Differential rates of inhibition of N2 fixation by sustained low concentrations of NH4+ and NO3- in northern ecotypes of white clover (Trifolium repens L.)

An experimental study in flowing solution culture compared threeNorwegian ecotypes (from Saerheim, Pasvik and Bod) and a commercialcultivar (Ac51) of white clover (Trifolium repens L.) and demonstrateddifferences with respect to the rate and intensity with whichN₂ fixation was affected when or were supplied at 20 mmol m^–3 over 14 d. Plants were nodulated and N₂ fixation wasestablished over 17 d prior to supplying mineral N, with shoottemperature 25/15 C day/night and root temperature adjustedprogressively to 12C. Control plants received no mineral Nthroughout, and did not grow as rapidly as those supplied with or . Mineral N generally depressed the total N₂ fixation per plantrelative to control plants, with two exceptions. The effectof on N₂ fixation was moresevere than that of and, over the 14 d, -fed plants fixed more N₂ than the comparable -fed plants. Interpolated daily rates of N₂ fixation per plant andnodule dry wts were used to calculate specific rates of N₂ fixation.These showed that (1) addition of either or initially stimulated fixation relative to control plants and to mineral N uptake;(2) this stimulation was greatest and was delayed by 1-2 d in-fed plants; and (3) nutrition sustained higher residualrates of N₂ fixation after 8 d compared with nutrition, under which fixation all but ceased after10 d in three of the genotypes. Ecotype Bode showed by far themost severe induced depression of N₂ fixation. Key words: N₂ fixation, nitrate, ammonium, white clover, northern ecotypes     

Effects of Light on Transport by Azolla pinnata

Effects of light flux density (LFD) during growth and uptakeassay on induction of transport system and kinetics of transport were studied using the Azolla pinnata-Anabaena azollae association (Azolla). Theinduction and uptake kinetics of the transport system were determined using an automated system that measuredthe NO₃ concentration in the growth medium as a function oftime, using an on-line high performance liquid chromatograph(HPLC) with a UV-VIS detector. Full induction of the transport system required about 1.5 to 2.0 h and occurred without any apparent lag phase,regardless of the LFD provided. The level of induction of transport of Azolla grown at 600 µmol m^–2s^–1 LFD was higher than for that grown at 100 µmolm^–2 s^–1. Similarly, 600 µmol m^–1 s^–1LFD during the assay resulted in a higher level of inductionthan did 100 umol m^–2 s^–1. An increase in the LFDeither during the growth or the assay period increased the uptake rate; however, an increase in LFD duringthe latter period had greater effect. Azolla grown and assayedat 600 umol m^–2 s^–1 had the highest uptake rate. The uptake rate at 50 cm³ m^–3ambient CO₂ concentration was initially higher than at 305 cm³m^–3, but the uptake rate decreased rapidly with time andeventually dropped below that at 305 cm³ m^–3 CO₂. Thesedata suggest that the energy required for transport in Azolla may bypass the photosynthetic CO₂ fixationand carbon-cycling. Key words: carbon dioxide, concentration dependence, light flux density, uptake     

Interactions Between the Uptake and Assimilation of Inorganic Nitrogen and Carbon in Amphidinium spp. (Dinophyceae)

The rate of exponential growth of Amphidinium carterae Hulburtwas the same (0.025 h^–1) with either or as sole N-source. Nevertheless, in short-term experiments, cells growing exponentially with as N-source took up added 5–6 times faster than . accumulated in the cells. Addition of inhibited, reversibly, disappearance of from the medium; prior N-deprivation of the cells did not affect this inhibition. N-repIetecells, grown with as N-source, took up for several hours both in light and darkness, butthe uptake by such cells soon ceased in darkness although it continued in light. When uptake ceased, the cells could still take up rapidly in darkness. Ammonium taken up was assimilated rapidly into organic-N includingglutamine, other amino acids and protein. Ammonium uptake in darkness was accompanied by the utilizationof cellular polysaccharide, mainly glucose polysaccharide. Mostof this carbon was unavailable for the dark assimilation of. Addition of the analogue, methylammonium, did not initiate polysacchande catabolism. Itis suggested that a control mechanism is in operation, througha product of assimilation, which operates on one or more of the enzymes concerned with polysaccharidebreakdown, for example, -amylase or phosphorylase. Uptake of was accompanied by a high rate of dark ¹⁴CO₂ fixation and with both Amphidintum klebsii Kof.et Swezy and A. carterae N-deprivation led to a marked increasein this rate following addition of ; addition of had much less effect. The possible implications of these findings for the eco-physiology of marine dinoflagellatesare discussed. Key words: Ammonium, nitrate, CO₂ fixation     

Carbon Metabolism in Seagrasses: I. THE UTILIZATION OF EXOGENOUS INORGANIC CARBON SPECIES IN PHOTOSYNTHESIS

Four species of seagrasses, Halophila stipulacea, Thalassodendronciliatum, Halodule uninervis, and Syringodium isoetifolium,were investigated for their ability to utilize and CO₂ as exogenous carbon sources for photosynthesis. Ratesof photosynthesis were measured as rates of O₂ evolution ina closed system in which the pH was continuously controlled.A computer program was written to calculate the concentrationsof different carbon species as a function of pH and other specifiedexperimental conditions. Bicarbonate as well as CO₂ were readily assimilated by all fourseagrass species. Saturating concentrations of , at saturating light intensities, were 0.5–1.8 mM dependingon the species. Rates of photosynthesis under such conditionswere 0.1–0.55 µmol O₂ min^–1 mg^–1 chlorophyll.At saturating CO₂ concentrations, i.e. 0.5–1.3 mM, ratesof photosynthesis were 0.22–1.4 µmol CO₂ min^–1mg^–1 chlorophyll. Photosynthetic rates in each specieswere considerably higher when CO₂ rather than was supplied at saturating concentrations. The concentration of in natural seawater was found to be saturating, and that of CO₂ insufficient forconsiderable photosynthetic rates in these plants under thegiven conditions It was thus concluded that is the major carbon source for photosynthesis in seagrasses.     

Salt-Stimulated Bicarbonate-Dependent Photosynthesis in the Marine Angiosperm Zostera muelleri

Millhouse, J. and Strother, S. 1986. Salt-stimulated bicarbonate-dependentphotosynthesis in the marine angiosperm Zostera muelleri.—J.exp. Bot. 37: 965–976. Photosynthetic oxygen evolution in the seagrass Zostera muelleriIrmisch ex Aschers. was inhibited in iso-osmotic sucrose. Theapparent affinity of the leaves for CO₂ in seawater increasesfrom pH 8?2 to 8?9 indicating that as well as CO₂ may act as a substrate for photosynthesis. Theaffinity for CO₂ was lower in iso-osmotic sucrose and was notaffected by pH. Under these conditions was not a substrate for photosynthesis. The differencebetween the photosynthetic rate in seawater and iso-osmoticsucrose at the same concentration of CO₂ was used to estimate assimilation. The Briggs-Maskell equation, which allows for an unstirred layer around the tissuewas more appropriate than the Michaelis-Menten theory for calculatingthe apparent affinity of the leaf slices for CO₂. The apparentK_m CO₂ was calculated as 116 mmol m^–3 at pH 8?2 by Michaelis-Mentenkinetics but only 8?10 mmol m^–3 by the Briggs-Maskellequation. The stimulation by various ions in Seawater of use was investigated. The cations,in decreasing order of effectiveness were Ca²⁺, Mg²⁺, K⁺ andNa⁺ Anions were ineffective. No single cation at its concentrationin seawater was capable of supporting use at the rate observed in seawater. Acetazolamide,an inhibitor of carbonic anhydrase, inhibited the use of for photosynthesis but had littleeffect on CO₂ photosynthesis. Thus, carbonic anhydrase activityis required for -dependent photosynthesis. Key words: Zostera muelleri, photosynthesis, salinity     

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

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

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     

Relative Uptake Rates of Inorganic Nutrients by NO3- and NH4+ Grown Ricinus communis and by two Plantago Species

Allen, S., Thomas, G. E. and Raven, J. A. 1986. Relative uptakerates of inorganic nutrients by and grown Ricinus communis and by two Plantago species.—J. exp. Bot. 37: 419–428. The relative rates of uptake and assimilation of C, N, P, S,Cl^–, K⁺ , Na⁺ Ca²⁺ and Mg²⁺ by and grown Ricinus conimunisand by NH₄NO₃- grown Plantago lanceolata and P. major were calculatedfrom data presented elsewhere. Results showed that for grown Ricinus the short term relativeuptake rates, for each nutrient X did not change significantly over the steady-state periodof exponential growth. The average gave , the mean relative uptake rate during exponential growth, for each nutrient. The amountof each nutrient taken up from a nutrient solution over a periodof time could, therefore, be calculated. For and -grown R. communis,the relative uptake rate of each nutrient was a constant fractionof the relative rate of carbon assimilation. It is suggestedthat this is typical of plants of cauline habit. For both Plantago spp., the relative rates of nitrogen uptakeand assimilation fell significantly during the exponential growthphase It is suggested that this could be a characteristic ofthe growth habit of the rosette plant. Key words: Relative uptake rates, Ricinus, Plantago, ammonium, nitrate, cauline, rosette     

Calcification in the Green Alga Halimeda: III. THE SOURCES OF INORGANIC CARBON FOR PHOTOSYNTHESIS AND CALCIFICATION AND A MODEL OF THE MECHANISM OF CALCIFICATION

Calcification and photosynthetic rates in Halimeda tuna weremeasured by the ¹⁴C method under conditions of differing pHand total inorganic carbon (CO₂) concentrations. The effectsof pH and CO₂ on photosynthesis and respiration were also monitoredwith a polarographic O₂ electrode. The results obtained indicatethat the intercellular pH and CO₂ differ from those of the externalmedium. Experiments carried out over a range of pH values show thatHalimeda can use for photosynthesis. Photosynthesis appears to stimulate calcification by removing CO₂ from theintercellular spaces. As these spaces are isolated from theexternal sea water by the layer of cell wall of the adpressedperipheral utricles, the removal of CO₂ results in a rise in[] and a rise in pH. This results in an increased rate of CaCO₃ precipitation. Respiratory CO₂ evolution has aninhibitory effect on calcification by decreasing the pH and[]. A model for calcification in Halimeda is proposed based on theresults of this and previous papers. Calcification in Halimedais seen to be a result of the anatomy of the thallus in whichthe sites of calcification are within a semi-isolated chamberwhere removal or addition of CO₂ due to photosynthesis or respirationcan effectively change [CO] thereby resulting in precipitation of CaCO₃. In the Appendix to this paper theoreticalcalculations illustrate the effects of CO₂, , and removal or addition in a closed system on the relative concentrations of the other inorganic carbonspecies.     

The Titrimetric Assay of OH- and HCO3- Excreted by Ricinus Cultivated on NO3--containing Nutrient Media: The Influence of Ionic Strength, End Point pH and CO2 Supersaturation

Allen, S. and Allen, C. R. 1987. The titrimetric assay of OH^–and excreted by Ricinus cultivatedon -containing nutrient media: the influence of ionic strength, end point pH and CO² supersaturation.–J. exp. Bot. 38: 607–617. When spent -containing nutrient media were titrated to the starting pH of 6.5 the titre wasequivalent to 50%orless of the base (i.e. ) excreted. Calculation of the total amount of baseexcreted could only be made from data obtained by titrationto pH 4–5. An accurate calculation of the amount of baserequired the inclusion of activity coefficients, estimated fromionic strength, in the calculations. Spent nutrient media contained from four to ten times the concentrationof CO₂ predicted from equilibrium values. It is probable that is very slow. Key words: Ricinus, nitrate-N nutrient medium, base excretion, ionic activity coefficients, carbon dioxide supersaturation     

Uptake of Methylammonium Ions by Hydrodictyon africanum

Methylamine influx into Hydrodictyon has been measured with[^l4C]methylamine. The influx increases with rising externalpH up to about pH 8. Between pH 8 and 9 influx remains quiteconstant, with a further increase above pH 9. Influx is light-dependent,temperature-sensitive, and is decreased by . During short-term influx (less than 4 h) metabolism of methylamineappears negligible. Prolonged influx results in CH₃ accumulation, and efflux of K⁺, Na⁺, and H⁺. There is no effecton Cl^– influx. Methylamine decreases the membrane electricp.d. by 60–120 mV at external concentrations of 0?2–1?0mM. The results indicate that, below pH 9, methylamine enters thecell almost entirely as CH₃. It is suggested that a passive electrogenic uniporter is involved, and thatby analogy uniport of may also be expected in Hydrodictyon. The results are discussed in relation to theevidence for uptake of CH₃ and by other plant cells.     

Compensatory Changes in Ion Fluxes into Barley (Hordeum vulgare L. cv. Betzes) Seedlings in Response to Differential Root/Shoot Growth Temperature

The effect of growth temperature on the short term influx of⁸⁶Rb⁺/K⁺, ³⁶Cl^–/Cl^–, , ⁴⁵Ca²⁺/Ca²⁺and into barley plants (Hordeum vulgareL. cv. Betzes) has been examined. When barley plants were grownwith a differential root: shoot temperature (15 ?C/25 ?C) therewas a marked stimulation of influx of all ions except Ca²⁺.Q₁₀ measurements were close to 2.0 for all ions except Ca²⁺,where Q₁₀ 1.0 was found. Kinetics of ion influx showed thatthere had been almost complete compensation for the lower growingtemperature over a range of concentrations. The regulation of uptake was affected by growth temperature. On exposure to 15?C efflux/influx fell initially but was restored after 5 d. Sensitivity of net uptake to was increased by growth at a lower temperature. The importance of these observations in relationto application to field experiments and the proposed mechanismfor uptake is discussed. Key words: Growth temperature, Nitrate, Ammonium     

The Effect of and Ions on the Reactions of Ribulose Bisphosphate Carboxylase

The effects of sulphite ion () and sulphate ion () on both the activation and the catalytic activities of ribulose- 1, 5-bisphosphate carboxylase (EC 4.1.1.39 [EC] )were studied and compared to those of other effectors of theenzyme, particularly inorganic phosphate (P₁). The activationby CO₂ and Mg²⁺ of a slow activating form of the carboxylasein the presence of the two anions produced high specific activitieswith significant lower concentrations of CO₂ than normally required.This was due to stabilization of the ternary complex betweenthe enzyme, CO₂ and Mg²⁺. With a rapidly activating speciesof enzyme, and caused only a small increase in activation with subsaturating CO₂. , and P₁, with saturatingconcentrations of CO₂ also enhanced the catalytic activity abovethat achieved with CO₂ and Mg²⁺ alone; P₁ was the most effectiveof the anions, producing a 50% increase in the specific activity,both with the slow and rapidly activating species. and were potent inhibitors of the carboxylase and oxygenase reactions of the enzyme. was a non-competitive inhibitor with respect to CO₂, and competitive/mixedwith respect to ribulose-1, 5-bisphosphate. The time courseof the carboxylase and oxygenase reactions in the presence of were biphasic with inhibition apparent only in the second phase. Key words: Ribulose bisphosphate carboxylase, Activation, SO₃^2-, SO₄^2-     

Mechanism of Inorganic Carbon Uptake in Chlorella saccharophila: The Lack of Involvement of Carbonic Anhydrase

The acid-tolerant green alga Chlorella saccharophila maintainedphotosynthesis and accumulated intracellular pools of inorganiccarbon over a a range of external pH from 4.0 to 7.5. This accumulationwas unaffected by treatment of cells with 10 mol m^–3 acetazolamide(AZA). Cells grown at alkaline pH had extracellular carbonicanhydrase (CA), but CA activity was repressed when cells weregrown at pH 5.0. Acid-grown cells retained a high affinity forCO₂, both at acid and alkaline pH, and the ability to accumulateinorganic carbon. Rates of photosynthesis of acid-grown cellsand alkaline-grown AZA-treated cells at pH 8.0 were 2.5-foldhigher than the rate of CO₂ supply from the uncatalysed dehydrationof , indicating that the cells can take up as a source of substrate for photosynthesis. Isotopic disequilibrium experiments with acid-grown cells maintainingsteady-state photosynthesis at pH 7.5 demonstrate that ¹⁴C from¹⁴CO₂ was taken up more rapidly than from H¹⁴. This uptake takes place against a concentration gradient. Theseresults demonstrate that C. saccharophila cells have activetransport systems for the uptake of both CO₂ and and both operate without the mediation of CA. Key words: Bicarbonate transport, carbon dioxide, carbonic anhydrase, Chlorella saccharophila, inorganic carbon accumulation     

Nitrate Metabolism in Alfalfa Root Nodules under Water Stress

Three-month-old plants (vegetative stage) of alfalfa (Medicagosaliva L cv. Aragon) were supplied for one week with 1.0dm³(uniformly distributed) nutrient solutions containing 0 or 20mol m^–3 . One week after initiation of treatment, the plants were subjected to drought by withholding water. Bacteroidsand cytosol of nodules were obtained at different stages ofstress, and used for enzyme assays and for determination of, and . Proteins of bacteroids were more stable than cytosolic proteinswith respect to the detrimental effects of water stress and. Protein contents of bacteroids and cytosol were inversely related to proteolytic activitiesagainst azocasein in both nodule fractions. Specific nitrate reductase activity (NRA) and nitrite reductaseactivity (NiRA) of bacteroids from -treated plants were inhibited by c. 70% and 45%, respectively, as leafwater potential (_w) declined from –0.5 MPa (control) to–1.8 MPa. At still lower _w both activities began to increase:NRA was doubled, whereas NiRA only returned to its control level.Cytosolic NRA was strongly inhibited by drought, but the correspondingNiRA remained constant. Ammonia concentration in bacteroids and nodule cytosol keptbasically constant, whereas accumulated in the cytosol at severe stress, due to the activationof bacteroid nitrate reductase. Results indicate that nitrate and nitrite reductases of thebacteroids and the nodule cytosol act in different form: assimilatory,the cytosolic enzymes; and dissimilatory, the enzymes of bacteroidsat low _w The possibility that assimilation of also occurs in bacteroids at control or mild waterstress conditions is suggested. Key words: Assimilatory and dissimilatory reduction, bacteroids, Medicago saliva, nodule cytosol, water stress     

Effects of Cl- on and Malate Fluxes and CO2 Fixation in Carrot and Barley Root Cells

Cl^– and ions interact apparently competitively during influx across the plasmalemma of carrot root cells. Cl^–,however, reduces influx much less than predicted from the effect of on Cl^– influx.Cl^– and plasmalemma influxes both increase with time after excision of carrot tissue. Cl^–and may therefore be transported by a common mechanism. The effect of pH changes on the influx of malate across theplasmalemma in barley roots shows that malate crosses the plasmalemmaas the singly charged anion. Stimulations of influx by bothK₂SO₄ and KCl suggest that the malate anion crosses in associationwith. K⁺. If malate entry is passive, P_mal- is about 2?10^–8cm s^–1, but it is thought that malate entry is partlyan active process. A slight, apparently competitive inhibition by Cl^– ofmalate flux into the vacuole of barley root cells suggests thatthe two anions may be transported by a common process at thetonoplast, but this is not thought to be physiologically significant. The accumulation of ¹⁴C from 1 mM is drastically reduced by 10 mM Cl^–. A quantitative analysis of the kineticsof ¹⁴C exchange shows that Cl^– directly inhibits the formationof malate from . The decreased influx of endogenously produced malate to the vacuole in the presenceof Cl^– is probably a secondary consequence of the fallin the cytoplasmic concentration. The nature of the Cl^–inhibition of malate formation is discussed. In KCl-loaded tissue the influx of external malate and the accumulationof ¹⁴C from external are reduced. The location of these effects is not certain, but the effects suggest thatregulation of malate synthesis and accumulation may be relatedto the negative-feedback regulation of Cl^– and transport.     

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     

ATP Synthesis Coupled to Nitrate Photoreduction in the Alga Hydrodictyon africanum

The presence of exogenous stimulates O₂evolution in illuminated H. africanum cells at the CO₂ compensationpoint, and it is likely that reduction in the light uses reductant produced in non-cyclic electron flow.When sources of ATP other than fermentation are absent, thepresence of , light, and a functional non-cyclic electron transport pathway stimulates active, ATP-dependentH₂ influx. This is consistent with non-cyclic electron flow associated with reduction being coupled to ATP synthesis. This -dependent ATP synthesis may be quantitatively important as a source ofATP during photolithotrophic growth with as N source in H. africanum and other algal unicells.     

Nitrate Uptake and Assimilation following Nitrate Deprivation

Upon first exposure to , the uptake and reduction capacities of dark-grown corn (Zea maysL.) roots are initially low, but increase markedly within 6h. The development of the accelerated uptake rate appears to be substrate ‘induced’ as is reductase (NR), the first enzyme in the assimilatory pathway. However, the ‘induction’of uptake is independent of NR induction. The effect of deprivation was studied to determine the role of endogenous on subsequent uptake and reduction. Corn roots were ‘induced’ for 24 h in 0–5 mol m^–3 nutrient solution and then exposed for 0 to 32 h to -free nutrient solution. Uptake and reduction of were determined periodically by exposing sets of roots to a1 h pulse of 0.5 mol m^–3 . Neither uptake (4.57 µmol root^–1 h^–1)nor the percentage of absorbed reduced (27%) was changed significantly (P 0.05) by exogenous deprivation. However, the estimated ‘induced’ componentof uptake decreased significantly (50% after 32 h). Concurrently, the ‘non-induced’ basal componentof uptake increased. Previously accumulated decreased from 23 to 4.5 µmol root^–1 after 32 h of exogenous deprivation. Nearly equivalent quantities of endogenous were used for translocation and reduction during deprivation. During each 1 h pulse, the amounts of translocation and net efflux of to the uptake solution were similar. Net efflux of was strongly correlated (r = 0.991) to the amount of endogenous . The remaining endogenous and its rate of utilization were apparently sufficient to minimize a rapid declineor complete loss in both the ‘induced’ uptake state and the rate of in vivo assimilation. Key words: reduction, translocation, efflux, root, Zea mays L     

Effect of Nitrogen Form and Counterion on Establishment of the Rhizobium trifolii-Trifolium repens Symbiosis

The effect of level of different counterion forms of or or both, on establishment of the Rhizobium trifolii-Trifolium repens symbiosis was evaluatedin plants cultured under bacteriologically controlled conditions. Ammonium had little effect on nodule formation, and even after exhaustion there was little compensatory nodule formation. Plant growth, nodule mass and acetylene reductionactivity all declined with increasing levels of . Except for , different counterion forms had little effect on plant growth, nodule numberand mass, and acetylene reduction activity. Ammonium markedlydecreased pH, the extent being dependent upon level and counterion present. Nitrate inhibited nodulation while levels remained in excessof plant uptake. Compensatory nodulation followed exhaustionof or at lower or levels. Nodule mass and acetylene reduction activity decreased, but plant growth increased withincreasing or levels. Nitrate raised the pH of the nutrient solution by one unit orless. Different or counterion forms had little effect on the symbiosis or plant growth. Ammonium nitrate severely inhibited nodulation but on exhaustionmarked compensatory nodulation occurred. The patterns of nodulemass, acetylene reduction activity and plant growth with increasingNH₄NO₃ levels were similar to or alone. Plants provided with NH₄NO₃ or no nitrogenwere similar in only slightly decreasing pH. Key words: Ammonium, Nitrate, Ammonium nitrate, Nitrogen fixation, Rhizobium trifolii, Trifolium repens, Symbiosis establishment, Nodulation