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     

Reciprocal Ammonium Transport Into and Out of Plant Roots: Modifications by Plant Nitrogen Status and Elevated Root Ammonium Concentration

Wheat and oat were grown for 20 d on a nitrate-containing solution(nitrogen-replete plants) or for the last 6 d of this periodon a nitrate-free solution (nitrogen-depleted plants). Exposureof the nitrogen-depleted plants on day 20 to nitrate-free solutionscontaining 500 mmol m^–3 ammonium (96 A% ¹⁵N) resultedin a cumulative net influx of ¹⁵N-ammonium over an 8 h periodthat was appreciably greater than that of the nitrogen-repleteplants. Both the initial rate and the more restricted rate afterthe first hour were enhanced by nitrogen deprivation. In thenitrogen-replete plants, cumulative net efflux of endogenous¹⁴N-ammonium was approximately equivalent to net ammonium uptakeduring the first hour, and was essentially complete after 1–2h. Pretreating nitrogen-depleted plants for 5 h in 500 mmolm^{–3 15}N-ammonium (99 A% ¹⁵N) resulted in root ammoniumconcentrations of 12.7?1.1 and 16.0?0.4 µmol for wheat and oat, respectively. Subsequent net efflux of ¹⁵N-ammoniumto 500 mmol m^–3 exogenous ¹⁴N-ammonium exceeded theseinitial amounts within 2 h. Increasing ambient ¹⁴N-ammoniumto 5000 mmol m^–3 increased net ¹⁵N-ammonium efflux suchthat net loss of the maximal original amount in the root tissuewas exceeded within 0.75 h. The data for both species indicatesubstantial reciprocal transfers of ammonium into and out ofroots of ammonium-treated plants and a significant degradationof recently synthesized products of ammonium assimilation concurrentwith ammonium assimilation. Key words: Accumulation, ammonium, efflux, oat, root, uptake, wheat     

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     

Inward and outward transport of ammonium in roots of maize and sorghum: contrasting effects of methionine sulphoximine

To examine the adaptive responses of ammonium efflux of nitrate-grownplants during first exposure to ammonium nutrition, isotopicflows of ammonium in and out of root tissue of maize (Zea maysL.) and sorghum (Sorghum bicolor L.) were delineated. Appreciableefflux of ¹⁴N-ammonium occurred during the first 2 min of exposure,suggesting that ammonium efflux is characteristic of plantsgrown with nitrate as the sole source of nitrogen. Treatmentwith methionine sulphoximine (MSX) enhanced the initial ¹⁴N-ammoniumefflux 5-fold in maize but less than 2-fold in sorghum. Increasingambient acidity from pH 6 to pH 4 had relatively little effecton 14N-ammonium efflux in either species. Initial influx ofexogenous ¹⁵N-ammonium was stimulated by MSX in maize (slightlyless at pH 4 than at pH 6), but either was not affected (atpH 6) or was inhibited (at pH 4) in sorghum. Assuming that therate of ammonium influx remained constant during the 2 h adaptationperiod, and that efflux of ¹⁵N-ammonium was negligible duringthe 0-2 min period, permitted estimates to be made of the unidirectionalinward and outward flow of the two isotopic ammonium species.The decline in efflux of endogenous ¹⁴N-ammonium after the 2min exposure was more than compensated for by an increase inefflux of the exo-genously-supplied 15N-ammonium. As a consequence,total ammonium efflux increased significantly during the adaptationperiod. By this analysis about 60% of the total efflux in maizewas as 1BN-ammonium within the 2-5 min period whereas in sorghumit was 20% to 40%. The data indicate that appreciable ammoniumefflux occurs in nitrate-grown seedlings in spite of low rootammonium concentrations, and that substantial futile cyclingof ammonium occurs in both maize and sorghum during net transportof ammonium into their root tissue. Key words: Ammonium, cycling, transport, maize, sorghum     

Synthesis and 15N-labelling of glutamine and glutamate in maize roots during early stages of 15N-ammonium assimilation

The concentrations of glutamine and glutamate, and the abundanceof ¹⁵N in these compounds, were measured in roots of intact,nitrate-grown maize plants fed with ¹⁵N-nitrate or ¹⁵N-ammoniumfor periods of 3–80 min. On supplying ¹⁵N-ammonium therewas a rapid and almost linear accumulation of glutamine, itsconcentration in the roots rising 3-fold over 1 h. Supplying¹⁵N-nitrate instead of ¹⁵N-ammonium did not increase root glutamine,and the concentration of glutamate was not affected by eithertreatment. The time-course of amide ¹⁵N-labelling seen in glutamine extractedfrom roots which had been supplied with ¹⁵N-ammonium could bestbe explained by a model in which (a) the ‘additional’glutamine which accumulates rapidly during ¹⁵N-ammonium feedingis heavily amide-labelled from the outset, and (b) of the glutaminealready present in the roots, only a small proportion (c. 10–15%)incorporates ¹⁵N during the initial 60–80 min of ¹⁵N-ammoniumfeeding, the remainder (c. 85–90%) remaining essentiallyunlabelled over this period. The latter is assumed to be locatedin the vacuoles. Even though prior N-starvation stimulated ammonium net uptakemarkedly, the data were not of sufficient quality to show whetherthe relative sizes of the extra-vacuolar and vacuolar glutaminefractions were altered by this treatment. For that reason itwas not possible to determine whether cytosolic glutamine hasa role in regulating N-absorption. Key words: Subcellular compartmentation, regulation, N-absorption     

Influx and Efflux of Nitrate and Ammonium in Italian Ryegrass and White Clover Roots: Comparisons Between Effects of Darkness and Defoliation

Seedlings of Italian ryegrass (Lolium multiflorum Lam. cv. RVP)and clonal stolon cuttings of white clover (Trifolium repensL. cv. Blanca) were grown for 19 d in flowing solution culture,with N supplied as either 250 mmol m^–3 NO₃^– or NH₃⁺.Rates of net uptake, influx and translocation of NO₃^–and NH₄⁺ were then determined using ¹⁵N and ¹³N labelling techniques:between 3–5 h into the photoperiod following 8 h darknessfor white clover (CL), and for ryegrass plants that were eitherentire (IL) or with shoots excised 90 min prior to ¹³N influx(IC); and 75 min into the photoperiod following 37–39h darkness for ryegrass (ID). Rates of net uptake, influx andefflux of NH₄⁺ exceeded those of NO₃^– in IL and IC ryegrassplants: the opposite occurred in white clover (CL). The decreasein net uptake following defoliation of ryegrass was greaterfor NH₄⁺ (62%) than NO₃^– (40%). For NH₄⁺ this was associatedwith a large decrease in influx from 110 to 6.0µmol h^–1g^–1 root fr. wt; but for NO₃^–, influx only decreasedfrom 42 to 37 µmol h^–1 g^–1. Prolonged exposureto darkness (ID plants) also lowered net uptake of NO₃^–and NH₄⁺ by, respectively, 86% and 95% of IL levels. For NH₄⁺this was characterized by a large decrease in influx and a smalldecrease in efflux; whilst for NO₃^– the effect of a largedecrease in influx was reinforced by a smaller increase in efflux. The data were used to estimate the translocatory fluxes of NO₃^–(03–20µmol h^–1 g^–1) and NH₄⁺ (003–0.4µmolh^–1 g^–1), assimilation in the roots of NO₃^–(02–26µmol h^–1 g^–1) and NH⁺⁴ (05–89 µmolh^–1 g^–1), and the concentrations of NO₃^– (9–15mol m^–3) in the cytoplasmic compartment of the roots.The relevance of variable influx and efflux to models for theregulation of N uptake is discussed. Key words: Lolium multiflorum, Trifolium repens, influx, efflux, nitrate, ammonium, ¹³N     

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     

The Effect of Root Temperature on Growth and Uptake of Ammonium and Nitrate by Brassica napus L. cv. Bien venu in Flowing Solution Culture: II. UPTAKE FROM SOLUTIONS CONTAINING NH4NO3

Macduff, J. H., Hopper, M. J. and Wild, A. 1987. The effectof root temperature on growth and uptake of ammonium and nitrateby Brassica napus L. CV. Bien venu in flowing solution culture.II. Uptake from solutions containing NH₄NO₃.—J. exp. Bot.38: 53–66 The effects of root temperature on uptake and assimilation ofNH₄⁺ and NO₃^– by oilseed rape (Brassica napus L. CV. Bienvenu) were examined. Plants were grown for 49 d in flowing nutrientsolution at pH 6?0 with root temperature decrementally reducedfrom 20?C to 5?C; and then exposed to different root temperatures(3, 5, 7, 9, 11, 13, 17 or 25?C) held constant for 14 d. Theair temperature was 20/15?C day/night and nitrogen was suppliedautomatically to maintain 10 mmol m^–3 NH₄NO₃ in solution.Total uptake of nitrogen over 14 d increased threefold between3–13?C but was constant above 13?C. Net uptake of NH₄⁺exceeded that of NO₃^– at all temperatures except 17?C,and represented 47–65% of the total uptake of nitrogen.Unit absorption rates of NH₄⁺ and of 1?5–2?7 for NO₃^–suggested that NO₃^– absorption was more sensitive thanNH₄⁺ absorption to temperature. Rates of absorption were relativelystable at 3?C and 5?C compared with those at 17?C and 25?C whichincreased sharply after 10 d. Tissue concentration of N in theshoot, expressed on a fresh weight basis, was independent ofroot temperature throughout, but doubled between 3–25?Cwhen expressed on a dry weight basis. The apparent proportionof net uptake of NO₃^– that was assimilated was inverselyrelated to root temperature. The results are used to examinethe relation between unit absorption rate adn shoot:root ratioin the context of short and long term responses to change ofroot temperature Key words: Brassica napus, oilseed rape, root temperature, nitrogen uptake     

Seasonal nitrogen assimilation and carbon fixation in a fjordic sea loch

Carbon (C) fixation and nitrogen (N) assimilation rates havebeen estimated from ¹⁴C and ¹⁵N techniques for a 12 month periodin a Scottish sea loch. The maximum rate of nitrogen assimilated(29.92 mmol N m^–2 day^–1) was in April at the mostseaward station; similar high rates were experienced duringMay at the other stations. Carbon fixation rates were maximal(488–4047 mg C m^–2day^–1) at the time of highphytoplankton biomass (maximum 8.3 mg m^–3 chlorophylla) during May, whilst nitrate concentrations remained >0.7µ.mol l^–1. C:N assimilation ratios suggest nitrogenlimitation only during the peak of the spring bloom, althoughat times nitrogen (nitrate and ammonium) concentration fellto 0.2 µmol l^–1 in the following months. The verticalstability of the water column, influenced by tidal and riverineflushing, varied along the axis of the loch, resulting in markeddifferences between sampling stations. Although ammonium waspreferentially assimilated by phytoplankton, >50% of productionwas supported by nitrate uptake and only during the summer monthswas the assimilation of ammonium quantitatively important.     

Growth and Preferences for Ammonium or Nitrate Uptake by Barley in Relation to Root Termperature

Barley plants (Hordewn vulgare L. cv. Atem) were grown fromseed for 28 d in flowing solution culture, during which timeroot temperature was lowered decrementally to 5?C. Plants werethen subjected to root temperatures of 3, 5, 7, 9, 11, 13, 17or 25 ?C, with common air temperature of 25/15 ?C (day/night).Changes in growth, plant total N, and NO₃^– levels, andnet uptake of NH₄⁺ and NO₃^– from a maintained concentrationof 10 mmol m^–3 NH₄NO₃ were measured over 14 d. Dry matterproduction increased 6-fold with increasing root temperaturebetween 3–25 ?C. The growth response was biphasic followingan increase in root temperature. Phase I, lasting about 5 d,was characterized by high root specific growth rates relativeto those of the shoot, particularly on a fresh weight basis.During Phase I the shoot dry weight specific growth rates wereinversely related to root temperature between 3–13 ?C.Phase 2, from 5–14 d, was characterized by the approachtowards, and/or attainment of, balanced exponential growth betweenshoots and roots. Concentrations of total N in plant dry matterincreased with root temperature between 3–25 ?C, moreso in the shoots than roots and most acutely in the youngestfully expanded leaf (2?l–6?9% N). When N contents wereexpressed on a tissue fresh weight basis the variation withtemperature lessened and the highest concentration in the shootwas at 11 ?C. Uptake of N increased with root temperature, andat all temperatures uptake of NH₄⁺, exceeded that of NO₃^–,irrespective of time. The proportions of total N uptake over14 d absorbed in the form of NH₄⁺ were (%): 86, 91, 75, 77,76, 73, 77, and 80, respectively, at 3, 5, 7, 9, Il, 13, 17,and 25 ?C. At all temperatures the preference for NH₄⁺ overNO₃^– uptake increased with time. An inverse relationshipbetween root temperature (3–11 ?C) and the uptake of NH₄⁺as a proportion of total N uptake was apparent during PhaseI. The possible mechanisms by which root temperature limitsgrowth and influences N uptake are discussed. Key words: Hordeum vulgare, root temperature, ammonium, nitrate, ion uptake, growth rate     

Ammonium and nitrate uptake by soybean during recovery from nitrogen deprivation

Soybean [Glycine max (L.) Merrill] plants that had been subjectedto 15 d of nitrogen deprivation were resupplied for 10 d with1.0 mol m^–3 nitrogen provided as NO₃^–, NH₄⁺, orNH₄⁺+NO₃^– in flowing hydroponic culture. Plants in a fourthhydroponic system received 1.0 mol m^–3 NO₃^– duringboth stress and resupply periods. Concentrations of solublecarbohydrates and organic acids in roots increased 210 and 370%,respectively, during stress. For the first day of resupply,however, specific uptake rates of nitrogen, determined by ionchromatography as depletion from solution, were lower for stressedthan for non-stressed plants by 43% for NO₃^- resupply, by 32%for NH₄⁺ + NO₃^– resupply, and 86% for NH₄⁺ resupply. Whenspecific uptake of nitrogen for stressed plants recovered torates for non-stressed plants at 6 to 8 d after nitrogen resupply,carbohydrates and organic acids in their roots had declinedto concentrations lower than those of non-stressed plants. Recoveryof nitrogen uptake capacity of roots thus does not appear tobe regulated simply by the content of soluble carbon compoundswithin roots. Solution concentrations of NH₄⁺ and NO₃^– were monitoredat 62.5 min intervals during the first 3 d of resupply. Intermittent‘hourly’ intervals of net influx and net effluxoccurred. Rates of uptake during influx intervals were greaterfor the NH₄⁺ -resupplied than for the NO₃^– -resuppliedplants. For NH₄⁺ -resupplied plants, however, the hourly intervalsof efflux were more numerous than for NO₃^– -resuppliedplants. It thus is possible that, instead of repressing NH₄⁺influx, increased accumulation of amino acids and NH₄⁺ in NH₄⁺-resupplled plants inhibited net uptake by stimulation of effluxof NH₄⁺ absorbed in excess of availability of carbon skeletonsfor assimilation. Entry of NH₄⁺ into root cytoplasm appearedto be less restricted than translocation of amino acids fromthe cytoplasm into the xylem. Key words: Ammonium, nitrate, nitrogen-nutrition, nitrogen-stress, soybean     

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     

Interactions of NH4+ and L-glutamate with NO3- transport processes of non-mycorrhizal Fagus sylvatica roots

The processes of NO₃^– uptake and transport and the effectsof NH₄⁺ or L-glutamate on these processes were investigatedwith excised non-mycorrhizal beech (Fagus sylvatica L.) roots.NO₃^– net uptake followed uniphasic Michaelis-Menten kineticsin a concentration range of 10µM to 1 mM with an apparentK_m of 9.2 µM and a V_max of 366 nmol g^–1 FW h^–1.NH₄⁺, when present in excess to NO₃^–, or 10 mM L-glutamateinhibited the net uptake of NO₃^– Apparently, part of NO₃^–taken up was loaded into the xylem. Relative xylem loading ofNO₃^– ranged from 3.21.6 to 6.45.1% of NO₃^– netuptake. It was not affected by treatment with NH₄⁺ or L-glutamate.¹⁶N/¹³N double labelling experiments showed that NO₃^–efflux from roots increased with increasing influx of NO₃^–and, therefore, declined if influx was reduced by NH₄⁺ or L-glutamateexposure. From these results it is concluded that NO₃^–net uptake by non-mycorrhizal beech roots is reduced by NH₄⁺or L-glutamate at the level of influx and not at the level ofefflux. Key words: Nitrate transport, net uptake, influx, efflux, ammonium, Fagus, Fagaceae     

Evidence of ammonium assimilation via the glutamine synthetase-glutamate synthase system in rice seedling roots

When rice seedling roots were fed ¹⁵N-ammonium for 1 hr, theamide nitrogen of glutamine showed the highest ¹⁵N abundance.Moreover, glutamine amino, glutamic acid, aspartic acid andalanine showed higher ¹⁵N abundance than ammonium did. In roots whose GS activity was inhibited with MS, both the amountof ammonium and its ¹⁵N abundance were increased. In contrast,both the amount of all examined amino acids containing glutamicacid and their ¹⁵N abundance decreased in roots whose GS activitywas inhibited. From these results, it could be concluded thatthe first step of ammonium assimilation in rice seedling rootswas mainly glutamine synthesis by GS and the second was glutamicacid formation by the GOGAT system. The results of an experiment using ¹⁵N glutamine also supportedthis conclusion. (Received February 23, 1977; )     

Nitrate Uptake and Reduction of Aseptically Cultivated Spruce Seedlings, Picea abies (L.) Karst

Spruce (Picea abies (L.) Karst.) seedlings were asepticallycultivated and the effects of different N-nutrition on net uptakeand reduction of nitrate were investigated. The characteristicsof nitrate uptake were calculated, K_s as 0?2 mol m^–3 andV_max as 18 µmol g^–1 d^–1. Low pH, and Al³⁺ in the medium caused adecrease in nitrate uptake rate. An in vivo assay was set upwhich allowed the measurement of NRA in both roots and needlesof spruce seedlings. The in vivo nitrate reductase activitywas repressed by ammonium and stimulated by nitrate. Nitratereduction was similar to nitrate uptake, negatively affectedby low pH and ammonium. Therefore, a limited N-supply to spruceseemed to occur when pH was low in the rhizosphere combinedwith the presence of Al³⁺ and . Key words: Spruce, nitrate uptake, nitrate reduction     

Diurnal regulation of NO3- uptake in soybean plants I. Changes in NO3- influx, efflux, and N utilization in the plant during the day/night cycle

The effect of light on NO₃^– utilization was investigatedin non-nodulated soybean (Clycine max L. Merr., cv. Kingsoy)plants during a 14/10 h light/dark period at a constant temperatureof 26C. A 30–50% decrease of net NO₃^– uptake ratewas observed 2–6 h after the lights were turned off. Thiswas specifically due to an inhibition of NO₃^– influx asmeasured by ¹⁵N incorporation during 5 min. The absolute valuesof NO₃^– efflux depended on whether the labelling protocolinvolved manipulation of the plants or not, but were not affectedby illumination of the shoots. Darkness had an even more markedeffect in lowering the reduction of ¹⁵NO₃^– in both rootsand shoots, as well as xylem transport of ¹⁵NO₃^– and reduced¹⁵N. Concurrently with this slowing down of transport and metabolicprocesses, accumulations of NO₃^– and Asn were significantlystimulated in roots during the dark period. These data are discussedin view of the hypothesis that darkness adversely affects NO₃^–uptake through specific feedback control, in response to alterationsin the later steps of N utilization which are more directlydependent on light. Key words: Glycine max, light/dark cycles, nitrate uptake, nitrate reduction     

Measurement of Ammonium Ions in Flowing Solution Culture and Diurnal Variation Uptake by Lolium perenne L

An existing system of flowing solution culture, in which pHand the concentration of several nutrient ions are automaticallymonitored and controlled, has been extended to include at 10 mmol N m^–3. A brief account is givenof the use of the equipment with a simulated sward of perennialryegrass (Lolium perenne L.). Uptake of measured over three successive days in June, varied with dailysolar radiation and exceeded 1000 mg N m^–2 d^–1.The uptake of showed a pattern of diurnal variation similar to the variation in solar radiation, but witha lag period for uptake of 5 h. Hourly uptake rates ranged from32 to 67 mg N m^–2h^–1 and solar radiation from 0to 2.8 MJ m^–2 h^–1. During a 24 h period, additionalmeasurements were made of K⁺ uptake and net H⁺ efflux, bothof which showed patterns of diurnal variation with lag periodsof 6 h hand 7 h, respectively. The stoichiometric ratio of thesum of and K⁺ to the net efflux of H⁺ was1.02: 1. Key words: Ion uptake, diurnal variation, Lolium perenne L.     

Plant Growth in Relation to the Supply and Uptake of NO3-: A Comparison Between Relative Addition Rate and External Concentration as Driving Variables

Two approaches to quantifying relationships between nutrientsupply and plant growth were compared with respect to growth,partitioning, uptake and assimilation of NO₃^– by non-nodulatedpea (Pisum sativum L. cv. Marma). Plants grown in flowing solutionculture were supplied with NO₃^– at relative addition rates(RAR) of 0·03, 0·06, 0·12, and 0·18d^–1, or constant external concentrations ([NO₃^–)of 3, 10, 20, and 100 mmol m^–3 over 19 d. Following acclimation,relative growth rates (RGR)approached the corresponding RARbetween 0·03–0.12 d^-1, although growth was notlimited by N supply at RAR =0.18 d^-1. Growth rates showed littlechange with [NO₃–] between 10–100 mmol m^–3(RGR=0·15 –0·16 d^-1). The absence of growthlimitation over this range was suggested by high unit absorptionrates of NO₃^–, accumulation of NO₃^– in tissues andprogressive increases in shoot: root ratio. Rates of net uptakeof NO₃^– from 1 mol m^–3 solutions were assessed relativeto the growth-related requirement for NO₃^–, showing thatthe relative uptake capacity increased with RGR between 0·03–0·06d^–1 , but decreased thereafter to a theoretical minimumvalue at RGR     

The Influence of NO-3 and NH+4 Nutrition on the Gas Exchange Characteristics of the Roots of Wheat (Triticum aestivum) and Maize (Zea mays) Plants

Respiratory oxygen consumption by roots was 1·4- and1·6-fold larger in NH⁺₄-fed than in NO^-₃-fed wheat (Triticumaestivum L.) and maize (Zea mays L.) plants respectively. Higherroot oxygen consumption in NH⁺₄-fed plants than in NO^-₃-fedplants was associated with higher total nitrogen contents inNH⁺4-fed plants. Root oxygen consumption was, however, not correlatedwith growth rates or shoot:root ratios. Carbon dioxide releasewas 1·4- and 1·2-fold larger in NO⁺3-fed thanin NH⁺₄-fed wheat and maize plants respectively. Differencesin oxygen and carbon dioxide gas exchange rates resulted inthe gas exchange quotients of NH^-₄-fed plants (wheat, 0·5;maize, 0·6) being greatly reduced compared with thoseof NO^-₃-fed plants (wheat, 1·0; maize, 1·1). Measuredrates of HCO^-₃ assimilation by PEPc in roots were considerablylarger in 4 mM NH⁺₄-fed than in 4 NO^-₃ plants (wheat, 2·6-fold;maize, 8·3-fold). These differences were, however, insufficientto account for the observed differences in root carbon dioxideflux and it is probable that HCO^-₃ uptake is also importantin determining carbon dioxide fluxes. Thus reduced root extension in NH⁺₄-fed compared with NO^-₃-fedwheat plants could not be ascribed to differences in carbondioxide losses from roots.Copyright 1993, 1999 Academic Press Triticum aestivum, wheat, Zea mays, maize assimilation, ammonium assimilation, root respiration     

Concurrent Rates of N2 Fixation, Nitrate and Ammonium Uptake by White Clover in Response to Growth at Different Root Temperatures

Nodulated white clover plants (Trifolium repens L. cv. Huia)were grown for 71 d in flowing nutrient solutions containingN as 10 mmol m_–3 NH₄NO₃, under artificial illumination,with shoots at 20/15°C day/night temperatures and root temperaturereduced decrementally from 20 to 5°C. Root temperatureswere then changed to 3, 7, 9, 11, 13, 17 or 25°C, and theacquisition of N by N₂ fixation, NH₄+ and NO₃^– uptakewas measured over 14 d. Shoot specific growth rates (d. wt)doubled with increasing temperature between 7 and 17°C,whilst root specific growth rates showed little response; shoot:root ratios increased with root temperature, and over time at11°C. Net uptake of total N per plant (N₂ fixation + NH₄++ NO₃^–) over 14 d increased three-fold between 3 and 17°C.The proportion contributed by N₂ fixation decreased with increasingtemperature from 51% at 5°C to 18% at 25°C. Uptake ofNH₄+ as a proportion of NH₄+ + NO₃^– uptake over 14 d variedlittle (55–62%) with root temperature between 3 and 25°C,although it increased with time at most temperatures. Mean ratesof total N uptake per unit shoot f. wt over 14 d changed littlebetween 9 and 25°C, but decreased progressively with temperaturebelow 9°C, due to the decline in the rates of NH₄+ and NO₃^–uptake, even though N₂ fixation increased. The results suggestthat N₂ fixation in the presence of sustained low concentrationsof NH₄+ and NO₄^– is less sensitive to low root temperaturethan are either NH₄+ or NO₃^– uptake systems. White clover, Trifolium repens L. cv. Huia, root temperature, nitrogen fixation, ammonium, nitrate