Depression of Nitrate and Ammonium Transport in Barley Plants with Diminished Sulphate Status. Evidence of Co-regulation of Nitrogen and Sulphate Intake

When barley plants were grown in a solution with nitrate asthe sole N-source but deprived of sulphate (–Splants)for 1 to 5 d, the capacity for sulphate transport by the rootsincreased very markedly; subsequent measurement of influx using³⁵S-labelled showed increases of > 10-fold compared to plants continuously supplied with sulphate (+S plants).There were only small effects on plant growth over a 5 d periodand yet the influx of , labelled with the short-lived tracer ¹³N, was diminished by approximately 30%.By contrast, the influx of phosphate was little affected bysulphate-deprivation. When a sulphate supply was restored to– S plants, the sulphate influx was quickly repressedover the subsequent 24 h and the nitrate influx was restoredto >90% of the value in +S plants. When plants were grown in a solution with a mixed nitrate andammonium supply and deprived of sulphate for 1 d or 5 d thedepression of nitrate influx was more strongly marked (up to55% depression). The influx of ammonium was also depressed after5 d of sulphate-deprivation, but not at 1 d, nor where the concentrationof ammonium in the uptake solution was lowered to 20 mmol m^–3or less. Additional measurements with ¹⁵N-labelled nitrate and ammoniumover longer periods were used to determine net uptake. Net uptakeof nitrate was depressed to a similar extent to efflux, butnet ammonium uptake was depressed only in unbuffered uptakesolution where the pH decreased to pH 4.9 during the uptakeperiod. The ¹⁵N-tracer experiments showed that the translocationof label to the shoot, from both nitrate and ammonium, was depressedto a greater extent than net uptake in –S plants. Thedepression of nitrate influx, caused by 5 d of sulphate deprivation,could be relieved almost completely by providing plants with1.0 mol m^–3 L-methionine during the day prior to influxmeasurement. This treatment substantially decreased sulphateand potassium (⁸⁶Rb-labelled) influx in both +S and –Splants, but greatly increased total S-status of the plants.This methionine treatment had no effect on ammonium influx ornet uptake in – S plants but increased influx significantlyin +S ones. When plants were grown with sulphate but deprived of nitratefor 4 d there was a marked depression of the sulphate influx(by 48–65%) but a smaller effect on phosphate influx (21–37%of +N). The results are discussed in relation to the effects of sulphate-deprivationon growth rate and the root: shoot weight ratio. It is concludedthat the effects on influx and net uptake of nitrogen are moresevere than could be accounted for by these factors. The decreasedtranslocation of either nitrate, or the products of nitrateand ammonium assimilation from the roots, is suggested as areason for the depression of influx. The restoration of nitrateinflux and net uptake by methionine suggests that, for thision at least, a shortage of S-amino acids within the plant maylead to the accumulation of inhibitory concentrations of non-Samino acids in the transport pool. Key words: ¹³N, sulphate, nitrate, ammonium, ion-uptake, barley     

Effects of Nitrogen Deficiency on the Absorption of Nitrate and Ammonium by Barley Plants

When young barley plants which had been supplied with nitratewere deprived of this source of N, an enhanced capacity forabsorption of either nitrate or ammonium ions developed, reachinga maximum in about 3 d under the particular experimental conditionsused. The net uptake rate of either nutrient was then approximatelythree times that in plants which had received nitrate throughout.Likewise, withholding external N from plants previously growingwith ammonium caused a 2.4-fold increase in their subsequentcapacity to absorb that ion, compared with control plants grownwith an uninterrupted ammonium supply. Accelerated nitrate uptakein N-starved plants was not accompanied by additional phosphateor sulphate absorption, but the plants had the capacity to absorbmore potassium, whether or not ammonium was also present inthe solution. Indirect evidence from analyses of root tissuesuggests that these responses to mild N-stress may depend onsome property of an N fraction which does not include nitrateor ammonium. Hordeum vulgare, barley, nitrogen, ammonium, nitrate, N-deficiency, absorption     

Selectivity and Kinetics of Ion Uptake by Barley Plants Following Nutrient Deficiency

Barley plants grown without an external supply of phosphorus,sulphur, chlorine or nitrogen subsequently absorbed these nutrients,as phosphate, sulphate, chloride and nitrate, more rapidly thandid nutrient, sufficient control plants under similar conditions.With phosphorus, sulphur and chlorine, increased absorptionwas restricted to the nutrient which had been deficient, orto close chemical analogues of it, the uptake of other anionsbeing unaffected or decreased. The selectivity of enhanced nitrateuptake by nitrogen-deficient plants was not examined. The differencesin the rates of phosphate, sulphate and chloride absorptionby plants of differing nutrient status were due principallyto changes in the maximum transport capacity for these anionsper unit weight of root, although in plants grown without externalchloride there was some evidence that the roots also developedan increased affinity for that ion. Hordeum vulgare, barley, mineral nutrient deficiency, ion absorption, kinetics of ion uptake, phosphate, sulphate, chloride, arsenate, bromide, selenate     

Nitrogen-13 Studies of Nitrate Fluxes in Barley Roots: II. EFFECT OF PLANT N-STATUS ON THE KINETIC PARAMETERS OF NITRATE INFLUX

Influx of nitrate into the roots of intact barley plants wasfollowed over periods of 1–15 min using nitrogen-13 asa tracer. Based on measurements taken over 15 min from a rangeof external nitrate concentrations (0·2–250 mmolm^–3), the kinetic parameters of influx, I_max and K_m, werecalculated. Compared with plants grown in the presence of nitrate throughout,plants that had been starved of N for 3 d showed a significantlygreater value ofI_max for ¹³N-nitrate influx (by a factor of1·4–1·8), but a similar value of K_m (12–14mmol m^–3). Pre-treating N-starved plants with nitratefor about 5 h further increased the subsequent rate of ¹³N-nitrateinflux, but had little effect in the unstarved controls. Allowingfor this induction of additional nitrate transport, the differencein rates of nitrate influx in control and N-starved plants wassufficient to account for the previously-observed differencein net uptake by the two groups of plants. In barley plants grown without any exposure to nitrate, butwith ammonium as N-source, both I_max and K_m for subsequent ¹³N-nitrateinflux were significantly decreased (by about one-half) comparedwith the corresponding nitrate-grown controls. The importance of changes in the rate of influx in the regulationof net uptake of nitrate is discussed. Key words: Ion transport, nitrate, influx, kinetic parameters, N-deficiency     

Nitrate and ammonium absorption by plants growing at a sufficient or insufficient level of phosphorus in nutrient solutions

Summary Absorption of nitrate and ammonium was studied in water culture experiments with 4 to 6 weeks old plants of barley (Hordeum vulgare L.), buckwheat (Fagopyrum esculentum L. Moench) and rape (Brassica napus L.). The plants were grown in a complete nutrient solution with nitrate (5.7±0.2 mM) or nitrate (5.6±0.2 mM) + ammonium (0.04±0.02 mM). The pH of the nutrient solution was kept at 5.0 using a pH-stat. It was found that phosphorus deficiency reduced the rate of nitrate uptake by 58±3% when nitrate was the sole N source and by 83±1% when both nitrate and ammonium were present. The reduction occurred even before growth was significantly impeded by P deficiency. The inhibition of the uptake of ammonium was less,i.e. ammonium constituted 10±1% of the total N uptake in the P sufficient plants and 30±5% in the P deficient plants. The reduction of nitrate absorption greatly decreased the difference between the uptake of anions and cations. It is suggested that P deficiency reduced the assimilation of NO ₃ ^– into the proteins, which might cause a negative feedback on NO ₃ ^– influx and/or stimulate NO ₃ ^– efflux.     

Nutrient-limited growth rates: quantitative benefits of stress responses and some aspects of regulation

Young sunflower plants (Helianthus annuus L.) under stress oflow nitrate or phosphate availability exhibited increases inroot: shoot ratio and in kinetic parameters for uptake. Theyshowed no significant changes in photosynthetic utilizationof either nutrient. Increases in root: shoot ratio were achievedby early and persistent suppression of shoot growth, but notroot growth. Affinity for phosphate uptake, 1/K_m(P), increasedwith phosphate stress, as did affinity for nitrate uptake, 1/K_m(N),with nitrate stress. Maximal uptake rate, V_max, for phosphateuptake increased with phosphorus stress; V_max for nitrate didnot increase with nitrogen stress. Phosphate V_max was relatedstrongly to root nutrient status. Decreases in V_max with plantage were not well explained by changes in age structure of roots.Estimated benefits of acclimatory changes in root: shoot ratioand uptake kinetics ranged up to 2-fold increases in relativegrowth rate, RGR. The relation of RGR to uptake physiology followedpredictions of functional balance moderately well, with somesystematic deviations. Analyses of RGR using growth models implyno significant growth benefit from regulating Vmax, specifically,not from down-regulating it at high nutrient availability. Quantitativebenefits of increases in root: shoot ratio and uptake parametersare predicted to be quite small under common conditions whereinnutrient concentrations are significantly depleted by uptake.The root: shoot response is estimated to confer the smallestbenefit under non-depleting conditions and the largest benefitunder depleting conditions. Even then, the absolute benefitis predicted to be small, possibly excepting the case of heterogeneoussoils. Depleting and non-depleting conditions are addressedwith very different experimental techniques. We note that atheoretical framework is lacking that spans both these cases,other than purely numerical formulations that are not readilyinterpreted. Key words: Nutrient stress, nutrient uptake, nutrient use efficiency, relative growth rate, Helianthus annuus     

Effects of Atrazine,Metolachlor, Carbaryl and Chlorothalonil on Benthic Microbes and Their Nutrient Dynamics

Atrazine, metolachlor, carbaryl, and chlorothalonil are detected in streams throughout the U.S. at concentrations that may have adverse effects on benthic microbes. Sediment samples were exposed to these pesticides to quantify responses of ammonium, nitrate, and phosphate uptake by the benthic microbial community. Control uptake rates of sediments had net remineralization of nitrate (−1.58 NO₃ µg gdm⁻¹ h⁻¹), and net assimilation of phosphate (1.34 PO₄ µg gdm⁻¹ h⁻¹) and ammonium (0.03 NH₄ µg gdm⁻¹ h⁻¹). Metolachlor decreased ammonium and phosphate uptake. Chlorothalonil decreased nitrate remineralization and phosphate uptake. Nitrate, ammonium, and phosphate uptake rates are more pronounced in the presence of these pesticides due to microbial adaptations to toxicants. Our interpretation of pesticide availability based on their water/solid affinities supports no effects for atrazine and carbaryl, decreasing nitrate remineralization, and phosphate assimilation in response to chlorothalonil. Further, decreased ammonium and phosphate uptake in response to metolachlor is likely due to affinity. Because atrazine target autotrophs, and carbaryl synaptic activity, effects on benthic microbes were not hypothesized, consistent with results. Metolachlor and chlorothalonil (non-specific modes of action) had significant effects on sediment microbial nutrient dynamics. Thus, pesticides with a higher affinity to sediments and/or broad modes of action are likely to affect sediment microbes'' nutrient dynamics than pesticides dissolved in water or specific modes of action. Predicted nutrient uptake rates were calculated at mean and peak concentrations of metolachlor and chlorothalonil in freshwaters using polynomial equations generated in this experiment. We concluded that in natural ecosystems, peak chlorothalonil and metolachlor concentrations could affect phosphate and ammonium by decreasing net assimilation, and nitrate uptake rates by decreasing remineralization, relative to mean concentrations of metolachlor and chlorothalonil. Our regression equations can complement models of nitrogen and phosphorus availability in streams to predict potential changes in nutrient dynamics in response to pesticides in freshwaters.     

Ammonium Nutrition Enhances Chlorophyll and Glaucousness in Kohlrabi

Kohlrabi (Brassica oleracea var.gongylodes) plants were grownin the greenhouse under autumn conditions and fertilized eitherwith pellets containing nitrogen as 40% ammonium sulphate and60% urea or with nutrient solution containing nitrogen predominantlyas nitrate. Plants given nitrogen as ammonium ions developedglaucous leaves compared to those supplied with nitrate whichformed glossy leaves. Ammonium-induced glaucousness was theresult of a two-fold increase in the amount of epicuticularwax and a markedly altered fine structure. Leaves from ammoniumfertilized kohlrabi plants also showed a 21% increase in chlorophyllcontent together with a reduction in the chlorophyll a:b ratioand decreased ground state fluorescence compared to plants suppliedwith nitrate. Photosynthesis and stomatal transpiration wereunaffected by the form of supplied nitrogen. Brassica oleracea ; chlorophyll; chlorophyll fluorescence; epicuticular wax; glaucousness; photosynthesis; transpiration     

Differences in steady-state net ammonium and nitrate influx by cold- and warm-adapted barley varieties

A flowing nutrient culture system permitted relatively rapid determination of the steady-state net nitrogen influx by an intact barley (Hardeum vulgare L. cv Kombar and Olli) plant. Ion-selective electrodes monitored the depletion of ammonium and nitrate from a nutrient solution after a single pass through a root cuvette. Influx at concentrations as low as 4 micromolar was measured. Standard errors for a sample size of three plants were typically less than 10% of the mean.     

Changes in Crassulacean Acid Metabolism of Kalanchoe blossfeldiana by Different Nitrogen Sources

Kalanchoe blossfeldiana Poelln. cv. Hikan (a Crassulacean acidmetabolism (CAM) plant) was grown in pots containing soil for6 months and then cultured in nutrient solution containing 10mM nitrate or ammonium as a sole nitrogen source for 2 or 3months, under a long-day (16 h) condition. Plant growth was better in the nitrate medium. Leaves of thenitrate-grown plants showed greater diurnal fluctuations intitratable acidity and malate content than those of the ammonium-grownplants. The diurnal patterns in CO₂ exchange of nitrate-grownplants were basically similar for both groups, but the amountof net CO₂ uptake at night was twice as large in the nitrate-grownplants. The leaves of the nitrate-grown plants had 1.3 to 2.5times higher activities of phosphoenolpyruvate carboxylase (PEPC),phosphofructokinase (PFK) and NAD glycelaldehyde-3-phosphatedehydrogenase (G3PDH). These results indicate that K. blossfeldianagrown in nitrate medium showed more CAM activity than thosein ammonium medium. (Received August 13, 1987; Accepted February 22, 1988)     

Rapid, Reversible Inhibition of Nitrate Influx in Barley by Ammonium

The rate of influx of nitrate into the roots of intact barleyplants was measured over a period of 3–5 min from externalnitrate concentrations of 1–150 mmol m^–3, using¹³N-labelled nitrate as tracer. Ammonium at external concentrationsof 0.005–50 mol m^–3 inhibited nitrate influx ina manner which did not conform to a simple kinetic model butincreased approximately as the logarithm of the ammonium concentration.At any particular ammonium concentration, inhibition of nitrateinflux reached its full extent within 3 min of the ammoniumbeing supplied and was not made more severe by up to 17 minpre-treatment with ammonium. On removing the external ammonium,nitrate influx returned to its original rate within about 3min. Potassium at 0.005–50 mol m^–3 did not reproducethe rapid effect of ammonium on nitrate influx. Net uptake of nitrate also decreased when ammonium was supplied,over a similar timescale and to a similar extent as nitrateinflux. The decrease in nitrate influx caused by ammonium wassufficient to account for the observed reduction in net uptake,without necessitating any acceleration of nitrate efflux. Key words: Hordeum vulgare, roots, ion transport, short-lived isotopes, ¹³N     

The effect of ammonium ions on membrane potential and anion flux in roots of barley and tomato

The effects of ammonium (0–5 mol m^?3) on root hair membrane potential and on the influx of nitrate and phosphate were investigated in roots of intact barley and tomato plants. In both species, addition of ammonium to the medium bathing the roots caused an almost immediate depolarization of the membrane potential; the depolarization was greater at higher concentrations of ammonium. Influx of ¹³NC₃^? and ³²Pi was inhibited over the same time scale and concentration range. In tomato roots, there was little further depolarization of the membrane potential or inhibition of anion influx at ammonium concentrations above 0.4 mol m^?3. In barley roots, the inhibition of nitrate influx and the depolarization of the membrane potential did not saturate below 5 mol m^?3 ammonium.     

The photosynthetic response of nutrient-depleted dilute cultures of Skeletonema costatum to pulses of ammonium and nitrate; the importance of phosphate

Experimental data on changes in carbon fixation rate causedby nutrient pulses in dilute cultures of nutrient-depleted Skeletonemacostatum are reported. Pulses contained single nutrients ormixtures of a nitrogen source and phosphate, concentrationsranging from 0 to 5 µM for ammonium and nitrate and from0 to 1 µM for phosphate. The cultures were incubated overnightafter pulsing to allow the rapid stage of nutrient uptake tocome to completion before measurement of carbon fixation thenext day. Increments in N:C ratios due to nitrogenous nutrientuptake depended upon the P:C ratio in the cells as well as theconcentration of the pulse. When P:C ratios were low (<0.005),increases in nitrogen repressed photosynthesis. Phosphate uptakewas independent of the absence or presence of a nitrogen sourceand had only a small stimulatory effect upon carbon fixation.When added jointly with ammonium or nitrate, however, largeincreases in the rate of photosynthesis resulted. These weredue mainly to rises in chlorophyll a concentrations resultingfrom higher N:C ratios in the cells. Chlorophyll-specific carbonfixation rates were hyperbolic functions of P:C ratios but exhibiteda C-shaped relationship to N:C ratios. The stimulatory and repressiveeffects of the nutrient pulses are shown to be consistent withthe view that the rate of photosynthesis at constant illuminationand carbon dioxide partial pressure is mainly controlled bythe chlorophyll a concentration and phosphate availability inthe phytoplankton.     

Nitrogen Utilization in Lemna: II. Studies of Nitrate Uptake Using NO(3)

¹³N-labeled nitrate was used to trace short-term nitrate influx into Lemna gibba L. G3 in experiments where disappearance of both radioactivity and total nitrate from the incubation medium was measured continuously and simultaneously. In plants performing net nitrate uptake from an initial nitrate concentration of 40 to 60 micromolar, there was no discrepancy between net uptake and influx, irrespective of the N status of the plants, indicating that concomitant nitrate efflux was low or nil. Plants treated with tungstate to inactivate nitrate reductase were able to take up nitrate following induction of the uptake system by exposure to a low amount of nitrate. Also, in this case, net uptake was equivalent to influx. In tungstate-treated plants preloaded with nitrate, both net uptake and influx were nil. In contrast to these observations, a clear discrepancy between net uptake and influx was observed when the plants were incubated at an initial nitrate concentration of approximately 5 micromolar, where net uptake is low and eventually ceases. It is concluded that plasmalemma nitrate transport is essentially unidirectional in plants performing net uptake at a concentration of 40 to 60 micromolar, and that transport is nil when internal nitrate sinks (vacuole, metabolism) are eliminated. The efflux component becomes increasingly important when the external concentration approaches the threshold value for net nitrate uptake (the nitrate compensation point) where considerable exchange between internal and external nitrate occurs.     

Uptake of ammonium and nitrate by carob (Ceratonia siliqua) as affected by root temperature and inhibitors

Seedlings of carob ( Ceratonia siliqua L. cv. Mulata) were grown in nutrient solution culture for 5 weeks, with or without nitrogen at different root temperatures (10, 16, 22, 30, 35 or 40deg;C) and with the air temperature kept between 20 and 24°C. The nitrogen was given as either ammonium or nitrate. At all root temperatures studied, nitrogen-depleted plants developed higher net uptake rates for nitrogen than plants grown in the presence of nitrogen. Temperature affected the kinetic parameters of nitrate uptake more than those of ammonium uptake. With increasing root temperature, the K_m of ammonium uptake decreased, but to a lesser extent than the K_m for nitrate. The increase in V_max of ammonium uptake with temperature was also less noticeable than that for nitrate uptake. Ammonium and nitrate uptakes were inhibited in a similar way by respiratory or protein synthesis inhibitors. It may be noted that ammonium uptake in the presence of inhibitors at 40°C was higher than uptake at 10°C without inhibitors. Some similarities between the transport mechanisms for nitrate and ammonium are underlined in the present work. Components of both transport systems displayed saturation kinetics and depended on protein synthesis and energy. The following components of nitrate uptake were distinguished: (a) a passive net influx into the apparent free space; (b) a constitutive active uptake and (c) active uptake dependent on protein synthesis. We may similarly define three ammonium uptake systems: (a) a passive influx into the apparent free space; (b) passive diffusion uptake at high temperature and (c) active uptake dependent on protein synthesis. The possible role of the ratio between mechanism (c) and mechanism (b) as determinant of ammonium sensitivity is discussed.     

Nitrogen Utilization in Lemna: II. Studies of Nitrate Uptake Using 13NO3?

¹³N-labeled nitrate was used to trace short-term nitrate influx into Lemna gibba L. G3 in experiments where disappearance of both radioactivity and total nitrate from the incubation medium was measured continuously and simultaneously. In plants performing net nitrate uptake from an initial nitrate concentration of 40 to 60 micromolar, there was no discrepancy between net uptake and influx, irrespective of the N status of the plants, indicating that concomitant nitrate efflux was low or nil. Plants treated with tungstate to inactivate nitrate reductase were able to take up nitrate following induction of the uptake system by exposure to a low amount of nitrate. Also, in this case, net uptake was equivalent to influx. In tungstate-treated plants preloaded with nitrate, both net uptake and influx were nil. In contrast to these observations, a clear discrepancy between net uptake and influx was observed when the plants were incubated at an initial nitrate concentration of approximately 5 micromolar, where net uptake is low and eventually ceases. It is concluded that plasmalemma nitrate transport is essentially unidirectional in plants performing net uptake at a concentration of 40 to 60 micromolar, and that transport is nil when internal nitrate sinks (vacuole, metabolism) are eliminated. The efflux component becomes increasingly important when the external concentration approaches the threshold value for net nitrate uptake (the nitrate compensation point) where considerable exchange between internal and external nitrate occurs.     

The Inducible Formation and Stability of Nitrate Reductase in Higher Plants: I. EFFECTS OF NITRATE AND MOLYBDENUM ON ENZYME ACTIVITY IN CAULIFLOWER (BRASSICA OLERACEA VAR. BOTRYTIS)

The enzyme nitrate reductase could not be detected in leaf tissuesof cauliflower plants grown in sterile cultures with glutamicacid or ammonium sulphate if nitrate was absent. Excised leaftissues from these plants formed the enzyme for several hoursat a steady rate when infiltrated with nitrate. Plants starvedof nitrate for short periods lost enzyme activity which wasrestored in excised tissues upon infiltration with nitrate butnot with ammonium sulphate or nitrite. Molybdenum-deficientplants grown with nitrate also lacked enzyme activity whichwas restored in excised tissues after infiltration with molybdenum.Both nitrate and molybdenum were required to produce maximalrates of enzyme formation in excised tissues of plants grownwith ammonium sulphate and no molybdenum. Apparent Michaelisconstants for nitrate and molybdenum were found to be about10^-5 and 10^-7 respectively. The capacity of excised tissuesto respond to the inducer varied with their age and leaf positionon the plant and was exercised under conditions where growthwas unlikely. Increases in specific activities were similar.There was no evidence of a lag in response to nitrate or molybdenumwith tissues of plants grown with ammonium sulphate or glutamicacid in sterile cultures but lag periods were observed withtissues from plants deprived of nitrate. Cell-free preparationswere unable to respond to either factor. The results are interpretedas evidence for induced enzyme formation in vivo in responseto the substrate or the constituent metal.     

Effects of Exposure to Ammonium and Transplant Shock upon the Induction of Nitrate Absorption

In barley (Hordeum vulgare L. cv Steptoe) seedlings, the time course for induction of root nitrate absorption varied significantly with pretreatment. Net nitrate uptake of nitrogen-deprived plants more than doubled during the 12 hours after first exposure to nitrate. For these plants, gentle physical disturbance of the roots inhibited net nitrate absorption for more than 6 hours and potassium absorption for 2 hours. Pretreatment with ammonium appeared sufficient to induce nitrate absorption; plants either grown for 2 weeks on or exposed for only 10 hours to a medium containing ammonium as a sole nitrogen source showed high rates of net nitrate uptake when first shifted to a medium containing nitrate. Gentle physical manipulation of these plants inhibited nitrate absorption for 2 hours and potassium absorption for more than 12 hours. These results indicate (a) that experimental protocols should avoid physical manipulation of the roots when-ever possible and (b) that ammonium or a product of ammonium assimilation can induce nitrate absorption.     

Inward and Outward Movement of Ammonium in Root Systems: Transient Responses during Recovery from Nitrogen Deprivation in Presence of Ammonium

Morgan, M. A. and Jackson, W. A. 1988. Inward and outward movementof ammonium in root systems: transient responses during recoveryfrom nitrogen deprivation in presence of ammonium.— J.exp. Bot. 39: 179-191. Net ammonium uptake by 20-d-old wheat (Triticum aestivum cv.Kleiber) and oat (Avena sativa cv. Tarok) seedlings was increased5- to 10-fold when the seedlings were deprived of nitrate duringthe 14-20 d period. The effect of nitrogen deprivation was toincrease net ¹⁵N-ammonium influx and decrease net ¹⁴N-ammoniumefflux during a 1 h assay period. The sizeable rate of net ¹⁵N-ammoniuminflux resulting from nitrogen deprivation was stimulated furtherby prior exposure of the seedlings to ¹⁴N-ammonium for 5 h.Additional exposure to ¹⁴N-ammonium caused the stimulated rateof ¹⁵N-ammonium influx to decline. During the 1 h assays in¹⁵N-ammonium, net ¹⁴N-ammonium efflux increased after 2 h exposureto ¹⁴N-ammonium, peaked at 5–10 h, and then declined.The consequence of the differential response of the influx andefflux processes in wheat was a marked decrease in net ammoniumuptake in the initial 2–5 h, followed by a recovery which,in turn, was followed by a slow decline. In oat, there was norecovery in net ammonium uptake after 2–5 h. Interference in ammonium assimilation by presence of methioninesulphoximine after 5 h did not inhibit expression of the ammonium-stimulatednet ¹⁵N-ammonium influx at 10 h but did substantially increasenet ¹⁴N-ammonium efflux. In nitrogen depleted seedlings, andin those exposed to ¹⁴N-ammonium for 2 h, subsequent net ¹⁴N-ammoniumefflux during 1 h in ¹⁵N-ammonium exceeded the quantity of ¹⁴N-ammoniuminitially in the roots. The increase in ¹⁵N-ammonium influx upon nitrogen deprivation,its further stimulation with 5-10 h exposure to ammonium andits subsequent decline, are discussed as possibly resultingfrom (a) the operation of two ammonium influx systems (b) theinterplay of tissue ammonium and a product of its assimilationrespectively acting as positive and negative effectors of asingle influx system and (c) variations in energy supply fromthe shoots. Key words: Net ammonium uptake, stimulated ammonium influx, ammonium efflux, tissue ammonium     

Nitrogen Assimilation in Barley (Hordeum vulgare L. cv. Mazurka) in Response to Nitrate and Ammonium Nutrition

Barley plants (Hordeum vulgare L. cv. Mazurka) were grown inaerated solution cultures with 2 mM or 8 mM inorganic nitrogensupplied as nitrate alone, ammonium alone or 1:1 nitrate+ammonium.Activities of the principal inorganic nitrogen assimilatoryenzymes and nitrogen transport were measured. Activities ofnitrate and nitrite reductases, glutamine synthetase and glutamatesynthase were greater in leaves than in roots but glutamatedehydrogenase was most active in roots. Only nitrate and nitritereductases changed notably (4–10 times) in response tothe different nitrogen treatments. Nitrate reductase appearedto be rate-limiting for nitrate assimilation to glutamate inroots and also in leaves, where its total in vitro activitywas closely related to nitrate flux in the xylem sap and wasslightly in excess of that needed to reduce the transportednitrate. Xylem nitrate concentration was 13 times greater thanthat in the nutrient solution. Ammonium nitrogen was assimilatedalmost completely in the roots and the small amount releasedinto the xylem sap was similar for the nitrate and the ammoniumtreatments. The presence of ammonium in the nutrient decreasedboth export of nitrate to the xylem and its accumulation inleaves and roots. Nitrate was stored in stem bases and was releasedto the xylem and thence to the leaves during nitrogen starvation.In these experiments, ammonium was assimilated principally inthe roots and nitrate in the leaves. Any advantage of this divisionof function may depend partly on total conversion of inorganicnitrogen to amino acids when nitrate and ammonium are givenin optimal concentrations. Hordeum vulgare L., barley, nitrate, ammonium, nitrate reductase, nitrite reductase, glutamine synthetase, glutamate synthase, glutamate dehydrogenase, nitrogen transport