The Distribution of Nitrate Assimilation Between Root and Shoot in Vicia faba L.

Nitrate assimilation was examined in two cultivars (Banner Winterand Herz Freya) of Vicia faba L. supplied with a range of nitrateconcentrations. The distribution between root and shoot wasassessed. The cultivars showed responses to increased applied nitrateconcentration. Total plant dry weight and carbon content remainedconstant while shoot: root dry weight ratio, total plant nitrogen,total plant leaf area and specific leaf area (SLA) all increased.The proportion of total plant nitrate and nitrate reductase(NR) activity found in the shoot of both cultivars increasedwith applied nitrate concentrations as did NO₃^–: Kjeldahl-Nratios of xylem sap. The cultivars differed in that a greaterproportion of total plant NR activity occurred in the shootof cv. Herz Freya at all applied nitrate concentrations, andits xylem sap NO₃^–: Kjeldahl-N ratio and SLA were consistentlygreater. It is concluded that the distribution of nitrate assimilationbetween root and shoot of V. faba varies both with cultivarand with external nitrate concentration. Vicia faba L., field bean, nitrate assimilation, nitrate reductase, xylem sap analysis     

Nitrate Effects on Growth of the First Four Main Stem Leaves of a Range of Temperate Cereals and Pasture Grasses

The effects of different applied NO₃^– concentrations onextension growth and final length and area of leaves 1–4of five cereals and six pasture grasses of temperate originwere examined. Increased applied NO₃^– in the range 0.1–50mol m^–3; caused decreased duration of growth but increasedgrowth rate and final length of leaves 2–4 of the cerealsAvena saliva, Hordeum vulgare, Secale cereale x Triticosecaleand Triticum aestivum. For all cereals, increased NO₃^–resulted in increased area of leaves 1-4. Pasture grasses weresupplied either 0.5 or 50 mol m^–3; NO₃^–. Increasedapplied NO₃^– (0.5–50 mol m^–3) resulted indecreased duration of growth and increased growth rate and finalarea of leaves 1–4 of Bromus willdenowii leaves 2–4of Festuca arundinaceae and leaves 3 and 4 of Lolium multiflorum.In addition, length of leaves 3 and 4 of B. willdenowii increasedwith increased NO₃. Increased NO₃ resulted in increased areaof leaves 2–4 of Daciylis glomerata and Lolium perenneand leaves 3 and 4 of Phalaris aquatica but had no effect onextension growth of all three species. Avena saliva L., oat, Hordeum vulgare L., barley, Secale cereaie L., rye, x Triticosecale Wittm, triticale, Triticum aestivum L., wheat, Bromus willdenowii Kunth, prairie grass, Dactylis glomerata L., cocksfoot, Festuca arundinaceae Shreb, tall fescue, Lolium multiflorum Lam, Italian ryegrass, Lolium perenne L, perennial ryegrass, Phalaris aquatica L, nitrate,, leaf extension, leaf expansion     

Modelling of the Partitioning, Assimilation and Storage of Nitrate within Root and Shoot Organs of Castor Bean (Ricinus communis L.)

An experimentally-based modelling technique was developed todescribe quantitatively the uptake, flow, storage and utilizationof NO₃-N over a 9 d period in mid-vegetative growth of sandcultured castor bean (Ricinus communis L.) fed 12 mol m^–3nitrate and exposed to a mean salinity stress of 128 mol m^–3NaCl. Model construction used information on increments or lossesof NO₃-N or total reduced N in plant parts over the study periodand concentration data for NO₃-N and reduced (amino acid) Nin phloem sap and pressure-induced xylem exudates obtained fromstem, petiole and leaf lamina tissue at various levels up ashoot. The resulting models indicated that the bulk (87%) of incomingnitrate was reduced, 51% of this in the root, the remainderprincipally in the laminae of leaves. The shoot was 60% autotrophicfor N through its own nitrate assimilation, but was oversuppliedwith surplus reduced N generated by the root and fed to theshoot through the xylem. The equivalent of over half (53%) ofthis N returned to the root as phloem translocate and, mostly,then cycled back to the shoot via xylem. Nitrate comprised almosthalf of the N of most xylem samples, but less than 1% of phloemsap N. Laminae of leaves of different age varied greatly inN balance. The fully grown lower three leaves generated a surplusof reduced N by nitrate assimilation and this, accompanied byreduced N cycling by xylem to phloem exchange, was exportedfrom the leaf. Leaf 4 was gauged to be just self-sufficientin terms of nitrate reduction, while also cycling reduced N.The three upper leaves (5–7) met their N balance to varyingextents by xylem import, phloem import (leaves 6 and 7 only)and assimilation of nitrate. Petioles and stem tissue generallyshowed low reductase activities, but obtained most of theirN by abstraction from xylem and phloem streams. The models predictedthat nodal tissue of lower parts of the stem abstracted reducedN from the departing leaf traces and transferred this, but notnitrate, to xylem streams passing further up the shoot. As aresult, xylem sap was predicted to become more concentratedin N as it passed up the shoot, and to decrease the ratio ofNO₃-N to reduced N from 0·45 to 0·21 from thebase to the top of the shoot. These changes were reflected inthe measured N values for pressure-induced xylem exudates fromdifferent sites on the shoot. Transfer cells, observed in thexylem of leaf traces exiting from nodal tissue, were suggestedto be involved in the abstraction process. Key words: Ricinus communis, nitrogen, nitrate, nitrate reduction, partitioning, phloem, xylem, flow models     

Nitrate Effects on Growth of the First Four Main Stem Leaves of a Range of Temperate Cereals and Pasture Grasses

The effects of different applied NO₃^– concentrations onextension growth and final length and area of leaves 1–4of five cereals and six pasture grasses of temperate originwere examined. Increased applied NO₃^– in the range 0.1–0.5.0mol m^–3 caused decreased duration of growth but increasedgrowth rate and final length of leaves 2–4 of the cerealsAvena saliva, Hordeum vulgare, Secale cereale, x Triticosecaleand Triticum aestivum. For all cereals, increased NO₃^–resulted in increased area of leaves 1–4. Pasture grasseswere supplied either 0.5 or 50 mol m^–3 NO₃^–. Increasedapplied NO₃^– (0.5–5.0 mol m^–3) resulted indecreased duration of growth and increased growth rate and finalarea of leaves 1–4 of Bromus wiltdenowii, leaves 2–4ofFestuca arundinaceae and leaves 3 and 4 of Lolium muitiflorum.In addition, length of leaves 3 and 4 of B. witidenowii increasedwith increased NO₃^–. Increased NO₃^– resulted inincreased area of leaves 2–4 of Dactylis gtomerata andLolium perenne and leaves 3 and 4 of Phalaris aquaiica but hadno effect on extension growth of all three species. Avena sativa L, oat, Hordeum vulgare L, barley, Secale cereale L, rye, x Triticosecale Wittm, triticale, Triticum aestivum L, wheat, Bromus willdenowii Kunth, prairie grass, Dactylis gtomerata L, cocksfoot, Festuca arundinaceae Shreb, tall fescue, Lolium multijlorum Lam, Italian ryegrass, Lolium perenne L, perennial ryegrass, Phalaris aquatica L, nitrate, leaf extension, leaf expansion     

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     

The influence of salinity on the utilization of root anaplerotic carbon and nitrogen metabolism in tomato seedlings

In hydroponically grown Lycopersicon esculentum (L.) Mill. cv.F144 the site of NO₃^– reduction and assimilation withinthe plant was shifted from the shoot to the root by salinity.Uptake of NO₃^– from the root solution was strongly inhibitedby salinization. Consequently, NO₃^– concentrations inthe leaf, stem and root tissues as well as the nitrate reductaseactivities of the leaves were lower in salinized than in controlplants. Lower NO₃^–, but higher reduced-N, concentrationswere observed in the xylem sap as a result of the enhanced participationof the root in NO₃^– reduction in salinized plants. Lowerstem K⁺ concentrations and leaf malate concentrations were foundin salinized compared to control plants which indicates reducedfunctioning of the K⁺–shuttle in the salinized plants. Incorporation of inorganic carbon by the root was determinedby supplying a pulse of NaH¹⁴CO₃ followed by extraction andseparation of the labelled products on ion exchange resins.The rate of H¹⁴CO₃^– incorporation was c. 2-fold higherin control than in salinized plants. In salinized plants theproducts of H¹⁴CO₃^– incorporation within the roots werediverted into amino acids, while the control plants divertedrelatively more ¹⁴C into organic acids. Products of inorganiccarbon incorporation in the roots of salinized plants providean anaplerotic source of carbon for assimilation of reducedNO₃^– into amino acids, while in control plants the productswere predominantly organic acids as part of mechanisms to maintainionic balance in the cells and in the xylem sap. Key words: Tomato, nitrate, PEPc, respiration, salinity     

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     

The Uptake and Flow of C, N and lons between Roots and Shoots in Ricinus communis L.

Seedlings of Ricinus communis L. cultivated in quartz sand weresupplied with a nutrient solution containing either 1 mol m^–3NO^–₃ or 1 mol m^–3 NH⁺₄ as the nitrogen source. Duringthe period between 41 and 51 d after sowing, the flows of N,C and inorganic ions between root and shoot were modelled andexpressed on a fresh weight basis. Plant growth was clearlyinhibited in the presence of NH⁺₄. In the xylem sap the majornitrogenous solutes were nitrate (74%) or glutamine (78%) innitrate or ammonium-fed plants, respectively. The pattern ofamino acids was not markedly influenced by nitrogen nutrition;glutamine was the dominant compound in both cases. NH⁺₄ wasnot transported in significant amounts in both treatments. Inthe phloem, nitrogen was transported almost exclusively in organicform, glutamine being the dominant nitrogenous solute, but theN-source affected the amino acids transported. Uptake of nitrogenand carbon per unit fresh weight was only slightly decreasedby ammonium. The partitioning of nitrogen was independent ofthe form of N-nutrition, although the flow of nitrogen and carbonin the phloem was enhanced in ammonium-fed plants. Cation uptakerates were halved in the presence of ammonium and lower quantitiesof K⁺, Na⁺ and Ca²⁺ but not of Mg²⁺ were transported to theshoot. As NH⁺₄ was balanced by a 30-fold increase in chloride in thesolution, chloride uptake was increased 6-fold under ammoniumnutrition. We concluded that ammonium was predominantly assimilated inthe root. Nitrate reduction and assimilation occurred in bothshoot and root. The assimilation of ammonium in roots of ammonium-fedplants was associated with a higher respiration rate. Key words: Ricinus communis, nitrogen nutrition (nitrate/ammonium), phloem, xylem, transport, partitioning, nitrogen, carbon, potassium, sodium, magnesium, calcium, chloride     

Carbon Flow from Nodulated Roots to the Shoots of Soybean (Glycine max L. Merr.) Plants: An Estimation of the Contribution of Current Photosynthate to Ureides in the Xylem Stream

Kouchi, H. and Higuchi, T. 1988. Carbon flow from nodulatedroots to the shoots of soybean {Glycine max L. Merr.) plants:An estimation of the contribution of current photosynthate toureides in the xylem stream.–J. exp. Bot. 39: 1015–1023. Well-nodulated, water-cultured soybean plants were allowed toassimilate ¹³CO₂ at a constant specific activity for 10 h andthe ¹³C-labelling of total carbon and ureides in xylem sap wasinvestigated. Labelled carbon appeared very rapidly in the xylem stream. Percentageof labelled carbon (relative specific activity, RSA) in xylemsap was 18% at 2 h after the start of ¹³CO₂ assimilation andreached 53% at the end of the 10 h assimilation. The amountof labelled carbon exported from nodulated roots to the shootsvia the xylem during the 10 h labelling period accounted for33% of total labelled carbon imported into the nodulated roots.Ureides (allantoin and allantoic acid) in xylem sap were stronglydependent on currently assimilated carbon. The RSA of ureidesin xylem sap had reached 83% at the end of the assimilationperiod. Labelled carbon in ureides accounted for 51% of totallabelled carbon returned from nodulated roots to the shootsvia the xylem during the 10 h assimilation period. A treatmentwith 20 mol m^–3 nitrate in the culture medium for 2 ddecreased the ureide concentration in the xylem sap slightly,but greatly decreased the RSA of ureides. By comparing the data with the results of analysis of the xylemsap of nodule-detached plants, it was concluded that the majorityof labelled carbon exported to the xylem stream from noduleswas in ureide form. A considerable amount of carbon was alsoreturned from roots to shoots via the xylem stream but it wasmore dependent on (non-labelled) carbon reserved in the roottissues. Key words: Soybean(Glycine max L.), root nodule, carbon partitoning, ¹³CO₂ assimilation, xylem     

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.     

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     

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     

Assimilation of 15N2 and 15NO3- by Partially Nitrate-Tolerant Nodulation Mutants of Soybean

Growth-chamber studies were conducted to evaluate nitrogen assimilationby three hypernodulated soybean [Glycine max (L.) Merr.] mutants(NOD1–3, NOD2–4, NOD3–7) and the Williamsparent. Seeds were inoculated at planting and transplanted atday 7 to nutrient solution with 1 mol m^–3 urea (optimizesnodule formation) or 5 mol m^–3 NO₃^– (inhibits noduleformation). At 25 d after planting, separate plants were exposedto ¹⁵NO₂ or ¹⁵NO₃^– for 3 to 48 h to evaluate N₂ fixationand NO₃^– assimilation. Plant growth was less for hypernodulatedmutants than for Williams with both NO₃^– and urea nutrition.The major portion of symbiotically fixed ¹⁵N was rapidly assimilated(30 min) into an ethanol-soluble fraction, but by 24 h aftertreatment the ethanolinsoluble fraction in each plant part wasmost strongly labelled. Distribution patterns of ¹⁵N among organswere very similar among lines for both N growth treatments aftera 24 h ¹⁵N₂ fixation period; approximate distributions were40% in nodules, 12% in roots, 14% in stems, and 34% in leaves.With urea-grown plants the totalmg ¹⁵N fixed plant^–1 24h^–1 was 1·18 (Williams), 1·40 (N0D1-3),107 (NOD2-4), and 0·80 (NOD3-7). The 5 mol m^-3 NO₃^- treatmentresulted in a 95 to 97% decrease in nodule mass and ¹⁵N₂ fixationby Williams, while the three mutants retained 30 to 40% of thenodule mass and 17 to 19% of the ¹⁵N₂ fixation of respectiveurea-grown controls. The hypernodulated mutants, which had restrictedroot growth, absorbed less ¹⁵NO₃^- than Williams, irrespectiveof prior N growthcondition. The ¹⁵N from ¹⁵NO₃^- was primarilyretained in the soluble fraction of all plant parts through24 h. The ¹⁵N incorporation studies confirmed that nodule developmentis less sensitive to external NO₃^- in mutant lines than in theWilliams parent, and provide evidence that subsequent metabolismand distribution within the plant was not different among lines.These results further confirm that the hypernodulated mutantsof Williams are similar in many respects to the hyper- or supernodulatedmutants in the Bragg background, and suggest that a common mutationalevent affectingautoregulatory control of nodulation has beentargeted. Key words: Glycine max (L.) Merr., soybean, N₂fixation, nitrate assimilation, nodulation mutants, ¹⁵N isotope     

Nitrate Inhibition of Chloride Influx in Barley: Implications for a Proposed Chloride Homeostat

Net accumulation of Cl^– by intact barley plants was virtuallyeliminated in roots and reduced by 40% in shoots when externalmedia (0.5 mol m^–3 CaSO₄ plus 0–5 mol m^–3KCI) were supplemented with 0.25 mol m^– Ca(NO₃)₂. Plasmalemma³⁶Cl^– influx (_oc) was shown to be insensitive to externalNO₃^- in plants which had previously been grown in solutionslacking ^–3, but _oc became sensitive to NO₃^-after a lagperiod of 3–6 h. Kinetic analyses revealed that the inhibitionof 36C1^– influx by external NO₃^- was complex. At 0.25mol m^–3 NO₃^- the V_max for Cl^– influx was reducedby greater than 50%, with insignificant effects upon K_m. At0.5 mol m^–3 NO₃^- there was no further effect upon V_maxbut K_m for influx increased from 38±5 mmol m^–3to 116±26 mmol m^–3. By contrast, Cl^– effluxwas found to be insensitive to external NO₃^-. A model for theregulation of Cl^– influx is proposed which involves bothnegative feedback effects from vacuolar NO₃^- +Cl^–) concentrationand (external) NO₃^- inhibition of Cl^– influx at the plasmalemma.These combined effects serve to discriminate against Cl^–accumulation, favouring NO₃^- accumulation, when the latter ionis available. Such observations are inconsistent with recentproposals for the existence of bona fide homeostats for chlorideaccumulation in higher plants. Key words: Nitrate inhibition, Chloride influx, Barley     

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

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

Xylem exudation is related to nitrate assimilation pathway in detopped maize seedlings: use of nitrate reductase and glutamine synthetase inhibitors as tools

The xylem exudation of detopped 7-d-old seedlings of Zea maysL. doubled when KCI was present in the root medium comparedto seedlings maintained on water. It was further enhanced whenKCI was replaced by nitrogen compounds such as nitrate, ammoniumand glutamine. The role of the nitrate assimilation pathwayon the enhancement of xylem exudation rate was investigatedusing tungstate, an inhibitor of nitrate reductase (NR) activity,and phosphinothricin or methionine sulphoximine, inhibitorsof glutamine synthetase (GS) activity. The sap levels of NO₃^–,NH₄⁺, glutamine, and asparagine was used to ascertain the invivo inhibition of both enzymes. The tungstate effects werealso checked by measuring leaf in vitro NA activity and NR proteincontent. Xylem exudation rate of detopped seedlings fed withKNO₃ decreased when the nitrate assimilation pathway was blockedeither at the NR or at GS sites. This decrease was preventedwhen urea (acting as NH₄⁺ supply) was given simultaneously withtungstate. KNO₃ does not act directly on exudation, but throughthe involvement of NH₄⁺. The involvement of glutamine was alsoshown since GS inhibition resulted in a cancellation of theenhancing effect of KNO₃ on exudation. As change of exudationrate was not linked to change in sap osmolarity, it is assumedthat the assimilation chain could modify root water conductance.The role of glutamine was discussed. Key words: Exudation, maize, nitrate, conductance, NR, GS     

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     

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

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

Effect of NaCI Salinity on Flows and Partitioning of C, N, and Mineral Ions in Whole Plants of White Lupin, Lupinus albusL.

Plants of Lupinus albus L., cv. Ultra, were grown hydroponicallywith NO₃^–-nutrition for 51 d under control (0.05 mol m^–3Na⁺ and 10 mol m^–3 Cl^–) and saline (40 mol m^–3NaCI) conditions. Plants were harvested 41 and 51 d after germinationand analysed for content and net increment of C, N and the mineralcations K⁺, Na⁺, Mg²⁺, and Ca²⁺ and the anions Cl^–, NOJ,malate, phosphate, and SO₄^2–. Roots, stem interaodes,petioles and leaflets were analysed separately. During the studyperiod net photosynthesis, respiratory losses of CO₂ from shootand root and the composition of the spontaneously bleeding phloemsap and the root pressure xylem exudate were also determined.Using molar ratios of C over N in the transport fluids, incrementsof C and N, and photosynthetic gains as well as respiratorylosses of C, the net flows of C and N in the xylem and phloemwere then calculated as in earlier studies (Pate, Layzell andMcNeill, 1979a). Knowing the carbon flows, the ratios of ionto carbon in the phloem sap, and ion increments in individualorgans, net flows of K⁺, Na⁺, and Cl^– over the study periodwere also calculated. Salt stress led to a general decrease of all partial componentsof C and N partitioning indicating that inhibitions were notdue to specific effects of NaCI salinity on photosynthesis oron NO₃ uptake. However, there were differences between variouslyaged organs, and net phloem export of nitrogenous compoundsfrom ageing leaves was substantially enhanced under saline conditions.In addition, NO₃^–reduction in the roots was specificallyinhibited. Uptake and xylem transport of K⁺ was more severelyinhibited than photosynthetic carbon gain or NO₃^– uptakeby the root. K⁺ transport in the phloem was even more severelyrestricted under saline conditions. Na⁺ and Cl^– flowsand uptake, on the other hand, were substantially increasedin the presence of salt and, in particular, there were thenmassive flows of Na^– in the phloem. The results are discussedin relation to the causes of salt sensitivity of Lupinus albus.The data suggest that both a restriction of K⁺ supply and astrongly increased phloem translocation of Na⁺ contribute tothe adverse effects of salt in this species. Restriction ofK⁺ supply occurs by diminished K⁺ uptake and even more by reducedK⁺ cycling within the plant. Key words: Lupinus albus, salt stress, phloem transport, xylem transport, partitioning, carbon, nitrogen, K⁺, Na⁺, CI^–     

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