. Phosphate uptake was inhibited by NH₄⁺ similarly to NO₃^? uptake, but only in P- and N-starved plants, not after mere P starvation. Influx of NO₃^? and H₂PO ₄^? into the negatively charged cells of Lemna is mediated by anion/H⁺ cotransport, but NH₄⁺ influx can follow the electrochemical gradient. Its saturating component may reflect a carrier-mediated NH₄⁺ uniport, the linear component diffusion of NH₄⁺ or NH₃. Inhibition of anion/H⁺ cotransport by high NH₄⁺ influx rates may be due to loss of the proton-driving force, Δμ?H⁺, across the plasmalemma. Reversible inhibition by NH₄⁺ of the H⁺ extrusion pump may contribute to the finding that Δμ?H⁺ cannot be reconstituted in the presence of higher NH₄⁺ concentrations.     

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.     

Ion currents and the nitrogen status of roots of Hordeum vulgare and non-nodulated Trifolium repens

Abstract. Profiles of self-generated ion currents associated with the growing primary root tips of intact Hordeum vulgare L. and Trifolium repens L. (nonnodulated) seedlings were measured using a highly sensitive vibrating electrode in media containing NH⁺₄ or NO^-₃, and compared to control roots growing in nitrogen free media. Under these three nutrient regimes, positive current entered the root at regions corresponding to the meristematic tissues and main elongation zones of root tips and left from the mature root tissues. Mapping the surface of the roots with a pH-sensitive microelectrode revealed regions of external alkalinity where positive electrical current entered the root, and external acidity where positive current exited. The correlation between pH-profile and the pattern of ion current generation in these experiments suggests that H⁺ ions were responsible for carrying the bulk of the root-generated current. Assimilation of NHJ results in net H⁺ extrusion while assimilation of NO^-₃, results in net OH^-₃ efflux. Growth on NH⁺₄, as compared to growth on NO^-₃, stimulated the magnitude of the electrical current but did not affect significantly the growth rate of the roots. However, despite the differing stresses on internal pH regulation that arise due to growth on the two exogenous forms of combined nitrogen, the current profiles were qualitatively similar under the different conditions that were examined. The role of the circulating proton current is not yet known; however, the constancy of the current profile under different nutrient regimes sustains the hypothesis that the current may have a role in the regulation of root polarity.     

NH(4)(+)-stimulated low-K(+) uptake is associated with the induction of H(+) extrusion by the plasma membrane H(+)-ATPase in sorghum roots under K(+) deficiency

The effect of external inorganic nitrogen and K⁺ content on K⁺ uptake from low-K⁺ solutions and plasma membrane (PM) H⁺-ATPase activity of sorghum roots was studied. Plants were grown for 15 days in full-nutrient solutions containing 0.2 or 1.4 mM K⁺ and inorganic nitrogen as NO₃^-, NO₃^-/NH₄⁺ or NH₄⁺ and then starved of K⁺ for 24, 48 and 72 h. NH₄⁺ in full nutrient solution significantly affected the uptake efficiency and accumulation of K⁺, and this effect was less pronounced at the high K⁺ concentration. In contrast, the translocation rate of K⁺ to the shoot was not altered. Depletion assays showed that plants grown with NH₄⁺ more efficiently depleted the external K⁺ and reached higher initial rates of low-K⁺ uptake than plants grown with NO₃^-. One possible influence of K⁺ content of shoot, but not of roots, on K⁺ uptake was evidenced. Enhanced K⁺-uptake capacity was correlated with the induction of H⁺ extrusion by PM H⁺-ATPase. In plants grown in high K⁺ solutions, the increase in the active H⁺ gradient was associated with an increase of the PM H⁺-ATPase protein concentration. In contrast, in plants grown in solutions containing 0.2 mM K⁺, only the initial rate of H⁺-pumping and ATP hydrolysis were affected. Under these conditions, two specific isoforms of PM H⁺-ATPase were detected, independent of the nitrogen source and deficiency period. No change in enzyme activity was observed in NO₃^--grown plants. The results suggest that K⁺ homeostasis in NH₄⁺-grown sorghum plants may be regulated by a high capacity for K⁺ uptake, which is dependent upon the H⁺-pumping activity of PM H⁺-ATPase.     

Effects of pH on ammonium uptake by Typha latifolia L.

The effects of solution pH on NH₄⁺ uptake kinetics and net H⁺ extrusion by Typha latifolia L. were studied during short-term (days) and long-term (weeks) exposure to pH in the range of pH 3.5–8.0. The NH₄⁺ uptake kinetics were estimated from depletion curves using a modified Michaelis-Menten model. T. latifolia was able to grow in solution culture with NH₄⁺ as the sole N source and to withstand a low medium pH for short periods (days). With prolonged exposure (weeks) to pH 3.5, however, the plants showed severe symptoms of stress and stopped growing. The solution pH affected NH₄⁺ uptake kinetics. The affinity for NH₄⁺, as quantified by the half saturation constant (K_1/2) and C_min (the NH₄⁺ concentration at which uptake ceases), decreased with pH. K_1/2 was increased from 7.1 to 19.2 mmol m^?3 and C_min from 2.0 to 5.7 mmol m^?3 by lowering the pH in steps from 8.0 to 3.5. V_max was, however, largely unaffected by pH (～22 μmol h^?1 g^?1 root dry weight). Under prolonged exposure to constant pH, growth rates were highest at PH 5.0 and 6.5. At pH 8.0 growth was slightly depressed and at pH 3.5 growth completely stopped. NH₄⁺ uptake kinetics were similar at pH 5.0, 6.5 and 8.0 whereas at pH 3.5 NH₄⁺ uptake almost completely stopped. The ratio between net H⁺ extrusion and NH₄⁺ uptake decreased significantly at low pH. The adverse effects of low pH on NH₄⁺ uptake kinetics are probably a consequence of a reduced H⁺-ATPase activity and/or an increased re-entry of H⁺ at low pH, and the associated decrease in the electrochemical gradient across the plasma membranes of the root cells.     

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.     

Differential effects of mineral and organic N sources,and of ectomycorrhizal infection by Hebeloma cylindrosporum,on growth and N utilization in Pinus pinaster

The effect of ectomycorrhizal association of Pinus pinaster with Hebeloma cylindrosporum was investigated in relation to the nitrogen source supplied as mineral (NH₄⁺ or NO₃^?) or organic N (L ‐glutamate) and at 5 mol m^?3. Plants were grown for 14 and 16 weeks with mineral and organic N, respectively, and samples were collected during the last 6 weeks of culture. Total fungal biomass was estimated using glucosamine amount and its viability was assessed using the glucosamine to ergosterol ratio. Non‐mycorrhizal plants grew better with NH₄⁺ than with NO₃^? and grew very slowly when supplied with L ‐glutamate. The presence of the fungus decreased the growth of the host plant with mineral N whereas it increased it with L ‐glutamate. Whatever the N source, most of the living fungal biomass was associated with the roots, whereas the main part of the total biomass was assayed outside the root. The form of mineral N did not significantly affect N accumulation rates over the 42 d in control plants. In mycorrhizal plants grown on either N source, the fungal tissues developing outside of the root were always the main N sink. The ectomycorrhizal association did not change ¹⁵NH₄⁺ uptake rate by roots, suggesting that the growth decrease of the host‐plant was related to the carbon cost for fungal growth and N assimilation rather than to a direct effect on NH₄⁺ acquisition. In contrast, in NO₃^?‐grown plants, in addition to draining carbon for NO₃^? reduction the fungus competed with the root for NO₃^? uptake. With NH₄⁺ or NO₃^? feeding, although mycorrhizal association improved N accumulation in shoots, we concluded that it was unlikely that the fungus had supplied the plant with N. In L ‐glutamate‐grown plants, the presence of the fungus increased the proportion of glutamine in the xylem sap and improved both N nutrition and the growth rate of the host plant.     

PHOTOSYNTHETIC RESPONSE OF LAKE PLANKTON TO COMBINED NITROGEN ENRICHMENT1

The complex interplay between photosynthesis and the uptake of nitrogen was investigated in samples from five lakes of different size and trophic state. When enriched with ¹⁵NH₄⁺, the photosynthetic rate was often reduced for 4–5 h in samples believed to be nitrogen deficient. This implies that energy was reallocated from photosynthesis to the uptake and assimilation of N. Stimulation in C uptake at low levels of NH₄⁺ enrichment was followed by a progressive decline with further NH₄⁺ enrichment. On other occasions when ambient NH₄⁺ was undetectable, nutrient regeneration by zooplankton supplied a significant fraction of the required nitrogen. At these times and when the plankton had sufficient available N, there usually was no change in photosynthetic rate with either NH₄⁺ or NO₃^?enrichment. Typically, little NO₃^? was taken up and no photosynthetic response was observed. On two occasions, however, the uptake of NO₃^? was significant due to high NO₃^? and low NH₄⁺ levels early in the season. At one of these times there was a reduction in photosynthesis with NO₃^? enrichment. A further complication was observed when photosynthesis decreased with NH₄⁺ enrichment but increased with NO₃^? enrichment despite negligible NO₃^? uptake. These observations illustrate that the complex metabolism of these two nitrogen sources is not fully understood. At optimum light intensity, C:N uptake ratios, even under NH₄⁺ enrichment, are only sufficient to maintain the cellular C:N ratio unless much of the fixed C is respired or excreted. Three observations suggest that photosynthesis and N uptake are not coupled, (i) Photoinhibition of C uptake, but not N uptake was observed when low light adapted populations are exposed to high light conditions, (ii) The light intensity for maximum N uptake was slightly less than that for carbon. (iii) Dark N uptake was always near 50% of the maximum rate in the light whereas the C uptake was near 2% of P_opt. Certainly, there is an interconnection because dark C uptake was enhanced by NH₄⁺ enrichment.     

Relationships among nitrogen accumulation, nitrogen assimilation and plant growth in chrysanthemums

Relationships among growth, N accumulation and assimilation were investigated in Chrysanthemum morifolium Ramat cv. Fiesta in experiments testing the effects of varying levels of NO^–3₃supply and of increasing NH⁺₄ added to a constant level of NO^–3₃ Flowing solution culture systems were used to provide NO^?₃at concentrations of 0.03 to 5.0 mol m^–3 and NH⁺₄ levels from 0.05 to 0.3 mmol m^–3 added to 0.1 mol m^–3NO^?₃. Rates of growth, N absorption, accumulation, distribution and utilization were estimated by regression analysis of data obtained from sequential plant harvests, and rates of NO^?₃ and NH^?₄ net uptake were estimated from solution depletion. A sustained ambient NO^?₃ concentration of 0.03 mol m^–3 was evidently adequate to support growth, since relative growth rates were not affected by increasing NO^?₃ supply from 0.03 to 1.0 mol m^–3, nor from 0.25 to 5.0 mol m^–3, in separate experiments. Shoot growth rates were stimulated by NH₄ added to NO^?₃ one experiment, but not when the experiment was repeated under ambient conditions less favorable to growth. Relative accumulation rates for total N increased with increasing NO^?₃ and with NH⁺₄added to NO^?₃ A constant proportion of NO^?₃ taken up was reduced when NO^?₃ alone was supplied. Both the proportion of total N taken up as NO^?₃ and the proportion of NO^?₃ reduced decreased with increasing NH⁺₃ added to NO^?₃ NH⁺₄ uptake apparently must exceed a threshold of about 30% of the total uptake to inhibit NO^?₃ uptake. Utilization of N in chrysanthemum was apparently limited by redistribution since relative accumulation rates for total N were equal to or greater than relative growth rates, in contrast to results reported for several other species. Results of this study and other information support the postulate that NH⁺₄ added to NO^?₃might stimulate growth by increasing transport of reduced N from roots to shoots, thus increasing the supply of reduced N available to support growth of shoot meristems.     

Measurement of net fluxes of ammonium and nitrate at the surface of barley roots using ion-selective microelectrodes

Neutral carrier-based liquid membrane ion-selective microelectrodes for NH₄⁺ and NO₃⁻ were developed and used to investigate inorganic nitrogen acquisition in two varieties of barley, Hordeum vulgare L. cv Olli and H. vulgare L. cv Prato, originating in cold and warm climates, respectively. In the present paper, the methods used in the fabrication of ammonium- and nitrate-selective microelectrodes are described, and their application in the study of inorganic nitrogen uptake is demonstrated. Net ionic fluxes of NH₄⁺ and NO₃⁻ were measured in the unstirred layer of solution immediately external to the root surface. The preference for the uptake of a particular ionic form was examined by measuring the net flux of the predominant form of inorganic nitrogen, with and without the alternative ion in solution. Net flux of NH₄⁺ into the cold-adapted variety remained unchanged when equimolar concentrations (200 micromolar) of NH₄⁺ and NO₃⁻ were present. Similarly, net flux of NO₃⁻ into the warm-adapted variety was not affected when NH₄⁺ was also present in solution. The high temporal and spatial resolution afforded by ammonium- and nitrate-selective microelectrodes permits a detailed examination of inorganic nitrogen acquisition and its component ionic interactions.