Effect of Ammonia on Dark CO2 Fixation by Anabaena Cells Treated with Methionine Sulfoximine

Dark CO₂ fixation by Anabaena cylindrica was stimulated aboutthree-fold by the addition of NH₄Cl to the cells. The ¹⁴CO₂incorporation experiments showed that ¹⁴C is most rapidly incorporatedinto aspartate and then glutamine by adding NH4CI. Glutamineaccumulated predominantly after the addition of NH₄Cl showingthat NH₄ is incorporated into glutamine by glutamine synthetase.The stimulating effect of NH4Cl on CO₂ fixation and amino acidsynthesis was suppressed by methionine sulfoximine, an inhibitorof glutamine synthetase. It was suggested that dark CO₂ fixationwas stimulated by the action of glutamine synthesis which isenhanced by ammonia. (Received February 10, 1981; Accepted April 2, 1981)     

Effect of Methionine Sulfoximine on Photosynthetic Carbon Metabolism in Wheat Leaves

Methionine sulfoximine (MSO) greatly reduced the carbon dioxideexchange rate (CER) of detached wheat (Triticum aestivvm L.cv Roland) leaves in 21% O₂, but only slightly reduced it in2% O₂. A supply of 50 mM NH₄Cl had little effect on the CERirrespective of the O₂ concentration. A simultaneous additionof glutamine and MSO protected against the inhibition of photosynthesisto a considerable extent and caused the accumulation of moreNH₃ than did the addition of MSO alone. Fixation of ¹⁴CO₂ in wheat leaves was inhibited by MSO treatmentin 22% O₂, and there was decreased incorporation of ¹⁴G intoamino acids and sugars and increased label into acid fractions.The addition of MSO and glutamine together eliminated the effectof MSO on the photosynthetic ¹⁴CO₂ fixation pattern. NH₄Cl stimulatedthe synthesis of amino acids from ¹⁴CO₂, especially the synthesisof serine in 22% O₂. Our observations show that factors other than the uncouplingof photophosphorylation by accumulated NH₃ may be responsiblefor the early stage of photosynthesis inhibition by MSO underphotorespiratory conditions. ¹Present address: Department of Agricultural Chemistry, KyushuUniversity, Fukuoka 812 Japan. ²Also at U.S. Department of Agriculture, Agricultural ResearchService, Urbana, Illionois 61801, U.S.A. (Received September 13, 1983; Accepted February 2, 1984)     

Effects of dark preincubation and chloramphenicol on blue light-induced CO2 incorporation in Chlorella cells

Chlorella cells incubated in the dark longer than 12 hr showedpronounced blue light-induced ¹⁴CO₂ fixation into aspartate,glutamate, malate and fumarate (blue light effect), whereasthose kept under continuous light showed only a slight bluelight effect, if any. 2) During dark incubation of Chlorellacells, phosphoenolpyruvate carboxylase activity and the capacityfor dark ¹⁴CO₂ fixation decreased significantly, whereas ribulose-1,5-diphosphatecarboxylase activity and the capacity for photosynthetic ¹⁴CO₂fixation (measured under illumination of white light at a highlight intensity) did not decrease. 3) In cells preincubatedin the dark, intracellular levels of phosphoenolpyruvate and3-phosphoglycerate determined during illumination with bluelight were practically equal to levels determined during illuminationwith red light. 4) The blue light effect was not observed incells incubated widi chloramphenicol, indicating that blue light-inducedprotein synthesis is involved in the mechanism of the effect. (Received April 9, 1971; )     

Translocation Profiles of [11C] and [13N]-Labelled Metabolites after Assimilation of 11CO2 and [13N]-Labelled Ammonia Gas by Leaves of Helianthus annuus L. and Lupinus albus L.

Tracer amounts of atmospheric [¹³N]-Iabelled ammonia gas, wereabsorbed by leaves of Lupinus albus and Helianthus annuus inboth the light and the dark. Exogenous [¹³N]-ammonia was onlyabsorbed in the dark when the feeding occurred shortly aftera period of illumination and the tissue was not depleted ofits carbohydrate reserves (e.g. starch). Incorporation of the[¹³N]-ammonia appeared to occur via the leaf glutamine synthetase/glutamatesynthase (GS/GOGAT) cycle since 2.0 mol m^–3 MSX, an inhibitorof the GS reduced uptake in both the light and dark. Photosyntheticincorporation of ¹¹CO₂ was not affected by this treatment Therate of movement of [¹³N]-assimilates in the petiole of attachedleaves of Helianthus and Lupinus was similar to that of the¹¹Cl-photo assimilates. Export of both [¹³N] and [¹¹C]-Iabelledassimilates from the leaf and movement in the petiole in boththe light and the dark was inhibited by source leaf anoxia (i.e.nitrogen gas). Translocation was re-established at the samerate when the feed leaf was exposed to gas containing more than2% O₂ which permitted dark respiration to proceed. After aninitial feeding of either ¹¹CO₂ or [¹³N]-ammonia at ambient(21%) O₂ exposure of the source leaf to 2% O2, or 50% O² didnot alter the rates of translocation, indicating that changesin photosynthetic activity in the source leaf due to photorespiratoryactivity need not markedly alter, at least during the shortperiod, the loading and translocation of either [¹¹C ] or [¹³N]-labelledleaf products. Key words: Translocation, CO₂, NH₃, Leaves, Helianthus annuus, Lupinus albus     

Effect of Ammonia on Cellular pH of Anabaena cylindrica Determined with 31P NMR Spectroscopy

The pH changes in the blue-green alga (cyanobacterium) Anabaenacylindrica caused by addition of ammonia were investigated using³¹P NMR spectroscopy. A pH shift of 0.9 or more was observedwhen 30 nM NH₄OH was added to the cell suspension, but no significantcellular pH change was observed with 50 mM NH₄CI, a concentrationhigh enough to stimulate dark CO₂ fixation of this alga. Thechange in cellular pH does not seem to cause ammonia-inducedstimulation of dark CO₂ fixation. (Received June 22, 1985; Accepted January 10, 1986)     

Transient Changes in the Energy State of Adenylates and the Redox States of Pyridine Nucleotides in Chlorella Cells Induced by Environmental Changes

The unicellular green algae Chlorella ellipsoidea was used tostudy transient changes in the energy state of adenylates andthe redox states of pyridine nucleotides induced by environmentalchanges. The transition from anaerobic to aerobic conditionsin the dark induced a sharp rise in the ATP ratio [ATP/(ATP+ADP+AMP)],a sudden decrease in the NADH ratio [NADH/(NAD⁺+NADPH)] anda transient drop in the NADPH ratio [NADPH/(NADP⁺+NADPH)]. Illuminationafter a dark period under anaerobic, CO₂-free conditions inducedsharp increases in the ATP and NADPH ratios and a slower decreasein the NADH ratio. Illumination under aerobic conditions, ineither the presence or absence of CO₂, caused a sharp increasein the NADPH ratio, a small increase in the ATP ratio and aslower increase in the NADH ratio. In the presence of CO₂, asubsequent large drop in the NADPH ratio occurred. Darkeningunder anaerobic, CO₂-free conditions induced a sudden decreasein the ATP ratio, a temporary fall in the NADPH ratio and aslow increase in the NADH ratio. Darkening under aerobic conditionsinduced transient drops in the ATP and NADPH ratios and a suddendrop in the NADH ratio. The addition of CO₂ to the atmospherewith illumination produced a decrease in all three parameters. These results are discussed in relation to current theoriesof the interaction between photosynthesis and respiration. Ourobservations indicate that the energy and reducing potentialsgenerated by photochemical processes are used for and controlother processes besides CO₂ fixation in photosynthetic cells. (Received December 3, 1981; Accepted May 4, 1982)     

Comparative Studies of Leaf Tissue 4: III. EFFECTS OF WALL-DEGRADING ENZYMES AND OSMOTIC STRESS

Commercially available cell wall-degrading enzymes frequentlyused for protoplast isolation inhibited CO₂ fixation and photosyntheticO₂ evolution, and stimulated dark respiration by leaf tissueand isolated mesophyll protoplasts of Nicotiana tabacum L. andAntirrhinum majus L. They also depolarized the membrane potentialof cells of leaf tissue, inhibited uptake of ⁸⁶Rb by tobaccoleaf tissue and isolated mesophyll protoplasts, and stimulated³⁶CI uptake by tobacco leaf tissue. Where studied, these effectswere found to be reversible. The depolarization effect on Antirrhinumleaf cells occurred even when the enzyme preparations had beendenatured, dialysed, or desalted, and the effect was greatestin those fractions of the enzyme preparation which showed thehighest cellulase activity. Plasmolysis of tobacco leaf tissue inhibited photosyntheticO₂ evolution, CO₂ fixation, and ⁸⁶Rb uptake to levels belowthose exhibited by isolated protoplasts in media of the samecomposition and osmolarity. The implications of these resultsfor work with leaf tissue and isolated protoplasts are discussed.     

N2 Fixation and the Respiratory Costs of Nodules, Nitrogenase Activity, and Nodule Growth and Maintenance in Fiskeby Soyabean

The respiratory effluxes of nodules and of roots of FiskebyV soyabean (Glycine max (L.) Merr.), grown in a controlled environment,were measured at intervals in air and 3% O₂ from shortly afterthe onset of N₂ fixation until plant senescence. The respiratoryburdens linked with nitrogenase plus ammonia metabolism, andnodule growth and maintenance, were calculated from gas exchangedata and related to the concurrent rates of N₂ fixation. The specific respiration rates of nodules increased to a maximumof 21 mg CO₂ g^–1 h^–1 at the time pods began development:the equivalent maximum for roots was c. 4.5 mg CO₂ g^–1h^–1. Maximum nodule and root respiration rates per plantwere attained about 25 d later at the time N₂ fixation peakedat 15 mg N d^–1 plant^–1. The relationship between nodule respiration and N₂ fixationindicated an average respiratory cost of 13.2 mg CO₂ mg^–1N until the last few days of plant development Separation ofnodule respiration into the two components: nitrogenase (+ NH₃metabolism) respiration and nodule growth and maintenance respiration,indicated that the latter efflux accounted for c. 20% of nodulerespiration while N₂ fixation was increasing and new noduletissue was being formed. When nodule growth ceased and N₂ fixationdeclined, this component of respiration also declined. The respiratorycost of nitrogenase activity plus the associated metabolismof NH₃ varied between 11 mg CO₂ mg^–1 N during vegetativeand early reproductive growth, to 12.5 mg CO₂ mg^–1 N duringthe later stages of pod development. Key words: N2 fixation, Respiration, Nodules, Nitrogenase     

Antagonism Between Malate and Ethylene in the Regulation of Avena sativa Coleoptile Elongation

Etiolated Avena sativa L. coleoptile sections were used to determinethe influence of C₂H₄ on in vivo and in vitro rates of CO₂ fixation,and to measure the influence of various permutations of C₂H₄,CO₂, and malate on growth. Whereas 1 mM malate or 320 µI^-1 CO₂ stimulated growth by approximately 100 per cent, inhibitionof growth by 10-8 µ I_-1 C₂H₄ was substantial only in thepresence of malate or CO₂ The increase in growth rate in responseto these two agents was eliminated by the simultaneous applicationof C₂H₄. The in vivo rate of dark [¹⁴C]bicarbonate fixationand in vitro enzymic assays of fixation were not measurablyinhibited by C₂H₄. These results are discussed in the lightof evidence which indicates that CO₂-stimulated growth is mediatedby dark fixation. The data do not support the view that C₂H₄inhibition of growth results from an inhibition of fixation,but suggests that C₂H₄ may inhibit some step in the processby which malate stimulates growth.     

Light-induced changes in membrane potential in Spirogyra

Spirogyra cells exhibited changes in membrane potential whenthey were exposed to light. Cells made chloroplast-free didnot show any light-induced potential change (LPC) upon illuminationwith white light and also monochromatic red (680 nm) and farred (720 nm) light. LPC was observed when the cell containedonly a small fragment of chloroplast, whether the cell had anucleus or not. The magnitude of LPC depended on the amountof chloroplast in the cell. DCMU at 10^–5 M, CCCP at 10^–5 M and DNP at 10^–4M at pH 5.5 suppressed LPC, while CCCP at 1–5 ? 10^–6M, NH₄Cl at 5 ? 10^–2 M and DNP at 10^–4 M at pH 7.0stimulated LPC. PMS at 10^–4 M stimulated LPC and couldinduce LPC which was completely inhibited by DCMU. These factssuggest that LPC is related to noncyclic and cyclic electronflows. The influences of light and dark conditions and various metabolicinhibitors (DCMU, DNP, CCCP, NH₄Cl) on ATP level have been investigated.No significant difference in the ATP level was observed betweencells in the light and dark. DNP at 10^–4 M (pH 5.5) andCCCP at 5 ? 10^–6 M decreased the ATP level significantly,while DCMU and NH₄Cl only slightly. Good correlation was notfound between the total ATP level and LPC in Spirogyra. LPC occurred even when the external medium contained only asingle salt such as KCl, NaCl or CaSO₄. LPC was also recorded in chloroplasts in situ and in vitro.The mode of LPC of chloroplasts was quite different from thatof the cell. On illumination, the chloroplast potential changedvery rapidly and transiently in the positive direction thenrecovered spontaneously to almost the original potential level. Possible causes of LPC are discussed in relation to the electrogenicion pump. ¹ Present address: Department of Botany, Faculty of Science,University of Tokyo, Hongo, Bunkyo, Tokyo 113, Japan. (Received November 9, 1977; )     

Involvement of the Accumulation of Glutamine in the Initiation of Adventitious Buds in Stem Segments of Torenia

Active meristematic divisions in stem segments of Torenia culturedin vitro can be induced in the epidermis by application of cytokininor the calcium ionophore A23187 [GenBank] , resulting in the differentiationof adventitious buds. Endogenous free glutamine accumulatedat a high concentration in the epidermal tissues during theearly stages of such cultures. The accumulation of glutaminewas caused by an increase in glutamine synthetase (GS) activity,and the increase of GS activity was suppressed by the applicationof some inhibitors of GS activity, mRNA synthesis, protein synthesis,or calmodulin. Incorporation of these inhibitors into the culturemedium also inhibited initiation of adventitious buds. The inhibitoryeffect of an inhibitor of GS, methionine sulfoximine (MSX),was apparent only at the very begining of the culture, and theeffect could be overcome by the simultaneous addition of glutamine.The inhibitory action of MSX on initiation of buds seemed tobe caused by an accumulation of ammonium ions. Reduction inlevels of NH₄NO₃ in or its elimination from the culture mediumstimulated the initiation of adventitious buds. Therefore, boththe accumulation of glutamine and the reduction in levels ofammonium ions seem to play a role in the initiation of adventitiousbuds in stem., segments of Torenia. ¹Present address: Faculty of Agriculture, University of Saga,Honjo-cho, Saga, Saga, 840 Japan. (Received October 3, 1988; Accepted March 9, 1989)     

Effects of Prolonged Darkness on the Sensitivity of Leaf Respiration to Carbon Dioxide Concentration in C3and C4Species

Predicting responses of plant and global carbon balance to theincreasing concentration of carbon dioxide in the atmosphererequires an understanding of the response of plant respirationto carbon dioxide concentration ([CO₂]). Direct effects of thecarbon dioxide concentration at which rates of respiration ofplant tissue are measured are quite variable and their effectsremain controversial. One possible source of variation in responsivenessis the energy status of the tissue, which could influence thecontrol coefficients of enzymes, such as cytochrome-c oxidase,whose activity is sensitive to [CO₂]. In this study we comparedresponses of respiration rate to [CO₂] over the range of 60to 1000 µmol mol^-1in fully expanded leaves of four C₃andfour C₄herbaceous species. Responses were measured near themiddle of the normal 10 h dark period, and also after another24 h of darkness. On average, rates of respiration were reducedabout 70% by the prolonged dark period, and leaf dry mass perunit area decreased about 30%. In all species studied, the relativedecrease in respiration rate with increasing [CO₂] was largerafter prolonged darkness. In the C₃species, rates measured at1000 µmol mol^-1CO₂averaged 0.89 of those measured at 60µmol mol^-1in the middle of the normal dark period, and0.70-times when measured after prolonged darkness. In the C₄species,rates measured at 1000 µmol mol^-1CO₂averaged 0.79 of thoseat 60 µmol mol^-1CO₂in the middle of the normal dark period,and 0.51-times when measured after prolonged darkness. In threeof the C₃species and one of the C₄species, the decrease in theabsolute respiration rate between 60 and 1000 µmol mol^-1CO₂wasessentially the same in the middle of the normal night periodand after prolonged darkness. In the other species, the decreasein the absolute rate of respiration with increase in [CO₂] wassubstantially less after prolonged darkness than in the middleof the normal night period. These results indicated that increasingthe [CO₂] at the time of measurement decreased respiration inall species examined, and that this effect was relatively largerin tissues in which the respiration rate was substrate-limited.The larger relative effect of [CO₂] on respiration in tissuesafter prolonged darkness is evidence against a controlling roleof cytochrome-c oxidase in the direct effects of [CO₂] on respiration.Copyright 2001 Annals of Botany Company Carbon dioxide, respiration, Abutilon theophrasti(L.), Amaranthus retroflexus(L.),Amaranthus hypochondriacus (L.), Datura stramonium(L.), Helianthus annuus(L.), Solanum melongena(L.), Sorghum bicolor(L. Moench), Zea mays     

Energy conversion and changes in physical parameters in chloroplasts and subchloroplast particles II. Energy conversion in chloroplasts and different types of subchloroplast particles

The light-induced absorbance change at 515 nm, light-inducedhydrogen ion uptake and ATP formation were compared in chloroplastsand different types of sonicated subchloroplast particles. Noparallel relationship among the activities for ATP formation,hydrogen ion uptake and the 515-nm change was observed in differenttypes of preparations. NH₄Cl inhibited ATP formation in chloroplastsbut had little effect on subchloroplast particles. In contrast,the light-induced hydrogen ion uptake was inhibited by NH₄Clin a similar manner. Tetraphenylboron (TPB), at 1 µM, inhibited ATP formationby about 30% in both chloroplasts and subchloroplast particles.In the presence of TPB, ATP formation in chloroplasts was stronglyinhibited by NHC₄Cl, but in subchloroplast particles the additionalinhibitory effect of NH₄Cl was small. A synergistic inhibitionof photophosphorylation by valinomycin plus NH₄Cl was much clearer.Although acceleration of the recovery of the 515-nm change byNH₄Cl or valinomycin was moderate, the 515-nm change virtuallydisappeared when NH₄Cl and valinomycin were added simultaneously. Although the membrane potential has a major role as the principaldriving force for ATP formation in subchloroplast particles,the simultaneous abolishment of the pH gradient and membranepotential may be required to uncouple ATP formation. ¹Present address: Fukuoka Women's University, Kasumigaoka, Fukuoka813, Japan. ²Present address: Ryukyu University, Naha, Okinawa 903, Japan. (Received February 5, 1974; )     

Environmental Influences on Carbon Recycling in a Terrestrial CAM Bromeliad, Bromelia humilis Jacq.

The effects of night-time temperature, leaf-to-air vapour pressuredeficit (VPD) and water stress on CO₂ recycling in Bromeliahumilis Jacq. grown under two light and nitrogen regimes wereinvestigated. At night-time temperatures above 30°C, integratednet dark CO₂ uptake was severely reduced and CO₂ for malatesynthesis was mainly derived from dark respiration. At 35°C,up to 84% of the CO₂ liberated by dark respiration was refixedinto malic acid. Below 30 °C only nitrogen deficient plantsshowed significant recycling. No significant differences wereobserved between high and low light grown plants in CO₂ recycling.A doubling of leaf-to-air VPD from 7-46 Pa kPa^–1 to 15.49Pa kPa^–1 resulted in a 2- to 20-fold decrease in leafconductance and about 50 to 65% reduction in integrated darkCO₂ uptake. However, about twice as much CO₂ was recycled atthe higher VPD as in the lower. Ten days of water stress resultedin 80 to 100% recycling of respiratory CO₂. Under high VPD andwater stress treatments, the amount of water potentially savedthrough recycling of CO₂ reached 2- to 6-fold of the actualtranspiration. In general, nitrogen deficient plants had higherper cent recycling of respiratory CO₂ in response to high night-timetemperature, increased VPD or water stress. The results emphasizethe ecological relevance of carbon recycling in CAM plants. Key words: Bromelia humilis, CAM, PPFD, dark respiration, temperature, VPD, water stress     

Ammonia Induces Starch Degradation in Chlorella Cells

When ammonia was added to cells of Chlorella which had fixed¹⁴CO₂ photo synthetically, ¹⁴C which had been incorporated intostarch was greatly decreased. A similar effect was observedwhen potassium nitrate and sodium nitrite were added. The ammonia-induceddecrease in ¹⁴C-starch was observed in all species of Chlorellatested. With cells of C. vulgaris 11h, most of the radioactivityin starch was recovered in sucrose, indicating that ammoniainduces the conversion of starch into sucrose. The percent of¹⁴C recovered in sucrose differed from species to species andpractically no recovery in sucrose was observed in C. pyrenoidosa.In most species tested, the enhancing effects of blue lightand ammonia on O₂ uptake as well as the ammonia effect on starchdegradation were greater in cells which had been starved inphosphate medium in the dark than in non-starved cells. In contrast,the enhancing effect of ammonia on dark CO₂ fixation was muchgreater in non-starved cells. C. pyrenoidosa was unique in thatblue light did not show any effect on its O₂ uptake. (Received August 15, 1984; Accepted November 16, 1984)     

The Respiration of Mature Field Bean (Vicia faba L.) Leaves During Prolonged Darkness

Dark respiration in attached and detached mature leaves of thefield bean (Vicia faba L.) was studied whilst leaves experiencedup to 60 h of darkness. The results showed: (1) the initialrespiration rate to vary according to the irradiance duringthe previous photoperiod; (2) the dark respiration rate (perunit area) of attached leaves to be essentially constant duringa normal 12 h night although there was a rapid loss in leafd. wt during this time; (3) after 12 h, the respiration rateof attached leaves decayed to an asymptotic value at about 36h; (4) the respiration rate of leaves detached at the end ofthe photoperiod and maintained in the dark on deionised water,decayed only after 36 h of darkness; (5) there was no differencebetween the respiration rate of attached and detached leavesduring the normal 12 h night. It is concluded that the dark respiration of attached fieldbean leaves is intially related to the synthesis and translocationof sucrose in addition to maintenance. After about 36 h, whenthe rate of CO₂ efflux is more or less steady, the CO₂ effluxreflects the intensity of maintenance processes only. The maintenancerespiration rate (determined after 60 h in the dark) rangedfrom 062 to 151 mg CO₂ (g d. wt)^–1 h^–1 but wasrelatively unaffected by several applied treatments. Vicia faba L., field bean, respiration, maintenance, nitrate, non-structural carbohydrate, export     

Non-Autotrophic CO2 Fixation and Drought Tolerance in Mosses

Cratoneuron filicinum, a drought-sensitive moss, and Tortularuralis, a drought-tolerant moss, fix CO₂ non-autotrophicallyat a rate of about 1.2 and 2.2 µmol h^–1 g^–1dry wt. respectively. During drying, T. ruralis fixes CO₂ atan undiminished rate until the tissue loses about 60% of theinitial fresh weight. Thereafter, CO₂ fixation rapidly declinesto zero. Dark CO₂ fixation by C.filicinum declines steadilyduring the dehydration period. On rehydration, dark CO₂ fixationis resumed immediately in T. ruralis but not in C.filicinum.When dried T. ruralis is equilibrated with an atmosphere ofnearly 100% relative humidity, its weight increases to about40% of the original fresh weight and dark CO₂ fixation resumesat a rate about 60% of the fresh moss. In C.filicinum thereis only a small increase in weight and little CO₂ fixation inthe dark. The non-autotrophically fixed carbon, in both mossesstudied, is incorporated into amino acids (more than 60% ofthe total, mainly into aspartate, alanine and glutamate) andorganic acids (less than 40% of the total, mainly into malate).It is suggested that on rehydration immediate availability ofNADPH, known to be produced by transhydrogenation from NADHduring dark CO₂ fixation, may be an important factor in therepair of drought-induced cellular damage by reductive biosynthesisof membrane components and other cellular constituents. Key words: Mosses, Dehydration, Rehydration, Dark CO₂ fixation, Amino acids, Organic acids, NADPH, Drought tolerance.     

The regulation of ammonia uptake in Chara australis

The regulation of ammonia uptake was investigated in internodalcells of the freshwater alga Chara australis. Ammonia uptakewas estimated by monitoring (i) its depletion from the bathingsolution, (ii) the uptake of radiolabelled methylamine, an analogueof ammonia, and (iii) depletion of ammonia in the unstirredlayer with the microelectrode ion-flux estimation technique(MIFE). Distribution of methylamine (¹⁴CH₃NH₃⁺) between thevacuole and cytoplasm was estimated with efflux analysis. Whencells were bathed continuously in solutions containing ammoniaor methylamine, the uptake rates of both amines decreased over12 to 48 h despite the continuing existence of a large electrochemicalgradient favouring influx of the NH⁺₄ and CH₃NH⁺₄ cations. Treatmentwith 1.0 to 10.0 mM MSX, an inhibitor of glutamine synthetase,caused the internal ammonia concentration to rise and reducedthe subsequent uptake of ammonia and methylamine by up to 70%within 2 h. These results suggest that the permease facilitatingNH⁺₄/CH₃NH⁺₄ influx is under feedback or kinetic regulationfrom either internal ammonia or an intermediate of nitrogenassimilation. Treatment with metabolic inhibitors (CCCP, azide and DCMU) andsome weak acids (DMO and butyric acid) for 30 to 60 min inhibitedmethylamine uptake, but the changes in the electrical potentialdifference across the plasma membrane could not account forthe magnitude of inhibition. The rate of cytopiasmic streaming,which is an indicator of the cellular ATP concentration in Chara,was inhibited by many of these treatments. However, under certainconditions of external pH and concentration, butyric acid couldreversibly inhibit ammonia and methylamine uptake without affectingcytoplasmic streaming, demonstrating that a decrease in cytoplasmicATP concentration was not responsible for the inhibition. Theeffect of butyric acid was rapid, causing a 60% inhibition ofuptake in 15 min. We conclude that weak acids can inhibit theNH⁺₄/CH₃NH⁺₄ permease by acidifying the cytoplasm and suggestthat this may also explain the effects of the metabolic inhibitorson ammonia and methylamine uptake. Key words: Ammonia, methylamine, uptake, regulation, Chara     

Reduction of Respiration by High Ambient CO2 and the Resulting Error in Measurements of Respiration Made with O2 Electrodes

Respiration rates of Lemna gibba fronds and Orobanche aegyptiacaand Lactuca sativa seedlings, were measured with a Clark typeoxygen electrode in the presence or absence of a carbon-dioxideabsorber (KOH) in the gas phase. Measured respiration ratesin the presence of KOH were 17-34% higher than in its absence.The suppression of respiration by high CO₂ concentrations, [CO₂],was confirmed by parallel studies of CO₂ efflux, made by infraredgas spectrometry. These results are consistent with other reportsof reduced rates of respiration at high [CO₂]. Measurements of respiration quotients of Lemna and Lactuca weremade at 0 and 100 Pa [CO₂]. Results did not support the possibilityof induced dark fixation of CO₂ at the ambient atmospheric [CO₂]predicted for the next century (35-100 Pa). It is concluded that the numerous reports of respiration measurementsmade with O₂ electrodes, in the absence of a CO₂ absorber, maycontain a significant errorCopyright 1993, 1999 Academic Press Lemna gibba, Lactuca sativa, Orobanche aegyptiaca, CO₂ accumulation, O₂ electrode, respiration, dark CO₂ fixation, respiration quotient, atmospheric CO₂     

Effects of Elevated Carbon Dioxide Concentration in the Dark on the Growth of Soybean Seedlings

Previous work has shown that elevated carbon dioxide (CO₂) concentrationsin the dark reversibly reduce the rate of CO₂ efflux from soybeans.Experiments were performed exposing soybean plants continuallyto concentrations of 350 or 700 cm³ m^-3 for 24 h d^-1, or to350 during the day and 700 cm³ m^-3 at night, in order to determinethe importance of the reduced rate of dark CO₂ efflux for plantgrowth. High CO₂ applied only at night conserved carbon andincreased dry mass during initial growth compared with the constant350 cm³ m^-3 treatment. Long-term net assimilation rate was increasedby high CO₂ in the dark, without any increase in daytime leafphotosynthesis. However, leaf area ratio was reduced by thedark CO₂ treatment to values equal to those of plants continuallyexposed to the higher concentration. From days 14-21, leaf areawas less for the elevated night-time CO₂ treatment than foreither the constant 350 or 700 cm³ m^-3 treatments. For the days7-21-period, relative growth rate was significantly reducedby the high night CO₂ treatment compared with the 350 cm³ m^-3continuous treatment. The results indicate that some functionallysignificant component of respiration was reduced by the elevatedCO₂ concentration in the dark.Copyright 1995, 1999 AcademicPress Glycine max L. (Merr.), carbon dioxide, plant growth, respiration