Endogenous light-on rhythm in respiration of a long-day duckweed, Lemna gibba G3 II. On basic and rhythmic components of the rhythm

Effects of respiratory substrates (glucose, malate, citrateand pyruvate) and inhibitors (fluoride, iodoacetate, azide andDNP) on the O₂-uptake rhythm in a long-day duckweed,Lemna gibbaG3 in continuous light period were examined. Rates of O₂-uptake at the starting point (6 hr after the beginningof a continuous light period) and at the time of the first peakof the rhythm (18 hr after the beginning of a continuous lightperiod) were equally increased by exogenous substrates. Sensitivityof respiration to fluoride or iodoacetate was almost the sameat the 6th and 18th hr. The O₂-uptake (at the 6th, 18th, 30thand 42nd hr) was increased by DNP by the same amount. Azideat lower concentrations than 5X10^–4 M did not affect O₂-uptakeat the 6th hr, but inhibited uptake at the 18th hr. In the presenceof 5 X 10^–4 M of azide the rates of O₂-uptake at the 18th,30th or 42nd hr were down to the rate at the 6th hr, which wasinsensitive to azide. These results suggest that the O₂-uptakerhythm consists of two components, i.e. the basic respirationwhich is promoted by exogenous substrate, sensitive to DNP andinsensitive to azide; and rhythmic respiration, which is sensitiveto azide, but is not influenced by exogenous substrate and DNP. (Received February 19, 1971; )     

External and Internal CO2 Transport in Lemanea: Interactions with the Kinetics of Ribulose Bisphosphate Carboxylase

The rate of photosynthesis by the freshwater alga Lemanea mamillosais proportional to CO₂ concentration, virtually to the pointof saturation, and inversely proportional to the radius of thethallus. By contrast, the CO₂ response curve of very thin slicesof the thallus is a rectangular hyperbola with a (lower) halfsaturation concentration of 10 mmol m^–3. For the intactplant, the kinetics of CO₂ fixation are strongly masked by internalCO₂ transport limitations, although the maximum rate of photosynthesisis probably determined by the rate of supply of ribulose bisphosphate(RuBP). The flow of water over the alga becomes turbulent atwater velocities greater than about 90 mm s^–1 and thethallus stretches significantly at higher water velocities.In its natural habitat, therefore, the external unstirred layerwill be thin (< 10 µm) and the thallus will be stretched,leading to rapid external and increased internal rates of CO₂transport from the bulk solution. The estimated maximum rateof CO₂ transport is commensurate with the maximum rate of photosynthesis(i.e. the rate of supply of RuBP). Key words: Transport limitations, Kinetics of CO₂ fixation     

Deterination of Effective Partition Coefficients of Alkali Chlorides by Measurements of Concentration Potential across the Nitella flexilis (L.) Ag. Isolated Cell Wall

We report a study in which we estimate by means of non-equilibriumprocesses the value of an effective partition coefficient, x,a parameter that we use to take into account the non-ideal behaviourof the electrolyte of the external medium in the cell wall space.This parameter is defined by the ratio of the salt partitioncoefficient to the activity of the non-diffusing charges. Tothat purpose, concentration potential in NaCl, LiCI and CuCl₂solutions was measured across an isolated cell wall of Nitellaflexilis at different pH. No significant difference appearedbetween the potential values measured in the presence of NaClor LiCl (pH 5.5, 4.0 and 3.0). At pH 3.0, in CuCl₂ solutions,the Nitella wall behaved as an uncharged membrane. Two methods—high concentrations and least-squares approximation-wereused to calculate from these results the value of the effectivepartition coefficient. Both gave similar values of x At a givenpH, the values of x are independent of the concentration ofthe external medium over a large range of concentration ratios.The activity of the exchange sites was estimated by the experimentallydetermined fraction of absorbed monovalent cations. The ionicconcentrations in the wall space were calculated at pH 5.5 and3.0 by introducing the values of x in the relations which correlatethem to the ionic concentrations in the wall for that givenvalue of the activity of the carboxylate sites. From their values,it appears that the negative fixed charges of the wall werefunctionally shaded by the condensation of a part of the adsorbedmonovalent cations. The activity of the former was, therefore,reduced to a value considerably lower than that estimated bythe cationic exchange capacity during equilibrium processes. Key words: Nitella flexilis (L.) Ag., cell wall, concentration potential, partition coefficient, ion condensation     

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     

Ionic Currents across the Plasmalemma of Chara inflata Cells: II. EFFECTS OF EXTERNAL Na+, Ca2+ AND Cl- ON K+ AND Cl CURRENTS

The electrical conductance of the plasmalemma of cells of Charainflata, due to the diffusion of ions, consists predominantlyof K⁺, Cl^– and leak components. When the membrane electricalpotential difference is stepped in a negative direction witha voltage-clamp, the resulting inward current has componentsI_K, I_Cl and I_L (leak). During such voltage-clamp steps I_K isinactivated, and Ic activated with voltage-dependent half-times.Increases in the external NaCl concentration reduce the magnitudeof I_K and increase the magnitude of I_c, but reduce the half-timeof inactivation or activation. The NaCl-induced changes in I_kand I_Cl and their kinetics were more pronounced at pH₀ =6.5than at pH₀ =9.5. When the concentration of external CaCl₂ wasincreased, I_k, I_Cl and the half-time of inactivation, (T_1/2),of I_k were all reduced. The half-time of activation of I_Cl wasincreased. The NaCI-induced changes could result from increases in bothexternal ion concentration and osmotic pressure. Previous experimentshave shown that an increase in external osmotic pressure alonealters the properties of the conductances. In this paper weattempt to separate the purely ionic effects from the osmoticones. Key words: Chara inflata, ionic effects, K⁺ and Cl^– currents     

Response of Photosynthesis, Stomatal Conductance and Water Use Efficiency to Elevated CO2 and Nutrient Supply in Acclimated Seedlings of Phaseolus vulgaris L.

Plants of Phaseolus vulgaris L were grown from seed in open-topgrowth chambers at present day (350 µmol mol^–1)and double the present day (700 µmol mol^–1) atmosphericCO₂ concentration with either low (L, without additional nutrientsolution) or relatively high (H, with additional nutrient solution)nutrient supply Measurements of assimilation rate, stomatalconductance and water use efficiency were started 17 d aftersowing on each fully expanded, primary leaf of three plantsper treatment Measurements were made in external CO₂ concentrations(C₂) of 200, 350, 450, 550 and 700 µmol mol^–1 andrelated to both C_a and to C₁, the mean intercellular space CO₂concentration Fully adjusted, steady state measurements weremade after approx 2 h equilibration at each CO₂ concentration The rate of CO₂ assimilation by leaves increased and stomatalconductance decreased similarly over the range of C_a or C₁ inall four CO₂ and nutrient supply treatments but both assimilationrate and stomatal conductance were higher in the high nutrientsupply treatment than in the low nutrient treatment The relationbetween assimilation rate or stomatal conductance and C₁ wasnot significantly different amongst plants grown in present-dayor elevated CO₂ concentration in either nutrient supply treatment,i e there was no evidence of down regulation of photosynthesisor stomatal response Increase in CO₂ concentration from 350to 700 µmol mol^–1 doubled water use efficiency ofindividual leaves in the high nutrient supply treatment andtripled water use efficiency in the low nutrient supply treatment The results support the hypothesis that acclimation phenomenaresult from unbalanced growth that occurs after the seed reservesare exhausted, when the supply of resources becomes growth limiting CO₂ enrichment, Phaseolus vulgaris L., net CO₂ assimilation rate, stomatal conductance, water use efficiency     

Uncoupling Properties of the Herbicide N-n-Pentyl-N-Methyl-N'-(3,4-Dichlorophenyl)-Urea

The herbicide D₅ (N-n-pentyl-N-methyl-N'-(3, 4-dichlorophenyl)-ureacan uncouple oxidative phosphorylation in isolated plant mitochondria.This paper confirms that D₅ is an uncoupler that catalyzes thecollapse of the transmembrane potential gradient by inducinga movement of protons across the membrane. However, D₅ is notitself capable of transporting protons. D₅ gives complete uncouplingat 40 µM, a lower concentration than that required foruncoupling by the n-butyl homologue ‘neburon’. Analysisof the shape of the state 4 stimulation curve suggests thatD₅ might act as a dimer in the membrane. Attempts to demonstrate binding of D₅ to a membrane target gaveambiguous results, binding is not evident at 10 °C and 25°C but might occur at 15 °C and 20 °C. The calculatedherbicide concentration in the membrane (40 µM of which4–0 µM is as the dimer) is high and similar to thatof the major phospholipids. The calculated partition coefficientbetween medium and membrane (3.8 x 103) is in agreement withthe lipophilicity of the substituted urea herbicides. In the presence of a substrate, D₃ blocks both influx and effluxcalcium movement through the mitochondrial membrane but in theabsence of substrate, D₅ induces binding of calcium. Bindingrequires Mg⁺⁺ but not K⁺ or phosphate and leads to a releaseof H⁺. Ruthenium Red causes a partial inhibition of bindingbut no other reagent or ionophore tested had any effect. Sincebinding does not occur in turnip mitochondria which are unableto transport Ca⁺⁺ it is concluded that the effect is not directlylinked to the uncoupling action. The mechanism of action of D₅ is discussed and it is concludedthat D₅ probably acts as a dimer and perturbs membrane structure.The site of action is probably the lipid components of the membrane. Key words: Plant mitochondria, Herbicide, Substituted ureas, Calcium, Uncoupling     

Some General Characteristics of Glycolysis in vitro and its Inhibition by High Concentrations of Chloride Salts

Glycolysis in vitro was studied, using extracts from germinatingpea seeds. Steady state rates of CO₂ evolution declined long before allsubstrate, either glucose or sugar phosphate, was convertedto CO₂ and ethanol. High rates of CO₂ evolution were restoredby addition of more substrate. Some factors which regulate glycolysis in vitro were investigated,using glucose as a substrate. High levels of P₁ and Mg²⁺ wererequired to obtain optimum rates of CO₂ evolution. Levels ofintermediates indicated that high P₁ acted via a stimulationof 6-phosphofructokinase. Glucose, in the presence of hexokinase, induced substantiallyhigher rates of CO₂ evolution than a number of sugar phosphates.However, evolution of CO₂, from glucose-6-phosphate or fructose-6-phosphate,was greatly stimulated by the addition of purified yeast pyruvatedecarboxylase, rates sometimes exceeding those attained whenglucose was used as a substrate. The reason for this stimulationby pyruvate decarboxylase is unknown. Effects of high KCl and NaCl were measured, using glucose asa substrate. KCl and NaCl, at 200 mM and higher concentrations,reduced steady state rates of glycolysis. The degree of inhibitionwas much greater at high than at low rates of glycolysis. Thiswas shown, for example, when using different concentrationsof P₁. High levels of KCl and NaCl greatly increased levels of fructose-1,6-diphosphateand triose phosphates, particularly at low P₁, and did not reducethe ‘net carbon movement past 6-phosphofructokinase’Thus high KCl and NaCl weakened the control of glycolysis by6-phos-phofructokinase.     

Localization and Kinetic Properties of {beta}-Glycerophosphatase in Barley Roots

The study of ß-glycerophosphatase activity in cell-wallpreparations and in excised root tips from barley seedlingssupports the view that the former, which constitutes about 20per cent of the activity of the whole homogenate, representsthe fraction located at the surface of the roots in vivo. Theactivities of the cell-wall suspension and intact roots arevirtually identical, and further show identical relations topH, substrate concentration (K_m), and competitive inhibitionby molybdate and inorganic phosphate (K_i). The enzyme must thereforebe freely exposed to the external solution without any permeabilitybarrier separating it from either substrate or inhibitors. Theabsence of any lag phase in the hydrolysis in excised root tipssuggests that the surface enzyme may be limited to the outermostlayers of the root. The solubilization of some of the activityof the cell-wall preparation by treatment with sodium chlorideand ammonium sulphate suggest that surface activity may havebeen lost from these preparations rather than adsorbed duringhomogenization and extraction. The K_m and pH-activity curveof the supernatant activity remaining after centrifugation ofthe cell-wall fraction indicate that only this enzyme and noother detectable glycerophosphatase exists in the roots.     

The Effect of Ethylene on Quantitative and Qualitative Aspects of Respiration During the Breaking of Dormancy of Spergula arvensis L. Seeds

The effect of ethylene on quantitative and qualitative aspectsof respiration in Spergula arvensis L. seeds was studied. Oxygenconsumption and CO₂ evolution from seeds increased 60 h afteraddition of ethylene to the dry seed; these increases were consideredto be a function of the number of seeds that were germination,rather than a direct effect of ethylene. The respiratory quotient(RQ) on day 1 (0.5) was lower than on subsequent days, indicatingan absence of anaerobic processes in the early stages of germinationand the use of a different respiratory substrate to that oxidizedon days 2–4. Measurements of C₄/C₁ ratios showed thatethylene did not break dormancy by initiating a switch fromthe Embden-Meyerhof-Parnas pathway (EMPP) to the pentose phosphatepathway (PPP). On the contrary, some evidence was obtained fora switch from the PPP to the EMPP after 3–6 h in pre-imbibedand non pre-imbibed seeds treated with ethylene. Spergula arvensis L., seed germination, ethylene, respiration, Embden-Meyerhof-Parnas pathway, pentose phosphate pathway, respiratory quotient     

Physiological Studies of the Sulpholipids of Diatoms

The sulpholipids of three species of freshwater and marine diatomNitzschia palae Kutz, Navicula muralis Lewin and Navicula incertaGrün, have been investigated under various culture conditions.The plant sulpholipid, sulphoquinovosyl diglyceride, was predominantlysynthesized in the light rather than in the dark while the unknownsulpholipids, designated as U₁ and U₂, were produced more inthe dark than in the light. It was found that cells starvedof carbon or sulphate utilized their sulpholipid reserve assources of these materials. Generally, cultures incubated inthe light and bubbled with air (with or without CO₂) showeda high level of incorporation of ³⁶S into sulpholipids. In culturesbubbled with oxygen-free nitrogen the incorporation of tracerwas very small. The photosynthetic and respiratory inhibitors,DCMU and DNP appreciably reduced the amount of tracer incorporatedinto the sulpholipids.     

Role of Coccoliths in the Utilization of Inorganic Carbon by a Marine Unicellular Coccolithophorid, Emiliania huxleyi: a Survey Using Intact Cells and Protoplasts

The utilization of inorganic carbon and role of the coccolithswere investigated in intact cells and protoplasts of a marineunicellular calcareous alga, Emiliania huxleyi. Protoplastswith high photosynthetic activity were obtained by artificialdecalcification with 50 mM MES-NaOH (pH5.5). (1) The kineticsof the photosynthetic evolution of O₂ at various concentrationsof externally added NaHCO₃ were the same for intact cells andprotoplasts, indicating that the kinetic properties with respectto dissolved inorganic carbon (DIC) were not affected by thepresence or absence of the coccoliths on the cell surface. Double-reciprocalplots and plots of the concentration of substrate divided byvelocity (s/v) against the concentration of substrate (s) werebiphasic in the case of both intact cells and protoplasts. TheCO₂-utilization reaction was, therefore, considered to involvetwo processes with different values of K_m and V_max. From thekinetic analyses, K_m and V_max [µmoles O₂ (ml PCV)^–1h^–1] were deduced to be 92 µM and 76.3 for a "low-K_m"reaction and 4.1 mM and 252 for a "high-K_m" reaction, respectively.(2) In short-term (40-min) experiments, time courses of thetotal uptake of ¹⁴C-DIC and the incorporation of ¹⁴C into acid-stableproducts of photosynthesis and the internal pool of DIC, determinedas acid-labile compounds, under CO₂-limiting conditions (80µM) were very similar for intact cells and protoplasts.However, incorporation of ¹⁴C into CaCO₃ apparently occurredmore slowly in protoplasts than in intact cells. (3) In longterm (24-h) experiments, patterns of incorporation of ¹⁴C werealmost same for intact cells and protoplasts, with the exceptionthat the amount of ¹⁴C incorporated into CaCO₃ was much smallerin the former than the latter. The production of Ca¹⁴CO₃ increasedduring the course of 10 h after a 4-h lag. However, after 10h the level of Ca¹⁴CCO₃ started to decrease. The decrease wasaccompanied by an increase in ¹⁴C in the products of photosynthesis,suggesting that CaCO₃ was reutilized for the photosyntheticfixation of CO₂ and, therefore, that the coccoliths functionas sites of storage of DIC. However, the internal level of DICremained at the same level even after the supply of externalDIC has been almost completely depleted. (Received July 25, 1995; Accepted December 11, 1995)     

HCO-3 as an Exogenous Carbon Source for Aquatic Bryophytes Fontinalis antipyretica and Fissidens grandifrons

The aquatic bryophytes Fontinalis antipyretica Hedw. and Fissidensgrandifrons Brid. were investigated for their ability to utilizeHCO^–₃ and CO₂ as exogenous carbon sources for photosynthesis.In NaHCO₃ solutions Fontinalis increased the pH to a maximumof 9.6 corresponding to a CO₂ compensation point of 1.1 mmolm^–3 CO₂. Measured photosynthetic rates cannot be explainedonly by uptake of CO₂. Net photosynthesis decreased at highpH but did not decline to zero until pH 10.10 in Fissidens andpH 11.8–12.0 in Fontinalis. Furthermore, photosynthesiswas increased by higher HCO_–3 concentrations at constantCO₂ concentration. It is concluded that Fontinalis antipyreticahas the capability to utilize HCO₃. Key words: Carbon source, Photosynthesis, Aquatic bryophytes     

The Role of Transmembrane Electrical Potential in Determining the Absorption Isotherm for Chloride in Potato

Measurements have been made of the electrical potential differencebetween the vacuoles of single potato tuber cells and externalCl^- solutions over the range 1–40 mM. With K⁺ as the counter-ion,the relationship between this transmembrane electrical potentialand external Cl^- concentration, for fresh cells at 20° C,was found to be one of decreasing negative polarity with increasingCl^- concentration (E at 1 mM Cl^- external = – 81 m V;change in E for a 10-fold change in external concentration,E₁₀ = 46 m V). The linearity of this relationship, apparenton a semi-logarithmic plot, was virtually unaltered by low temperature(0.5–2.5° C) or by previous ageing of the cells forperiods up to four days (indicating that metabolic ion absorptionis not an electrogenic process). When the counter-ion was maintainedat a constant high concentration (40 mM K⁺), the change in potentialover the Cl^- concentration range was only 4 m V, polarity becomingmore negative with increasing Cl^- concentration. With Ca⁺⁺ asthe counter-ion, the potential to external Cl^- concentrationrelationship was similar to that found in KCl solutions, exceptthat E₁₀ was only about 20 m V. Curves for the influx of C1^- to be expected on the basis ofthese electrochemical data alone have been shown to run closelyparallel to Cl^– absorption isotherms previously determinedexperimentally. This confirms the opinion, already formed onthe basis of theoretically derived values for passive Cl^- influx,that Cl^- uptake by both fresh and one-day-aged potato tissue,from KCl solutions and Cl^- solutions with a fixed high K⁺ concentration,is rate-determined at o° C by passive movement across theplasmalemma. Uptake of Cl^- by fresh tissue at 20° C appearsto be similarly regulated. No such parallelism was found between observed and expectedpatterns of Cl^- uptake from CaCl₂ solutions, or from KCl bytwo-day-aged tissue, and here factors in addition to the electrochemicalones must determine low temperature Cl^-uptake.     

The role of photosynthesis and respiration in the light-induced recovery of "giant" cells of the Emerson strain of Chlorella

The light-induced recovery of cell division and chloroplastdevelopment in "giant", "bleached" cells of the Emerson strainof Chlorella is unaffected by treatments (atrazine. CMU, incubationin a CO₂-free atmosphere) which interfere with photosynthesis.Anaerobic conditions or the presence of respiratory inhibitors(DNP, KCN, NaN₃) markedly suppress recovery. Recovery is accompaniedby a mobilization of the reserve starch which follows a linearcourse over the first 9 hr. Chloramphenicol (50 µg/ml),which inhibits chlorophyll synthesis and the development ofa photosynthetic capacity, is without effect on the early rateof starch mobilization. Evidence is presented that the contributionof photosynthesis towards recovery is only significant whenthe reserve starch has been depleted. Recovery does not requirecontinuous light; the critical light-stimulated processes apparentlytaking place during the first 9 hr. The possible nature of thelight stimulation of recovery is discussed. (Received June 18, 1973; )     

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; )     

The Effect of External pH on the Apparent CO2 Affinity of Chlorella saccharophila

The carbon dioxide compensation point of the unicellular greenalga, Chloretla saccharophila, was determined in aqueous mediumby a gas chromatographic method. Compensation points decreasedmarkedly from 63 cm³ m^–3 at an external pH of 4.0 to 3.2cm³ m^–3 at pH 8.0 and were not affected by the O₂ concentrationof the medium. The calculated CO² concentration required tosupport the half-maximum photosynthetic rate of the algal cellsranged from 6.0 mmol m_–3 at an external pH of 60 to 1.5mmol m^–3 at pH 8.0 and these values were not affectedby O₂ concentration. The K_m(CO₂) of nbulose-l,5-bisphosphatecarboxylase isolated from cells grown either at pH 4.0 or pH8.0 was determined to be 64 mmol m^–3. These results indicatethat loss of CO₂ by photorespiration does not occur in C. saccharophilacells at acid pH and the disparity between the apparent affinityfor CO₂ of the intact cells and that of the carboxylase indicatesthe operation of a ‘CO₂ concentrating mechanism’in this alga at acid pH. Key words: Acidophilic alga, bicarbonate transport, Chlorella saccharophila, compensation point, CO₂ affinity, PH, RuBP carboxylase     

Carbohydrate Metabolism of Rice Seedlings Grown in Oxygen Deficient Solution

Atwell, B. J. and Greenway, H. 1987. Carbohydrate metabolismof rice seedlings grown in oxygen deficient solution.—J.exp. Bot. 38: 466–478. Rice seedlings (Oryza sativa L.) were grown in the dark forup to 4 d in solutions containing various concentrations ofO₂. The rate of depletion of the endosperm was most rapid inaerated solution (0·25 mol O₂ m^–3), largely dueto the inhibition of growth of seedlings at very low O₂ concentrations.Earlier suggestions that there is a deficit of sugars for growthand energy generation in O₂ deficient coleoptiles were tested. Coleoptiles, shaking in aerated solution, respired about one-thirdof the endogenous sugars to CO₂ and incorporated the rest intostructural compounds. In contrast, the proportion of carbonwhich went to growth in anoxia was very low. Consistent withthese results, endogenous sugar levels were generally highestat low O₂ concentrations. Even so, coleoptiles grown and testedas low as 0·03 mol O₂ m^–3 showed appreciable metabolismof exogenous ¹⁴C-glucose to CO₂, soluble and insoluble compounds,suggesting that a minimal O₂ supply was sufficient to sustainsome growth. Furthermore, glucose feeding caused little or norise in O₂ uptake or tissue sugar levels. Similarly, the specificactivity of the evolved CO₂ was not markedly different in coleoptilesgrowing at 0·03 and 0·25 mol O₂ m^–3 Further evidence was obtained to show that endogenous substrateswere adequate for growth and respiration at both low and highO₂ concentrations. Exogenous glucose and malate did not stimulateO₂ uptake at any stage of growth in aerated coleoptiles. Therewas sufficient endogenous substrate to sustain a 35–45%rise in O₂ uptake induced by uncoupling and enrichment withO₂. Exogenous glucose did not stimulate growth of intact seedlingsat any O₂ concentration. Key words: Rice seedlings, carbohydrate metabolism, oxygen deficient solution     

Calcification in the Green Alga Halimeda: IV. THE ACTION OF METABOLIC INHIBITORS ON PHOTOSYNTHESIS AND CALCIFICATION

The effects of a number of metabolic inhibitors on calcificationand photosynthesis in Halimeda tuna, H. discoidea, and H. macrolobaare described. The inhibitors used are CCCP, DNP, DCMU, azide,cyanide, chloramphenicol, cycloheximide, and Diamox. The effectsof these inhibitors, although complex, are consistent with ourmodel of calcification in Halimeda. Inhibition of photosyntheticCO₂ uptake inhibits calcification as does stimulation of respiratoryCO₂ evolution (i.e. uncoupling). There is also indirect evidencefor the presence of a possible light stimulated H⁺ efflux whichinhibits calcification. The observed calcification rate is thereforethe result of a number of factors which affect the concentrationof CO^–and the pH in the intercellular space of the Halimedathallus. The results obtained with the carbonic anhydrase inhibitor Diamoxprovide further evidence for the effective separation of theintercellular space from the external medium by the appressedperipheral utricles.     

Nitrate Transport into Chara corallina Cells using 36ClO-3 as an Analogue for Nitrate: I. INTERACTION BETWEEN 36ClO-3 AND NO-3 AND CHARACTERIZATION OF 36CIO-3/NO-3 INFLUX

The use of chlorate as an analogue for NO^–₃ during nitrateuptake into Chara corallina cells has been investigated. NO^–₃inhibits ³⁶C1O^–₃ influx into Chara over the concentrationrange 0–1000 mmol m^–3. Lineweaver-Burke plots ofthe data are characteristic of competitive inhibition by NO^–3in the low concentration range (0–300 mmol m^–3 ClO^–₃)and apparent K_INO3 is 140 mmol m^–3 which is of a similarorder of magnitude as apparent K_mCIO3- 180 mmol m^–3. Athigher substrate concentrations the inhibition by NO^–₃was not characteristic of competitive or uncompetitive inhibition. ³⁶C1O^–₃/NO^–₃ influx was dependent on K⁺ and Ca²⁺in the external medium and inhibited by FCCP. NO^–₃ pretreatmentor N starvation increased subsequent ³⁶C1O^–₃/NO^–₃influx into Chara. A comparison between rates of net NO^–₃uptake and ³⁶C1O^–₃/NO^–₃ influx supported the previoushypothesis that NO^–₃ efflux is an important componentin the determination of overall uptake rates. Key words: Nitrate, Chara, ³⁶CIO^–₃