Development of Accelerated Net Nitrate Uptake

A method for determination of nitrate concentration and estimation of kinetic parameters of nitrate uptake by spectroscopy based on absorbances at multiple wavelengths has been developed to estimate nitrate uptake by barley (Hordeum vulgare L. cv. Steptoe) seedlings. Nitrate concentration in the nutrient solution was determined from the slope of the linear regression line of the absorbances to nitrate absorption coefficients at 12 wavelengths. Interference by root exudates was only due to an absorption component changing with wavelength in correlation with nitrate absorption. The standard error of the determination decreased in reverse proportion to the square root of the number of the wavelengths. A linear form of the net uptake equation, NUR = ImaxC/Km+C‐E, could be expressed as NUR = (Imax ‐ E) ‐ Km NUR/C ‐ KmE I/C where NUR is net uptake rate, Imax is maximum influx, C is concentration, Km is the Michaelis constant, and E is an efflux constant. The method described here was used to determine the time course of nitrate depletion by barley seedlings from their nutrient solution. The isotherm of net nitrate uptake rates derived from the time course was analyzed after modifications based on the linear form of the net uptake equation. The analysis yielded highly significant results (P<0.0001).

Net nitrate uptake, 36ClO−3/NO−3 influx and 36Cl− influx into Pisum sativum L. cv. Feltham First seedlings have been examined following growth in culture medium containing different combinations of chloride and nitrate. When young (6 days old) seedlings, that had been grown in the absence of N were used, nitrate accumulation stimulated net nitrate uptake and 36ClO−3/NO−3 influx (r2= 0.99) while chloride accumulation inhibited nitrate uptake and 36ClO−3/NO−3 influx (r2= 0.65). When nitrate was provided during growth there was no effect of chloride pretreatment on net nitrate uptake and there was little effect of total [NO−3+ Cl−]i on 36ClO−3/NO−3 influx (r2= 0.26). A direct effect of Cl− on 36ClO−3/NO−3 influx was only found when seedlings had been starved of N for more prolonged periods (14 days).When moderate chloride was supplied during growth, 36Cl− influx was insensitive to nitrate or chloride accumulated, but significantly correlated with loge [NO−3+ Cl−]i (r2= 0.75). When trace amounts of Cl− were supplied during growth 36Cl− influx was inhibited by (a) NO−3 in the external medium and (b) Cl− pretreatment, but was insensitive to NO−3 pretreatment. The sensitivity of 36Cl− influx to external nitrate was not found following Cl− pretreatment in the absence of nitrate. The possibility that there are two populations of chloride carriers which differ in their sensitivity to external nitrate is discussed. Tentative schematic models to account for the regulation of nitrate and chloride uptake are proposed in the context of current hypotheses for regulation of ion transport and control systems theory.

Varietal differences in net nutrient uptake rate and transport efficiency in the presence of aluminium have seldom been investigated in rice. Therefore, effects of Al on growth, uptake and transport of macronutrients (K, P, Ca, and Mg) and micronutrients (Fe, Zn, Cu, and Mn) were evaluated in 3 rice cultivars (BG35, DA14 and IR45) with different Al sensitivity. The plants were grown in nutrient solution at pH 4.1. An initial growth was completed in the time interval 1 to 5 days immediately before the addition of Al. The final growth period with Al (0, 140, 280 or 560 μM) was completed on day 26. With Al, a comparatively high P accumulation occurred in shoots and roots of the Al tolerant cultivar BG35. In contrast, the Al sensitive cultivar IR45 maintained a relatively high Ca accumulation during the Al treatment. A reduced total net uptake rate of P and Ca by IR45 in the time period 5 to 26 days was due to both a reduced root fresh weight and a reduced net uptake rate per g fresh weight of root. Moreover, net Ca transport to the shoots higher than net uptake rate in DA14 and IR45 at &gt; 140 μM Al during the test period suggests restricted Ca uptake by the roots in combination with a continuous net loss of Ca from the roots to the shoots as time proceeds. In the case of Mg and Mn, there was a general reduction of net uptake rates, irrespective of Al sensitivity of cultivars. With Al treatment, comparatively high accumulation of Fe, Zn and Cu occurred in the roots of IR45, concomitant with a high net Zn and Cu uptake rate. It is concluded that differences in Al sensitivity among rice cultivars BG35, DA14 and IR45 are not primarily linked to the depressed internal Mg or Mn status of the plants but rather to changes in the uptake and distribution of Ca and P.

Root respiration rates of Lolium multiflorum supplied with nitrate or ammonium were measured continuously during several days (Exp. A). Net uptake rate of nitrate was similarly measured by an ion selective nitrate electrode in a system of flowing nutrient solution (Exp. B). Diurnal variation of in vitro nitrate reductase activity and nitrate content of tops and roots were determined (Exp. C). Two levels of irradiance were applied throughout, with day:night of 16:8 h.Root respiration rates showed diurnal patterns, most pronounced in the nitrate treatment, with two peaks appearing about 6 and 16 h after commencement of the photoperiod. Respiration rates were highest in the nitrate treatment and at high irradiance. Respiration rates fell after removal of nitrogen, particularly in the nitrate supplied plant and at high irradiance. Net uptake rate of nitrate exhibited diurnal patterns, often with two peaks occurring at the same times as those of respiration rates. In vitro nitrate reductase activity of tops increased steeply 16 h after commencement of the photoperiod and remained at the high level during the following 8 h of darkness. Nitrate content of tops was highest during the 8 h dark period and fell at the start of the photoperiod. Possible controlling systems of the apparent coincidences of diurnal variation rates, net nitrate uptake and nitrate reduction are discussed.

Long-term warming and acidification interaction drives plastic acclimation in the diatom Pseudo-nitzschia multiseries.

Background aqueous chemistry and 15Nnitrate tracer injection methods were used to calculate in‐stream nitrate uptake metrics at Red Canyon Creek, a third‐order stream in the Rocky Mountains in the state of Wyoming, United States. ‘Net’ nitrate uptake lengths, which reflect both nitrate uptake and regeneration, and ‘gross’ nitrate uptake lengths, which exclude re‐mineralization, were quantified separately from background nitrate chemistry and 15N labelling tracer data, respectively. Gross nitrate uptake lengths, from tracer injections of 15N labelled nitrate, ranged from 502 to 3140 m. Net nitrate uptake lengths, from background nitrate chemistry downstream of a point source, ranged from 1170 to 4330 m. Diurnal changes in uptake lengths suggest the importance of nitrate utilization by autotrophs in the stream and benthic zone. The differences between net and gross nitrate uptake lengths along lower reaches of Red Canyon Creek allowed us to estimate the nitrate regeneration rate, which was 0·056–0·080 µmol m−2 s−1 during the day and 0·0062–0·0083 µmol m−2 s−1 at night. Spatial patterns of streambed pore water chemistry indicate those areas of the hyporheic zone where denitrification was likely occurring. Permanent log dams generated stronger redox gradients in the hyporheic zone than areas with transient beaver dams. By combining isotopically labelled nitrate additions, estimates of uptake from background aqueous nitrate chemistry and characterization of redox conditions in the hyporheic zone, we were able to determine the nitrate regeneration rate and the redox processes responsible for nitrogen cycling in the hyporheic zone. Copyright © 2010 John Wiley &amp; Sons, Ltd.

van der Werf, A., Kooijman, A., Welschen, R. and Lambers, H. 1988. Respiratory energy costs for the maintenance of biomass, for growth and for ion uptake in roots of Carex diandra and Carex acutiformis. ‐ Physiol. Plant. 72: 483–491.The respiratory characteristics of the roots of Carex diandra Schrank and Carex acutiformis Ehrh. were investigated. The aims were, firstly to determine the respiratory energy costs for the maintenance of root biomass, for root growth and for ion uptake, and secondly to explain the higher rate of root respiration and ATP production in C. diandra.The three respiratory energy components were derived from a multiple regression analysis, using the relative growth rate and the net rate of nitrate uptake as independent variables and the rate of ATP production as a dependent variable. Although the rate of root respiration and ATP production was significantly higher in C. diandra than in C. acutiformis, the two species showed no significant difference in their rate of ATP production for the maintenance of biomass, in the respiratory energy coefficient for growth (the amount of ATP production per unit of biomass produced) and the respiratory energy coefficient for ion uptake (amount of ATP production per unit of ions absorbed). It is concluded that the higher rate of root respiration of C. diandra is caused by a higher rate of nitrate uptake. At relatively high rates of growth and nitrate uptake, the contribution of the rate of ATP production for ion uptake to the total rate of ATP production amounted to 38 and 25% for C. diandra and C. acutiformis, respectively. At this growth rate, the respiratory energy production for growth contributed 37 and 50%, respectively, to the total rate of ATP production. The relative contribution of the rate of ATP production for the maintenance of biomass increased from 25 to 70% with increasing plant age for both species. The results suggest that ion uptake is one of the major sinks for respiratory energy in roots.These experimentally derived values for the rate of ATP production for the maintenance of biomass, the respiratory energy coefficient for growth and the respiratory energy coefficient for ion uptake are discussed in relation to other experimentally and theoretically derived values.

The decrease in inorganic phosphate concentration in bean (Phaseolus vulgaris L. cv. Zlota Saxa) roots induced decrease in respiration rate. The decrease observed in ATP pool in phosphate deficient (-P) roots was greater than it would result from the decline in respiration and possible involvement of alternative pathway, suggesting an increased energy utilization for growth and ion uptake. Indeed, relative growth rate was higher in -P plants until 12 d of culture and later dropped to the rate similar to the control. Net nitrate uptake rate was higher in -P plants than in +P plants at the beginning of phosphate starvation, then during the prolonged culture it decreased rapidly in -P plants and after 19 d it was 8 times lower than that in the control. The decline in ATP production during prolonged phosphate starvation influenced NO3 - uptake more than root growth.

Barley seedlings (Hordeum vulgare L.) were grown hydroponically with (induced) or without (uninduced) nitrate in a light/dark cycle with high photon flux density to determine the effects of light on time courses, induction and kinetics of net nitrate uptake. Nitrate uptake was induced by external nitrate in both light and dark and was prevented by 1 mol m–3 p‐fluorophenylalanine. In high light, nitrate uptake was about 2‐fold higher than in low light. During time course experiments the uptake rates oscillated due to daily light–dark changes. Rates of nitrate uptake also increased at about 2200 h during continuous darkness. This increase coincided approximately with the time at which the dark period started during the previous culture of the plants, indicating that it was due to a mechanism associated with an endogenous diurnal rhythm. When calculating the kinetics of nitrate uptake, a model with two saturable systems, including a high‐affinity system (HATS) and a low‐affinity system (LATS), gave the best fit to data in all treatments. The apparent affinity of the HATS ranged from 7·7 to 12·2 mmol m–3 in induced plants in all light conditions. The effect of light on the HATS was mainly an increase of apparent Vmax in the step from low to high light. In uninduced plants the HATS operated at a very low activity which was strongly enhanced during induction. Interpretation of the calculated kinetics of the LATS was much more difficult on the basis of net uptake data. The apparent affinity of the LATS increased from 24·3 mol m–3 in low light up to 0·17 mol m–3 after acceleration in high light. These extreme changes in apparent affinity of the LATS could not be explained satisfactorily, and the nature of this system is also discussed with respect to the method used.

Prince Rupprecht’s Larch from the Loess Plateau takes up and assimilates a greater proportion of N as NO , particularly at neutral pH, whereas Chinese Fir assimilates a greater proportion of N as NH , particularly at low pH levels. The effects of pH on nitrate and ammonium uptake and assimilation in two coniferous species were compared. Prince Rupprecht’s Larch (Larix principis-rupprechtii Mayr) grows on the loess plateau in alkaline soils with low available nitrogen (N), whereas Chinese Fir (Cunninghamia lanceolata) grows in acidic soils. In the present study, the net fluxes in ammonium (NH4 +) and nitrate (NO 3 ) were measured using a non-invasive microelectrode ion flux measurement system, and the expression of NH4 + and NO 3 transporters (AMTs and NRTs, respectively) as well as H+-ATPase was examined to provide insights into the N uptake mechanisms in Prince Rupprecht’s Larch and Chinese Fir. The enzyme assays involved in N assimilation were also determined. For Prince Rupprecht’s Larch, low pH (pH 4) resulted in a decrease in net ammonium uptake, which remained unchanged in Chinese Fir. Net nitrate uptake in Prince Rupprecht’s Larch and Chinese Fir was much lower in soils with pH 4 relative to those with pH 7. Low pH significantly decreased the H+-ATPase activity and the expression level of NRTs in roots of Prince Rupprecht’s Larch. However, the expression level of AMTs in Prince Rupprecht’s Larch was significantly higher at pH 7 than at pH 4. The H+-ATPase activity in roots of Chinese Fir remained unaltered in response to changes in pH, and the transcript abundances of AMTs and NRTs were down-regulated by low pH. Low pH decreased N assimilation in both conifer species with the exception of NH4 + assimilation in Chinese Fir, which displayed higher glutamine synthetase (GS) and glutamate synthetase (GOGAT activities) at low pH. Prince Rupprecht’s Larch from the Loess Plateau takes up and assimilates a greater proportion of N as NO 3 , particularly at neutral pH, whereas Chinese Fir assimilates a greater proportion of N as NH4 +, particularly at low pH levels. This study contributes to our understanding of nitrogen metabolism mechanisms in response to pH changes.

Primary productivity and nitrogenous nutrient assimilation dynamics during the Subarctic Pacific Iron Experiment for Ecosystem Dynamics Study

Concentration dependence of net ammonium and nitrate uptake was monitored for a cultivar of tomato, Lycopersicon esculentum, and two accessions of a neotropical wild relative, L. hirsutum. The kinetics of net NH 4+ uptake differed among these taxa and were not dependent on the ionic composition of the nutrient solution. The kinetics of net NO 3- uptake were dependent on the composition of the nutrient solution; the presence of NH 4+ or Cl- enhanced net NO 3- uptake for the cultivated species and for a highland accession of the wild species. The capacity for net NO 3- uptake was greater than the capacity for net NH 4+ uptake in all three taxa; the proportion of NO 3- to NH 4+ absorbed was much greater for the wild taxa. Our data suggest that NO 3- may be a more important source of mineral nitrogen than NH 4+ for these tropical taxa.

Efficient utilization of fertilizer-nitrogen (N) by turfgrasses is probably related to N uptake efficiency of roots and metabolic efficiency of absorbed N in roots and shoots. This study evaluated Kentucky bluegrass (Poa pratensis L.) cultivars for potential differences in nitrate uptake rate (NUR), temporal variation in NUR, and the relationship between NUR and N use efficiency (NUE), defined as grams dry matter per gram N. Six cultivars were propagated from tillers of seeded plants, grown in silica sand, mowed weekly, and watered daily with a complete nutrient solution containing 1.0 mm nitrate. A nutrient depletion method from an initial nitrate concentration of 0.5 mm was used to determine NUR of 5-month-old plants. NUR (μmol·h-1 per plant) of the six cultivars ranked as follows: `Blacksburg' &gt; `Conni' &gt; `Dawn' &gt; `Eclipse' = `Barzan' &gt; `Gnome'. When NUR was based on root weight, `Conni' ranked highest; when NUR was based on root length, surface, or volume, `Eclipse' ranked highest. Averaged across cultivars, NUR on the second day was greater than NUR for the first day of nitrate exposure. Temporal variation was greatest in `Blacksburg', while none was noted in `Conni' or `Eclipse'. Cultivar differences in NUE were significant in fibrous roots, rhizomes, and leaf sheaths, but not in leaf blades and thatch. Total nitrate uptake was positively related to total N recovered and total plant dry matter, but NUR based on root weight was negatively correlated with NUE of the whole plant.

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, Km and Vmax, 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.

Short-term dynamics of nano- and picoplankton production in an embayment of the southern Benguela upwelling region