NITRATE UPTAKE IN MARINE PHYTOPLANKTON: (NITRATE,CHILORIDE)-ACTIVATED ADENOSINE TRIPHOSPHATASE FROM SKELETONEMA COSTATUM (BACILLARIOPHYCEAE)1,2

A membrane-bound (nitrate, chloride)-activated AT Pase was found in the neritie diatom, Skeletonema costatum (Grev.) Cleve. The enzyme is suggested to translocate NO₃⁻ across the plasmalemma, against a concentration gradient. The K_m of the enzyme with respect to NO₃⁻ is ca. 0.9 × 10⁻⁶ M, and is in close agreement with the reported K₈ for NO₃⁻ uptake by the whole cell.     

UPTAKE OF INORGANIC NITROGEN BY CODIUM FRAGILE SUBSP. TOMENTOSOIDES (CHLOROPHYTA)1

The uptake of nitrate, nitrite and ammonium by Codium fragile subsp. tomentosoides (van Goor) Silva was measured at different combinations of temperature (6–30 C) and irradiance (0–140 μEin.m-^2. s^-1). Uptake of all three forms of N was greater at 12–24 C than at 6 and 30 C. Although uptake was stimulated by light, saturation occurred at relatively low irradiance (7–28 μEin m^-2 s^-1, depending on the N source and temperature). The Michaelis-Menten uptake constants (V_max K)varied with temperature. V_max was greatest at intermediate temperatures and K was lowest at lower temperatures. The V_maxfor NH₄⁺ was higher and the K, for NH₄⁺was lower than those for NO₃^-_- and NO₂^-_-. Codium was capable of simultaneously taking up all three forms of inorganic N although the presence of NH₄⁺ reduced the uptake of both NO₃^-_- and NO₂^-_-. The results of this study indicate that part of the ecological success of Codium in a N-limited environment may be due to its N uptake capabilities.     

Growth,pigment, and chromophoric dissolved organic matter responses of tropical Chattonella subsalsa (Raphidophyceae) to nitrogen enrichment

Blooms of the raphidophyte Chattonella subsalsa have been associated with massive fish‐kill events in several parts of the world. However, there have been few studies into physiological responses of tropical strains that could contribute to bloom outcomes. Such knowledge could provide insight into the C. subsalsa blooms recently documented within tropical coastal waters (e.g., 2010 and 2012 events in Singapore). Strains used in this study were isolated from the East Johor Straits (EJS), Singapore, an enclosed water channel frequently subjected to high levels of eutrophication. These cells were classified within the ‘global’ clade (and distinct from the ‘Adriatic Sea’ clade) based on morphology. The present study examined cellular responses to varying inputs of different forms of nitrogen (N), specifically nitrate, ammonium, and urea. Results from the study indicated that cells were unable to utilize urea as an N‐source, but grew well on a nitrate (V_max = 0.73 day^?1) and ammonium (V_max = 0.81 day^?1) supply. These growth rates were high compared to other strains from around the world, indicating that tropical C. subsalsa could exhibit elevated bloom potential within frequently eutrophic environments such as the EJS. Six pigments were detected in all cultures. These pigments were chlorophylls a and c; fucoxanthin; diadinoxanthin; violaxanthin; and β‐carotene. Chlorophyll‐a and fucoxanthin were the dominant pigments under both nitrate and ammonium regimes. Measurements of chromophoric dissolved organic matter generally increased both in molecular weight and in total content across the N‐concentration ranges. Such outcomes could have consequences for the chemical and optical conditions of the coastal environment.     

Nutrient uptake and alkaline phosphatase (ec 3:1:3:1) activity of emiliania huxleyi (PRYMNESIOPHYCEAE) during growth under n and p limitation in continuous cultures

Emiliania huxleyi (strain L) expressed an exceptional P assimilation capability. Under P limitation, the minimum cell P content was 2.6 fmol P·cell^?1, and cell N remained constant at all growth rates at 100 fmol N·cell^?1. Both, calcification of cells and the induction of the phosphate uptake system were inversely correlated with growth rate. The highest (cellular P based) maximum phosphate uptake rate (V_max^P) was 1400 times (i.e. 8.9 h^?1) higher than the actual uptake rate. The affinity of the P‐uptake system (dV/dS) was 19.8 L·μmol^?1·h^?1 at μ = 0.14 d^?1. This is the highest value ever reported for a phytoplankton species. V_max and dV/dS for phosphate uptake were 48% and 15% lower in the dark than in the light at the lowest growth rates. The half‐saturation constant for growth was 1.1 nM. The coefficient for luxury phosphate uptake (Q_maxt/Q_min) was 31. Under P limitation, E. huxleyi expressed two different types of alkaline phosphatase (APase) enzyme kinetics. One type was synthesized constitutively and possessed a V_max and half‐saturation constant of 43 fmol MFP·cell^?1·h^?1 and 1.9 μM, respectively. The other, inducible type of APase expressed its highest activity at the lowest growth rates, with a V_max and half‐saturation constant of 190 fmol MFP·cell^?1·h^?1 and 12.2 μM, respectively. Both APase systems were located in a lipid membrane close to the cell wall. Under N‐limiting growth conditions, the minimum N quotum was 43 fmol N·cell^?1. The highest value for the cell N‐specific maximum nitrate uptake rate (V_max^N) was 0.075 h^?1; for the affinity of nitrate uptake, 0.37 L·μmol^?1·h^?1. The uptake rate of nitrate in the dark was 70% lower than in the light. N‐limited cells were smaller than P‐limited cells and contained 50% less organic and inorganic carbon. In comparison with other algae, E. huxleyi is a poor competitor for nitrate under N limitation. As a consequence of its high affinity for inorganic phosphate, and the presence of two different types of APase in terms of kinetics, E. huxleyi is expected to perform well in P‐controlled ecosystems.     

Kinetics of nitrate uptake by different species from nutrient-rich and nutrient-poor habitats as affected by the nutrient supply

The species Urtica dioica L., Plantago major ssp. major L., Plantago lanceolata L., Hypochaeris radicata L. ssp. radicata and Hypochaeris radicata ssp. ericetorum Van Soest were grown under high and low nutrient conditions (1/4 Hoagland and 2% of 1/4 Hoagland further called the 100% and 2% treatment, containing 3.75 mM NO^-₃ and 0.075 mM NO^-₃, respectively). After a certain period half of the plants were transferred from low to high or high to low nutrients, yielding the 100%/2% and the 2%/100% treatments. The kinetics of nitrate uptake in the range of system I of the five species grown under the different nutrient conditions were measured during a three week experimental period. The nitrate uptake of all the species showed the characteristic features of Michaelis-Menten kinetics. Under low nutrient conditions the apparent V_max of U. dioica expressed per g dry root was lower than under high nutrient conditions. For H. radicata ssp. radicata and for H. radicata ssp. ericetorum the reverse was found. The V_max values of P. major ssp. major were almost the same for the two treatments. The apparent V_max in young plants of P. lanceolata was higher in the 100% treatment than in 2%; whereas the reverse was found in mature plants. The results are explained in relation to the relative growth rate, the shoot to root ratio and the natural environment of the species. The apparent K_m values were not influenced by the different treatments. Differences in K_m between the species, if any, were very small. It is suggested that the V_max is a more important parameter for the distribution of plant species in the field than the K_m. The rate of nitrogen accumulation was calculated from growth data and the contents of nitrate and reduced nitrogen. It is concluded that the V_max of system I for nitrate uptake in most cases was sufficient to explain the observed growth rates.     

Kinetic parameters of nitrate uptake by different catch crop species: effects of low temperatures or previous nitrate starvation

The pollution of aquifers by NO^?₃ in temperate environments is aggravated by farming practices that leave the ground bare during winter. The use of catch crops during this time may decrease nitrate loss from the soil. Nitrate uptake by several catch crop species (Brassica napus L., Sinapis alba L., Brassica rapa L., Raphanus sativus L., Trifolium alexandrinum L., Trifolium incarnatum L., Phacelia tanacetifolia Benth., Lolium perenne L., Lolium multiflorum Lam. and Secale cereale L.) was here studied in relation to transpiration rate and low temperatures applied to the whole plant or to roots only. The Michaelis constant (K_m), maximum uptake rate (V_max), time of induction and contributions of inducible and constitutive mechanisms were estimated from measurements of NO^?₃ depletion in the uptake medium. There were large differences between species, with K_m (μM) values ranging between 5.12 ± 0.64 (Trifolium incarnatum) and 36.4 ± 1.97 (Lolium perenne). Maximum NO^?₃ uptake rates expressed per unit root weight were influenced by ageing, temperature and previous NO^?₃ nutrition. They were also closely correlated with water flow through the roots and with shoot/root ratio of these species. The combined results from all species and treatments showed that V_max increased with shoot/root ratio, suggesting a regulatory role for the shoots in NO^?₃ uptake. Overall, the results showed a great diversity in NO^?₃ uptake characteristics between species in terms of kinetic parameters, contribution of the constitutive system (100% of total uptake in ryegrass, nil in Fabaceae) and time of induction.     

Constitutive and inducible aspects of nitrate-nitrogen uptake by Chlamydomonas reinhardtii

Similarly to higher plant root systems, Chlamydomonas reinhardtii Dangeard (UTEX 90) cells exhibited biphasic NO₃^? uptake kinetics. The uptake pattern was similar in cells cultured in 10 mM NO₃^? (NO₃^?-grown), 0.25 mM NO₃^? (N-limited) or 10 mM NO₃^? followed by an 18-h period of N-deprivation (N-starved). In all cell types there was an apparent phase transition in uptake at 1.1 mM NO₃^?, although there were variations in the uptake V_max of both isotherms. The rate of uptake via isotherm 0 ([NO₃^?]<1.1 mM) in N-limited cells was higher than that of either NO₃^?-grown or N-starved cells. In contrast, NO₃^?-grown and N-limited cells exhibited comparable V_max values when supplied with 1.1 to 1.8 mM NO₃^? (isotherm 1). When supplied with 1.6 mM NO₃^?, both N-limited and N-starved cells exhibited enhanced linear uptake after 60 min of incubation. We ascribed this to an induction phenomenon. This trend was not observed when NO₃^?-grown cells were supplied with 1.6 mM NO₃^?, or when N-limited and N-starved cells were supplied with 0.6 mM NO₃^?. The ‘inducible’ aspect of uptake by N-limited cells was blocked by cycloheximide (10 mg l^?1), but not by actinomycin D (5 mg l^?1), thus indicating the involvement of a translational or post-translational event. To investigate this phenomenon further, we analysed the cell proteins of N-limited cells supplied with either 0.6 or 1.6 mM NO₃^? for 90 min, using two-dimensional gel electrophoresis. Comparison of protein profiles enabled the identification of a single cell membrane-associated polypeptide (21 kDa, pI ca 5.5) and ten soluble fraction polypeptides (17–73 kDa, pI ca 5.0 to 7.1) unique to the high NO₃^? treatment. We propose that the ‘inducible’ portion of NO₃^? uptake may provide the means by which C. reinhardtii cells regulate uptake in accordance with assimilatory capacity.     

Seasonal pattern of ammonium (methylamine) uptake by phytoplankton in an oligotrophic lake

Our primary objective was to determine if a relationship existed between seasonal change in phytoplankton and high affinity for (K _m) or uptake rates (V _maX) of ammonium which might explain seasonal phytoplankton succession in oligotrophic ecosystems. We measured ammonium uptake using [¹⁴C]-methylamine and estimatedK _m andV _max using Hanes Plots at 2-week intervals during 6 months of thermal stratification in Mountain lake, Virginia (37° 22 N, 80° 32 W). Community composition, nutrient levels, and other variables were determined in all uptake experiments. A second objective was to determine if ammonium was preferentially utilized over nitrate and to characterize further the ammonium transport system.V _max increased steadily from May until the end of July, each increase coinciding with major changes in the phytoplankton community. Cryptophyceans dominated in May, chlorophyceans in June and July, and cyanophyceans from the end of July to late October. With cyanophycean dominance,V _max declined until chlorophyceans reestablished dominance in late October. By contrast,K _m values increased from May to the end of July, but thereafter showed no correlation. Acetylene reduction experiments showed no nitrogen fixation during late summer and fall when blue-green algae were present. Preference for ammonium was implied also by negative nitrate reductase assays. Overall, the coincidence ofV _max andK _m values for [¹⁴C]-methylamine uptake and changing phytoplankton community structure suggests the possibility that successive algal communities may be changing as a result of specific species differences in ammonium affinity and uptake rates.     

Humic acid differentially improves nitrate kinetics under low‐ and high‐affinity systems and alters the expression of plasma membrane H+‐ATPases and nitrate transporters in rice

Humic acids (HAs) have a major effect on nutrient uptake, metabolism, growth and development in plants. Here, we evaluated the effect of HA pretreatment applied with a nutrient solution on the uptake kinetics of nitrate nitrogen (N‐NO₃^?) and the metabolism of nitrogen (N) in rice under conditions of high and low NO₃^? supply. In addition, the kinetic parameters of NO₃^? uptake, N metabolites, and nitrate transporters (NRTs) and the plasma membrane (PM) H⁺‐ATPase gene expression were examined. The plants were grown in a growth chamber with modified Hoagland and Arnon solution until 21 days after germination (DAG), and they were then transferred to a solution without N for 48 h and then to another solution without N and with and without the addition of HAs for another 48 h. After this period of N deprivation, the plants received new nutrient solutions containing 0.2 and 2.0 mM N‐NO₃^?. Treatment of rice plants with HA promoted the induction of the genes OsNRT2.1‐2.2/OsNAR2.1 and some isoforms PM H⁺‐ATPase in roots. The application of HAs differentially modified the parameters of the uptake kinetics of NO₃^? under both concentrations. When grown with 0.2 mM NO₃^?, the plants pretreated with HA had lower K_m and C_min values as well as a higher V_max/K_m ratio. When grown with 2 mM NO₃^?, the plants pretreated with HA had a higher V_max value, a greater root and shoot mass, and a lower root/shoot ratio. The N fractions were also altered by pretreatment with HA, and a greater accumulation of NO₃^? and N‐amino was observed in the roots and shoots, respectively, of plants pretreated with HA. The results suggest that pretreatment with HA modifies root morphology and gene expression of PM H⁺‐ATPases and NO₃^? transporters, resulting in a greater efficiency of NO₃^? acquisition by high‐ and low‐affinity systems.     

NITRATE UPTAKE KINETICS AND CLONAL VARIABILITY IN THE NERITIC DIATOM BIDDULPHIA AURITA1

Nitrate uptake experiments were conducted at 18 and 25 C on subtropical and temperate isolates of the neritic diatom Biddulphia aurita (Lyngb.) Breb. & Godey. Two clones (STX-88 and B1) were grown in a 12:12 LD cycle in NO₃-limited continuous cultures used for uptake experiments. The half-saturation constant (K_s) and the maximum NO₃- uptake rate (V_m) were measured under conditions in which external NO₃- concentration controlled uptake rate. The kinetic constants for both clones were higher at the higher temperature. The subtropical clone (STX-88) had lower kinetic values than temperate clone (B1) at both temperatures. The differences appear to be of adaptive significance since STX-88 has a NO₃- uptake system, described by its lower K_s value, which minimizes the double stress effects of high temperature-low NO₃- concentration conditions characteristic of its habitat. Cytoplasmic enzyme electrophoretic analyses were conducted on the clones. Differences in banding patterns and mobilities showed evidence of allelic substitution. The results suggest genetic variation at the intraspecific level.     

AMMONIUN AND NITRATE UPTAKE BY THE MARINE MACROPHYTES HYPNEA MUSVUFORMIS (RHODOPHYTA) AND MACROCYSTIS PYRIFERA (PHAEOPHYTA)1, 2

NH₄⁺ and NO₃^? uptake were measured by continuous sampling with an autoanalyzer. For Hypnea musciformis (Wulfen) Lamouroux, NO₃^?up take followed saturable kinetics (K₂=4.9 μg-at N t^?1, V_max= 2.85 μg- at N, g(wet)^?1. h^?1. The ammonium uptake data fit a trucatd hyperbola, i.e., saturation was not reach at the concentrations used. NO₃^? uptake was reduced one-half in the presence of NH₄⁺, but presence of NO₃^? had no effect on NH₄⁺ uptake. Darkness reduced both NO₃^? and NH₄⁺ uptake by one-third to one-half. For Macrocystis pyrufera (L) C. Agardh, NO₃^? uptake followed saturable kinetices: K₂=13.1 μg-at N. l^?1. V_max=3.05 μg-at N. g(wet)^?1. h^?1.NH₄⁺ uptake showed saturable kinetics at concentration below 22 μg-at N l -1 (K₂=5.3 μg-at N.1–1, V_max= 2.38 μg-at N G (wet)^?1.h^?1: at higher concentration uptake increased lincarly with concentrations. NO₃^?and NH₄⁺ were taken up simulataneously: presence of one form did not affect uptake of the other.     

NITRATE UPTAKE BY LAMINARIA LONGICRURIS (PHAEOPHYCEAE)1,2

Laminaria longicrucis De la Pylaie took up exogenous nitrate under both summer and winter conditions. During July and August no NO₃^- was detected in the ambient water or in algal tissues although it was present in both in February. Discs (2.3 cm diam.) of thin blade tissue were incubated with NO₃^- at four temperatures, with and without illumination. Similar values Jor NO₃^- uptake were found for both summer and winter collected plants when measured in light at 0 C. An apparent K of 4–6 μM was recorded for both types of plants; the V_max ranged from 7 to 10 μmol h^-1 g^-1 dry wt measured in ca. 1800 μW cm^-2 of cool-white fluorescent light. Uptake rates at 5 C were 66%, and at 0 C 30% of those for controls run at 15 C. The alga scavenged NO₃- from solutions <0.5 μM. Ammonia did not inhibit NO₃- uptake. Antibiotic pretreatment reduced NO₃- uptake by a maximum of 12%. Nitrite uptake was inhibited in proportion to the concentration of NO₃- in the medium.     

Direct measurements of nanomolar nitrate uptake by the marine diatom Phaeodactylum tricornutum (Bohlin). Implications for studies of oligotrophic ecosystems

A new automated procedure for nanomolar nitrate analysis was applied to the study of uptake of low nitrate concentrations (< 100 ngat l^–1) by phytoplankton. The precision of this analytical method (± 3 ngat l^–1) made it possible to monitor the absorption of very low quantities of nitrate over short term periods by a low cell-density culture of the marine diatom Phaeodactylum tricornutum, where the levels of particulate nitrogen and chlorophyll were equivalent to those found in oligotrophic areas (0.5 µgat N l^–1 and 0.4 µg l^–1 respectively). By continuous monitoring of nitrate disappearance from the culture medium, we are able to describe accurately the transient uptake responses of the diatom after a spike addition of trace quantities of nitrate and thus to provide new information on the still largely unknown small-scale phenomenon of pulsed nitrate supply in the upper layer of stratified oceans and rapid uptake of these nitrate patches by phytoplankton.The results show that a N-limited culture of Phaeodactylum tricornutum is immediately capable of taking up trace quantities of nitrate (< 100 ngat l^–1) at high rates (0.10–0.14 h^–1) . These initial rates are one order of magnitude higher than the theoretical rates calculated from the Michaelis-Menten equation and are close to the level of V _max (0.15 h^–1) obtained when cells are exposed to saturating nitrate concentrations. This rapid initial uptake would be a considerable advantage in oligotrophic areas where nanomolar nitrate supply is thought to be episodic. The present results suggest that phytoplankton evolve adaptations to utilize the available nitrate at the spatial and temporal scales at which it occurs. On the other hand, we can consider this physiological adaptation as evidence of nitrate pulses in the field which would invalidate the steady-state approach to the oligotrophic ecosystems.     

Nitrate assimilatory properties of barley grown under long-term N limitation: Effects of local nitrate supply in split-root cultures

The effect of nitrate availability on characteristics of the nitrate assimilatory system was investigated in N-limited barley (Hordeum valgare L. cv. Golf), grown with the seminal root system split into initially equal-sized halves. The cultures were continuously supplied with nitrate-N at a relative addition rate (RA) of 0.09 day^?1, which resulted in relative growth rates (RG) that were ca 85% of those observed under surplus nitrate nutrition. The total N addition was divided between the subroots in ratios of 100:0, 80:20, 70:30, 60:40, and 50:50. For comparison, standard cultures were grown at RAs ranging from 0.03 to 0.18 day^?1. Initially, biomass and N partitioning to the subroots responded strongly and proportionally to the nitrate distribution ratio. After 12-14 days no further effect was observed. The V_max for net nitrate uptake and in vitro nitrate reductase (NR) activity were measured in acclimated plants, i.e., after > 14 days under a certain nitrate regime. In subroots fed from 20 to 100% of the total N addition, V_max for net nitrate uptake increased slightly, whereas NR activity was unaffected. Uptake and NR activities were insignificant in the 0%-subroot. Uneven nitrate supply to individual subroots had negligible effect on the whole-plant ability for nitrate uptake, and the relative V_max (unit N taken up per unit N in whole plant tissue and time) remained about 7-fold in excess of the demand set by growth. Balancing nitrate concentrations (the resulting external nitrate concentrations at a certain RA) generally ranged between 2 and 10 μM at growth-limiting RA, both when predicted from uptake kinetics and when actually measured. When comparing split root and standard cultures when acclimated, it appears that uptake and NR activities in roots respond more strongly to over-all nitrate availability than to nitrate availability to individual subroots.     

Oligotrophic waters of the Northwest Atlantic support taxonomically diverse diatom communities that are distinct from coastal waters

Diatoms are important components of the marine food web and one of the most species-rich groups of phytoplankton. The diversity and composition of diatoms in eutrophic nearshore habitats have been well documented due to the outsized influence of diatoms on coastal ecosystem functioning. In contrast, patterns of both diatom diversity and community composition in offshore oligotrophic regions where diatom biomass is low have been poorly resolved. To compare the diatom diversity and community composition in oligotrophic and eutrophic waters, diatom communities were sampled along a 1,250 km transect from the oligotrophic Sargasso Sea to the coastal waters of the northeast US shelf. Diatom community composition was determined by amplifying and sequencing the 18S rDNA V4 region. Of the 301 amplicon sequence variants (ASVs) identified along the transect, the majority (70%) were sampled exclusively from oligotrophic waters of the Gulf Stream and Sargasso Sea and included the genera Bacteriastrum, Haslea, Hemiaulus, Pseudo-nitzschia, and Nitzschia. Diatom ASV richness did not vary along the transect, indicating that the oligotrophic Sargasso Sea and Gulf Stream are occupied by a diverse diatom community. Although ASV richness was similar between oligotrophic and coastal waters, diatom community composition in these regions differed significantly and was correlated with temperature and phosphate, two environmental variables known to influence diatom metabolism and geographic distribution. In sum, oligotrophic waters of the western North Atlantic harbor diverse diatom assemblages that are distinct from coastal regions, and these open ocean diatoms warrant additional study, as they may play critical roles in oligotrophic ecosystems.     

Spatial and temporal patterns of net nitrate uptake regulation and kinetics along the tap root of <Emphasis Type="Italic">Citrus aurantium</Emphasis>

Spatial–temporal variation of the regulation and the kinetics of net nitrate (NO₃ ⁻) uptake rate (NNUR) along the tap root of Citrus aurantium L. were analysed. Suberin incrustation in the peripheral cell layers and plasma membrane (PM) H⁺-ATPase localisation, anatomical and physiological factors involved in NO₃ ⁻ uptake were also investigated. The results clearly indicated a spatially uniform distribution of the regulation process, accompanied by a temporal heterogeneous pattern of the kinetics of NO₃ ⁻ uptake along citrus tap root. In particular, kinetic analysis had a biphasic pattern, saturating (high affinity transport system) and linear (low affinity transport system), in response to increasing external NO₃ ⁻ concentrations in each root region, where 200 μM NO₃ ⁻ represented the threshold separating these two systems. Kinetic parameters, K _m and V _max, clearly indicated that apical segments reached the maximum value of induction before basal segments. Hence, the apical root zones, early exhibiting the maximum of potential capacity to absorb the NO₃ ⁻, could be considered more efficient than basal root segments for acquiring NO₃ ⁻ from external solution. Suberin incrustations in the hypodermal cell layer, characterised by uniform fluorescence intensity among the root segments, could be responsible for the unchanged NNUR, while the PM H⁺-ATPase could explain the temporal pattern of NNUR.     

Growth and nitrate uptake properties of plants grown at different relative rates of nitrogen supply. II. Activity and affinity of the nitrate uptake system in Pisum and Lemna in relation to nitrogen availability and nitrogen demand

Abstract Net nitrate uptake rates were measured and the kinetics calculated in non-nodulated Pisum sativum L. cv. Marma and Lemna gibba L. adapted to constant relative rates of nitrate-N additions (R_A), ranging from 0.03 to 0.27 d^?1 for Pisum and from 0.05 to 0.40 d^?1 for Lemna, V_max of net nitrate uptake (measured in the range 10 to 100 mmol m^?3 nitrate, i.e. ‘system I’) increased with R_A in the growth limiting range but decreased when R_A exceeded the relative growth rate (RGR), K_m was not significantly related to changes in R_A. On the basis of previous ¹³N-flux experiments, it is concluded that the differences in V_max at growth limiting R_A are attributable to differences in influx rates. Linear relationships between V_max and tissue nitrogen concentrations were obtained in the growth limiting range for both species, and extrapolated intercepts relate well with the previously defined minimal nitrogen concentrations for plant growth (Oscarson, Ingemarsson & Larsson, 1989). Analysis of V_max for net nitrate uptake on intact plant basis in relation to nitrogen demand during stable, nitrogen limited, growth shows an increased overcapacity at lower R_A values in both species, which is largely explained by the increased relative root size at low R_A. A balancing nitrate concentration, defined as the steady state concentration needed to sustain the relative rate of increase in plant nitrogen (R_N), predicted by R_A, was calculated for both species. In the growth limiting range, this value ranges from 3.5 mmol m^?3 (R_A 0.03 d^?1) to 44 mmol m^?3 (R_A 0.21 d^?1) for Pisum and from 0.2 mmol m^?3 (R_A 0.05 d^?1) to 5.4 mmol m^?3 (R_A 0.03 d^?1) for Lemna. It is suggested that this value can be used as a unifying measure of the affinity for nitrate, integrating the performance of the nitrate uptake system with nitrate flux and long term growth and demand for nitrogen.     

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

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

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.     

NH4 : NO3 nutrition influence on biomass productivity and root respiration of poplar and willow clones

Nitrogen fertilization often improves the yield of intensively managed, short‐rotation coppices. However, information of N nutrition form on the growth of common species and clones used for biomass production is limited. Thus, this study aims at evaluating N form effects on the growth of two Salicaceae clones. Cuttings of the poplar clone Max 4 (Populus maximovizcii × P. nigra) and the willow clone Inger (Salix triandra × S. viminialis) were fertilized in a pot experiment with four ratios of nitrate (NO₃^?) to ammonium (50%, 62.5%, 75% and 87.5% NO₃^? balanced with ammonium (NH₄⁺) to constant total N) for one growing season and under stable soil pH. Plants were harvested for analysis of biomass and morphology of leaves, stem and roots. Respiration of fine and coarse roots (RR) was determined and related to biomass growth. Salix cv. Inger accumulated more total dry matter than Populus cv. Max 4. In both Salicaceae clones, the total biomass was significantly influenced by the nitrate ratio and greatest in plants fertilized with 50% NO₃^? of the total N supply. Both clones possess a different leaf and root morphology, but no significant influence of the NO₃^? ratio on the morphology was found. Fine RR rates differed significantly between clones, with significantly greater fine RR in Max 4; 87.5% NO₃^? fertilization increased the fine RR. Fine RR and total accumulated plant biomass were closely related. Our study is the first to show the tremendous influence of fine root respiration, especially including the carbon‐intensive reduction of NO₃^? to NH₄⁺, on the aboveground growth of Salicaceae clones. Ways to improve yield in SRC are thus to lower the assimilate consumption by fine roots and to match fertilization regimes to the used clones or vice versa.