Effect of Ions on the Orientation of Cortical Microtubules in Spirogyra Cells

Effects of ions on the orientation of cortical micro-lubules(MTs) in Spirogyra cells were studied. After depo-lymerizalionwith amiprophos-methyl (APM), MTs were allowed to reorganizein NaCI solutions of various concentrations. As the concentrationof NaCI increased, the frequency of cells that had oblique MTsincreased. When cells in NaCI solution were transferred intoartificial pond water (APW) and incubated for 6 h, all the MTschanged to become transverse to the longitudinal axis of thecell. KC1 and MgCl₂ also had effects on the orientation of MTs.However, NH₄Cl, CaCl₂;, CoCl₂, and Co(NO₃)₂ did not show anyeffect. These results suggest that Na⁺, K⁺, and Mg²⁺have effectson MT orientation and that NH⁺₄, Ca²⁺, Co²⁺, Cl^–, andNO^–₃ have little effect. When MTs were reorganized ineither NaCl or KCl solutions, all the oblique MTs were organizedinto an S-helix. In contrast, some of the oblique MTs were foundas a Z-helix in the cells incubated in MgCl₂ or mannitol solutions.These results suggest that effects of Na⁺ and K⁺ on the orientationof MTs are not the same as those of Mg²⁺ and mannitol. Theseresults provide the first evidence that ions are involved inthe orientation of MTs in algae. (Received January 27, 1998; Accepted August 10, 1998)     

Calcium-Sensitivity of Cortical Microtubules in the Green Alga Mougeotia

Membrane ghosts were prepared from protoplasts of the greenalga Mougeotia, and the Ca²⁺-sensitivity of microtubules onthe ghosts was examined. Microtubules on the protoplast ghosts were not depolymerizedby 3 min treatment with 1 mM Ca²⁺. As the treatment was prolonged,some depolymerization of microtubules became evident, but evenafter 10 min about 50% of the ghosts showed no depolymerization.Ca²⁺ introduced into intact protoplasts seemed to be ineffectivein depolymerizing microtubules; abundant microtubules were presenton membrane ghosts prepared from protoplasts which had beentreated with 2x10^–5M Ca²⁺-ionophore A23187 [GenBank] plus 1 mM Ca²⁺for 20 or 30 min. Neither 3 min treatment with 0.2% Triton X-100 nor with 1 mMCa²⁺ solution containing 5 min MgSO₄ and 100 mM KCl caused depolymerisationof microtubules on protoplast ghosts. However, when given successively,these treatments caused complete depolymerization of microtubules. These results suggest that Mougeotia microtubules are stableto Ca²⁺ and that the stability is conferred by a microtubule-associatedfactor which can easily be removed by Triton X-100 treatment. (Received July 19, 1985; Accepted October 25, 1985)     

Transport and Fixation of Inorganic Carbon during Photosynthesis in the Cells of Anabaena Grown under Ordinary Air. II. Effect of Sodium Concentration during Growth on the Induction of Active Trasport System for IC

The requirement of sodium for growth of Anabaena variabilisM3 was investigated under low (0.04%) and high (1.5 or 5%) CO₂conditions. The growth rates under both conditions were stronglyaffected by NaCl concentrations up to 0.5 mM in the medium.In the presence of 40 µM NaCl, the cells were not ableto grow under a low CO₂ condition, but were able to grow undera high CO₂ condition. The sodium requirement for growth wasdependent on pH: in the Na⁺-deficient condition, cells couldgrow at pH6.8, while no growth occurred at pH 8.2, suggestingthat the requirement of Na⁺ for growth observed in the low CO₂condition can be substituted for by a lower pH. In the presence of 20 mM NaCl at pH 7.8, ¹⁴CO₂ as well as H¹⁴CO₃^–were actively transported into the cells which had been grownin air. In contrast, the transport of both of these inorganiccarbon (IC) species was suppressed under the Na⁺-deficient condition.These results suggest that sodium is required for the stimulationof transport of IC during photosynthesis. This is one of thereasons why Na⁺ is required for the growth of Anabaena underordinary air and alkaline conditions. (Received September 27, 1986; Accepted March 26, 1987)     

Effects of Na2SO4, K2SO4 and KCl on Growth and Ion Uptake of Callus Cultures of Vaccinium corymbosum L. cv. Blue Crop

Non-selected and Na₂SO-, K₂SO₄- or KCl-selected callus culturesof Vaccinium corymbosum L. cv. Blue Crop were grown on mediasupplemented with 0, 25 and 50 mM Na₂SO₄ (non-selected and Na₂SO(-selectedonly), 0, 25 and 50mMK₂SO₄ (non-selected and K₂SO₄-selectedonly) or 0, 50 and 100 mM KCl (non-selected and KCl-selectedonly). On all media, growth of selected callus (on a fresh-weightor dry-weight basis) was greater than that of non-selected callus,and selected callus grew optimally on the level and type ofsalt on which it was selected. Selected callus was friable andmaintained a higher f. wt:d. wt ratio. Tissue water potentialin selected callus was more negative than in non-selected callus. Flame photometry and chloridometry showed Na⁺, K⁺ and Cl^–accumulated in callus to concentrations equal to or greaterthan the initial concentration in the medium. Turbidometry showedthat tissue SO₄^2- concentration was lower than the concentrationin the medium. In most cases selected callus accumulated moreNa⁺, K^sup, SO₄^2– or Cl^– than non-selected callus.Vacuolar ion concentration was measured by electronprobe X-raymicroanalysis, and on most media selected callus had highervacuolar ion concentrations than non-selected callus. SO₄^2–and Cl^– were accumulated in the vacuoles at concentrationshigher than the external medium, but vacuolar Na⁺ concentrationdid not reach external concentration on Na₂SO₄ and on potassiumsalts was maintained between 12 and 17 mM. Vacuolar K⁺ concentration(approx. 142–191 mM on no salt) decreased on Na₂SO₄ andincreased on K₂SO₄ and KCl. There was no precise correlation between total or specific ionaccumulation (Na⁺, K⁺, SO₄^2– and Cl^– and fresh-weightyield. Results suggest that selection results in adaptationin response to decreased water potential of the medium. Vaccinium corymbosum, blueberry, electronprobe X-ray microanalysis, callus, in vitro selection, salt tolerance, KCl, K₂SO₄, Na₂SO₄     

Effect of chronically elevated CO2 on CA1 neuronal excitability

To study the effect of chronically elevated CO₂ on the excitability and function of neurons, we exposed mice to 7.5–8% CO₂ for 2 wk (starting at 2 days of age) and examined the properties of freshly dissociated hippocampal neurons. Neurons from control mice (CON) and from mice exposed to chronically elevated CO₂ had similar resting membrane potentials and input resistances. CO₂-exposed neurons, however, had a lower rheobase and a higher Na⁺ current density (580 ± 73 pA/pF; n = 27 neurons studied) than did CON neurons (280 ± 51 pA/pF, n = 34; P < 0.01). In addition, the conductance-voltage curve was shifted in a more negative direction in CO₂-exposed than in CON neurons (midpoint of the curve was –46 ± 3 mV for CO₂ exposed and –34 ± 3 mV for CON, P < 0.01), while the steady-state inactivation curve was shifted in a more positive direction in CO₂-exposed than in CON neurons (midpoint of the curve was –59 ± 2 mV for CO₂ exposed and –68 ± 3 mV for CON, P < 0.01). The time constant for deactivation at –100 mV was much smaller in CO₂-exposed than in CON neurons (0.8 ± 0.1 ms for CO₂ exposed and 1.9 ± 0.3 ms for CON, P < 0.01). Immunoblotting for Na⁺ channel proteins (subtypes I, II, and III) was performed on the hippocampus. Our data indicate that Na⁺ channel subtype I, rather than subtype II or III, was significantly increased (43%, n = 4; P < 0.05) in the hippocampi of CO₂-exposed mice. We conclude that in mice exposed to elevated CO₂, 1) increased neuronal excitability is due to alterations in Na⁺ current and Na⁺ channel characteristics, and 2) the upregulation of Na⁺ channel subtype I contributes, at least in part, to the increase in Na⁺ current density. sodium ion channels; oxygen deprivation     

Transport and Fixation of Inorganic Carbon during Photosynthesis in Cells of Anabaena Grown under Ordinary Air: III. Some Characteristics of the HCO3--Transport System in Cells Grown under Ordinary Air

In cells of cyanobacterium Anabaena variabilis grown under ordinaryair (low-CO₂ cells), the transport of both CO₂ and HCO₃^–was significantly enhanced by Na⁺. This effect was pronouncedas the external pH increased. When low-CO₂ cells were treatedwith an inhibitor of carbonic anhydrase (CA), only CO₂ transportbut not HCO₃^– transport, was inhibited. The initial rateof photosynthetic carbon fixation as a function of the concentrationof internal inorganic carbon (IC) was practically the same irrespectiveof whether CO₂ or HCO₃^– was externally supplied. Theseresults suggest that IC is actively transported through theplasma membrane in a form of HCO₃^– probably by some transporterand that the transmembrane Na⁺ gradient is involved in thisIC transport system. Free CO₂ may be hydrated by CA to HCO₃^–and then transported to the cells by this transporter. On the other hand, CO₂ is actively taken up by cells grown withair containing 5% CO₂ (high-CO₂ cells) though the enhancingeffect of Na⁺ was much smaller in high- CO₂ cells than in low-CO₂cells. The initial rate of fixation as a function of internal IC concentrationindicated that the rate of the carboxylation reaction of accumulatedIC is higher in I0W-CO₂ cells than in high-CO₂ cells. The studieswith ethoxyzolamide indicated that even in low-CO₂ cells, CAdoes not function inside Anabaena cells. These results suggestthat inside the low-CO₂ cells of Anabaena, some mediator(s)facilitates the transport of IC to RuBPCase. (Received January 23, 1987; Accepted April 24, 1987)     

Photolytic Decarboxylation of Carboxyl-14C-labelled Indol-3yl-acetic Acid in Leaves of the Apple Tree

The nature and rate of degradation of carboxyl-¹⁴C-labelledindol-3y1-acetic acid (IAA-[l-¹⁴C]) were studied in apple leaves.The labelled auxin was applied to the cut surface of the growingshoot after the apical part had been removed. The respiratoryCO₂ absorbed by chromatographic paper as Na₂CO₃ then freed byphosphoric acid was quantitatively measured by an internal gascounter. It was found that the concentration of ¹⁴CO₂ evolvedby leaves was 77 times higher in daylight than in darkness.The ratio of ¹⁴CO₂/CO₂ obtained from respiration from the uppersurface of leaf blades was two and seven times higher than thatfrom the lower surface after 15 and 30 h of daylight, respectively.No such differences were noticed in darkness. Similarly, thetotal radioactivity of leaf tissues tripled in daylight, presumablybecause of photosynthetic incorporation of radioactive CO₂ evolvedduring decomposition of LAA. These facts demonstrate the photolyticcharacter of auxin decarboxylation in apple leaves. Prolongeddarkness seemed to provoke a large metabolite withdrawal fromleaves and, to some extent, to protect auxin against decarboxylation.     

Changes in Photosystem Stoichiometry in Response to Environmental Conditions for Cell Growth Observed with the Cyanophyte Synechocystis PCC 6714

Changes in photosystem stoichiometry in response to shift ofenvironments for cell growth other than light regime were studiedwith the cyanophyte Synechocystis PCC 6714 in relation to thechange induced by light-quality shift. Following two environment-shiftswere examined: the shift of molecular form of inorganic carbonsource for photosynthesis from CO₂ to HCO₃^– (CO₂ stress)and the increase in salinity of the medium with NaCl (0.5 M)(Na⁺ stress). Both CO₂ and Na⁺ stresses induced the increasein PSI abundance resulting in a higher PSI/PSII stoichiometry.CO₂ stress was found to elevate simultaneously Cyt c oxidaseactivity (V_max). The feature was the same as that caused bylight-quality shift from preferential excitation of PSI to PSII(light stress) though the enhancement by either stress was smallerthan that by light stress. Under our experimental conditions,PSI/PSII stoichiometry appeared to increase at a fairly constantrate to the basal level even when the basal level had been differentlydetermined by the light-quality. Enhancing rates for PSI/PSIIstoichiometry and for Cyt c oxidase activity were also similarto each other. Since the two stresses affect the thylakoid electrontransport similarly to the shift of light-quality, we interpretedour results as follows: three environmental stresses, CO₂, Na⁺,and light stresses, cause changes in electron turnover capacityof PSI and Cyt c oxidase under a similar, probably a common,mechanism for monitoring redox state of thylakoid electron transportsystem. ¹On leave from Department of Biology, College of Natural Science,Kyngpook National University, Taegu 702-701, Korea. ₂Present address: Department of Marine Bioscience, Fukui Pre-fecturalUniversity, Obama, Fukui, 917 Japan.     

Effect of Sudden Salt Stress on Ion Fluxes in Intact Wheat Suspension Cells

Although salinity is one of the major problems limiting agriculturalproduction around the world, the underlying mechanisms of highNaCl perception and tolerance are still poorly understood. Theeffects of different bathing solutions and fusicoccin (FC),a known activator of plasma membrane ATPase, on plasma membranepotential (E_m) and net fluxes of Na⁺, K⁺and H⁺were studied inwheat suspension cells (Triticum aestivum) in response to differentNaCl treatments. E_mof cells in Murashige and Skoog (MS) mediumwas less negative than in cells exposed to a medium containing10 mM KCl + 0.1 m M CaCl₂(KSM) and to a basic salt medium (BSM),containing 1 m M KCl and 0.1 m M CaCl₂. Multiphasic Na⁺accumulationin cells was observed, peaking at 13 min after addition of 120m M NaCl to MS medium. This time scale was in good agreementwith net Na⁺flux changes measured non-invasively by moving ion-selectivemicroelectrodes (the MIFE system). When 120 m M NaCl was addedto all media studied, a quick rise of Na⁺influx was reversedwithin the first 20 min. In both 120 and 20 m M NaCl treatmentsin MS medium, net Na⁺efflux was observed, indicating that activeNa⁺transporters function in the plant cell response to saltstress. Lower external K⁺concentrations (KSM and BSM) and FCpre-treatment caused shifts in Na⁺fluxes towards net influxat 120 m M NaCl stress. Copyright 2000 Annals of Botany Company Sodium, potassium, proton, membrane potential, fusicoccin, salt stress, wheat, Triticum aestivum     

Beta-subunit of cardiac Na+-K+-ATPase dictates the concentration of the functional enzyme in caveolae

Previous studies showed the presence of a significant fraction of Na⁺-K⁺-ATPase -subunits in cardiac myocyte caveolae, suggesting the caveolar interactions of Na⁺-K⁺-ATPase with its signaling partners. Because both - and -subunits are required for ATPase activity, to clarify the status of the pumping function of caveolar Na⁺-K⁺-ATPase, we have examined the relative distribution of two major subunit isoforms (₁ and ₁) in caveolar and noncaveolar membranes of adult rat cardiac myocytes. When cell lysates treated with high salt (Na₂CO₃ or KCl) concentrations were fractionated by a standard density gradient procedure, the resulting light caveolar membranes contained 30–40% of ₁-subunits and 80–90% of ₁-subunits. Use of Na₂CO₃ was shown to inactivate Na⁺-K⁺-ATPase; however, caveolar membranes obtained by the KCl procedure were not denatured and contained 75% of total myocyte Na⁺-K⁺-ATPase activity. Sealed isolated caveolae exhibited active Na⁺ transport. Confocal microscopy supported the presence of ,-subunits in caveolae, and immunoprecipitation showed the association of the subunits with caveolin oligomers. The findings indicate that cardiac caveolar inpocketings are the primary portals for active Na⁺-K⁺ fluxes, and the sites where the pumping and signaling functions of Na⁺-K⁺-ATPase are integrated. Preferential concentration of ₁-subunit in caveolae was cell specific; it was also noted in neonatal cardiac myocytes but not in fibroblasts and A7r5 cells. Uneven distributions of ₁ and ₁ in early and late endosomes of myocytes suggested different internalization routes of two subunits as a source of selective localization of active Na⁺-K⁺-ATPase in cardiac caveolae. cardiac myocyte; caveolin; oligomer; ouabain; sodium pump     

Characterization of Photosynthetic 14Carbon Assimilation by Potamogeton lucens L.

Photosynthetic assimilation of exogenous ¹⁴CO₂ and H¹⁴CO₃^–by the aquatic angiosperm Potamogeton lucens L. is reported.Equivalent maximum rates of assimilation (1.5 µmol s^–1m^–2) were obtained in the presence of saturating levelsof ¹⁴CO₂ (1.0 mol m^–3, pH 5.3) or H¹⁴CO₃^– (1.5 molm^–3, pH, 9.2). Under subsaturating ¹⁴CO₂ levels, bothgaseous diffusion and H¹⁴CO₃^– transport were shown tooperate simultaneously, such that maximal photosynthetic rateswere established. An induction lag of approximately 3 min was observed when exogenous¹⁴CO₂ was assimilated. A longer lag of approximately 12 minwas required, however, before linear assimilation rates wereestablished when H¹⁴CO₃ acted as the carbon source. The light-activatedH¹⁴CO₃^– transport system was found to be quite labile.A brief (5 min) dark treatment returned the system to the inactivestate. Bicarbonate transport was shown to be competitively inhibitedby CO₃^2–ions. The possibility is discussed that this formof inhibition may be common to many HCO₃^– assimilators. Preliminary polar cation transport studies (from lower to upperleaf surface) indicated an almost exact one to one relationshipbetween the rates of Na⁺ influx and efflux and H¹⁴CO₃^–assimilation. The possible relationship(s) between these transportprocesses and the requirement for electrical neutrality is brieflydiscussed.     

A Comparative Study of the Influence of Salt Type and Concentration on 14CO2 Fixation in Potato Slices at 25{degrees} C and 0{degrees} C

Dark fixation of ¹⁴CO₂ was followed in potato disks under varyingsalt treatments at 0° C and 25° C. It is shown thatthe specific activity of the ¹⁴CO₂ supplied is heavily dilutedby endogenously produced CO₂ and that the apparently greaterfixation of ¹⁴CO₂, at 0° C as compared with that at 25 °C is due to the lower respiration rate at 0° C, with consequentlyless dilution of the ¹⁴CO₂. supplied. At 25° C organic acidformation in response to different salt treatments fulfils thecommon expectation, ¹⁴CO₂ fixation increasing in the presenceof K₂SO₄ and decreasing in CaCl₂ relative to that in KCl. Therole of organic acids in maintaining ionic balance within thecell at 25° C is thereby indicated but at 0° C organicacid adjustments did not follow the normal pattern. At 25°C but not at o° C increasing external concentration of KCIresulted in an increased level of ¹⁴CO₂ fixation.     

Na+-dependent HCO3- uptake into the rat choroid plexus epithelium is partially DIDS sensitive

Several studies suggest the involvement of Na⁺ and HCO₃^– transport in the formation of cerebrospinal fluid. Two Na⁺-dependent HCO₃^– transporters were recently localized to the epithelial cells of the rat choroid plexus (NBCn1 and NCBE), and the mRNA for a third protein was also detected (NBCe2) (Praetorius J, Nejsum LN, and Nielsen S. Am J Physiol Cell Physiol 286: C601–C610, 2004). Our goal was to immunolocalize the NBCe2 to the choroid plexus by immunohistochemistry and immunogold electronmicroscopy and to functionally characterize the bicarbonate transport in the isolated rat choroid plexus by measurements of intracellular pH (pH_i) using a dual-excitation wavelength pH-sensitive dye (BCECF). Both antisera derived from COOH-terminal and NH₂-terminal NBCe2 peptides localized NBCe2 to the brush-border membrane domain of choroid plexus epithelial cells. Steady-state pH_i in choroidal cells increased from 7.03 ± 0.02 to 7.38 ± 0.02 (n = 41) after addition of CO₂/HCO₃^– into the bath solution. This increase was Na⁺ dependent and inhibited by the Cl^– and HCO₃^– transport inhibitor DIDS (200 µM). This suggests the presence of Na⁺-dependent, partially DIDS-sensitive HCO₃^– uptake. The pH_i recovery after acid loading revealed an initial Na⁺ and HCO₃^–-dependent net base flux of 0.828 ± 0.116 mM/s (n = 8). The initial flux in the presence of CO₂/HCO₃^– was unaffected by DIDS. Our data support the existence of both DIDS-sensitive and -insensitive Na⁺- and HCO₃^–-dependent base loader uptake into the rat choroid plexus epithelial cells. This is consistent with the localization of the three base transporters NBCn1, Na⁺-driven Cl^– bicarbonate exchanger, and NBCe2 in this tissue. bicarbonate metabolism; BCECF; cerebrospinal fluid; acid/base transport; ammonium prepulse     

Enzymatic Inactivation of Extracellular Carbonic Anhydrase and its Effect on K1/2 (CO2) for Photosynthesis in Chlorella ellipsoidea C-27

The ratio of the extracellular to the intracellular activityof carbonic anhydrase (CA) in cells of Chlorella ellipsoideaC-27, adapted to low levels of CO₂ for 24 h (low-CO₂ cells),was about one to one. Treatment of intact cells with PronaseP inactivated about one-half of the extracellular CA activitywithout affecting photosynthetic activity. The CA activity incell homogenates and in cell-wall ghosts liberated during celldivision was completely inactivated by the same treatment. Pretreatmentwith Glycosidase mix, Chitosanase and Macerozyme enhanced theinactivation of the CA activity in intact cells. These resultssuggest that extracellular CA is evenly distributed throughoutthe whole cell-wall region. The apparent K_1/2 for dissolved inorganic carbon (DIC) in low-CO₂cells doubled when extracellular CA was inactivated by treatmentwith Pronase P, but the K_1/2 obtained was still one-half ofthat in high-CO₂ cells. Photosynthetic ¹⁴CO₂-fixation in low-CO₂cells was enhanced by acetazolamide, whereas H¹⁴CO₃^–-fixationwas suppressed. The results suggest that CO₂ is a dominant substrateutilized by cells and that HCO₃^– is utilized after conversionto CO₂. The present results show that both intracellular andextracellular CA contribute to the increase in affinity forDIC during photosynthesis in low-CO₂ cells of Chlorella ellipsoideaC-27. (Received May 7, 1990; Accepted July 18, 1990)     

Active Uptake of CO2 by the Diatom Navicula pelliculosa

Mass spectrometry has been used to investigate the transportof CO₂ in the freshwater diatom Navicula pelliculosa. The timecourseof CO₂ formation in the dark after addition of 100 mmol m^–3dissolved inorganic carbon (DIC) to cell suspensions showedthat no external carbonic anhydrase (CA) was present in thesecells. Upon illumination, cells pre-incubated at pH 75 with100 mmol m^–3 DIC, removed almost all free CO₂ from themedium at an initial rate of 285 µmol CO₂ mg^–1Chl h^–1. Equilibrium between HCO₃^– and CO₂ in themedium occurred rapidly upon addition of bovine CA, showingthat CO₂ depletion resulted from a selective uptake of CO₂ ratherthan an uptake of all inorganic carbon species. However, photosyntheticO₂ evolution rate remained constant after CO₂ had been depletedfrom the medium indicating that photosynthesis is sustainedprimarily by active HCO₃^– uptake. Treatment of cells with2-iodoacetamide (83 mol m^–3) completely inhibited CO₂fixation but had little effect on CO₂ transport since initialrates of CO₂ depletion were about 81% that of untreated cells.Transfer of iodoacetamide-treated cells to the dark caused arapid increase in the CO₂ concentration in the medium largelydue to the efflux of the unfixed intracellular DIC pool whichwas found to be about 194 times the concentration of that inthe external medium. These results indicate that Navicula pelliculosaactively takes up molecular CO₂ against a concentration gradientby a process distinct from HCO₃^– transport. Key words: Dissolved inorganic carbon, carbonic anhydrase, bicarbonate transport, CO₂ transport, mass spectrometry     

Membrane Potential Measurement of Protoplasts Isolated from Vigna mungo Hypocotyl Using a Fluorescent Probe, diS-C3-(5)

Membrane potentials of protoplasts isolated from Vigna mungohypocotyl segments were measured using the fluorescent probediS-C₃-(5). The fluorescence intensity changed in response tothe external K⁺ concentration. Membrane potential was estimatedto be inside negative (–85?8 mV at 0.1 mM KCl) from theNernst equation for K₊. The membrane potential was not affectedby DCCD (50 µM) or low temperature (5?C). Addition of0.5 mM Ca₂₊ to the protoplast suspension markedly depolarizedthe membrane potential, and subsequent EDTA treatment repolarizedit to the initial level. The Ca₂₊ effect on the membrane potentialmay be due to change in the permeability ratio of Cl^–to K₊. (Received December 16, 1986; Accepted April 22, 1987)     

Competitive inhibition of the response to Na+ by Ca2+ in water fibers of the frog glossopharyngeal nerve

Single water fibers of the frog glossopharyngeal nerve respondto low concentrations of CaCl₂ (1–2 mM) and to relativelyhigh concentrations of NaCl(>80 mM). However, stimulationby a mixture with a low concentration of CaCl₂ and relativelyhigh concentration of NaCl gives rise to only a small response,suggesting that the effects of Ca²⁺ and Na⁺ are mutually antagonistic.It has been reported that Na⁺ inhibits the response to Ca²⁺by competing with Ca²⁺ for a calcium receptor site (X_Ca; Kitadaand Shimada, 1980). In the present study, it was found tha Ca²⁺inhibited the response to Na⁺. Therefore, the sodium receptorsite (X_Na) responsible for the response to Na is different fromX_Ca. The inhibition of the response to Na⁺ by Ca²⁺ was examinedquantitatively on the assumption that the magnitude of the neuralresponse is proportinal to the amount of NaX_Na complex minusa constant (the threshold concentration of the NaX_Na complex).The results obtained indicate that Ca²⁺ competes with Na₊ forX_Na. The apparent dissociation constants for the NaX_Na complexand the CaX_Na complex obtained from the present study were 1.0M and 1.2 x 10^-3 M, respectively, X_Na as proposed here, doesnot represent simply a binding site for cations since therecan be competition for X_Na by an antagonistie cation. The highaffinity of X_Na for Ca²⁺ suggests that X_Na is a specific receptorsite involved in salt-taste reception. Since Mg²⁺ did not affectthe response to Na⁺, the affinity of X_Na for cations is notcharge-specific but is, rather, chemically specific. The presentresults indicate that both Ca²⁺ and Na⁺ have a dual action,being involved both in excitation and in inhibition, in waterfibers of the frog glossopharyngeal nerve.     

Utilization Modes of Inorganic Carbon for Photosynthesis in Various Species of Chlorella

Rates of CO₂ and HCC₃^– fixation in cells of various Chlorellaspecies in suspension were compared from the amounts of ¹⁴Cfixed during the 5 s after the injection of a solution containingonly ¹⁴CO₂ or H¹⁴CO₃^–. Results indicated that irrespectiveof the CO₂ concentration during growth, Chlorella vulgaris 11h and C. miniata mainly utilized CO₂, whereas C. vulgaris C-3,C. sp. K. and C. ellipsoidea took up HCO₃^– in additionto CO₂. Cells of C. pyrenoidosa that had been grown with 1.5%CO₂ (high-CO₂ cells) mainly utilized CO₂, whereas those grownwith air (low-CO₂ cells) utilized HCO₃^– in addition toCO₂. Cells that utilized HCO₃^– had carbonic anhydrase(CA) on their surfaces. The effects of Diamox and CA on the rates of CO₂ and HCO₃^–fixation are in accord with the inference that HCO₃^– wasutilized after conversion to CO₂ via the CA located on the cellsurface. CA was found in both the soluble and insoluble fractions;the CA on the cell surface was insoluble. Independent of the modes of utilization, the apparent K_m (NaHCO₃)for photosynthesis was much lower in low-CO₂ cells than in high-CO₂ones. The fact that the CA in the soluble fraction in C. vulgarisC-3 was closely correlated with the K_m(NaHCO₃) indicates thatsoluble CA lowers the K_m. ¹ Dedicated to the late Professor Joji Ashida, one of the foundersand first president of the Japanese Society of Plant Physiologists. ⁴ On leave from Research and Production Laboratory of Algology,Bulgarian Academy of Sciences, Sofia. (Received September 14, 1982; Accepted March 1, 1983)     

不同浓度NaCl和Na2CO3处理对菊芋幼苗光合及叶绿素荧光的影响

 以砂培菊芋(Helianthus tuberosus)幼苗作为试验材料,分别进行不同浓度NaCl (50、 100、150、200、250 mmol&;#8226;L^－1)和Na₂CO₃ (25、50、 75、100、125 mmol&;#8226;L^－1)胁迫处理,以1/2全营养液作为对照,处理7 d后研究NaCl和Na₂CO₃胁迫处理对菊芋幼苗叶片光合作用及叶绿素动力学 参数的影响。结果表明：1)在NaCl处理下,当浓度小于150 mmol&;#8226;L^－1时,增加了菊芋的叶绿素含量、净光合速率(Net photosynthetic rate, P_n)和气孔导度(Stomatal conductivity, G_s),对荧光参数PSⅡ的电子传递情况( F_m/F_o)、PSⅡ原初光能转换效率(F_v/F_m)、PSⅡ量子效率 (Actual quantum yield of PSⅡ under actinic irradiation,φPSⅡ)和光化学猝灭系数(Photochemical quenching coefficient, qP)和非 光化学猝灭系 数(Non-photochemical quenching coefficient, NPQ)没有显著影响,随着浓度的增加,各项生理指标与对照相比除了NPQ显著 增加,其余均显著降低;2)在Na₂CO₃胁迫处理下,随着Na₂CO₃浓度的增加,与对照相比菊芋幼苗叶绿素含量、P_n、G_s以及叶绿素a荧光诱导动力 学参数F_m/F_o、F_v/F_m、φPSⅡ和qP均显著降低,NPQ显著增加;3)就NaCl和Na₂CO₃相比而言,在相同Na⁺浓度情况下,处于Na₂CO₃胁迫下的菊芋 幼苗的叶绿素含量、P_n、G_s以及叶绿素a荧光诱导动力学参数F_m/F_o、F_v/F_m、φPSⅡ和qP下降幅度和NPQ的增加幅度均显著大于NaCl,这说明 NaCl和Na₂CO₃胁迫均对菊芋幼苗造成不同程度的伤害,但在相同Na⁺浓度情况下,Na₂CO₃的伤害程度大于NaCl。由此说明菊芋对盐的忍耐程度高 于碱。     

Transport and Fixation of Inorganic Carbon during Photosynthesis of Anabaena Grown under Ordinary Air. I. Active Species of Inorganic Carbon Utilized for Photosynthesis

Inorganic carbon transport during photosynthesis of cyanobacteriumAnabaena variabilis grown under ordinary air was investigatedby supplying ¹⁴CO₂ or H¹⁴CO₃^– solution to three differentstrains. Both CO₂ and HCO₃^– were accumulated within thealgal cells. In the cell suspension from which dissolved inorganiccarbon had been depleted by pre-illumination, CO₂ was transportedand accumulated faster than HCO₃^–. When the concentrationof HCO₃^– injected into the cell suspension of A. variabilisM3 was 25 times as high as that of CO2 (the expected ratio atequilibrium at pH 7.8), the initial rates of fixation of bothinorganic carbon species were practically the same. On the otherhand, when ¹⁴CO₂ or H¹⁴CO₃^– was added under steady statephotosynthetic conditions, both carbon species were transportedat similar rates. The ratio of fixed to transported carbon measuredafter the initial 5 s was only 23–27% regardless of thecarbon species supplied. This percentage is much lower thanthat reported for Chlorella cells. ¹ To whom reprint requests should be addressed (Received June 30, 1986; Accepted December 16, 1986)