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     

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     

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     

Inhibition of Nodule Development in Soybean by Nitrate or Reduced Nitrogen

Imsande, J. 1986. Inhibition of nodule development in soybeanby nitrate or reduced nitrogen.—J. exp. Bot. 37: 348–355. Nodulation of hydroponically grown soybean plants [Glycine max(L.) Merr.] is inhibited by continuous growth in the presenceof 4· mol m^–3 KNO₃ The presence of 4·0 molm^–3 ‘starter nitrate’ for 3-6 d during noduledevelopment, however, subsequently stimulates nodule dry weightaccumulation and nitrogenase activity. These stimulations occureven though 4· mol m^–3 nitrate temporarily delaysnodule development, i.e. the late steps of nodule developmentare reversibly inhibited by a short-term exposure to 4·0mol m^–3 nitrate. On the other hand, treatment with 4·0mol m^–3 nitrate in excess of 14 d significantly reducesnodule dry weight Thus, extended growth in the presence of 4·0mol m^–3 KNO₃ seems to block both early and late stepsof nodule development. Nodulation of hydroponically grown soybeansis also inhibited by continuous growth in the presence of 2·0mol m^–3 (NH₄)₂SO₄ This inhibition is not caused by acidityof the growth medium. On the other hand, nodule development6 d after inoculation with Rhizoblum japonicum is not delayedby a 7-d exposure to 2·0 mol m^–3 (NH₄)₂SO₄ butis partially inhibited by a prolonged exposure to (NH₄)₂SO₄Because repression of nodulation by 4·0 mol m^–3KNO₃ is more severe than that by 2·0 mol m^–3 (NH₄)₂SO₄and because ammonium taken up by the soybean plant is not activelyoxidized to nitrate, it is suggested that there are at leasttwo mechanisms by which nitrate utilization represses noduleformation in soybean. Key words: Glycine max, nitrogen, nitrogen fixation, nodulation     

Nitrate-Ammonium Ratio Required for pH Homeostasis in Hydroponically Grown Soybean

Imsande, J. 1986. Nitrate-ammonium ratio required for pH homeostasisin hydroponically grown soybean.—J. exp. Bot. 37: 341–347. Plant acid-base homeostasis is achieved when the mmoles of hydroxylions produced in the plant equal the mmoles of protons. Reductionof nitrate to ammonia is the major source of hydroxyl ions whereasammonium uptake-assimilation and the metabolism of neutral sugarsto organic acids are the primary sources of protons. Soybean[Glycine max (L.) Merr plants were grown hydroponically on mediumsupplemented with 3·0 mol m^–3 nitrogen providedas various combinations of KNO₃ and NH₄NO₃ Plant growth consumedessentially all available nitrogen in each case; however, onlyin flasks supplemented with approximately 1·8 minolesof KNO₃ plus 0·6 mmole of NH₄NO₃ was the pH of the mediumunchanged. Thus, for every mmole of nitrogen assimilated, approximately0·6 mmole of dissociable protons must have been producedby the conversion of neutral sugars to carboxylic acids. Also,it was shown that a plant obtaining all of its nitrogen fromnitrate must neutralize or excrete approximately 0·5mmole of hydroxyl ion d^–1. Conversely, the plant derivingall of its nitrogen from dinitrogen must excrete or neutralizeat least 0·8 mmole of hydrogen ion d^–1 whereasthe plant deriving all of its nitrogen from aminonium must excreteor neutralize approximately 2·1 mmoles of hydrogen iond^–1. Nevertheless, plants grown on medium supplementedwith 2·4 mol m^–1 nitrate plus 0·6 mol m^–3ammonium did not achieve a higher growth rate than plants grownon 3·0 mol m^–3 nitrate. Key words: Glycine max, nitrogen fixation, nitrate utilization     

Influence of Elevated CO2 and Temperature on the Photosynthesis and Respiration of White Clover Dependent on N2 Fixation

Single clonal plants of white clover (Trifolium repens L) grownfrom explants in a Perlite rooting medium, and dependent fornitrogen on N₂ fixation in root nodules, were grown for severalweeks in controlled environments which provided two regimesof CO₂, and temperature 23/18 °C day/night temperaturesat 680 µmol mol^–1 CO₂, (C680), and 20/15 °Cday/night temperatures at 340 µmol mol^–1 CO₂ (C340)After 3–4 weeks of growth, when the plants were acclimatedto the environmental regimes, leaf and whole-plant photosynthesisand respiration were measured using conventional infra-red gasanalysis techniques Elevated CO₂ and temperature increased ratesof photosynthesis of young, fully expanded leaves at the growthirradiance by 17–29%, despite decreased stomatal conductancesand transpiration rates Water use efficiency (mol CO₂ mol H₂O^–1)was also significantly increased Plants acclimated to elevatedCO₂, and temperature exhibited rates of leaf photosynthesisvery similar to those of C340 leaves ‘instantaneously’exposed to the C680 regime However, leaves developed in theC680 regime photosynthesised less rapidly than C340 leaves whenboth were exposed to a normal CO₂, and temperature environmentIn measurements where irradiance was varied, the enhancementof photosynthesis in elevated CO₂ at 23 °C increased graduallyfrom approx 10 % at 100 µmol m^–1 s^–1 to >27 % at 1170 µmol m^–2 s^–1 In parallel, wateruse efficiency increased by 20–40 % at 315 µmolm^–2 s^–1 In parallel, water use efficiency increasedby 20–40 % at 315 µmol m^–2 s^–1 In parallel,water use efficiency increased by 20–40 % at 315 µmolm^–2 s^–1 In parallel, water use efficiency increasedby 20–40 % at 315 µmol m^–2 s^–1 to approx100 % at the highest irradiance Elevated CO₂, and temperatureincreased whole-plant photosynthesis by > 40 %, when expressedin terms of shoot surface area or shoot weight No effects ofelevated CO₂ and temperature on rate of tissue respiration,either during growth or measurement, were established for singleleaves or for whole plants Dependence on N₂, fixation in rootnodules appeared to have no detrimental effect on photosyntheticperformance in elevated CO₂, and temperature Trifolium repens, white clover, photosynthesis, respiration, elevated CO₂, elevated temperature, water use efficiency, N₂ fixation     

Potassium-Ammonium Uptake Interactions in Tobacco Seedlings

Short-term (< 12 h) uptake experiments were conducted with6–7-week-old tobacco (Nicotiana tabacum L. cv. Ky 14)seedlings to determine absorption interactions between K⁺ andNH₄⁺. At equal solution concentrations (0.5 mol m^–3) netK⁺ uptake was inhibited 30–35% by NH₄⁺ and NH₄⁺ uptakewas decreased 9–24%. Removal of NH₄⁺ resulted in completerecovery in K⁺ uptake rate, but NH⁴⁺ uptake rate did not recoverwhen K⁺ was removed. In both cases, inhibition of the uptakerate of one cation saturated as the concentration of the othercation was increased up to 0.5 mol m^–3. The relative effectof K⁺-NH₄⁺ interactions was not altered when Cl- was replacedwith SO₄^2–, but the magnitudes of the uptake rates wereless in the absence of Cl-. The V_max for NH₄⁺ uptake was reducedfrom 128 to 105 µmol g^–1 dry wt. h^–1 in thepresence of 0.5 mol m^–3 K⁺ and the K_m for NH₄⁺ doubledfrom 12 to 27 mmol m^–3 in the presence of K⁺. The resultsof these K⁺-NH₄⁺ experiments are interpreted as mixed-noncompetitiveinteractions. However, an enhanced efflux of K⁺ coupled to NH₄⁺influx via an antiporter cannot be ruled out as contributingto the decrease in net K⁺ uptake. Key words: Nicotiana tabacum, K⁺, NH₄⁺, Uptake interactions     

Ammonium Nutrition in Ricinus communis: Its Effect on Plant Growth and the Chemical Composition of the Whole Plant, Xylem and Phloem Saps

Allen, S. and Smith, J A. C. 1986. Ammonium nutrition in Ricinuscommunis: its effect on plantgrowth and the chemical compositionof the whole plant, xylem and phloem saps.—J. exp. Bot.37: 1599–1610. The growth and chemical composition of Ricinus communis cultivatedhydroponically on 12 mol m ^{– 3} NO₃^–-N were comparedwith plants raised on a range of NH₄⁺-N concentrations. At NH₄⁺-Nconcentrations between 0·5 and 4·0 mol m^–3,fresh- and dry-weight yields of 62-d-old plants were not significantlydifferent from those of the NO₃^–-N controls. Growth wasreduced at 0·2 mol m^–3 NH₄⁺-N and was associatedwith increased root. shoot and C: organic N ratios, suggestingthat the plants were N-limited. At 8·0 mol m^–3NH₄⁺-N, growth was greatly restricted and the plants exhibitedsymptoms of severe ‘NH₄⁺ toxicity’. Plants growingon NH₄⁺-N showed marked acidification of the rooting medium,this effect being greatest on media supporting the highest growthrates. Shoot carboxylate content per unit dry weight was lower at mostNH₄⁺-N concentrations than in the NO₃^–-N controls, althoughit increased at the lowest NH₄⁺-N levels. Root carboxylate contentwas comparable on the two N sources, but also increased substantiallyat the lowest NH₄⁺-N levels. N source had little effect on inorganic-cationcontent at the whole-plant level, while NO₃^– and carboxylatewere replaced by Cl as the dominant anion in the NH₄⁺-N plants.This was reflected in the ionic composition of the xylem andleaf-cell saps, the latter containing about 100 mol m^–3Cl^– in plants on 8·0 mol m^–3 NH₄⁺. Xylem-saporganic-N concentration increased more than threefold with NH₄⁺-N(with glutamine being the dominant compound irrespective ofN source) while in leaf-cell sap it increased more than 12-foldon NH₄⁺-N media (with arginine becoming the dominant species).In the phloem, N source had little or no effect on inorganic-cation,sucrose or organic-N concentrations or sap pH, but sap fromNH₄⁺-N plants contained high levels of Cl^– and serine. Collectively, the results suggested that the toxic effects ofhigh NH₄⁺ concentrations were not the result of medium acidification,reduced inorganic-cation or carboxylate levels, or restrictedcarbohydrate availability, as is commonly supposed. Rather,NH₄⁺ toxicity in R. communis is probably the result of changesin protein N turnover and impairment of the photorespiratoryN cycle. Key words: Ricinus, ammonium nutrition, nitrate, whole-plant composition, xylem, Phloem, amino acids, carboxylate     

The Inhibition of Phloem Translocation by Ammonia

Bean plants (Phaseolus vulgaris L. cv. Fardenlosa Shiny) werelabelled with carbon-11 via their first trifoliate leaves when3-weeks-old and the transient inhibitions of translocation causedby the application of ammonium chloride solutions (10 mol m^–3)to a peeled region of stem were studied. At pH 6·5 theammonium was without effect. At pH 11·0 even a briefapplication inhibited translocation for many minutes, whilelonger applications inhibited translocation for considerablylonger. Solutions of 10 mol m^–3 sodium chloride were withouteffect at either pH. At pH 6·5 ammonium chloride solution contains predominantlyammonium ions (NH₄⁺) and at pH 11·0 predominantly dissolvedammonia gas (NH₃). Hence we conclude that phloem transport withinbean stems is inhibited by dissolved ammonia gas but not ammoniumions. Key words: Phloem translocation, transient inhibition, ammonia, ammonium ion     

Salinity-induced Malate Accumulation in Chara

Ion absorption by Chara corallina from solutions containingpredominantly KC1 or RbCl at up to 100 mol m^–3 resultedin accumulation of salts and turgor regulation. Turgor regulationdid not occur in solutions containing Na⁺ or Li⁺salts. Duringion absorption from various salts of K⁺ and Rb⁺ vacuolar cationconcentration exceeded Cl^– concentration. This differencewas shown to be balanced by the synthesis and accumulation ofmalate. Vacuolar malate concentration reached 48 mol m₃^–,with accumulation occurring at rates of up to 0.45 mol m^–3h^–1. Malate accumulation was inhibited by low externalpH and was dependent upon external HCO₃^– concentration.The synthesis of malic acid and its subsequent dissociationimposed a severe acid load on the cell. Biophysical regulationof cellular pH was achieved by a H⁺efflux at a rate of about40 nmol m^–2 s^–1from the cell. The results presentedargue against cytoplasmic Cl^–, HCO₃^– or pH regulatingmalate accumulation in Chara and it is suggested that malatetransport across the tonoplast may regulate malate accumulation. Key words: Malate, Chara corallina, pH regulation, salinity     

Further Characteristics of Salt-Dependent Bicarbonate Use by the Seagrass Zostera muelleri

Millhouse, J. and Strother, S. 1987. Further characteristicsof salt-dependent bicarbonate use by the seagrass Zostera muelleri.—J.exp. Bot. 38: 1055–1068. The contribution of HCO^–₃to photosynthetic O₂ evolutionin the seagrass Zostera muelleri Irmisch ex Aschers. increasedwith increasing salinity of the bathing seawater when the inorganiccarbon concentration was kept constant. K_1/2 (seawater salts)for HCO^–₃ -dependent photosynthesis was 66% of seawatersalinity. Both short- and long-term pretreatment at low salinitiesstimulated photosynthesis in full strength seawater. Twentyfour hours pre-incubation of seagrass plants in 3·0 molm^–3 NaHCO₃ resulted in increased photosynthesis at allsalinities, apparently due to stimulation of HCO^–₃ use(K_1/2 (seawater salts) = 26%). V_max (HCO^–₃) was not affectedby low salinity pretreatment. The kinetics of HCO^–₃ stimulationby the major seawater cations was investigated. Ca²⁺ was themost effective cation with the highest V_max (HCO^–₃) andwith K_1/2(Ca²⁺) = 14 mol m^–3. Mg²⁺ was also very effectiveat less than 50 mol m^–3 but higher concentrations wereinhibitory. This inhibition cannot be accounted for solely byprecipitation of MgCO₃. Na⁺ and K⁺ were both capable of stimulatingHCO^–₃ use. Stimulation was in two distinct parts. Up to500 mol m^–3, both citrate and chloride salts gave similarresults (K_1/2(Na⁺) 81 mol m^–3, V_max(HCO^–₃) 0·26µmol O₂ mg^–1 chl min^–1), but use of citratesalts above 500 mol m^–2 caused a second stimulation ofHCO^–₃ use (K_1/2(Na⁺) 830 mol m^–3, V_max(HCO^–₃)0·68 µmol O₂ mg^–1 chl min^–1). V_max(HCO^–₃)for the second-phase Na⁺ or K⁺ stimulation was of the same orderas for Ca²⁺-stimulated HCO^–₃ use. To further characterizesalt-dependent HCO^–₃ use, the sensitivity of photosynthesisto Tris and TES buffers was investigated. The effects of Trisappear to be due to the action of Tris⁺ causing stimulationof HCO^–₃ -dependent photosynthesis in the absence of salt,but inhibition of HCO^–₃ use in saline media. TES has noeffect on photosynthesis. External carbonic anhydrase, althoughimplicated in salt-dependent HCO^–₃ use in Z. muelleri,could not be detected in whole leaves. Key words: Zostera muelleri, HCO^–₃ use, salinity     

The Influence of the Concentration of Gaseous Ammonia on its Uptake by the Leaves of Italian Ryegrass, with and without an Adequate Supply of Nitrogen to the Roots

Whitehead, D. C. and Lockyer, D. R. 1986. The influence of theconcentration of gaseous ammonia on its uptake by the leavesof Italian ryegrass, with and without an adequate supply ofnitrogen to the roots.—J. exp. Bot. 38: 818–827. Plants of Italian ryegrass (Lolium multiflorum Lam.) were grownin pots of soil with two rates of ¹⁵N-labclled nitrate, oneproviding adequate, and the other less than adequate, N formaximum growth. After 25 d in a controlled environment cabinet,the plants were transferred to chambers and exposed for 33 dto NH₃in the air at one of nine concentrations ranging from14 to 709 µg NH₃ m^–3. Increasing the concentrationof NH₃ in the air increased the dry weight of the shoots ofplants grown at the lower but not the higher rate of nitrate.The content of total N in the plant shoots (% dry weight) waslinearly related to NH₃ concentration; at 709 µg NH₃ andin both sets of plants it was more than double the content at14 µg NH₃ m^–3. Calculations, based on ¹⁵N enrichment,indicated that the amount of N taken up from the NH₃ per unitleaf area increased linearly with increasing concentration ofNH₃ in the air uptake (µg dm^–2 h^–1) = 0.1009xat the lower rate of nitrate and 0-0829x at the higher rateof nitrate, where x is the concentration of NH₃ in the air expressedas µg NH₃m^–3. The proportion of the total plant N that was derived from theNH₃ ranged from 4?0% at a concentration of 14 µg NH₃ m^–3with the higher rate of nitrate addition to 77?5% at a concentrationof 709 µg m^–3 with the lower rate of nitrate addition.The proportions of the total N in the water-insoluble proteinof the leaf tissue that were derived from nitrate and gaseousNH₃ were similar to the proportions in the whole leaf material. Key words: Ammonia, nitrogen, leaf sorption, Lolium multiflorum     

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 Uptake of Gaseous Ammonia by the Leaves of Italian Ryegrass

Lockyer, D. R. and Whitehead, D. C. 1986. The uptake of gaseousammonia by the leaves of Italian ryegrass.—J. exp. Bot.37: 919–927. Plants of Italian ryegrass (Lolium multiflorum Lam.) grown insoil with two rates of added ¹⁵N-labelled nitrate were exposed,in chambers, for 40 d to NH₃ in the air at concentrations of16, 118 and 520 µg m^–3. At the highest concentrationof NH₃, this source provided 47?3% of the total nitrogen inplants grown with the lower rate of nitrate addition (100mgN kg^–1 dry soil) and 35?2% with the higher rate (200mgN kg^–1 dry soil) At the intermediate concentration ofNH₃, the contributions to total plant N were 19?6% and 10?8%,respectively, at low and high nitrate while, at the lowest concentrationof NH₃, they were 5?1% and 32%. Most of the N derived from theNH₃ remained in the leaves, but some was transported to theroots. The amount of N derived from the NH³ that was presentin the leaves was not reduced by washing the leaves in waterat pH 5?0 before harvesting, indicating that the N was assimilatedby the plant and not adsorbed superficially. Rates of uptakeof NH₃ per unit leaf area ranged from 1?7 µg dm^–2h^–1 at a concentration of 16 µg m^–3 to 29?0µg dm^–2 h^–1 at a concentration of 520 µgm^–3 and with the lower rate of nitrate addition. Increasingthe supply of nitrate to the roots slightly reduced the rateof uptake of NH₃ per unit leaf area. Uptake of N from the higherrate of nitrate was reduced at the highest concentration ofNH₃ in the air. Key words: Ammonia, nitrogen, leaf sorption, Lolium multiflorum     

Purification and Characterization of Soluble and Membrane-Associated DNA Polymerases from a Virus PBCV-1 Infected Green Alga Chlorella NC64A

DNA polymerases were purified several hundred-fold from the10 000 x g soluble (polymerase I) and particulate (polymeraseIII) fractions prepared from virus PBCV-1 infected ChlorellaNC64A extracts. Both DNA polymerases exhibited optimal activitywith activated calf thymus DNA at pH 8.5. DNA polymerase I required3.0 mol m^–3 MgSO₄ and 150 to 250 mol m^–3 KCl foroptimum activity whereas, DNA polymerase III required 2.0 molm^–3 MgSO₄ and 150 mol m^–3 KCl. Both enzymes wereinhibited by pyrophosphate, actinomycin D, ethidium bromide,dideoxythymidine triphosphate, and N-ethylmaleimide but wererelatively insensitive to aphidicolin. DNA polymerase I differedfrom DNA polymerase III in its response to cations (particularlyNH₄Cl), elution from a DEAE cellulose column, and molecularweight. Key words: Algal virus, DNA polymerase, Chlorella     

Effect of Salinity on Growth, Nodulation and Nitrogen Yield of Chickpea (Cicer arietinum L.)

Chickpea cultivar ILC 482 was inoculated with salt-tolerantRhizobium strain Ch191 in solution culture with different saltconcentrations added either immediately with inoculation or5 d later. The inhibitory effect of salinity on nodulation ofchickpea occurred at 40 dS m^–1 (34.2 mol m^–3 NaCl)and nodulation was completely inhibited at 7 dS m^–1 (61.6mol m^–3 NaCl); the plants died at 8 dS m^–1 (71.8mol m^–3 NaCl). Chickpea cultivar ILC 482 inoculated with Rhizobium strain Ch191^spcstrwas grown in two pot experiments and irrigated with saline water.Salinity (NaCl equivalent to 1–4 dS m^–1) significantlydecreased shoot and root dry weight, total nodule number perplant, nodule weight and average nodule weight. The resultsindicate that Rhizobium strain Ch191 forms an infective andeffective symbiosis with chickpea under saline and non-salineconditions; this legume was more salt-sensitive compared tothe rhizobia, the roots were more sensitive than the shoots,and N₂ fixation was more sensitive to salinity than plant growth. Key words: Cicer arietinum, nodulation, N₂ fixation, Rhizobium, salinity     

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     

Influence of nitrate on uptake of ammonium by nitrogen-depleted soybean: is the effect located in roots or shoots?

In non-nodulated soybean [Glycine max (L.) Merrill cv. Ransom]plants that were subjected to 15 d of nitrogen deprivation inflowing hydroponic culture, concentrations of nitrogen declinedto 1.0 and 1.4mmol Ng^–1 dry weight in shoots and roots,respectively, and the concentration of soluble amino acids (determinedas primary amines) declined to 40µmol g^–1 dry weightin both shoots and roots. In one experiment, nitrogen was resuppliedfor 10 d to one set of nitrogen-depleted plants as 1.0 mol m^–3NH₄⁺ to the whole root system, to a second set as 0.5 mol m^–3NH₄⁺ plus 0.5 mol m^–3 NO₃^– to the whole root system,and to a third set as 1.0 mol m^–3 NH₄⁺ to one-half ofa split-root system and 1.0 mol m^–3 NO₃^– to theother half. In a second experiment, 1.0 mol m^–3 of nitrogenwas resupplied for 4 d to whole root systems in NH₄⁺ : NO₃^–ratios of 1:0, 9:1, and 1:1. Nutrient solutions were maintainedat pH 6.0. When NH₄⁺ was resupplied in combination with NO₃^– to thewhole root system in Experiment I, cumulative uptake of NH₄⁺for the 10 d of resupply was about twice as great as when NH₄⁺was resupplied alone. Also, about twice as much NH₄⁺ as NO₃^–was taken up when both ions were resupplied to the whole rootsystem. When NH₄⁺ and NO₃^– were resupplied to separatehalves of a split-root system, however, cumulative uptake ofNH₄⁺ was about half that of NO₃^–. The uptake of NH₄⁺,which is inhibited in nitrogen-depleted plants, thus is facilitatedby the presence of exogenous NO₃^–, and the stimulatingeffect of NO₃^– on uptake of NH₄⁺ appears to be confinedto processes within root tissues. In Experiment II, resupplyof nitrogen as both NH₄⁺ and NO₃^– in a ratio of either1:1 or 9:1 enhanced the uptake of NH₄⁺. The enhancement of NH₄⁺uptake was 1.8-fold greater when the NH₄⁺: NO₃^–-resupplyratio was 1:1 than when it was 9:1; however, only 1.3 timesas much NO₃^– was taken up by plants resupplied with the1 :1 exogenous ratio. The effect of NO₃^– on enhancementof uptake of NH₄⁺ apparently involves more than net uptake ofNO₃^– itself and perhaps entails an effect of NO₃^–uptake on maintenance of K⁺ availability within the plant. Theconcentration of K⁺ in plants declined slightly during nitrogendeprivation and continued to decline following resupply of nitrogen.The greatest decline in K⁺ concentration occurred when nitrogenwas resupplied as NH₄⁺ alone. It is proposed that decreasedavailability of K⁺ within the NH₄⁺-resup-plied plants inhibitedNH₄⁺ uptake through restricted transfer of amino acids fromthe root symplasm into the xylem. Key words: Ammonium, Glycine max, nitrate, nitrogen-nutrition, nitrogen stress, split-root cultures     

The Combined Effects of Salinity and Root Anoxia on Growth and Net Na+ and K+-accumulation in Zea mays Grown in Solution Culture

The effects of excess salinity and oxygen deficiency on growthand solute relations in Zea mays L. cv. Pioneer 3906 were examinedin greenhouse experiments. The roots of plants 14 d old growingin nutrient solution containing additions of NaCl in the range1.0–200 mol m^–3 were either exposed to a severedeficiency of O₂ by bubbling with nitrogen gas (N₂ treatment),or maintained with a supply of air (controls), for a periodof 1–7 d. The threshold NaCl concentration resulting inappreciable inhibition of leaf extension, and shoot f. wt gainin controls was between 10 and 25 mol m^–3. At 25 mol m^–3NaCl the ratio of Na+/K+ transported to shoots was about 20times greater than in plants in 1.0 mol m^–3 NaCl. Theeffect of addition of NaCl to the nutrient solution was to enhanceNa+ movement but simultaneously depress the rate of K+ transportto shoots (per g f. wt roots). Interactions between NaCl levels and aeration treatment wereshown by analyses of variance to be statistically significantfor leaf extension, shoot and root f. wt gains, Na+ and K+ concentrationsin shoots and roots. When roots were N₂-treated, shoot and rootgrowth were depressed, the effect of aeration treatment beinggreatest at NaCl concentrations of 50 mol m^–3 or less.Additionally, N₂-treatment greatly accelerated Na- transportto shoots while depressing K+ transport still further, so thatat 10 mol m^–3 NaCl the ratio Na+/K+ acquired by the shootswas 230 times greater than in controls. Over the concentrationrange 1.0 to 50 mol m^–3 NaCl, the ratio Na+/K+ transportedto shoots by anoxic roots increased by a factor of 860. Mechanisms controlling changes in solute flux to the shoot,and the significance in relation to plant tolerance of excesssalts or oxygen deficiency are discussed. Anaerobic, corn, flooding, maize, oxygen-deficiency, salinity     

Growth of White Clover, Dependent on N2 Fixation, in Elevated CO2 and Temperature

Clonal plants of white clover (Trifolium repens L ), whollydependent on N₂ fixation, were grown for 6 weeks in controlledenvironments providing either (C680 regime) 23/18 °C day/nighttemperatures and a CO₂, concentration of 680 µmol mol^–1,or (C340 regime) 20/15 °C day/night temperatures and a CO₂,concentration of 340 µmol mol^–1 During the firsthalf of the experimental period the C680 plants grew fasterthan their C340 counterparts so that by week 3 they were twicethe weight this 2 1 superiority in weight persisted until theend of the experiment The faster initial growth of the C680plants was based on an approx 70 % increase in leaf numbersand an approx 30 % increase in their individual area Initially,specific leaf area (cm2 g^–1 leaf) was lower in C680 thanin C340 leaves but became similar in the latter half of theexperiment Shoot organ weights, including petioles and stolons,reflected the C680 plant's better growth in terms of photosyntheticsurface Throughout, C680 plants invested less of their weightin root than C340 plants and this disparity increased with timeAcetylene reduction assays showed that nitrogenase activityper unit nodule weight was the same in both C680 and C340 plantsBoth groups of plants invested about the same fraction of totalweight in nodules Nitrogen contents of plant tissues were similarirrespective of growth regime, but C680 expanded leaves containedslightly less nitrogen and their stolons slightly more nitrogenthan their C340 counterparts However, C680 leaves containedmore non-structural carbohydrate Young, unshaded C680 leavespossessed larger palisade cells, packed more tightly withinthe leaf, than equivalent C340 leaves The reason for the C680regime's loss of superiority in relative growth rate duringthe second half of the experiment was not clear, but more accumulationof non-structural carbohydrate, constriction of root growthand increased self-shading appear to be the most likely causes Trifolium repens, white clover, elevated CO₂, elevated temperature, growth, N₂ fixation, leaf structure