The Effects of Temperature and Form of Nitrogen Supply on the Relative Contribution of Root Uptake and Remobilization in Supplying Nitrogen for Laminae Regrowth of Lolium perenne L.

Plants of Lolium perenne L. cv. S23 were grown in sand culturesupplied with either ammonium (NH₄⁺) or nitrate (NO₃^–)in an otherwise complete nutrient solution at 12°C or 20°C.Three weeks after germination, plants were clipped weekly tosimulate grazing. After 10 weeks growth all nitrogen (N) wassupplied enriched with ¹⁵N to quantify the effects of form ofN supply and temperature on the relative ability of currentroot uptake and remobilization to supply N for laminae regrowth. The form of N supply had no effect on the dry matter partitioning,while at 20°C more dry weight was allocated to laminae regrowthand less to the remaining plant material. The current root uptakeof N, which subsequently appeared in the laminae regrowth, wassimilar for plants supplied with NH₄⁺ or NO₃^–, and bothwere equally reduced at the lower temperature of growth. Remobilizationof N to laminae regrowth was greater for plants receiving NH₄⁺than NO₃^–; remobilization with either form of N supplywas reduced at the lower temperature of growth. Remobilizationwas reduced to a lesser extent at 12°C than current rootuptake. It was concluded that remobilization became relativelymore important in supplying N for regrowth of laminae at lowertemperatures. Key words: Lolium perenne, ammonium, nitrate, temperature, remobilization     

Increased Defoliation Frequency Depletes Remobilization of Nitrogen for Leaf Growth in Grasses

Plants ofLolium perenneandFestuca rubrawere grown in sand culturereceiving all nutrients as a complete nutrient solution containing1.5 mMNH₄NO₃, and subjected to one of three defoliation treatments:undefoliated, defoliated on one occasion, or defoliated weekly.¹⁵Nlabelling was used to determine the rate of N uptake, allowingthe amount of N remobilized from storage for the growth of thetwo youngest leaves (subsequently referred to as ‘newleaves’) growing over a 14 d period after defoliationto be calculated. The total plant N uptake by both species wasreduced, compared with undefoliated plants, by both a singleand repeated defoliation, although neither caused complete inhibitionof uptake. Regularly defoliatedL. perennehad a greater reductionin root mass, concomitant with a greater increase in N uptakeper g root than did regularly defoliatedF. rubra. In both species,the amount of N derived from uptake recovered in the new leaveswas unaffected by the frequency of defoliation. BothL. perenneandF.rubramobilized nitrogen to the new leaves after a single defoliation,mobilization being sufficient to supply 50 and 41%, respectively,of the total nitrogen requirement. In regularly defoliated plants,no significant nitrogen was mobilized to the new leaves inL.perenne, and only a small amount was mobilized inF. rubra. Plantsachieved greater leaf regrowth when only defoliated once. Weconclude that increasing the frequency of defoliation of bothL.perenneandF. rubrahad little effect on the uptake of nitrogenby roots which was subsequently supplied to new leaves, butdepleted their capacity for nitrogen remobilization, resultingin a reduction in the rate of growth of new leaves. Lolium perenne; Festuca rubra; defoliation frequency; mobilization; root uptake; nitrogen     

Effect of Nitrogen Supply on the Grass and Clover Components of Simulated Mixed Swards Grown under Favourable Environmental Conditions II. Nitrogen Fixation and Nitrate Uptake

Simulated mixed swards of Perennial Ryegrass (Lolium perenneL.) cv. S23 and White clover (Trifolium repens L.) cv. S100were grown from seed under a constant 20 °C day/15 °Cnight temperature regime and harvested at intervals over and88 d growht period. The swards received a nutrient solutiondaily, which was either High (220 mg l^–1) or Low (10 mgl^–1) in nitrate N. The nitrate was labelled with the ¹⁵Nisotope. An acetylene reduction assay was carried out on eachsward just prior to harvest. Rates of acetylene reduction agreed qualitatively with the ^l5Nanalyses but absolute values did not match (assuming a 4:1 C₂H₄:N₂ratio) and errors in the acetylene assay are discussed. In theLow-N swards clover relied almost entirely on symbioticallyfixed N₂, fixing more than ten times as much as the High-N cloverplants. In the Low-N treatment the grass was N-deficient despiteobtaining much more nitrate per unit root dry weight than clover.In the High-N swards, however, clover took up more nitrate perunit root weight than grass. The High-N clover plants also fixedsome N₂ and maintained a higher total-N content than grass throughoutthe period. There was no evidence of transfer of symbioticallyfixed N from the clover to the grass in either treatment. Trifolium repens, Lolium perenne, nitrate, nitrogen fixation, ¹⁵N, acetylene reduction     

Nitrate Nutrition of Grasses from Steady-State Supplies in Flowing Solution Culture following Nitrate Deprivation and/or Defoliation: II. ASSIMILATION OF AND SHORT-TERM EFFECTS ON UPTAKE

Plants of two genotypes of Lolium perenne L. cv. S23 and a L.perenne ? L. multiflorum Lam. hybrid cv. Augusta were grownin flowing solution culture. N was suppled in one treatmentat 10 mmol m^–3 NO^–3 throughout (HN), and in another(LN) the N supply was terminated after 10 d for 11 d. When was re-supplied both LN and HN plants were leftentire or defoliated. The two genotypes showed similar responsesto all treatments. The concentration of N in shoot dry matterdeclined from 4.4% to 2.0% and in the root from 2.8% to 1.0%over the 11 d of N deprivation, with 95% of the initially present being assimilated during this period. LN plantsassimilated 10% more of their total uptake than did HN plants. The in vitro nitrate reductase activity(NRA) was 10- to 50-fold higher in the youngest fully-expandedleaves than in roots and declined in the leaves during N deprivation.Between 2–6 d after defoliation, there was a large increasein NRA in leaves of HN (but not LN) plants. After defoliationof HN plants, net uptake from 10 mmol m^–3 declined to negligible levels within 15 h, but in defoliatedLN plants it increased to levels similar to those of entireHN plants (15–20 µmol h^–1 g^–1 fr. wt.root) within 8 h. When was re-supplied to entire LN plants, uptake of increased to levels similar to those of entire HN plants within 2.3 h, butdid not markedly exceed that of HN plants for at least 10 h.Net uptake of by LN plants during depletion of stirred static nutrient solutions containing 1.0 mol m^–3 lagged behind that by HN plants by 70–100 min, but the maximum unit absorption rate was similar for LNand HN plants (5–7 µmol h^–1 g^–1 fr.wt. root). The nature of the short-term demand for uptake following recovery from the stresses of defoliation andN starvation is discussed. Key words: Lolium perenne, Lolium multiflorum, N-deficiency, defoliation, nitrate uptake, nitrate reductase, N-assimilation     

Diurnal variation in the simultaneous uptake and 'sink' allocation of NH4+ and NO3- by Lolium perenne in flowing solution culture

Perennial ryegrass (Lolium perenne L.) was grown in flowingsolution culture under artificial illumination with a 10 h lightperiod. The diurnal fluctuations in the net uptake of and from a 20 mmol m^–3 NH₄NO₃ supply were measured (i) fromthe amounts of these ions supplied automatically to maintainconcentrations in solution, and (ii) by ¹⁵N pulse-labellingthroughout the diurnal cycle. Diurnal variation in leaf extension,translocation and allocation of assimilated C (¹⁴CO₂ pulse-labelling)and N (¹⁵N pulse and steadystate labelling) were also followed.The apparent ‘sinkstrength’ of different organsfor recently assimilated C and N was calculated from the concentrationof label recovered in the tissues. Ammonium and showed qualitatively similar diurnal patterns ofnet uptake, with minimum and maximum rates, respectively, atthe start and end of the photoperiod; but uptake showed a proportionately steeper declineduring the dark period. This trend was mirrored by the decreasein translocation of N from roots to shoots during the dark,concurrent with an increase in the relative allocation of -N to expanding leaves and young tillers,and in -N to older expanded leaves. Overall the apparent ‘sink-strength’ ofexpanding leaves for N declined during the dark period. Pulse-chasesof ¹⁴CO₂ fed to the youngest fully expanded leaf at the startand end of the photoperiod showed that translocation of ¹⁴Cto the roots continued throughout the dark period, but thatthe apparent rate was halved after 9 h of darkness. The resultswere interpreted as contrary to the ‘carbohydrate supplylimitation’ hypothesis for the dark-related decline inN uptake, but compatible with regulation by ‘sink-strength’for N. Key words: Nitrate, ammonium, diurnal variation, perennial ryegrass, translocation     

Nitrate Transport Characteristics in Hordeum vulgare L. Seedlings using Three Different Tracer Techniques

Deane-Drummond, C. E. and Thayer, J.R. 1986. Nitrate transportcharacteristics in Hordeum vulgare L. seedlings using threedifferent tracer techniques.—J. exp. Bot. 37: 429–439. and have been used to investigate various properties of nitrate uptake and translocation intoHordeum vulgare L seedlings. Short term / influx into seed lings grown in CaSO⁴ was stimulated by after a lag of 2 h. The apparent kinetics of shortterm / influx over the concentration range 0?0–0?7mol m fitted Michaelis-Menten equations The apparent V_max didnot change when seedlings were used that had been pretreatedin 10 or 100 mmol m^–3 and V^max=3.77 and 3?56µmol g^–1 fr. wt. h^–1respectively. The apparent Michaelis constants were also similarand K_m=0?47 and 0?45 mol m^–3 respectively. Longer term pulse chase experiments with the heavy isotope ¹⁵Nhave shown that feeding roots with resulted in the preferential appearance of ¹⁵N labelled aminoacids in the xylem sap. Pulse chase experiments with the radioisotope¹³N have shown that feeding shoots with resulted in a radial pattern of distnbution of labelin the leaf veins, which can be detected using autoradiography. The limitations and advantages of all three techniques are comparedby reference to other known experimental data. Key words: ³⁶Chlorate, ¹³nitrate, ¹⁵nitrate, Hordeum vulgare     

Evidence for N feedback regulation of N2 fixation in Alnus glutinosa L.

Treatments were applied to vary C and N availability in Alnusglutinosa L. and plant growth, nodule activity (including acetylenereduction) and amino acid composition of the xylem sap weremeasured. Removing the buds, a sink for N, caused a decreasein nodule activity. Flushing root systems daily with 100% O₂destroyed nitrogenase activity and substantially decreased theamount of citrulline in the xylem sap. The amino acid compositionof xylem saps also altered according to the mode of N nutrition.In plants fed , xylem sap composition was similar to N₂-fixing plants, however, when plants were fed, citrulline content increased. The assimilation and subsequent distribution of nitrate wasfollowed in an experiment in which labelled ¹⁵ was added to the base of plant pots. After 12 h7% of root N was from applied ¹⁵ and this increased to 75% at 7 d; substantial enrichment ofN from ¹⁵ also occurred in stems, buds and leaves. After 7 d, 3.5% of nodule N was from¹⁵, consistent with some N being supplied by recycling of shoot N. Xylem saps were alsocollected and after 12 h, glutamate and aspartate were enrichedwith ¹⁵N to 53% and 37% increasing after 7 d to 80% and 49%,respectively. Citrulline content of the xylem sap increasedfrom 3 to 9 µmol cm^–3 following addition of ¹⁵ and at 7 d 80% of the N in the citrullinehad been derived from ¹⁵N. It is hypothesized that the growthand activity of A. glutinosa root nodules is sensitive to theN status of the plant and that the level of citrulline (or otheramino acids) returning to the nodules may feed back to regulatenodule growth and activity. Key words: Alnus glutinosa, citrulline, nitrate, feedback mechanism, N₂-fixation.     

The Role of Long-distance Transport in Intracellular pH Regulation in Phaseolus vulgaris Grown with Ammonium or Nitrate as Nitrogen Source, or Nodulated

Charge balance and intracellular pH regulation were studiedin Phaseolus vulgaris grown in water culture on or as N source, or nodulated and in N-free medium. Seedlings and 36-d-old plants were analysed for total P, C, organic N and S and ash alkalinity;xylem sap was analysed for mineral ion content and amino acids,amides and dicarboxylates. Both water uptake and H⁺ or baseexcreted during water culture were measured. It was shown that in -grown plants, H⁺ excretedwas related directly to the uptake and assimilation of ; there was no cation uptake associated with netH⁺ exchange. All shoot N was supplied as organic N, mainly glutamine,so that shoot pH perturbation would be limited to uronate productionin situ. This could be regulated by both shoot reduction and the transport of OH^–-generating carboxylatesin the xylem. In plants on , the great majority of reduction occurred in the root and most of the OH^–produced was converted to -COO^–, transported up the xylemand stored, so that the leaves in particular had a high ashalkalinity (20-fold that in the roots). Some OH^–, however,was excreted. Nodulated plants had a low N content, compared with the others,indicating a low rate of N₂-fixation for the earlier periodof water culture. At harvest, however, the xylem sap organicN content was comparable to that of plants. H⁺ excretion was in excess of that required for N assimilation.It was concluded that in these plants some net H⁺/cation exchangehad occurred which may have been a response to low nutrient(i.e. nitrogen) status. Key words: Phaseolus vulgaris, pH regulation, nitrate nutrition, ammonium nutrition, dinitrogen fixation, xylem sap composition.     

Nitrate Nutrition of Grasses from Steady-State Supplies in Flowing Solution Culture following Nitrate Deprivation and/or Defoliation: I RECOVERY OF UPTAKE AND GROWTH AND THEIR INTERACTIONS

Two Lolium genotypes, i.e. Lolium perenne cv. S23 and a L. perenne? L. multiflorum hybrid cv. Augusta were, in one treatment,grown in flowing solution culture with a maintained supply of at 10 mmol m–3 throughout. In another treatment, the plants were deprived of for 11 d, was then re-supplied and both control and N-deprived plants were either defoliatedor left entire. Effects of the stresses imposed by N deprivationor defoliation on subsequent net uptake of , shoot and root growth and their interactions were examined.There were no major differences in the behaviour of the twogenotypes throughout. The removal of NO3 supply did not altertotal dry matter production during the 11 d period of deprivationbut the rate of root growth accelerated markedly. There waslittle effect of N deprivation on the specific growth ratesof the recovering defoliated shoots. Cumulative net uptake of closely paralleled the pattern of growth and there were obvious and marked effects of both N deprivationand defoliation. Effects on uptake were more distinctly demonstrated when uptake was expressed on aper unit root or shoot basis, i.e. specific unit absorptionrates (S_NO3). In the control plants with sustained supply, there was a progressive decline with time in S_NO3, inspite of an increasing shoot: root ratio. After both N deprivationand defoliation treatments there was a rapid initial increasein S_NO3, followed by an oscillatory cycle of increasing anddecreasing rates with a 10–11 d period. The relationshipsbetween growth and uptake were complex and did not conform to a previously predicted linear relationshipbetween S_NO3, and the ratio total fresh plant weight: root freshweight. The results are discussed in relation to the possiblemechanisms involved. It is suggested that the oscillations inunit absorption rate indicated that the roots of N-deprivedor defoliated plants retained a high capacity for absorptionimmediately after re-supply or defoliation and that the stressedplants were unable to assimilate or utilize taken up; net uptake then decreased through a shift in favourof efflux over influx until the influx mechanism was able todominate and the pattern was then repeated. Key words: Lolium, genotypes, N deprivation, nitrate uptake, defoliation, unit absorption, growth     

A comparison of the impacts of various nitrogen sources on acid-base balance in C3 Triticum aestivum L. and C4 Zea mays L. plants

This work aimed to study the impacts of acquisition and assimilationof various nitrogen sources, i.e. NO₃^–, NH₄⁺ or NH₄NO₃,in combination with gaseous NH₃ on plant growth and acid-basebalance in higher plants. Plants of C₃ Triticum aestivum L.and C₄ Zea mays L. grown with shoots in ambient air in hydroponicculture solutions with 2 mol m^–3 of nitrogen source asNO₃^–, NH₄⁺ or NH₄NO₃ for 21 d and 18 d, respectively,had their shoots exposed either to 320 µg m^–3 NH₃or to ambient air for 7 d. Variations in plant growth (leaves,stubble and roots), and OH^– and H⁺ extrusions as wellas the relative increases in nitrogen, carbon and carboxylatewere determined. These data were computed as H⁺/N, H⁺/C, (C-A)/N,and (C-A)/C to analyse influences of different nitrogen sourceson acid-base balance in C₃ Triticum aestivum and C₄ Zea maysplants. Root growth in dry weight gain was significantly reduced bytreatment with 320 µg m^–3 NH₃ in Triticum aestivumand Zea mays growing with different N-forms, whereas leaf growthwas not significantly affected by NH₃. In comparison with C₃Triticum aestivum, non-fumigated C₄ Zea mays had low ratiosof OH^–/N in NO₃^–3-grown plants and of H⁺/N in NH₄⁺- and NH ₄NO₃-grown plants. Utilization of NH₃ from the atmospherereduced both the OH^–N ratios in NO₃^– -grown plantsand the H⁺/N ratio in NH₄⁺ - and NH₄NO₃ -grown plants of bothspecies. Furthermore, Zea mays had higher ratios of (C-A)/Nin NH₄⁺ - and NH₄NO₃-grown plants than Triticum aestivum. Thismeans that C₄ Zea mays had synthesized more organic anion perunit increase in organic N than C₃ Triticum aestivum plants.Within both species, different nitrogen sources altered theratios of (C-A)/N in the order: NH₄NO₃>NH₄⁺>NO₃^–.Fumigation with NH₃ increased organic acid synthesis in NO₃^–- and NH₄⁺ - grown plants of Triticum aestivum, whereas it decreasedorganic acid synthesis in Zea mays plants under the same conditions.Furthermore, these differences in acid-base regulation betweenC₃ Triticum aestivum and C₄ Zea mays plants growing with differentnitrogen sources are discussed. Key words: Acid-base balance, ammonia, ammonium, nitrate, ammonium nitrate, C₃ Triticum aestivum L., C₄ Zea mays L.     

Growth and 2, 4-Diaminobutyric Acid Composition of Flatpea (Lathyrus sylvestris L.) as Influenced by Mineral- and Symbiotically Fixed-Nitrogen

Flatpea (Lathyrus sylvestris L. cv. ‘Lathco’) plants,grown hydroponically, were inoculated with rhizobia (Rhizobiumleguminosarum strain 92F2), supplied with nitrate nitrogen (2meq 1^–1), or provided with both rhizobia and . Nodulated plants supplied with had the highest biomass and the highest level of free amino acids in leavesand roots. Nitrogen fixation was depressed 80% in inoculatedplants supplemented with . Compared to plants whose nitrogen requirement was satisfied either partially orfully by , plants that were totally dependent upon biological nitrogen fixation had fewer lateral shoots andlower concentrations of nitrogen, soluble protein, and totalfree amino acids in the shoots, especially in the leaves. Thesymbiotic association between plants and rhizobia for nitrogenfixation was not essential to the production and accumulationof 2,4-diaminobutyric acid (A₂bu) and other non-protein aminoacids in flatpea. Levels of A₂bu, and most of the other freeamino acids quantified, were significantly lower in the nodulatedplants grown in the absence of than in the inoculated or noninoculated plants supplied with . Arginine and glutamic acid were exceptions in that levels ofthese two amino acids were higher in roots bearing effectivenodules. Plant responses to the different treatments are attributedto the amount of nitrogen available to the plant, rather thanthe method or form of nitrogen supplied. Key words: 2,4-Diaminobutyric acid, Lathyrus, nitrogen     

Interactions between Root Temperature and Nitrogen Deficiency Influence Preferential Uptake of NH+4 and NO-3 by Oilseed Rape

Oilseed rape (Brassica napus L. cv. Bien venu) plants were grownin a system of flowing solution culture and pre-treated at roottemperatures of 3 ?C or 13 ?C for 7 d with or without N suppliedas 10 mmol m^–3 NH₄NO₃. Subsequently, N was re-suppliedand root temperatures were reversed for 7 d. Shoot temperatureswere common at 20/15 ?C day/night. Net uptake of , and K⁺, leaf area, root length, and transpirationwere measured and compared with control plants having root temperaturesof 3 ?C and 13 ?C throughout. Plants that were continuouslysupplied with N and pre-treated at 3 ?C showed a 50% increasein total uptake of , and measured at 13 ?C over 7 d compared with control plants at 3?C,but uptake of was 28% lower and uptake of was 43% higher than that shown by control plants at 13 ?C. Pre-treatment at 3 ?C did not enhance the subsequentuptake of total N or of K⁺ at 13 ?C relative to the 13 ?C control.Transpiration rates at 3 ?C were on average 70% of those at13 ?C. The concentration of total N in plants was halved after7 d without a supply of N, but total dry matter production wasnot significantly affected. N starvation also increased thetemperature sensitivity of subsequent uptake relative to that of uptake. After N starvation at 13 ?C the uptake of and measured at 13 ?C was 50% higher over 7 d than that measuredunder continuous N supply. In contrast, after N starvation at3 ?C the uptake of at 3 ?C was 70% less, whilst uptake was 50% more than the respective totals absorbed by plants that were continuously supplied withN at 3 ?C. After N starvation the uptake of was generally 40–60% of the daily total N uptake, comparedwith 60–80% in plants continuously supplied with N. Key words: Brassica napus, oilseed rape, root temperature, nitrate, ammonium, potassium, N-deficiency, ion uptake rate, transpiration     

Proliferation of Maize (Zea mays L.) Roots in Response to Localized Supply of Nitrate

Maize (Zea mays L.) plants with two primary nodal root axeswere grown for 8 d in flowing nutrient culture with each axisindependently supplied with . Dry matter accumulation by roots was similar whether 1.0 mol m^–3 was supplied to on( or both axes. When was supplied to only one axis, however, accumulationof dry matter within the root system was significantly greaterin the axis supplied with . The increased dry matter accumulation by the +N-treated axis was attributableentirely to increased density and growth of lateral branchesand not to a difference in growth of the primary axis. Proliferation of lateral branches for the + N axis was associatedwith the capacity for in situ reduction and utilization of aportion of the absorbed , especially in the apical region where lateral primordia are initiated. Althoughreduced nitrogen was translocated to the –N axis, concentrationsin the –N axis remained significantly lower than in the+N axis. The concentratio of reduced nitrogen, as well as invitro reductase activity, was greater in apical than in more basal regions of the +N axis. The enhancedproliferation of lateral branches in the + N axis was accompaniedby an increase in total respiration rate of the axis. Part ofthe increased respiration was attributable to increased massof roots. The specific respiration rate (umol CO₂ exolved perhour per gram root dry weight) was also greater for the +N thanfor the –N axis. If respiration rate is taken as representativeof sink demand, stimulation of initiation and growth of lateralsby in situ utilization of a localized exogenous supply of establishes an increased sink demand through enhancedmetabolic activity and the increased partitioning of assimilatesto the + N axis responds to the difference in sink demand between+N and –N axes. Key words: NO₃^- reduction, NO₃^- uptake nitrogen partitioning, root respiration, sink demand     

Potassium Transport Across the Membranes of Chara: I. THE RELATIONSHIP BETWEEN RADIOACTIVE TRACER INFLUX AND ELECTRICAL CONDUCTANCE

Smith, J. R., Smith, F. A. and Walker, N. A. 1987. Potassiumtransport across the membrane of Chara. I. The relationshipbetween radioactive tracer influx and electrical conductance.—J.exp. Bot. 38:731–751. The ⁴²K influx () and the electrical conductance (G_m) were measured simultaneously for the ‘membrane’of internodal cells of Chara australis as a function of theexternal [KCl] (K?. In bathing solutions of pH = 5?0, progressively increased from 20?5to 430?60 nmol m^–2 s^–1 and G_m increased from 0?36?0?02to 3?8?0?8 S m^–2 when K? was increased from 0?1 to 10mol m^–3. The resting membrane potential difference (p.d.)was approximately -135 mV for low K? and approached the expectedNernst equilibrium p.d. for K⁺ ions when K? > 1?0 mol m^–3.Measurements of ³⁶Cl influx suggested that the ⁴²K influx waspredominantly electrogenic. The equivalent Goldman permeabilityto K⁺ ions (P_k) was approximately 20–30 nm s^–1 anddid not vary significantly with increasing K?. The equivalentconductance attributable to the electrogenic transport of K⁺ ions was calculated from assuming passive, independent diffusionof K⁺ ions and the ratio was found to be typically close to one. It was also found that themagnitudes of and G_m measuredsimultaneously for each individual cell were also well correlatedfor K? 1?0 mol m^–3, and that the slope of the line ofbest fit was close to one. For each K? it was found that theconductance not attributable to K⁺ translocation and presumablyassociated primarily with the transport of protons or theirequivalents was typically 0?2–0?5 Sm^–2. For K? >1?0 mol m^–3 the results indicated that the transport ofK⁺ ions was essentially independent, i.e. there was no evidencefor flux interactions. The results also indicated that the equivalentconductance derived from the measured ⁴²K influx could usefullyindicate the fraction of the electrical conductance attributableto the translocation of K⁺ ions. Key words: Potassium, conductance, influx     

Development of the Xylem Ureide Assay for the Measurement of Nitrogen Fixation by Pigeonpea (Cajanus cajan (L.) Millsp.)

Quantification of N₂ fixation by pigeonpea (Cajanus cajan (L.)Millsp.) in the field has proved difficult using techniquessuch as ¹⁵N isotope dilution, acetylene reduction and N difference.We report experiments to develop the ureide assay of N₂ fixationbased on extraction and analysis of xylem exudate. Plants ofpigeonpea cv. Quantum, inoculated with effective Rhizobium spp.CB756, were grown in a temperature-controlled glasshouse inlarge pots filled with a sand: vermiculite mixture, in waterculture and in a slightly acidic, red-brown earth in replicatedfield plots. Xylem exudate was collected as bleeding sap fromboth nodulated and unnodulated roots, and from detached nodules.Exudate was extracted also from detached shoots and stems ofpigeonpea using a mild vacuum (60–70 kN m^–2). Largedifferences in the composition of N solutes exported from rootsof N₂-dependent and nitrate-dependent plants suggested thatshifts in plant dependence on N₂ fixation may be reflected byconcomitant changes in N solutes. Thus, nodulated plants weresupplied throughout growth with either N-free nutrients or nutrientssupplemented with 1, 2, 5, 5, 10, or 20 mol m^{–3 15}. Plants were harvested at regular intervals fordry matter and vacuum-extracted exudate. The relative abundanceof ureides ([ureide-N/ureide-N + nitrate-N + -amino-N] ? 100)in the exudate was highly correlated with the proportion ofplant N (calculated using a ¹⁵N isotope dilution technique)derived from N₂ fixation. Two distinct phases of plant growthwere recognized and standard curves were prepared for each.The relationship between proportional dependence of plants onN₂ and xylem relative ureides was unaffected by mineral-N source,i.e. nitrate or ammonium. This result is discussed in relationto interpretation of material from field-grown plants. The effectsof plant genotype, strain of rhizobia, section of stem extracted,removal of leaves, time delay between shoot detachment and extraction,and diurnal characteristics were examined in order to identifypotential sources of error and to optimize sampling procedures. Key words: Ureides, allantoin, allantoic acid, N₂ fixation, pigeonpea, Cajanus cajan     

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

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

The Effect of SO2 on Lolium perenne L. Grown at Different Levels of Sulphur and Nitrogen Nutrition

The effects of exposure to SO₂ (55 m^–3) on the growthand sulphur nutrition of perennial ryegrass (Lolium perenneL.) cv. S23 were examined using a system of specially constructedgrowth chambers. The plants were grown in soil with two ratesof added nitrogen, either with or without added sulphate. Theywere harvested twice during the exposure period of 85 d. The high rate of added nitrogen gave a four-fold increase inthe yield of plants given added sulphate but resulted in sulphurdeficiency, with reduced yield and number of tillers, when nosulphate was given. Exposure to SO₂ alleviated the effects ofdeficiency but was without effect on the yield of plants givensulphate. With the low rate of added nitrogen, the yield ofplants and the number of tillers were unaffected by the additionof sulphate to the soil or of SO₂ to the air. The only indicationof possible injury through exposure of these plants to SO₂ wasa small increase in the proportion of dead leaves at the secondharvest. Analysis of the leaves for total S and sulphate-S gave valuesconsistent with sulphur deficiency and its alleviation throughexposure to SO₂; the contents of 0?045% S and 42 parts 10^–6sulphate-S in deficient plants were raised to 0?102% and 153parts 10^–6 respectively. The organic-S content of theleaves was always increased through exposure to SO₂. There wasno evidence that exposure to SO₂ increased the ‘transpirationcoefficient’ for plants having an adequate supply of sulphurfrom the soil.     

Respiratory Utilization of 13C-Labelled Photosynthate in Nodulated Root Systems of Soybean Plants

Kouchi, H., Akao, S. and Yoneyama, T. 1986. Respiratory utilizationof ¹³C-labelled photosynthate in nodulated root systems of soybeanplants.—J. exp. Bot. 37: 985–993. An improved method for the measurement of respiratory utilizationof current photosynthate in the nodulated root system of water-culturedsoybean (Glycine max L.) plants was developed using a steady-state¹³CO₂ labelling technique. Well-nodulated plants at the latevegetative stage were allowed to assimilate ¹³CO₂ for 10 h incontinuous light at a constant CO₂ concentration with a constant¹³C abundance. The respiratory evolution of ¹³CO₂ from rootsand nodules was measured continuously throughout the periodof ¹³CO₂ assimilation and during a subsequent 36 h chase periodby using a differential infrared ¹³CO₂ analyser. The plantswere grown with nitrogen-free or (15 mmol dm^–3)-containing culture solution for 3 d before¹³CO₂ assimilation. In plants grown without , nodule respiration averaged 69% of the total respiration of the undergroundparts over the full experimental period and the CO₂ respiredreached an apparent isotopic equilibrium at 80–85% labellingafter initiating ¹³CO₂ assimilation. By contrast, the CO₂ respiredfrom the roots did not reach an isotopic equilibrium and labellingwas only 56% at the end of exposure to ¹³CO₂ These findingsdemonstrated that nodule respiration is strongly dependent onrecently assimilated carbon compared with root respiration. Plants supplied with in the culture solution showed a decreased rate of nodule respirationand a slightly increased rate of root respiration. The extentsand time courses of labelling of respired CO₂ from both theroots and nodules were similar in the presence and absence of except that the maximum level of labelling of CO₂ derived from nodule respiration in plantswith was significantly higher (about 91%) than for plants growing without . Key words: Soybean (Glycine max L.), nodule respiration, ¹³CO₂, assimilation, carbon partitioning     

Expression of Characteristics of Ammonium Nutrition as Affected by pH of the Root Medium

To study the effect of root-zone pH on characteristic responsesof -fed plants, soybeans (Glycine max {L.}Merr. cv. Ransom) were grown in flowing solution culture for21 d on four sources of N (1.0 mol m^–3 , 0.67 mol m^–3 plus 0.33 mol m^–3, 0.33 mol m^–3 plus 0.67 mol m^–3 , and 1.0 mol m^–3) with nutrient solutions maintained at pH 6.0, 5.5, 5.0, and 4.5. Amino acid concentration increased inplants grown with as the sole source of N at all pH levels. Total amino acid concentration in the rootsof -fed plants was 8 to 10 times higher than in -fed plants, with asparagine accounting for more than 70% of the total in the roots of these plants.The concentration of soluble carbohydrates in the leaves of-fed plants was greater than that of -fed plants, but was lower in roots of -fed plants, regardless of pH. Starch concentration was only slightlyaffected by N source or root-zone pH. At all levels of pH tested,organic acid concentration in leaves was much lower when was the sole N source than when all or part of theN was supplied as . Plants grown with mixed plus N sources were generally intermediate between - and -fed plants. Thus, changes in tissue compositioncharacteristic of nutrition when root-zone pH was maintained at 4.5 and growth was reduced, still occurredwhen pH was maintained at 5.0 or above, where growth was notaffected. The changes were slightly greater at pH 4.5 than athigher pH levels. Key words: Ammonium, nitrogen nutrition, root-zone pH, soybean, tissue composition     

White Clover N2-Fixation in Response to Root Temperature and Nitrate: I GROWTH AND UPTAKE OF NITRATE FROM FLOWING NUTRIENT SOLUTIONS

Established, nodulated white clover plants (Trifolium repensL. cv. Blanca) were transferred to eight plant culture unitsof a system of flowing solution culture. Solution temperatureswere 5, 11, 17, and 25 ?C (two units per temperature), withshoot temperature of 25/15 ?C day/night and light regime commonto all plants. After 7 d, was supplied at 10 mmol m^–3 to one of each pair of culture units and thenet uptake of was monitored over 14 d. The remaining four culture units served as controls in which plantswere entirely dependent on N₂ fixation, as estimated by sequentialharvesting and the change in the amount of N in plants. Totalfresh and dry weights increased exponentially with time andwith increasing root temperature, between 5–25 ?C in nitrateplants and 5–17 ?C in control plants, respectively, byfactors of 2.9 and 1.8. Nodule dry weight of nitrate plantsshowed little increase after 6 d compared with control plants.Dry weight shoot: root ratios increased with time in all treatmentsexcept 5 ?C control plants. Total net uptake of over 13 d was 0.41, 4.27, 11.92, and 12.77 mmol plant-1, respectively,at 5, 11, 17, and 25 ?C. At all temperatures except 5 ?C, plantsaccumulated high concentrations of (10–40 mol m^–3) in leaflets and roots, within 2 or 3 days ofsupplying . Daily mean unit absorption rates of increased 12-fold with increasing temperature between 5 and 25 ?C, and showed little change with time at 5,17, and 25 ?C. The total N₂ fixed by nitrate plants over 14d increased 5-fold with temperature between 5 and 17 ?C, butwas always less than the amount fixed by control plants. Thepercentage contributed by N2 fixation to total N acquisitionby nitrate plants over 14 d decreased with increasing temperature,from 77% at 5 ?C to 11 % at 25 ?C. Mean daily rates of N₂ fixationper unit dry weight of nodule were lower in nitrate plants thanin control plants throughout treatment at 5 ?C and 25 ?C, butat 11 ?C and 17 ?C the rates for nitrate plants increased progressivelywith time and exceeded the rates for control plants after 8d. In both nitrate and control plants the effect of temperatureon N₂ fixation per nodule dry weight was proportionately lessthan that on unit absorption rate of . The results are discussed in terms of the overall regulation ofN accumulation by white clover and the adaptive significanceof differences in the sensitivities of uptake and N₂ fixation to root temperature. Key words: Trifolium repens, white clover, root temperature, N₂ fixation, nitrate uptake