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     

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     

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     

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     

An analysis of the physiological basis of commonality between diurnal patterns of NH4+, NO3- and K+ uptake by Phleum pratense and Festuca pratensis

Commonality in diurnal variation in net uptake of by the grasses Phleum pratense L. cv. Bodin (timothy),and Festuca pratensis Huds. cv. Salten (fescue) was evaluatedin flowing solution culture under a semi-natural light regime.Hourly uptake rates from constant 20 mmol m^–3 concentrationsof each ion were measured concurrently over 7 d, without physicaldisturbance. The light period was 11 h, natural light supplementedby constant artificial illumination, with a step-transitionto a 13 h dark period. Uptake of all three ions showed a broadlysimilar pattern of diurnal variation, rates increasing duringthe light period and decreasing during the darkness to a minimumwithin 2 h of the end of the dark period. The amplitude wasgreatest during high-irradiance days, and both proportionatelyand absolutely greater for (mean min:max uptake?0.24) than (min:max=0.41) or K⁺ (min:max?0.34). There were significantdifferences between and the other two ions whose behaviour coincided in almost all respects, in timing of maximum and minimumrates, and acceleration in uptake during the light period. Preferentialuptake of over increased sharply during the first half of the lightperiod; in relative terms uptake accelerated twice as fast as uptake. Fescue always absorbed more than but timothy showed a preference for during part of the dark period. The results are interpreted in terms ofthe hypothesis that diurnally fluctuating ‘sink-strength’for nutrients is the primary determinant of nutrient transportrates, although uptake may become temporarily uncoupled fromnutrient demand during periods of physiological stress or perturbation,such as towards the end of the dark period, during which uptakerates are determined by factors (e.g. carbohydrate supply) otherthan current nutrient demand. Key words: Ammonium, diurnal variation, Gramineae, ion uptake, nitrate, potassium, regulation     

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     

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     

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     

Pumpkin (Cucurbita sp.) seed globulin IV. Terminal sequences of the acidic and basic peptide chains and identification of a pyroglutamyl peptide chain

Pumpkin seed globulin is composed of heterogeneous polypeptidechains, acidic and chains and basic ₁ and ₂ chains (12). This study showed that the basicchains had similar N-terminal sequences, Gly-Leu-Asp-Glu-Thr-Ile-for the ₁ chain and Gly-Leu-Glu-Glu-Thr-Ile- for ₂. On the contrary,the N-terminal sequences of the acidic and chains were dissimilar, Ile-Gln-Gly-Tyr- for the chain and no N-terminal residue for the chain, according to routine terminal analysis. Pyrrolidonylpeptidase digestion of the chain and its thermolysin digestion followed by Edman degradationsrevealed that the N-terminal sequence of the chain was < Glu-Ile-Glu-Gln-Gln-Glu-Pro(Trp,Ser)-. The N-terminal sequences and the C-terminal residuesindicated that the acidic and chains were more heterogeneous than the basic ₁ and ₂ chains.A preliminary study on the degradation of storage globulin isalso presented. (Received November 9, 1979; )     

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     

Stage of development is an important determinant in the effect of nitrate on photoassimilate (13C) partitioning in chicory (Cichorium intybus)

The effect of nitrogen supply to chicory plants on carbon partitioningbetween shoot, root and tuberized root was studied at differentstages of vegetative growth, using long-term ¹³CO₂ labelling-chaseexperiments. This approach was complemented by measurement ofstorage carbohydrates and activities of enzymes involved inroot sucrose metabolism (sucrose-sucrose fructosyl transferase(SST), sucrose synthase, invertase). In both young and matureplants, low resulted in a 30–35% decrease in ¹³C assimilation. However, the partitioningof ¹³C between shoot and root was affected differently at differentstages of development. In young plants, in which carbohydrateswere being used for structural root and shoot growth, neither¹³C shoot/¹³C root ratio nor root activities of the above enzymeswere modified by supply. In contrast, in mature plants storing large amounts of carbohydratesas fructan in the tuberized root, low caused the ratio to decrease from 0.6 to 0.2, despiteunchanged net flux of ¹³C from shoot to root. The extractableactivity of SST was elevated in mature plants, compared to youngplants, at both low and high , consistent with its role in fructan synthesis. However, matureplants grown at low exhibited SST activity double that of plants grown at high . From these results, it is concluded that the observeddecrease in shoot/root dry weight ratio at low supply is caused by increased utilization of carbohydratesfor storage due to elevated root SST activity. Key words: Chicory, nitrate, ¹³C, shoot/root ratio, fructans, SST     

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.     

Intracellular pH Regulation in Ricinus communis Grown with Ammonium or Nitrate as N Source: The Role of Long Distance Transport

Allen, S. and Raven, J. A. 1987. Intracellular pH regulationin Ricinus communis grown with ammonium or nitrate as N source:the role of long distance transport.—J. exp. Bot. 38:580–596. Charge balance and intracellular pH regulation were studiedin Ricinus communis grown in water culture with 1–0 molm^–3 or l-0 mol m ^–3 as sole N source. Seedling and 70-d-old plant parts were analysed for total P, C, organic N and S, and ash alkalinity;xylem and phloem saps were analysed for mineral ion content,and amino acids, amides, and dicarboxylates. Excreted H⁺ andbase were also measured. It was shown that in -N plants, H ⁺ produced and excreted directly bythe roots accounted for all net —COOH produced in theplant, but not for cation uptake by net H⁺ exchange. IntracellularpH perturbation in the shoot was regulated partly by reduction and partly by the transportof OH^- -generating dicarboxylates in the xylem. Phloem sap hadthe capacity to transport organic N and carboxylates excessto shoot requirements back to the root. In -N plants, 60% of total reduction occurred in the root, and 70% of all OH^- produced by root and reduction was excreted directly as base. Very little —COO^–from root reduction was stored in the root: most was transported to the shoot as xylem dicarboxylateand stored in the shoot. Of the OH^- produced from shoot and reduction, 40% was stored as shoot carboxylate: the phloem saphad the capacity to transport the rest back to the roots whereit was excreted as base. Key words: Intracellular pH, ammonium nutrition, nitrate nutrition, xylem dicarboxylates, phloem dicarboxylates, charge balance     

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.     

Evidence for Bicarbonate Transport in Species of Red and Brown Macrophytic Marine Algae

Cook, C. M., Lanaras, T. and Colman, B. 1986. Evidence for bicarbonatetransport in species of red and brown macrophytic marine algae.—J.exp. Bot. 37: 977–984. The capacity of 17 species of marine macrophytes to take up has been examined by comparing the rate of photosynthetic O₂-evolution with the photosyntheticrate which could be supported solely by CO₂ arising from theuncatalysed dehydration of . No external carbonic anhydrase was detected by potentiometricassay in any of the species used. At pH 8?0, the rates of photosyntheticO₂-evalution exceeded the CO₂ supply rate 6 to 24-fold in 15species of red algae, and 7 to 11-fold in 2 species of brownalgae. The ratio of photosynthetic O₂-evalution to the CO₂ supplyrate was even higher (19 to 101: 1) at pH 9?0. It is evidentfrom this data that the rate of CO₂ supply from the spontaneousbreakdown of cannot support the observed rates of photosynthesis in these algae. Thus, thedata provide substantive evidence that is taken up by these marine macrophytes as a sourceof substrate for photosynthesis. Key words: Macrophytic marine algae, bicarbonate transport     

Nitrate transport in Zea mays cell suspension cultures: the effect of solution composition and cell age

Nitrate transport characteristics of an amino acid-grown Zeamays P3377 cell culture line were studied. Age (days after subculturing)of the cells was shown to have a significant effect on transport; older stationary phasecultures did not absorb nitrate from the medium as rapidly asyounger growing cultures. Solution composition had a pronouncedimpact on induction of accelerated nitrate transport and transportrates. Maximum uptake rates required fresh culture media ratherthan simple solutions. Differences in ionic strength among uptakesolutions of equal concentration were shown to affect the apparent uptake rates by changing theactivity coefficient of . The uptake kinetics were established by following uptake for 24h in a wide range of nitrate concentrations. Uptake patternsof cells in solutions ranging from 0.02 to 2 mM were as typicallyreported for plants. The kinetic constants for the Zea mayscell suspension cultures concurred with reports of other solution-culturedcells. When cells were placed in solutions containing greater than 2 mM, uptake patternssuggested a significant passive uptake component. Passive diffusionof was estimated by Nernst analysis and indicated to be an important component of nitrateuptake in maize cell suspension cultures grown in the absenceof nitrate then transferred into nitrate-containing media. Key words: Cell suspension culture, nitrate, passive uptake, Zea mays     

Plant response to nitrate starvation is determined by N storage capacity matched by nitrate uptake capacity in two Arabidopsis genotypes

In a low-input agricultural context, plants facing temporalnutrient deficiencies need to be efficient. By comparing theeffects of NO-starvation in two lines of Arabidopsis thaliana (RIL282 and 432 from the Bay-0xShahdarapopulation), this study aimed to screen the physiological mechanismsallowing one genotype to withstand NO-deprivation better than another and to rate the relative importance of processessuch as nitrate uptake, storage, and recycling. These two lines,chosen because of their contrasted shoot N contents for identicalshoot biomass under N-replete conditions, underwent a 10 d nitratestarvation after 28 d of culture at 5 mM NO. It was demonstrated that line 432 coped betterwith NO-starvation, producing higher shoot and root biomass and sustaining maximal growthfor a longer time. However, both lines exhibited similar featuresunder NO-starvation conditions. In particular, the nitrate pool underwent the same drastic andearly depletion, whereas the protein pool was increased to asimilar extent. Nitrate remobilization rate was identical too.It was proportional to nitrate content in both shoots and roots,but it was higher in roots. One difference emerged: line 432had a higher nitrate content at the beginning of the starvationphase. This suggests that to overcome NO-starvation, line 432 did not directly rely on theN pool composition, nor on nitrate remobilization efficiency,but on higher nitrate storage capacities prior to NO-starvation. Moreover, the higher resistanceof 432 corresponded to a higher nitrate uptake capacity anda 2–9-fold higher expression of AtNRT1.1, AtNRT2.1, andAtNRT2.4 genes, suggesting that the corresponding nitrate transportersmay be preferentially involved under fluctuating N supply conditions. Key words: Arabidopsis thaliana, genetic variability, N partitioning, N recycling, N use efficiency, nitrate deficiency, nitrate remobilization rate, nitrate transporter gene expression, nitrogen reserves, plant development Received 12 July 2007; Revised 21 November 2007 Accepted 17 December 2007     

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     

Translocation of Some Anions, Cations, and Acids in Turnip (Brassica napus L.) Mitochondria

The translocation of various anions, cations, and acids acrossthe membrane system of turnip mitchondria has been investigatedby light scattering techniques. The results imply that the membranesof turnip mitochondria are not permeable to choline, K⁺, Na⁺,, , , and . It has been established that Cl^–,, and SCN– passively permeate the mitochondrial membrane. It is probable that permeates as NH₃ and that and cross the membrane either as the corresponding acid or on ananion/ hydroxide antiporter. The alkali metals Na⁺ and K⁺ equilibrateacross the membrane via specific H⁺/Na⁺ and H⁺/K⁺ antiporters,having similar activity at neutral pH.     

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.