Effect of Plant Growth Regulators on Nitrate Utilization by Roots of Nitrogen-Depleted Dwarf Bean

We examined the effect of pretreatments (18 h at 5 µmoldm^–3) with abscisic acid, the ethylene-releasing substance‘Ethephon’, gibberellic acid, indoleacetic acid,kinetin and zeatin on nitrate uptake and in vivo nitrate reductaseactivity (NRA) in roots of nitrogen-depleted Phaseolus vulgarisL. Nitrate uptake showed an apparent induction pattern witha steady state after about 6 h, in all treatments. The nitrateuptake rate after 6 h was unaffected or at most 30% lower aftertreatments with the plant growth regulators. Gibberellic acid, kinetin and zeatin induced substantial NRAin roots in the absence of nitrate, whereas Ethephon enhancedNRA only during nitrate nutrition. Kinetin-induced NRA (Ki-NRA)was maximal after a pretreatment at 1 µmol dm^–3,and showed a lag phase of 6–8 h. Ki-NRA was additive tonitrate-induced NRA (NO^–₃-NRA) for at least 24 h, independentof the induction sequence. After full induction, Ki-NRA approximated20% of NO-₃-NRA. Abscisic acid counteracted the developmentof Ki-NRA, but not of NO^–₃-NRA. Cycloheximide and tungstatewere equally effective to suppress the development of nitratereductase activity after supply of kinetin or NO^–₃. Our data are consistent with the operation of two independentenzyme fractions (Ki-NRA and NO^–₃-NRA) with apparentlyidentical properties but with separate control mechanisms. Theabsence of major effects of plant growth regulators on the time-courseand rate of nitrate uptake suggests that exogenous regulators,and possibly endogenous phytohormones are of minor importancefor initial nitrate uptake. The differential effect of someregulators on nitrate uptake and root NRA furthermore indicatesthat the processes of uptake and reduction of NO^–₃ arenot obligatory or exclusively coupled to each other.     

Nitrate Transport into Chara corallina Cells using 36ClO-3 as an Analogue for Nitrate: I. INTERACTION BETWEEN 36ClO-3 AND NO-3 AND CHARACTERIZATION OF 36CIO-3/NO-3 INFLUX

The use of chlorate as an analogue for NO^–₃ during nitrateuptake into Chara corallina cells has been investigated. NO^–₃inhibits ³⁶C1O^–₃ influx into Chara over the concentrationrange 0–1000 mmol m^–3. Lineweaver-Burke plots ofthe data are characteristic of competitive inhibition by NO^–3in the low concentration range (0–300 mmol m^–3 ClO^–₃)and apparent K_INO3 is 140 mmol m^–3 which is of a similarorder of magnitude as apparent K_mCIO3- 180 mmol m^–3. Athigher substrate concentrations the inhibition by NO^–₃was not characteristic of competitive or uncompetitive inhibition. ³⁶C1O^–₃/NO^–₃ influx was dependent on K⁺ and Ca²⁺in the external medium and inhibited by FCCP. NO^–₃ pretreatmentor N starvation increased subsequent ³⁶C1O^–₃/NO^–₃influx into Chara. A comparison between rates of net NO^–₃uptake and ³⁶C1O^–₃/NO^–₃ influx supported the previoushypothesis that NO^–₃ efflux is an important componentin the determination of overall uptake rates. Key words: Nitrate, Chara, ³⁶CIO^–₃     

Nitrate Transport into Chara corallina Cells using as an Analogue for Nitrate: III. INTERACTIONS BETWEEN CHLORIDE AND NITRATE TRANSPORT PROCESSES

The effects of Cl^– and pretreatment on ³⁶Cl^–influx and influx into Characorallina cells were examined. Both treatments reduced ³⁶Cl^–influx into C. corallina cells in the acid pH range (4.5–7.0). pretreatment stimulated influx into C. corallina cells, but Cl^– pretreatment hadno effect. There was a direct inhibitory effect of CI on influx into C. corallina cells, but no apparenteffect on net NO uptake. The time course of ³⁶Cl accumulation into cytoplasmic and vacuolarcompartments during incubation of the cells with showed that significant radioactivity appeared in the vacuolarsap after 30 min. There was a linear increase in the percentageof total ³⁶CI which crossed the tonoplast (_c-v). There was noeffect of Cl^– or pretreatment on accumulation of radioactivity in the vacuole. Thin layer chromatography ofthe vacuolar sap showed that after 2 h only one component waspresent which had an R_F which was similar to ³⁶CI^–. Therate of accumulation of ³⁶C1^– in the vacuole could beused to estimate rates of reduction. Key words: Chloride, Chlorate, Chara, Nitrate     

Nitrogen utilization by plant species from acid heathland soils: I. Comparison between nitrate and ammonium nutrition at constant low pH

Seven heathland species, four herbaceous plants and three dwarfshrubs, were tested for their capacity to utilize NH₄⁺ or NO₃^–. When cultured in solution at pH 4.0 with 2mol m^–3 N,all species showed similar growth responses with respect toN source. Nitrate was assimilated almost equally well as ammonium,with relative growth rate generally averaging 5–8% lowerfor NO₃^– grown plants, albeit not always significantly.However, N source was significantly and consistently correlatedwith biomass partitioning, as NH₄⁺-fed plants allocated moredry matter to shoots and less to roots when compared to NO₃^–-fed plants. The strong difference in biomass partitioning mayrelate to the relative surplus of carbon per unit plant N (or,alternatively, the relatively suboptimal rate of N assimilationper unit plantC) in NO₃^–-fed plants Inherently slow-growing dwarf shrubs accumulated virtually nofree nitrate in their tissues and reduction of nitrate was strictlyroot-based. Faster-growing herbaceous plants, however, partitionedthe assimilation of nitrate over both shoots and roots, therebyaccumulating relatively high tissue NO₃^– levels. Ion uptakerates depended clearly on the ‘relative shoot demand’.At similar shoot demands, especially in the herbaceous species,specific uptake rates for N and total inorganic (non-N) anionswere higher in NH₄⁺ -fed plants, whereas the uptake rate fortotal (non-N) cations was higher in NO₃^–-fed plants. Rateof P uptake was enhanced with increasing plant demand, but wasindependent of the N source. Net H⁺ extrusions ranged from 1.00to 1.34 H⁺ per NH₄⁺, and from –0.48 to –0.77 H⁺per NO₃^– taken up. Key words: Ammonium, biomass partitioning, heathland plants, low pH, nitrate, nitrate reductase activity, relative shoot demand, specific absorption rate     

Phosphate Regulation of Nitrate Assimilation in Soybean

It is known that phosphorus deficiency results in alterationsin the assimilation of nitrogen. An experiment was conductedto investigate mechanisms involved in altered ¹⁵NO^–₃ uptake,endogenous ¹⁵N translocation, and amino acid accumulation insoybean (Glycine max L. Merrill, cv. Ransom) plants deprivedof an external phosphorus supply for 20 d in solution culture.Phosphorus deprivation led to decreased rates of ¹⁵NO^–₃uptake and increased accumulation of absorbed ¹⁵N in the root.Both effects became more pronounced with time. Asparagine, theprimary transport amino acid in soybean, accumulated in largeexcess in roots and stems. In roots of phosphorus-deprived plants,concentrations of ATP and inorganic phosphate declined rapidly,but dry weight accumulation was similar to or above that ofthe control even after 20 d of treatment. Arginine accumulationin leaves was greatly enhanced, even though ¹⁵N partitioninginto the insoluble reduced-N fraction of leaves was unaffected.The results suggest that decreases in NO^–₃ uptake in lowphosphorus plants could be caused by feedback control factorsand by limited ATP availability. The decline in endogenous Ntransport from the root to the shoot may be associated withchanges in membrane properties, which also result in paralleleffects on hydraulic conductance and the upward flow of waterthrough the plant. Key words: Phosphorus stress, nitrate uptake, nitrate translocation, arginine     

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     

Studies of the Uptake of Nitrate in Barley: III. COMPARTMENTATION OF NO3-

Compartmental analyses of intact roots of barley (Hordeum vulgareL. cv. Klondike) plants, grown with different levels of NO₃^–(up to 1·0 mol m^–3) in the external media, wereundertaken using ¹³NO₃^–. Two additional treatments, namelysodium dodecyl sulphate (SDS) or brief exposure to high temperature,designed to investigate the identity of the three NO₃^–compartments revealed by compartmental analyses, provided supportfor the identification of the latter as corresponding to superficialsolution, apoplasm, and cytoplasm. Half-lives for exchange ofthese compartments, 3 s, 30 s, and 7 mm, were unaffected bythe level of NO₃^– provided during growth. Independentestimates of ¹³NO₃^– fluxes obtained by direct methodsagreed well with values of fluxes calculated from the compartmentalanalyses. Cytoplasmic [NO₃^–], estimated from the compartmental analyses,were in the range from 1–37 mol m^–3, and increasedwith increasing [NO₃^–] of the medium. Such values forcytoplasmic [NO₃^–] are inconsistent with an earlier proposal(Siddiqi, Glass, Ruth, and Rufty, 1990; Glass, Siddiqi, Ruth,and Rufty, 1990) of passive NO₃^– uptake in the concentrationrange above 10 mol m^–3. A model, based upon localizeddistribution of nitrate reductase activity in epidermal cells,is proposed in which the proposed passive low affinity NO uptakeat high external [NO₃^–] is restricted to epidermal cells. During loading periods with ¹³NO₃^–, significant amountsof ¹³N were translocated to the shoot. Two pools of ¹³N, onebeing the root symplasm, appear to participate in the transferof labelled N to the shoot. Key words: Barley, compartmentation, nitrate, nitrate reductase, ¹³N     

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     

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     

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     

N2 Fixation and Nitrate Uptake by White Clover Swards in Response to Root Temperature in Flowing Solution Culture

Nodulated white clover plants (Trifolium repens L. cv. Huia)were grown as simulated swards for 71 d in flowing nutrientsolutions with roots at 11 C and shoots at 20/15 C, day/night,under natural illumination. Root temperatures were then changedto 3, 5, 7, 11, 13, 17 or 25 C and the total N₂, fixation over21 d was measured in the absence of a supply mineral N. Alltreatments were subsequently supplied with 10 mmol m^–2NO₂ ^– in the flowing solutions for 14 d, and the relativeuptake of N by N₂, fixation and NO₃ ^– uptake was compared.Net uptake of K⁺ was measured on a daily basis. Root temperature had little effect on root d. wt over the 35-dexperimental period, but shoot d. wt increased by a factor of3.5 between 3 and 25 C, with the sharpest increase occurringat 7–11 C. Shoot: root d. wt ratios increased from 25to 68 with increasing temperature at 7–25 C. N₂-fixationper plant (in the absence of NO₂ ^–) increased with roottemperature at 3–13C, but showed little change above13 C. The ratios of N₂ fixation: NO₂ ^– uptake over 14d (mol N: mol N) were 0.47–0.77 at 3–7 C, 092–154at 11–17 C, and 046 at 25 C, reflecting the dominanceof NO₃ ^– uptake over N₂ fixation at extremes of high andlow root temperature. The total uptake of N varied only slightlyat 11–25 –C (095–110 mmol N plant^–1),the decline in N₂ fixation as root temperature increased above11 C was compensated for by the increase in NO^– ₃ uptake.The % N in shoot dry matter declined with decreasing root temperature,from 32% at 13 C to 15% at 3 C. In contrast, concentrationsof N expressed on a shoot water content basis showed a modestdecrease with increasing temperature, from 345 mol m^–3at 3 C to 290 mol m^–3 at 25 C. Trifolium repens L, white clover, root temperature, N₂ fixation, potassium uptake, nitrate uptake, flowing solution culture     

Nodulation and Nitrogen Fixation of Faba Bean (Vicia faba L.) as Affected by Removal of the Cotyledons and Nitrate Supply

Cotyledons of faba bean (^{Vicia faba} L. cv. Fiord) were removedto determine whether an apparent delay in nodulation of thiscultivar could be attributed to an inhibitor from these organs.Cotyledons were left intact or excised from seedling plants14 and 18 d after sowing and plants grown with or without 2·5mm NO₃^–. Seedling growth was depressed when cotyledons were removed onday 14 but not when removed on day 18. Removal of the cotyledonsat day 14 reduced nodule number and nodule weight in the absenceof NO₃^–, but in the presence of NO₂^–, nodule numberwas unaffected and only nodule weight was reduced. Cotyledonremoval at day 18 increased both nodule number and nodule weightwith +NO₃^– but not with –NO₂^–. Acetylene reduction(AR) was markedly depressed by NO₃^–. Cotyledon removalat day 14 decreased AR but removal at day 18 resulted in anincrease in AR. We suggest from these results that faba beancotyledons have an inhibitory effect on nodule activity andon nodulation and this interacts with NO₃^–. This can beexplained through a ‘feed-back’ regulation of N₂fixation by soluble N in the seedling. Vicia faba, faba bean, nodule number, nodulation, nodule activity, acetylene reduction, N₂ fixation, cotyledon removal, nitrate     

Nitrate Stimulation of Mobilization of Seed Reserves in Temperate Cereals: Importance of Water Uptake

Relationships between nitrate (NO^-₃) supply, uptake and assimilation,water uptake and the rate of mobilization of seed reserves wereexamined for the five main temperate cereals prior to emergencefrom the substrate. For all species, 21 d after sowing (DAS),residual seed dry weight (d.wt) decreased while shoot plus rootd.wt increased (15–30%) with increased applied NO^-₃concentrationfrom 0 to 5–20 mM . Nitrogen (N) uptake and assimilationwere as great with addition of 5 mM ammonium (NH⁺₄) or 5 mMNO^-₃but NH⁺₄did not affect the rate of mobilization of seedreserves. Chloride (Cl^-) was similar to NO^-₃in its effect onmobilization of seed reserves of barley (Hordeum vulgare L.).Increased rate of mobilization of seed reserves with additionalNO^-₃or Cl^-was associated with increases in shoot, root and residualseed anion content, total seedling water and residual seed watercontent (% water) 21 DAS. Addition of NH⁺₄did not affect totalseedling water or residual seed water content. For barley suppliedwith different concentrations of NO^-₃or mannitol, the rate ofmobilization of seed reserves was positively correlated (r >0.95)with total seedling water and residual seed water content. Therate of mobilization of seed reserves of barley was greaterfor high N content seed than for low N content seed. Seed watercontent was greater for high N seed than for low N seed, 2 DAS.Additional NO^-₃did not affect total seedling water or residualseed water content until 10–14 DAS. The effects of seedN and NO^-₃on mobilization of seed reserves were detected 10and 14 DAS, respectively. It is proposed that the increasedrate of mobilization of seed reserves of temperate cereals withadditional NO^-₃is due to increased water uptake by the seedlingwhile the seed N effect is due to increased water uptake bythe seed directly. Avena sativa L.; oat; Hordeum vulgare L.; barley; Secale cereale L.; rye; xTriticosecale Wittm.; triticale; Triticum aestivum L.; wheat; nitrate; seed; germination; seed reserve mobilization     

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 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     

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     

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     

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     

Nitrate Transport into Chara corallina Cells using 36C1O3 as an Analogue for Nitrate: II. COMPARISON WITH 14C METHYLAMINE FLUXES AT DIFFERENT pH0 AND NH+4sol;NO3 INTERACTIONS

³⁶C1O₃/NO₃ influx into Chara cells was found to be sensitiveto pH_o and a maximum was found at pH_o = 4.5. By contrast ¹⁴Cmethylamine influx into Chara showed a maximum at pH_o = 8.5,and at this pH_o influx rates were about 150 times higher thanrates of ³⁶C1O₃/NO₃ influx. However, at pH_o = 4.5, ³⁶C1O₃/NO₃influx rates were, in some cases, comparable with rates of ¹⁴Cmethylamine influx. ³⁶C1O₃/NO₃ influx into Chara cells was stimulatedby Rb ⁺, K⁺, Na ⁺, and NH₄⁺, but not by Cs⁺ or Li ⁺. NO₃ andCl reduced ¹⁴C methylamine influx into Chara by 30%. NH₄⁺ causedvery considerable inhibition of ¹⁴C methylamine influx intoChara, but had no effect on ³⁶C1O₃/NO₃ influx in the presenceof K ⁺. Net NO³ uptake into Chara was completely prevented byNH₄⁺ even at relatively low NH₄⁺ concentrations (25 mmol m ^–3).This latter effect was reversed by diethylstilbestrol (DES).Evidence is presented for the stimulation of NO₃ efflux by NH⁴⁺as the mechanism responsible for the immediate effects of NH₄⁺on net NO₃ uptake into Chara cells. Key words: Chara, ¹⁴C methylamine, ³⁶ClO^–₃, pH     

Photosynthetic Nitrogen Metabolism in High and Low CO2-adapted Scenedesmus: I. INORGANIC CARBON-DEPENDENT O2 EVOLUTION, NITRATE UTILIZATION AND NITROGEN RECYCLING

Larsson, M., Larsson, C.-M. and Guerrero, M. G. 1985. Photosyntheticnitrogen metabolism in high and low CO₂-adapted Scenedesmus.I. Inorganic carbon-dependent O2 evolution, nitrate utilizationand nitrogen recycling.—J. exp Bot. 36: 1373–1386 Scenedesmus obtusiusculus Chod. was grown on an inorganic mediumflushed with either air or air supplemented with 3% CO₂. Inair-grown cells, O₂ evolution dependent on low, but not high,HCO^–₃ concentrations was strongly inhibited by the carbonicanhydrase inhibitor acetazolamide. Cells grown with 3% CO² exhibitedlow rates of O2 evolution at low external inorganic C; however,after 30 min in air O2 evolution rates at low inorganic C approachedthose of air-grown cells. These results are compatible withthe view that Scenedesmus develops a ‘CO² concentratingmechanism’ in air, with carbonic anhydrase as an importantconstituent When 3% CO²-grown cells were subjected to air-level of CO²,just a transient decline in NO^–₃ utilization was observed,but in the presence of acetazolamide the rate of the processdecreased drastically in response to the decrease in the CO²level. In CO²-free air NO₃^– was taken up at high ratesbut in a deregulated manner, leading to release of NH₄⁺. A portionof the NO₃^– taken up in the absence of CO² was apparentlyassimilated Cellular nitrate reductase (NR) activity initially decreasedbut subsequently recovered after a transition from 3% CO² toair. In the presence of acetazolamide, a persistent decreasein NR activity was observed. Cellular glutamine synthetase (GS)activity increased after transition from 3% CO² to air, theactivity increase being unaffected by acetazolamide. NH₄⁺ releaseto the medium in the presence of L-methionine-D, L-sulphoximine(MSO) transiently increased in 3% CO²-grown cells in responseto a transfer to air. MSO-induced NH₄⁺ release was in fact higherin air-grown cells than in 3% CO²-grown cells. Glycollate wasinitially released after transition from 3% CO² to air, butthere was no difference in glycollate release between MSO-treatedand untreated cells. In air-adapted Scenedesmus, N recyclingseems to be of minor importance in comparison to primary N assimilation Key words: CO²-fixation, N recycling, nitrate uptake, Scenedesmus