Photosynthetic Characteristics of Chloroplasts Isolated from Euglena gracilis Z Grown Photoautotrophically

Photosynthetically competent chloroplasts were isolated fromcells of Euglena gracilis Z grown photoautotrophically in 1.5%CO₂. The isolated chloroplasts were intact and substantiallyfree from cytosolic, mitochondrial and microbody materials.The effects of some compounds on the activity of photosynthetic¹⁴CO₂ fixation were examined. The optimal pH and sorbitol concentrationwere 8.0 and 0.33 M, respectively. The chloroplasts requireda high level of P, (5 to 20 mM) for the maximal rate of photosynthesis.They were insusceptible to 10 mM of free Mg²⁺. ATP, ADP andAMP at 1 to 5 mM notably stimulated photosynthesis, althoughhigh concentrations of AMP were unfavorable. In the assay mediumdeveloped for this study, the chloroplasts exhibited photosyntheticactivity of 120µmoles-mg^–1 Chl-h^–1 at 30?C. Chloroplasts could also be isolated from cells grown under ordinaryair. The rate of photosynthetic ¹⁴CO₂ fixation at 1 mM NaH^l4CO₃was higher in these chloroplasts than in those isolated fromcells grown in 1.5% CO₂, whereas at 10 mM NaH^l4CO₃, the ratesof the two types of chloroplasts were nearly the same. Theseresults suggest that the CO₂ concentration given during growthof the algal cells affects the affinity for dissolved inorganiccarbon at the chloroplast level. (Received March 30, 1987; Accepted August 17, 1987)     

Rates of Photosynthesis in Characean Cells: I. PHOTOSYNTHETIC 14CO2 FIXATION BY NITELLA TRANSLUCENS

A study has been made of photosynthetic ¹⁴CO₂ fixation by isolated‘mature’ internodes of Nitella translucens. Experimentalconditions were similar to those used in studies of the ionicrelations of these cells. Maximum rates of photosynthesis were33–40µµmoles CO₂, fixed per cm² of surfacearea per second (equivalent to 12–15 /xmoles fixed permg chlorophyll per hour). ^l4CO₂ fixation was inhibited to thedark level by 3(3,4,dichlorophenyl)-1, 1-dimethylurea (at 0-6µM or 10µM) and by the uncoupler carbonyl cyanide-m-chlorophenylhydrazone(SµM). The presence of imidazole or ammonium sulphate(both of which uncouple ATP production in vitro) did not resultin an inhibition of 14CO2 fixation. These results are discussedin relation to published work on solute uptake by Nitella translucens.During photosynthesis there was rapid movement of ¹⁴C-labelledorganic compounds out of the chloroplasts. ¹⁴C-labelled sucrose,ammo-acids, and sugar phosphates were found in samples of vacuolarsap.     

Stomatal Responses to Applied ABA and CO2 in Epidermis Detached from Well-Watered and Water-Stressed Plants of Commelina communis L.

Epidermal strips from either well-watered or water-stressedplants of Commelina communis L. were subjected to a range ofABA concentrations (10^–6–10^–3 mol m^–3)in the presence (330 parts 10^–6 in air) or virtual absence(3 parts 10–6 in air) of CO₂. The stomatal response toCO₂ was greater in epidermis from water-stressed plants, althoughthere was a distinct CO₂ response in epidermis from well-wateredplants. Additions of ABA via the incubation medium had littleeffect on the relative CO₂ response. Stomata responded to ABAboth in the presence and virtual absence of CO₂, but the relativeresponse to ABA was greatest in the high CO₂ treatment. Whenwell-watered plants were sprayed with a 10^–1 mol m^–3ABA solution 1 d prior to use, the stomatal response of detachedepidermis to both CO₂ and ABA was very similar to that of epidermisdetached from water-stressed leaves. It is hypothesized thata prolonged exposure to ABA is necessary before there is anymodification of the CO₂ response of stomata.     

The Rate of Photosynthesis in Isolated Chloroplasts

Chloroplasts were isolated using aqueous and nonaqueous procedures.Aqueous chloroplasts lost approximately 50 per cent, of theirsoluble proteins during isolation. Nonaqueous chloroplasts retainedall their soluble enzymes, but lost their ability to performthe light reactions of photosynthesis. It was possible to reconstitutea chloroplast system of higher activity by adding soluble enzymesfrom nonaqueous chloroplasts to protein-deficient aqueous chloroplasts.The properties of the reconstituted chloroplast system wereas follows: 1. The CO₂ fixation rate of the reconstituted chloroplast system( 4 µM./. chlorophyll/hr.) was 3–4 times that ofthe aqueous chloroplasts ( I µM./. chlorophyll/hr.). Thefixation of aqueous chloroplasts isapparently limited in partby lack of soluble enzymes. 2. During light-fixation, the reconstituted chloroplast systemaccumulated PGA. This indicates that the reduction of PGA totriosephosphate is a rate-limiting step in this system. 3. It was possible to increase the CO₂ fixation to 12 µM.CO₂/mg. chlorophyll/ hr. by addition of ATP and TPNH to thesystem, but the reduction of PGA was still rate-limiting. 4. Further increase in the fixation rate was obtained by concentratingthe reaction mixture. Part of the striking differences of theCO₂-fixing capabilities of chloroplasts in vivo and in vitrois caused by dilution effects. Extrapolation of the dilutioneffect to the protein concentration which exists in chloroplastsyields a CO₂ fixation rate of approximately 30 µM./mg.chlorophyll/hr. 5. Inhibitors which are located in vivo outside the chloroplastsaffect the CO₂ fixation in vitro. 6. Under consideration of the examined factors which influencethe CO₂ fixation of isolated chloroplasts, it is possible toraise the fixation from approximately 1 per cent, to at least15 per cent, of the fixation in vivo.     

Protoplasts as a tool for isolating functional chloroplasts from leaves

Leaf protoplasts from various grasses can be used for isolatingchloroplasts with high photosyndietic activity. The protoplastswere stable for more than 20 hr during which time chloroplastscould be isolated from protoplasts without any loss of originalCO₂ fixation capacity (100–157 µmoles/mg chl-hr).Using Triticum aestiuum to optimize assay conditions, the pHoptimum for CO₂ fixation by the chloroplasts isolated from protoplastswas between 8.2 and 8.6. Magnesium (0.75 mM) was required formaximum CO₂ fixation by the isolated chloroplasts and sodiumascorbate in the medium allowed a more linear increase in CO₂fixation with time. Based upon ¹⁴CO₂ fixation and ferricyanide-dependentoxygen evolution as criteria of intactness, chloroplasts fromprotoplasts exhibited a high degree of intactness compared tothose obtained by mechanical grinding. Chloroplasts isolatedfrom grass leaves by mechanical grinding had a relatively lowcapacity for endogenous CO₂ fixation and required addition ofribose-5-phosphate and ADP for maximum activity. (Received September 8, 1975; )     

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; )     

Stomatal Functioning of In Vitro and Greenhouse Apple Leaves in Darkness, Mannitol, ABA, and CO2

Leaves from in vitro and greenhouse cultured plants of Malusdomestica (Borkh.) cv. Mark were subjected to 4 h of darkness;4 h of 1 M mannitol induced water stress; 1 h of 10^–4M to 10^–7 M cis-trans abscisic acid (ABA) treatment; 1h of 0.12% atmospheric CO₂. Stomatal closure was determinedby microscopic examination of leaf imprints. In all treatments,less than 5% of the stomata from leaves of in vitro culturedplants were closed. The diameter of open stomata on leaves fromin vitro culture remained at 8 µm. In contrast, an averageof 96% of the stomata on leaves of greenhouse grown plants wereclosed after 4 h in darkness; 56% after 4 h of mannitol inducedwater stress; 90% after 1 h of 10^–4 M ABA treatment; 61%after 1 h in an atmosphere of 0.12% CO₂. Stomata of in vitroapple leaves did not seem to have a closure mechanism, but acquiredone during acclimatization to the greenhouse environment. Thelack of stomatal closure in in vitro plants was the main causeof rapid water loss during transfer to low relative humidity.     

Delay in the Response of Stomata to Abscisic Acid in CO2-free Air

Abscisic acid (10^–5 M) was fed via their petioles to leavesdetached from well watered plants of Xanthium strumartum, whilethe intercellular spaces were flushed with air of known CO₂content. A closing response to ABA occurred in the presenceor absence of CO₂, and the stomata responded to CO₂ whetheror not ABA was supplied to the leaves. A factorial experimentrevealed no interaction between CO₂ and ABA, and suggested thattheir effect on the rate of closure was purely additive. Theonly evidence of interdependence between the two corn poundswas a delay in the response to ABA in C0 air, which was moremarked in a high light intensity. A hypothesis which is consistentwith the data is that ABA induces stomatal closure by interferingwith the energy supply required for the active transport processeson which guard cell turgor depends. The inhibitory action ofABA takes longer in CO₂-free air because, in the absence ofCO₂ fixation, energy is available from chioroplasts as wellas mitochondria.     

Photosynthesis by developing embryos of oilseed rape (Brassica napus L.)

The aim of this study was to assess the photosynthetic potentialof developing seeds of oilseed rape (Brassica napus L.) andto compare photosynthetic properties of embryo plastids withthose of leaf chloroplasts from the same species. Measurementsof CO₂-dependent O₂ evolution show that developing seeds ofB. napus are photosynthetically active in vitro. Essentially,all of the photosynthetic activity of the developing seed isaccounted for by the embryo. The rate of photosynthesis by developingembryos increased until the onset of desiccation, after whichit declined, so that by maturity embryos were no longer photosyntheticallyactive. Photosynthetic activity was positively correlated withchlorophyll content throughout development. Comparison of thephotosynthetic characteristics of leaf and embryo chloroplastsrevealed that rates of uncoupled electron transport were 2.5-foldgreater in those from the embryo. Light-saturated rates of CO₂-dependentO₂ evolution, per unit chlorophyll, and CO₂ saturation pointswere similar for chloroplasts from both tissues. However, light-saturationpoints and chlorophyll a/b ratios were lower for embryo thanfor leaf choroplasts. Embryos and embryo chloroplasts also containedconsiderably less ribulose 1,5-bisphosphate carboxylase/oxygenaseprotein per unit total protein, than leaves. Although excisedembryos were capable of high rates of CO₂-dependent O₂ evolution(90–100 mol mg^–1 chlorophyll h^–1) under asaturating photosynthetic photon flux density (PPFD), low transmittanceof light through the silique wall (30%), together with the highPPFD required to achieve light compensation points in developingseeds (500 mol m^–2 s^–1), suggests that photosynthesisin vivo is unlikely to make a net contribution to carbon economyunder normal environmental conditions. Key words: Embryo, development, photosynthesis, chloroplast, Brassica napus L.     

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)     

Chloroplast ultrastructure of sugar beet (Beta vulgaris L.) cultivated in normal and elevated CO2 concentrations with two contrasted nitrogen supplies

Sugar beet (Beta vulgaris L., cultivar Celt) plants were grownunder simulated field conditions in pots and supplied with adequateor deficient nitrogen (HN and LN, respectively) combined withtwo CO₂ concentrations, ambient (c. 350µmol mol^–1C0₂—AC), or elevated CO₂ (c. 600 µmol mol^–1CO₂—HC). Chloroplast structure in mesophyll palisade cellsof mature leaves (leaf number 19 in HN and 9 in LN), sampledat midday on 16 August 1993 was studied by transmission electronmicroscopy and quantified stereologically. The ultrastructureof palisade parenchyma chloroplasts was affected by the elevatedCO₂ concentration and strikingly affected by nitrogen supply.Chloroplast diameter (cross-sectional length) was slightly,but not significantly, greater in HC than AC treatments withinan N treatment, but was smaller in LN than HN; chloroplast cross-sectionalarea also increased with HC in both N treatments, but only significantlyso in LN. Elevated CO₂ reduced the proportion of total thylakoids(significant at 5% and 0.1% in HN and LN, respectively) dueto decreased granal thylakoids, but the proportion of inter-granal(stromal) thylakoid membranes was not affected compared to chloroplastsfrom plants grown with ambient CO₂. Chloroplast stroma increasedas a proportion of chloroplast volume with elevated comparedto ambient CO₂ with HN but not LN. Starch inclusions were notsignificantly different with elevated compared to ambient CO₂at HN, but the proportion of starch increased considerably atelevated compared to ambient C0₂ at LN, indicating an over-productionof assimilates. Plastoglobuli in chloroplasts increased withdeficient N, but decreased with elevated CO₂. Larger chloroplastswith a greater proportion of stroma, but a smaller proportionof granal thylakoids, suggest increased CO² assimilating capacityand decreased light harvesting/PSII capacity with elevated CO₂. Key words: Chloroplast, ultrastructure, elevated CO₂ concentration, nitrogen deficiency, sugar beet, Beta vulgaris     

Rates of Photosynthesis in Characean Cells: II. PHOTOSYNTHETIC 14CO2 FIXATION AND 14C-BICARBONATE UPTAKE BY CHARACEAN CELLS

Photosynthetic ¹⁴C fixation by Characean cells in solutionsof high pH containing NaH¹⁴CO₃ gave a measure of the abilityof these cells to take up bicarbonate (H¹⁴CO₃^–). Whereascells of Nitella translucens from plants collected and thenstored in the laboratory absorbed bicarbonate at 1–1.5µµmoles cm^–2 sec^–1, rates of 3–8µµmoles cm^–2 sec^–1 were obtained withN. translucens cells from plants grown in the laboratory. Influxesof 5–6 µµmoles cm^–2 sec^–1 wereobtained with Chara australis, 3–8 µµmolescm^–2 sec^–1 with Nitellopsis obtusa, and 1–5µµmoles cm^–2 sec^–1 with Tolypella intricata.It is considered that these influxes represent the activityof a bicarbonate pump, which may be an electrogenic process. In solutions of lower pH, H¹⁴CO₃^– uptake would be maskedby rapid diffusion of ¹⁴CO₂ into the cells: the four Characeanspecies fixed ¹⁴CO₂ at maximum rates of 30–40 µµmolescm^–2 sec^–1 (at 21° C).     

Effects of sulfite and pH on abscisic acid-dependent transpiration and on stomatal opening

In rice, alday, wheat and tobacco (Nicotiana tabacum L. Samsunand Samsun NN) plants which contained large amounts of ABA,the transpiration rate decreased rapidly with 2 ppm SO₂ fumigationand reached 20 to 65% of the initial level after 5- to 30-minexposure depending on their ABA contents. In the cases of broadbean and tobacco (N. glutinosa L.) with low ABA contents, therate slightly increased for 20 and 40 min, respectively, afterthe start of the fumigation and then decreased gradually. Thetranspiration rates of corn and sorghum, in spite of their extremelylow ABA contents, pronouncedly decreased with SO₂ fumigationand reached 65 and 50%, respectively, of the initial levelsafter 40-min exposure. Foliar application of 0.04 N HC1 to N.tabacum L. Samsun NN leaves remarkably depressed the transpirationrate, while the application of 0.04 M Na₂SO₃ decreased the rateonly to the same level as water treatment. Foliar applicationof either HCl or Na₂SO₃ to N. glutinosa L. leaves exerted littlechange in the transpiration rate. When 10^–4M ABA was appliedto broad bean leaves prior to HCl and Na₂SO₃ treatment, theirtranspiration rate was decreased by HCl, but not by Na₂SO₃ application.In sonicated epidermal strips peeled from broad bean leaves,Na₂SO₃ produced a slight increase in the stomatal aperture sizein the absence of ABA, but showed no effect in the presenceof ABA. The aperture size was identical in the pH range of 3.0to 7.0 in the incubation medium. In the presence of ABA in themedium, the aperture size was small in the acidic region ofpH with a minimal value at pH 4.0. ABA decreased the aperturesize at concentrations above 10^–9 M at pH 4.0 and 10^–6M at pH 7.0 in the medium. [2_–14C] ABA uptake by epidermalstrips was large in the acidic region, especially at pH 4.0. (Received February 28, 1980; )     

The Biological Properties of Cyclopenin and Cyclopenol

Cyclopenin (C₁₇H₁₄O₃N₂) and cyclopenol (C₁₇H₁₄O₄N₂), isolatedfrom an abberent strain of Penicillium cyclopium (NRRL 6233),significantly inhibited the growth of etiolated wheat (Triticumaestivum) coleoptile segments. The former inhibited at 10^–3and 10^–4 M, the latter at 10^–3 M. Cyclopenin producedmalformation of the first set of trifoliate leaves in bean (Phaseolusvulgaris) at 10^–2 M and necrosis and stunting in corn(Zea mays) at 10^–2 M. Cyclopenol induced no apparent effectsin bean or corn plants. Neither compound changed the growthor morphology of tobacco (Nicotiana tabacum) plants. Cyclopenininduced intoxication, prostration and ataxia in day-old chicksat 500 mg/kg, but they recovered within 18 hours. Cyclopenolwas inactive against chicks when dosed at levels up to 500 mg/kg. (Received October 11, 1983; Accepted December 15, 1983)     

Effect of Oxygen on Photosynthetic 14CO2 Fixation in Chroomonas sp. (Cryptophyta) II. Effects of Inhibitors, Uncouplers and an Artificial Electron Mediator on the Inhibition of 14CO2 Fixation by Anaerobiosis

In "air-grown" Chroomonas sp. cells, low concentrations of DCMU(less than 0.1 µM) could prevent the inhibition of ¹⁴CO₂fixation by anaerobiosis under light-saturating conditions (morethan 40 W.m^–2), with phenazine methosulfate showing asimilar effect. Antimycin A, carbonyl cyanide m-chlorophenylhydrazone(CCCP), and N,N'-dicyclohexylcarbodiimide strongly inhibitedanaerobic photosynthesis at concentrations which did not significantlyinhibit the rate under 2% O₂ at high light intensity (200 W.m^–2),although 0.2 µM CCCP stimulated the rate under 2% O₂ tosome extent. On the other hand, KCN inhibited the rate muchmore strongly under 2% O₂ than N₂, although it inhibited therate very strongly at concentrations above 5 µM both underN₂ and 2% O₂. These results suggest that the inhibition of photosynthetic¹⁴CO₂ fixation by anaerobiosis in this alga result from ATPdeficiency caused by over-reduction of electron carriers ofthe cyclic electron flow and that oxygen can prevent the over-reduction.Cyclic electron flow seems to be necessary to provide additionalATP for CO₂ reduction under anaerobic conditions, although itseems to be less necessary under aerobic conditions. (Received July 21, 1983; Accepted January 23, 1984)     

Uptake and Accumulation of Inorganic Carbon by a Freshwater Diatom

Colman, B. and Rotatore, C. 1988. Uptake and accumulation ofinorganic carbon by a freshwater diatom.—J. exp Bot 39:1025–1032. The mechanism of uptake of inorganic carbon and its accumulationhas been studied in the freshwater diatom Navicula pelliculosa.No external carbonic anhydrase could be detected, although itwas detected in cell extracts. The rate of photosynthetic O₂evolution, in media in the range pH 7.5–8.5, exceededthe calculated rate of CO₂ supply 2- to 5-fold, indicating thatHCO^–₃ was taken up by the cells. At an external pH of7.5, the internal pH, measured by ¹⁴C-dimethyloxazolidine-2,4-dione distribution between the cells and the medium, was pH7.6 in the light and pH 7.4 in the dark. Accumulation of inorganiccarbon was determined by the silicone oil centrifugation methodand inorganic carbon pools of 23.5 mol m^–3 were found,a concentration 21.6-fold that in the external medium. The resultsindicate an active accumulation of inorganic carbon againstpH and concentration gradients in this diatom, probably by activeHCO^–₃ uptake. Key words: Bicarbonate transport, carbon dioxide, carbonic anhydrase, CO₂ affinity, CO₂ concentrating mechanism, internal pH, Navicula pelliculosa     

Carbon Dioxide Fixation and Ribulose-1, 5-bisphosphate Carboxylase Activity in Intact Leaves of Sugar Beet Plants Exposed to Salinity and Water Stress

The influence of salinity in the growing media on ribulose-1,5-bisphosphate (RuBP) carboxylase and on CO₂ fixation by intactsugar beet (Beta vulgaris) leaves was investigated. RuBP carboxylase activity was mostly stimulated in young leavesafter exposure of plants for 1 week to 180 mM NaCl in the nutrientsolution. This stimulation was more effective at the higherNaHCO₂ concentrations in the reaction medium. Salinity also enhanced CO₂ fixation in intact leaves mostlyat rate-limiting light intensities. A 60 per cent stimulationin CO₂ fixation rate was obtained by salinity under 450 µEm^–2 s^–1. At quantum flux densities of 150 µEm^–2 s^–1 (400–700 nm) this stimulation was280 per cent. Under high light intensities no stimulation bysalinity was found. In contrast, water stress achieved by directleaf desiccation or by polyethylene glycol inhibited enzymeactivity up to fourfold at –1.2 MPa. Beta vulgaris, sugar beet, ribulose-1, 5-bisphosphate carboxylase, salt stress, water stress, carbon dixoide fixation, salinity     

Regulation of ACC-Dependent Ethylene Production by Excised Leaves from Normal and Albino Zea mays L. Seedlings

Dunlap, J. R. 1988. Regulation of ACC-dependent ethylene productionby excised leaves from normal and albino Zea mays L. seedlings.—J.exp. Bot. 39: 1079–1089. Albino corn (Zea mays L.) seedlings lacking natural leaf pigmentswere obtained by germinating seeds treated with fluridone, aninhibitor of carotenoid biosynthesis. Basal rates of ethyleneproduction were less than 2.0 nl g^–1 fr. wt h^–1in both treated (albino) and untreated (normal) leaves but increasedby 10- to 20-fold in the presence of added ACC. ACC-dependentethylene production (ADEP) was inhibited by cobalt or cyanideions and stimulated by NaHCO₃, CO₂ and light. ADEP in both tissueswas stimulated by glucose, fructose, galactose and sucrose.The accumulation of respiratory CO₂ did not account for thecarbohydrate response. The decline in the ADEP characteristicof albino leaf tissue was slowed by incubation in the presenceof sucrose. IAA and ABA stimulated ADEP in normal leaves butinhibited ADEP in albino leaves. Sucrose-stimulated ADEP wasinhibited in albino leaf tissue treated with IAA or ABA indicatinga possible role for the chloroplast in carbohydrate-facilitatedADEP. However, results from this study suggest that chloroplastsperform a function in the regulation of ethylene productionby leaf tissue that extends beyond merely influencing internallevels of CO₂. In the absence of detectable ACC, EFE was responsiblefor the entire series of responses expressed in regulation ofethylene biosynthesis by corn seedling leaf tissue. Key words: Corn, ethylene, sugars, phytohormones     

Role of carbonic anhydrase in photosynthesis in Chlorella derived from kinetic analysis of 14CO2 fixation1

Time courses of photosynthetic ¹⁴CO₂ fixation and its simulationare presented for Chlorella cells grown under low CO₂ concentration(low-CO₂ cells) and subsequently exposed to 0.2 mM NaH¹⁴CO₃or 130 ppm ¹⁴CO₂ in the presence or absence of carbonic anhydrase(CA) in the suspending medium. It was shown that Chlorella cells utilized only free CO₂ whenNaHCO₃ was given in the presence or absence of CA, or when CO₂was bubbled in the absence of CA. However, the present simulationindicated that both CO₃ and HCO₃^– were utilized when CO₂was given in the presence of CA. Based on these results, weconcluded that 1) Chlorella cells absorb only free CO₂ and 2)this gas is provided to algal cells in two ways, i.e., by directand indirect CO₂ supply. Usually, the dissolved CO₂ is directlyutilized by the algal cells (direct supply of CO₂). However,when the concentration of dissolved CO₂ is extremely low andwhen there is CA, CO₂ reconverted from HCO₃^– is also utilizedby Chlorella cells (indirect supply of CO₂). The utilizationof HCO₃^– indicated by the above simulation was explainedby the indirect supply of CO₂. We further assumed that the indirectsupply of CO₂ to ribulose 1,5-bisphosphate carboxylase occursmainly in the chloroplasts of low-CO₂ cells containing highCA. Thus, under low CO₂ concentrations, low-CO₂ cells can carryout more efficient CO₂ fixation than high-CO₂ cells, resultingin the lower apparent Km(CO₂). ³Department of Biology, Faculty of Science, Niigata University,Niigata, Japan. (Received April 2, 1980; )     

Abscisic Acid and Apical Dominance in Phaseolus coccineus L. The Role of Tissue Age

Abscisic acid (ABA) applied to the decapitated second internodeof runner bean plants enhanced outgrowth of lateral buds onlywhen internode stumps were no longer elongating. Applied toelongating internodes of slightly younger plants, ABA causesinhibition of bud outgrowth. Together with 10^–4 M indol-3-ylacetic acid (IAA), ABA stimulated internode elongation and interactedadditively in the inhibition of bud outgrowth. A mixture of10^–5 M ABA and 10^–6 M gibberellic acid (GA₃ ) causedsimilar effects on internode growth as IAA + ABA, but was mutuallyantagonistic in effect on growth of the lateral buds. Abscisic acid, apical dominance, gibberellic acid, indol-3yl acetic acid, Phaseolus coccineus, bean