Nitrogen utilization by plant species from acid heathland soils: II. Growth and shoot/root partitioning of NO3- assimilation at constant low pH and varying NO3-/NH4+ ratio

The growth of four heathland species, two grasses (D. flexuosa,M. caerulea) and two dwarf shrubs (C. vulgaris, E. tetralix),was tested in solution culture at pH 4.0 with 2 mol m^–3N, varying the N0₃^–/NH₄⁺ ratio up to 40% nitrate. In addition,measurements of NRA, plant chemical composition, and biomassallocation were carried out on a complete N0₃^–/NH₄⁺ replacementseries up to 100% nitrate. With the exception of M. caerulea, the partial replacement ofNH₄⁺ by NO₃^– tended to enhance the plant's growth ratewhen compared to NH₄⁺ only. In contrast to the other species,D. flexuosa showed a very flexible response in biomass allocation:a gradual increase in the root weight ratio (RWR) with NO₃^–increasing from 0 to 100%. In the presence of NH₄⁺, grassesreduced nitrate in the shoot only; roots did not become involvedin the reduction of nitrate until zero ambient NH₄⁺. The dwarfshrubs, being species that assimilate N exclusively in theirroots, displayed an enhanced root NRA in the presence of nitrate;in contrast to the steady increase with increasing NO₃^–in Calluna roots, enzyme activity in Erica roots followed arather irregular pattern. Free nitrate accumulated in the tissuesof grasses only, and particularly in D. flexuosa. The relative uptake ratio for NO₃^– [(proportion of nitratein N uptake)/(proportion of nitrate in N supply)] was lowestin M. caerulea and highest in D. flexuosa. Whereas M. caeruleaand the dwarf shrubs always absorbed ammonium highly preferentially(relative uptake ratio for NO₃^– <0.20), D. flexuosashowed a strong preference for NO₃^– at low external nitrate(the relative uptake ratio for N0₃^– reaching a value of2.0 at 10% NO₃^–). The ecological significance of thisprominent high preference for NO₃^– at low NO₃^–/NH₄⁺ratio by D. flexuosa and its consequences for soil acidificationare briefly discussed. Key words: Ammonium, heathland lants, N0₃^–/NH₄⁺ ratio, nitrate, nitrate reductase activity, soil acidification, specific absorption rate     

The Apparent Induction of Nitrate Uptake by Chara corallina Cells following Pretreatment with or without Nitrate and Chlorate

Nitrate provision has been found to regulate the capacity forChara corallina cells to take up nitrate. When nitrate was suppliedto N sufficient cells maximum nitrate uptake was reached after8 h. Prolonged treatment of the cells in the absence of N alsoresulted in the apparent ability of these cells to take up nitrate.Chlorate was found to substitute partially for nitrate in the‘induction’ step. The effects on nitrate reductionwere separated from those on nitrate uptake by experiments usingtungstate. Tungstate pretreatment had no effect on NO^–₃uptake ‘induced’ by N starvation, but inhibitedNO^–₃ uptake associated with NO^–₃ pretreatment. Chloridepretreatment similarly had no effect on NO^–₃ uptake ‘induced’by N deprivation, but inhibited NO^–₃ uptake followingNO^–₃ pretreatment. The data suggest that there are atleast two mechanisms responsible for the ‘induction’of nitrate uptake by Chara cells, one associated with NO^–₃reduction and ‘induced’ by CIO^–₃ or NO^–₃and one associated with N deprivation. Key words: Nitrate, Chlorate, Chara corallina, Induction     

The Partitioning of Nitrate Assimilation Between Root and Shoot of a Range of Temperate Cereals and Pasture Grasses

Nitrate reductase activity (NRA, in vivo assay) and nitrate(NO^-₃) content of root and shoot and NO^-₃ and reduced nitrogencontent of xylem sap were measured in five temperate cerealssupplied with a range of NO^-₃ concentrations (0·1–20mol m^–3) and three temperate pasture grasses suppliedwith 0·5 or 5 0 mol m^–3 NO^-₃ For one cereal (Hordeumvulgare L ), in vitro NRA was also determined The effect ofexternal NO^-₃ concentration on the partitioning of NO^-₃ assimilationbetween root and shoot was assessed All measurements indicatedthat the root was the major site of NO3 assimilation in Avenasatwa L, Hordeum vulgare L, Secale cereale L, Tnticum aestivumL and x Triticosecale Wittm supplied with 0·1 to 1·0mol m^–3 NO^-₃ and that for all cereals, shoot assimilationincreased in importance as applied NO^-₃ concentration increasedfrom 1.0 to 20 mol m^–3 At 5.0–20 mol m^–3 NO3,the data indicated that the shoot played an important if notmajor role in NO^-₃ assimilation in all cereals studied Measurementson Lolium multiflorum Lam and L perenne L indicated that theroot was the main site of NO^-₃ assimilation at 0.5 mol m^–3NO^-₃ but shoot assimilation was predominant at 5.0 mol m^–3NO^-₃ Both NRA distribution data and xylem sap analysis indicatedthat shoot assimilation was predominant in Dactylis glomerataL supplied with 0.5 or 5.0 mol m^–3 NO^-₃ Avena sativa L., oats, Hordeum vulgare L., barley, Secale cereale L., rye, x Triticosecale Wittm., triticale, Triticum aestivum L., wheat, Dactylis glomerata L., cocksfoot, Lolium multiflorum Lam., Italian ryegrass, Lolium perenne L., perennial ryegrass, nitrate, nitrate assimilation, nitrate reductase activity, xylem sap     

Seedling Nitrate Reductase Activity and Nitrate Partitioning in Maize (Zea mays L.) Strains Divergently Selected For Post-anthesis Leaf-Lamina Nitrate Reductase Activity

Two experiments were conducted to evaluate the effects of phenotypicrecurrent selection for high and low post-anthesis leaf-laminain vivo NRA on nitrate uptake, nitrate partitioning and in vitroNRA of seedling roots and leaves. In Experiment 1, intact plantsof cycle 0, 4, and 6 of the high and low NRA strains were grownon NH₄-N for 11 d, then exposed to 1.0 mol m^–3 KNO₃, andcultures sampled at 6 h and 28 h (induction and post-inductionperiods). Nitrate uptake, tissue nitrate concentration and invitro NRA were determined. The pattern of response to selectionin seedling leaf NRA was similar to that observed for in vivoNRA of field grown plants. Leaf NRA increased between 6 h and28 h. Root NRA was not affected by selection or sampling time.Treatments differed in total fresh weight but not in reductionor uptake of nitrate per unit weight, indicating a lack of correspondencebetween NRA and reduction and supporting the idea that concomitantreduction by NR is not obligatorily linked to nitrate influxin the intact plant. In Experiment 2, dark-grown plants of cycle 0, and 6 of thehigh and low NRA strains were cultured without N, detopped onday 6, transferred the following day to 0-75 mol m^–3 KNO₃and sampled at 6 h and 28 h. In contrast to Experiment 1, selectionpopulations differed in nitrate reduction and root NRA, whichby 28 h reached higher average levels than root NRA of intactplants. Translocation and reduction were inversely related amongstrains within each sampling time. The high level of translocationin detopped plants of the low NRA strain was difficult to reconcilewith its low leaf NRA level of Experiment 1. It is suggestedthat nitrate transport in detopped roots is altered relativeto the intact system in a way which permits greater NRA inductionand nitrate reduction. The results indicate that nitrate partitioningby detopped root systems should be interpreted with caution. Key words: Zea, nitrate reductase activity, nitrate uptake, nitrate reduction, nitrate partitioning, selection     

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     

Nitrate Effects on Pre-emergence Growth and Emergence Percentage of Wheat (Triticum aestivum L.) from Different Sowing Depths

The effects of a range of applied nitrate (NO₃^–) concentrations(0–20 mol m3) on germination and emergence percentageof Triticum aestivum L. cv. Otane were examined at 30, 60, 90and 120 mm sowing depths. Germination percentage was not affectedby either sowing depth or applied NO₃^– concentration whereasemergence percentage decreased with increased sowing depth regardlessof applied NO₃^– concentration. Nitrate did not affectemergence percentage at 30 mm sowing depth, but at 60 to 120mm depth, emergence percentage decreased sharply with an increasedapplied NO₃^– concentration of 0 to 1·0 mol m^–3then decreased only slightly with further increases in appliedNO₃^– of about 5·0 mol m^–3. Root and shoot growth, NO₃^– accumulation and nitrate reductaseactivity (NRA) of plants supplied with 0, 1·0 and 1·0mol m^–3 NO₃^– at a sowing depth of 60 mm were measuredprior to emergence. The coleoptile of all seedlings opened withinthe substrate. Prior to emergence from the substrate, shootextension growth was unaffected by additional NO₃^– butshoot fr. wt. and dry wt. were both greater at 1·0 and1·0 mol m^–3 NO₃^– than with zero NO₃^–.Root dry wt. was unaffected by NO₃^–. Nitrate concentrationand NRA in root and shoot were always low without NO₃^–.At 1·0 and 10 mol m3 NO₃^–, NO₃^– accumulatedin the root and shoot to concentrations substantially greaterthan that applied and caused the induction of NRA. Regardlessof the applied NO₃^– concentration, seedlings which failedto emerge still had substantial seed reserves one month afterplanting. Coleoptile length was substantially less for seedlingswhich did not emerge than for seedlings which emerged, but wasnot affected by NO₃^–. It is proposed that (a) decreasedemergence percentage with increased sowing depth was due tothe emergence of leaf I from the coleoptile within the substrateand (b) decreased emergence percentage with additional NO₃^–was due to the increased expansion of leaf 1 within the substrateresulting in greater folding and damage of the leaf. Key words: Triticum aestivwn L., nitrate, sowing depth, seedling growth, seedling emergence     

Function and contribution of the root tip in the induction of NO3- uptake along the barley root axis

The seminal roots of N-free-grown barley seedlings were ableto take up NO₃^– immediately upon initial exposure; theuptake rate in the tip was half of that in the older root zones(middle and base). A lag of 60 min was required in all rootzones before the uptake rates started to increase during inductionwith external NO₃^–. This increase could be prevented bythe addition of pFPA; we thus assume that additional NO₃^–transport proteins were synthesized during NO₃^– induction.During the time-course of NO₃^– induction different uptakerates were measured in morphologically different regions ofthe tip (1 mm segments) indicating a regulation of NO₃^–induction on a narrow local scale. In NO₃^– grown plants, NO₃^– uptake as well as NO₃^–content increased basipetally along the root axis concomitantlywith increasing vacuolization of the cells. Although NO₃^–uptake into the tip was only half of that into the older rootzones, this NO₃^– uptake was very important for the entireroot. Firstly, it provided the substrate for protein biosynthesisin the meristematic region: nitrate reductase activity and totalsoluble protein were highest in the first apical mm of the tip.Secondly, 3% of the NO₃^– taken up by the tip was foundin the base where it induced NO₃^– uptake: NO₃^– wastranslocated almost exclusively basipetally and as little as20nmolg^–1 root fr. wt. translocated from the tip weresufficient for acceleration of NO₃^– induction in the rootbase of N-free-grown plants. This clearly shows that the inductionof NO₃^– uptake does not depend exclusively on the availabilityof external NO₃^–, but can be mediated also with internallytranslocated NO₃^–.The root tip, therefore, may be consideredthe NO₃^– sensing region of the root. Key words: Barley, Hordeum vulgare L, internal NO₃^–, NO₃^– uptake, root zones     

Effect of Nitrate, Ammonium and Some Amino Acids on Growth and Nitrate Reductase Activity in Suspension Cultures of Atropa belladonna

Growth and nitrate reductase activity (NRA) of Atropa belladonnacells were studied in medium supplemented with NaNO₃, NH₄NO₃,and amino acid precursors to tropane alkaloids. Growth and NRAwere stimulated by NH₄⁺ and by proline, by proline plus ornithine,but not by glutamate, in NO₃^–-containing medium. Testedamino acids inhibited neither utilization of inorganic nitrogennor growth. (Received September 30, 1988; Accepted August 28, 1989)     

f-Ratio and its relationship to ambient nitrate concentration in coastal waters

The relationship between thef-ratio [NO₃^– uptake/(NO₃^–+ NH₄⁺) uptake] and ambient nitrate concentration was evaluatedfor eight data sets from coastal waters. The f-ratio increasedasymptotically with increase in nitrate concentration in mostdata sets. However, the rate at which f-ratio increased at lownitrate concentration (slope = m) and the maximum attained f-ratio(f_max) varied among regions; the initial slope varied most withvalues ranging in excess of an order of magnitude. The datawere analyzed in relation to environmental factors and methodologicalconsiderations known to influence the f-ratio. Ambient ammoniumconcentration was important in accounting for regional differencesin the f versus NO₃^– relationship. A further analysisof the data, relating f-ratio to the ratio of NO₃^–/(NO₃^–+ NH₄⁺) concentrations yielded a much more regionally consistentand approximately linear relationship; slopes varied by lessthan a factor of two in the extreme cases. Inclusion of knownalternative (aside from NH₄⁺) sources of reduced-N (e.g. urea)and correction for methodological/computational errors (isotopedilution) systematically reduce f-ratio estimates. Other factors,e.g. reduced-N uptake by microheterotrophs, may systematicallyincrease the f-ratio.     

Growth of Actinorhizal Plants as Influenced by the Form of N with Special Reference to Myrica gale and Alnus incana

Growth of two actinorhizal species was studied in relation tothe form of N supply in water culture. Non-nodulated bog myrtle(Myrica gale) and grey alder (Alnus incana) were grown withNH₄⁺, NH₄NO₃^– or NO₃^– (4 mol m^–3 N). A nodulatedseries of bog myrtle was also cultivated in N-free medium. Relative growth rate (RGR), utilization rate of N, and shoot/rootratio were highest for the two species with the N completelysupplied as NH₄⁺. In both species, nitrate was largely reducedin the roots and the presence of NO₃^– in combined-N supplyalways affected the RGR and N utilization rate negatively. BothN₂ fixation and complete NO₃^– nutrition represented conditionsof relative N-deficiency resulting in relatively low tissue-Nconcentrations and a greater allocation of dry mass to the roots.The physiological N status of nodulated M. gale plants was comparativelygood, as indicated by a normal nodule weight ratio and a relativelyhigh N₂-fixing rate per unit nodule mass. However, whole-plantN₂-fixing capacity remained relatively low in comparison withacquisition rates of N in combined-N plants. The anion charge from the nitrate reduction was largely directlyexcreted as an OH^– efflux. H ⁺ /N ratios generally agreedwith the theory. In comparison with NH₄⁺ nutrition, carboxylateconcentrations were higher in N₂-fixing M. gale plants and theH ⁺ /N ratio in nodulated plants was less than unity below thevalue for ammonium plants as previously found for other actinorhizalspecies. Therefore, NH₄⁺ should be an energetically more attractiveN source for actinorhizal plants than N₂. The results agree with commonly accepted views on energeticsof N uptake and assimilation in higher plants and support theconcept of a basically similar physiological behaviour betweennon-legumes and legumes. Key words: Actinorhizal symbioses, ammonium, H⁺/OH^– efflux, nitrate, N₂ fixation, NRA     

Developmental and Biochemical Regulation of 'Constitutive' Nitrate Reductase Activity in Leaves of Nodulating Soybean

The developmental profile of ‘constitutive’ nitratereductase activity (cNRA) in leaves of soybean (Glycine max(L.) cv. Bragg) plants at different ages is described. The youngestleaves had most cNRA and the activity dropped off as a newerleaf developed above it. Each leaf had its distinct active periodof in vivo cNRA. This pattern was different in urea-grown andsymbiotically-grown plants (inoculated with Bradyrhizobium japonicumstrain USDA 110), where the latter had no detectable in vivocNRA in older leaves. Urea-grown plants maintained considerablein vivo NRA in such older leaves. When symbiotically-grown plantshad their nodules removed, in vivo cNRA reappeared in olderleaves within 1 d of removal, nearly reaching levels of youngleaves at 3 d after nodule excision. Allantoic acid (ALL), oneof the known transport ureides of soybeans, was implicated asa possible signal molecule from nodules to leaves. Allantoicacid (100 µM) inhibited in vitro c₁ NRA significantly,with 400 µM ALL resulting in complete inhibition. In contrast,allantoin (ALN) had no inhibitive effect on NRA. Inhibitionof c₁NRA by ALL was by a competitive process, judging from Lineweaver-Burkeplots against nitrate. Kinetics showed a constant Vmax of around105 nmol NO₂ mg^–1 protein h^–1 and a K_m for nitrateof 15 mM, which increased to 60 mM in the presence of 200 µMallantoic acid. Non-specific (ionic and pH-related) influenceswere eliminated. Allantoic acid also had a slight stimulatingeffect of in vitro NRA (up about 25% at 400 µM). Thesefindings suggest that c1NRA may be involved in ureide metabolism,rather than in vivo nitrate metabolism. Key words: Root-shoot interaction, nitrogen metabolism, nodulation, symbiosis     

Orthophosphate Relations of Root: NO-3 Effects on Orthophosphate Influx, Accumulation and Secretion into the Xylem

Lamaze, T., Sentenac, H. and Grignon, C. 1987. Orthophosphaterelations of root: NO^–₃effects on orthophosphate influx,accumulation and secretion into the xylem.—J. exp. Bot.38: 923–934. Orthophosphate (Pi) accumulation by barley (Hordeum vulgareL.) roots was specifically inhibited by NO^–₃ as comparedto Cl^– and SO₄^{2 –}, and Pi secretion into the xylemwas stimulated. The inhibition of Pi accumulation by NO^–₃was also observed in roots of intact photosynthesizing horsebean(Vicia faba L.), rice (Oryza sativa L.) and soybean (Glycinemax L.) plants. NO^–₃ effects on Pi transport by rootswere more thoroughly investigated with corn (Zea mays L.). Theywere due to intracellular NO^–₃. Pi secretion was stillstimulated by NO^–₃ after Pi withdrawal from the absorptionsolution. ³²Pi influx decreased during Pi accumulation, supportingthe hypothesis that this ion allosterically regulated its owntransport system by feedback control. This control was modulatedby other anions: the decrease was more pronounced in the presenceof nitrate. Chronologically, the depressive effect of NO^–₃on ³²Pi influx appeared after the inhibition of Pi accumulation.Furthermore, under conditions where Pi accumulation was notaffected by NO^–₃, ³²Pi influx and Pi secretion into thexylem became insensitive to the presence of nitrate. Our hypothesisis that the stimulative effect of NO^–₃ on Pi secretionand the depressive one on ³²Pi influx are the repercussionsof an increase in the Pi cytosolic concentration due to an NO^–₃-induced decrease in Pi uptake by the vacuoles. Key words: Root, orthophosphate fluxes, orthophosphate accumulation, nitrate, ionic interaction     

Influence of abscisic acid on nitrogen partitioning, sucrose metabolism and nitrate reductase activity of chicory suspension cells

Batch suspension cultures of chicory cells (Cichorium intybusL. var. Witloof) possess a NADH-specific nitrate reductase activitythat peaks on day 3 of a 10 d growth cycle. When both nitrateand ammonium are used as nitrogen sources, chicory cells absorbnitrate irst. Ammonium uptake becomes predominant at day 3,even though NO₃^– was still present in the medium. Althoughabscisic acid impairs growth as well as ¹⁵NO₃^– uptakeand reduction, it promotes nitrate reductase activity as measuredboth in vivo and in vitro. Specific activity is 50% higher inABA-treated cells than in controls. These conflicting data maybe explained either in erms of nitrate reductase levels or bythe availability of reducing power and energy. Since NRA isgenerally controlled by the availability of the reducing power,the energy status of the cell, the adenylate nucleotide pools,were measured simultaneously with the carbohydrate levels withinthe cell and the growth medium. The energy charge was not modifiedduring the growth cycle, regardless of the rowth conditions.Yet ABA modified the intracellular carbohydrate metabolism andinhibited the acidic invertase, the sucrose synthase and thesucrose phosphate synthase activities. Modified assimilationrates of nitrate in chicory cells grown in the presence of ABA,were probably correlated to modified carbohydrate metabolismpathways leading to increased availability of reducing power,energy and C-skeletons. Key words: Abscisic acid, Cichorium intybus L, nitrate reductase, reductase, invertase, sucrose synthase, sucrose phosphate synthase     

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     

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     

Diurnal regulation of NO3-uptake in soybean plants II. Relationship with accumulation of NO and asparagine in the roots

Experiments were performed with soybean plants to test the hypothesisthat the inhibition of NO₃^– uptake in darkness is dueto feedback control by NO₃^– and/or Asn accumulating inthe roots. Xylem export of N compounds was shown to depend onwater flux in both excised root systems and ¹⁵N-labelled intactplants, suggesting that the shortage of transpiration in darknessmay be responsible for the retention of NO₃^– and Asn inthe roots. This was verified in experiments where the light/darkpattern of transpiration was modulated in intact plants by changingthe relative humidity of the atmosphere. Any decrease of transpirationat night was associated with a concurrent stimulation of NO₃^–and Asn accumulations in the roots. However, the light/darkrhythmicity of NO₃^– uptake was only marginally affectedby these treatments, and thusappeared quite independent fromtranspiration and root NO₃^– or Asn levels. Typically,the maintainance of a constant transpiration during the day/nightcycle did not suppress the inhibition of NO₃^– uptake indarkness, whereas it almost prevented the dark increase in rootNO₃^– and Asn contents. These data strongly support theconclusion that the effect of light on NO₃^– uptake isnot mediated by changes in translocation and accumulation ofN compounds. Key words: Glycine max, light/dark, cycles, nitrate uptake, transpiration, transport of N compounds, accumulation of N compounds     

Nitrate transport in intact wheat roots:II. Long-term effects of NO3- concentration in the nutrient solution on NO3- unidirectional fluxes and distribution within the tissues5

We have examined the long-term effects of NO₃^– concentrationson NO₃^– (¹⁵NO₃^–) fluxes and cellular pool sizesin roots of intact 30-d-old wheat (Triticum aestivum cv. Courtot)grown hydroponically. Compartmental analysis was performed understeady-state conditions at five different levels of NO₃^–concentration (from 0.1 up to 5 mol m^–3 taking into accountmetabolism and secretion into the xylem (Devienne et al., 1994).Nitrate and reduced nitrogen levels in the tissues were largelyindependent of external NO₃^– concentration although below1.5 mol m^–3 NO₃^–; concentration limited plant growth.In the chamber, marked diurnal variations in net uptake occurredand, in the light, higher NO₃^– concentrations yieldedhigher NO₃^– uptake rates. After transfer of the plantsto the laboratory, the increase in net uptake linked to elevationof NO₃^–; concentrations was even larger (from 0.1 to 8.8µmolh^–1 g^–1 FW) as a result of a marked increase (x10–11) in the unidirectional influx at the plasmalemmawhile NO₃^– efflux was less enhanced (x 4–5). Underthese conditions, influx into the vacuole was also higher (x2–4) while efflux from the vacuole was little affected(x 1–3). NO₃^– concentrations within the cell compartmentswere estimated under the clas sical assumptions. The vacuolarconcentration was a little modified by NO₃^– availabilitywhereas that in the cytosol increased from about 10 mol m^–3to about 20 mol m^–3 indicating that (1) the absolute valuefor the cytosol was high and (2) it displayed only a small increasedespite very large changes in NO₃^– fluxes. NO₃^–distribution within the cells did not seem to involve an activeaccumulation of NO₃^– in the vacuole. Key words: Wheat, ion transport, nitrate, ¹⁵N, compartmentation     

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     

Effects of nitrogen nutrition on salt-stressed Nicotiana tabacum var. Samsun in vitro plantlets

Tobacco shoots were grown in vitro for 35 d, in MS culture mediummodified to include various sources (nitrate-N, ammonium-N ora mixture) and levels (0–120 mM) of N, and in the presenceof 0–180 mM NaCI or iso-osmotic concentrations of mannitol.Growth of control plantlets was significantly inhibited whenNH₄⁺-N was the sole N source, and at high (120 mM) NO^–₃-N supply. Under conditions of salt stress (90 and 180 mM NaCI)growth was repressed, with roots being more severely affectedthan shoots. Salinity also inhibited root emergence in vitro.The only alleviation of the salt stress by nitrate nutritionobserved in this study was on shoot growth parameters of plantletsgrown on 60 mM NO^–₃-N and 90 mM NaCI. Although both weresignificantly inhibited by NaCI, nitrate reduc-tase activitywas more severely affected than nitrate uptake. When mannitolreplaced NaCI in the culture medium, similar Inhibition of growth,nutrient uptake and enzyme activity were recorded. These observations,together with the relatively low recorded values for Na⁺ andCI^– uptake, indicate that under in vitro salt stress conditionsthe negative effects of NaCI are primarily osmotic. Key words: Growth, nitrogen metabolism, osmotic stress, salinity     

Acclimation of NO-3 fluxes to low root temperature by Brassica napus in relation to NO-3 supply

Acclimation of NO^–₃ transport fluxes (influx, efflux)in roots of oilseed rape (Brassica napus L. cv. Bien venu) andtheir sensitivity to growth at low root temperature was studiedin relation to external NO^–₃ supply, defined by constantconcentrations ranging from sub- to supra-optimal with respectto plant growth rate. Plants were grown from seed in flowingnutrient solutions containing 250 mmol m^–3 NO^–₃at 17°C for 20d, and solution temperature in half the cultureunits was then lowered decrementally over 3 d to 7°C. Threedays later plants were supplied with NO^–₃ at 1, 10, 100or 1000 mmol m^–3 maintained for 18 d. Dry matter productionwas decreased more by low root zone temperature than low [NO^–₃]_e. Root specific growth rates were inversely related to [NO^–₃]_eand shoot:root ratios increased with time at [NO^–₃]_e between10–1000 mmol m^–3. Net uptake of NO^–₃ at 17°Cwas twice that at 7°C, and at both temperatures it doubledwith increasing [NO^–₃]_e between 1–10 mmol m^–3with further small increases at higher [NO^–₃]_e. Mean unitabsorption rates of NO^–₃ between 0–6 d and 6–14d were linearly related (r² of 0.79–0.99) to log₁₀[NO].Steady-state Q₁₀ (7–17°C) for uptake between 0–6d were 0.91, 1.62, 1.27, and 1.10, respectively, at [NO^–₃]_eof 1, 10, 100, and 1000 mmol m^–3, compared with correspondingvalues of 0.98, 1.38, 1.68, and 1.89 between 6–14 d. Thedata indicated that net uptake rates at 7 and 17°C divergedover time at high [NO^–₃]_e. Short-term uptake rates from1 mol m^–3 NO^–₃ measured at 17°C were higherin plants grown with roots at 7°C than at 17°C; for7°C plants there was a strong inverse linear relationship(r²=0.94) between uptake rate and treatment log₁₀ [NO^–₃]_ewhilst rates in 17°C plants were independent of prior [NO^–₃]_e. Rates of NO^–₃ influx and efflux under different steady-stateconditions of NO^–₃ supply and root temperature were calculatedfrom dilution of ¹⁵N added to culture solutions. Efflux wassubstantial relative to net uptake in all treatments, and wasinversely related to [NO^–₃]_e at 17°C but not at 7°C.Ratios of influx: efflux ranged from 1.6–2.9 at 17°Cand 1.3–1.8 at 7°C, indicating the proportionatelygreater impact of efflux at low root temperature. Ratios ofefflux: net uptake were 0.53–1.56 at 17°C and 1.21–3.58at 7°C. The apparent sensitivities of influx and effluxto steady-state root temperature varied with [NO^–₃]_e.Both fluxes were higher at 17°C than 7°C in the presenceof 100–1000 mmol m^–3 NO^–₃ but the trend wasreversed at 1–10 mmol m^–3 NO. Concentrations oftotal N measured in xylem exudate were at least 2-fold higherat 7°C compared with 17°C, attributable mainly to higherconcentrations of NO^–₃ glutamine and proline. The resultsare discussed in terms of acclimatory and other responses shownby the NO^–₃ transport system under conditions of limitingNO^–₃ supply and low root temperature. Key words: Brassica napus, nitrate supply, efflux, influx, root temperature, xylem exudate