The Effect of Carbon Dioxide Concentratioin, light Intensity and Isonicotinyl Hydrazide on the Photosynthetic Production of Glycollic Acid by Chlorella

Glycollic acid production by Chlorella was measured by colorimetricdetermination of the acid excreted into the medium. It was foundthat glycollic acid production showed a maximum at a low concentrationof carbon dioxide but tended toward zero as the rate of photosynthesisapproached carbon dioxide saturation. Glycollic acid productionbecame measurable at light intensities approaching that requiredto saturate photosynthesis and increased steadily with furtherincrease in intensity. Treatment with isonicotinyl hydrazideresulted in an approximately threefold stimulation of glycollicacid concentration over the range of conditions used. It issuggested that the precursor of glycollic acid is ribulose diphosphate,and that isonicotinyl hydrazide acts by inhibiting the furthermetabolism of glycollic acid.     

CATALYTIC EFFECTS OF CARBON DIOXIDE IN CARBON DIOXIDE ASSIMILATING CELLS

It has been confirmed that absence of carbon dioxide may decreasethe rate of oxygen production which accompanies the photochemicalreduction of p-benzoquinone in algae and chloroplasts. Thisinfluence of carbon dioxide partial pressure does not applyto the overall oxygen yield. In the blue-green alga Anacystisnidulans the initially small carbon dioxide deficiency effectincreases with time spent in the dark. The deterioration ofreaction rates is counteracted by light. There seems to be nodirect connection or interdependence between the photosyntheticreduction of carbon dioxide and the sensitivity of some partof the photochemical mechanism to loss of carbon dioxide. Notonly does addition of quinone to living cells in these experimentsdestroy their capacity for photosynthesis, but mutant cellsthat never had this capacity still retain the sensitivity towardslack of carbon dioxide when tested for their ability to reducequinone. Many different metabolic reactions have been seen topossess such dependency on traces of carbon dioxide, also innon-photosynthetic cells and tissues. The explanation for "catalytic"effects of carbon dioxide ought to be a general one–suchas an influence on the efficiency of certain phosphorylationswhich occur everywhere in the living world. ¹ Dedicated to Prof. H. TAMIYA on the occasion of his 60th birthday.These studies were aided by contract NONR 988 (10) between theOffice of Naval Research, Department of the Navy, and the FloridaState University, respectively. ² Present address: Charles F. KETTERING Research Laboratories,Yellow Springs, Ohio. (Received December 7, 1962; )     

Some Effects of Temperature and Substrate Content upon Respiration and the Carbon Balance of Field Beans (Vicia faba L.)

The effects of substrate content and temperature upon the productionof carbon dioxide in the dark were investigated in Vicia fabaand Sorghum vulgare, using the time courses for carbon dioxiderelease. No endogenous rhythms were found. With V. faba at highsubstrate contents, a steady rate of respiration was measuredat low temperatures, suggesting that the rate of respirationis limited by a third factor such as enzyme activity. This steadyrate eventually decreased rapidly. There was a first order rateof decrease with time at higher temperatures. Low substratecontents gave a complex decay. The rate of decrease of the rateof respiration was affected by the initial state of the plantsover the same range of rates. Reasons for this are discussed.The temperature sensitivity of respiration was investigated.The respiration of plants with high substrate contents had alower temperature sensitivity than those with low substratecontents. This was further investigated by measuring the stoichiometryof carbon dioxide production in the dark from total solublecarbohydrate (as hexose equivalent). It is likely that incompletehexose respiration, which occurs at high substrate contents,is less temperature sensitive than complete hexose respirationor the respiration of some other substrate. Vicia faba, Sorghum oulgare, carbon dioxide, respiration, temperature, substrate content     

Examination of a Model and Data Describing the Effect of Temperature on the Respiration Rate of Crop Plants

Some of the published evidence used in the synthesis and maintenancemodel of plant respiration is discussed in relation to the effectof temperature. Recalculations from the data of de Vries (1975b) give different results from those claimed by him. The modelis considered in terms of the use of substrate in the dark andits production in the light. It is suggested that starvationestimates of maintenance are not valid. The most reliable methodof observing synthesis respiration in whole plants appears tobe by following a discrete pool of substrate, as is possiblewith labelled carbon. Triticum aestivum L., Zea mays L., Helianthus annuus L., Vicia faba L., carbon dioxide, respiration, temperature, substrate content     

Short- and Long-Term Inhibition of Respiratory Carbon Dioxide Efflux by Elevated Carbon Dioxide

Dark carbon dioxide efflux rates of recently fully expandedleaves and whole plants of Amaranthus hypochondriacus L., Glycinemax (L.) Merr., and Lycopersicon esculentum Mill. grown in controlledenvironments at 35 and 70 Pa carbon dioxide pressure were measuredat 35 and 70 Pa carbon dioxide pressure. Harvest data and whole-plant24-h carbon dioxide exchange were used to determine relativegrowth rates, net assimilation rates, leaf area ratios, andthe ratio of respiration to photosynthesis under the growthconditions. Biomass at a given time after planting was greaterat the higher carbon dioxide pressure in G. max and L. esculentum,but not the C₄ species, A. hypochondriacus. Relative growthrates for the same range of masses were not different betweencarbon dioxide treatments in the two C₃ species, because highernet assimilation rates at the higher carbon dioxide pressurewere offset by lower leaf area ratios. Whole plant carbon dioxideefflux rates per unit of mass were lower in plants grown andmeasured at the higher carbon dioxide pressure in both G. maxand L. esculentum, and were also smaller in relation to daytimenet carbon dioxide influx. Short-term responses of respirationrate to carbon dioxide pressure were found in all species, withcarbon dioxide efflux rates of leaves and whole plants lowerwhen measured at higher carbon dioxide pressure in almost allcases. Amaranthus hypochondriacus L., Glycine max L. Merr., Lycopersicon esculentum Mill., soybean, tomato, carbon dioxide, respiration, growth     

The Interactive Effects of Atmospheric Carbon Dioxide and Light on Stem Elongation in Seedlings of Four Species

Four species,Sinapis albaL.,Medicago sativaL.,Gypsophila paniculataL.andPicea abies(L.) Karsten, were grown in three light regimes:darkness, low light (25 µmol m^-2s^-1for 10 min d^-1) andhigh light (120 µmol m^-2s^-1for 12 h  d^-1) and fourlevels of carbon dioxide: 0, 350, 700 and 1400±50 µll^-1. Germination was not affected by any of the treatments.The effects of carbon dioxide on stem elongation were identicalin low and high light: stem length increased at a decreasingrate with level of carbon dioxide in all species. Level of carbondioxide also affected stem elongation in complete darkness,but the pattern was more complex and varied among species. Totalweight did not vary with level of carbon dioxide to any significantextent in either darkness or low light, but increased with levelof carbon dioxide at high light in all four species. Due tothe absence of any effect of carbon dioxide on growth in darknessand low light, we suggest the effects of carbon dioxide on stemelongation are independent of effects on growth and may be dueto a direct interaction with developmental processes. In contrast,level of carbon dioxide had little effect on allocation patternsin the dark and low light experiments, but had marked effectsin high light. Therefore, the effect of carbon dioxide on allocationwas probably due to the effects of carbon dioxide on growthrather than to any direct interaction between carbon dioxideand development. An understanding of the mechanisms by whichcarbon dioxide affects development may help us understand theoften variable effects of carbon dioxide upon plants.Copyright1998 Annals of Botany Company Sinapis albaL.;Medicago sativaL.;Gypsophila paniculataL. andPicea abies(L.) Karsten; elevated carbon dioxide; stem elongation; germination; allocation; phytochrome.     

The Effect of Isonicotinyl Hydrazide on the Photosynthetic Incorporation of Radioactive Carbon Dioxide into Ethanol-Soluble Compounds of Chlorella

The effect of 10^-2M. isonicotinyl hydrazide (isoniazid) on theincorporation of radioactive carbon dioxide by Chlorella duringphotosynthesis has been studied under steady-state conditionsat two carbon dioxide concentrations. Isoniazid treatment resultsin increased radioactivity in sucrose, glycollic acid, and glycineand decreased radioactivity in sugar monophosphates, serine,and alanine. An unidentified compound which is strongly radioactiveafter short-term exposures to ¹⁴CO₂ is present in isoniazid-treatedcells. It is suggested that isoniazid pre-dominantly inhibitsthe conversion of glycine to serine.     

Ventilation in Plant Tissue Cultures and Effects of Poor Aeration on Ethylene and Carbon Dioxide Accumulation, Oxygen Depletion and Explant Development

A simple technique for comparing and quantifying the ventilationcapacity of vessels used for plant tissue culture is described.Ethylene was injected into culture vessels and its rate of lossmonitored by gas chromatography. From the resulting exponentialdecay curves, the time in hours for half the ethylene to belost (t₅₀) was calculated and used to compare different containersand sealing methods. Cultures of Ficus lyrata Warb. and Gerberajamesonii Bolus grown for up to 28 d in plastic vessels sufficientlywell-sealed to generate t₅₀ values of approx. 16 h, accumulatedethylene and carbon dioxide in association with depleted oxygen.The relationship between carbon dioxide accumulation and oxygendepletion within culture vessels indicated little if any anaerobicrespiration. Gerbera explants did not appear to be affectedby these gaseous environments. However, in Ficus, leaf expansionwas approximately halved, although fresh and dry mass of wholeshoots was not decreased. The smaller leaf size is attributedto the action of accumulated ethylene, because when the gaswas absorbed with 'Ethysorb' granules or its action inhibitedby 2,5–norbornadiene, leaf growth was normal. The removalof carbon dioxide with potassium hydroxide did not enhance theethylene effect, indicating little if any antagonism of ethyleneaction by carbon dioxide. Shoots of potato (Solanum tuberosumL. cv. Red Craig's Royal) were shortened in sealed culture vessels,in association with swelling, diageotropism and miniaturizationof the leaves. When tuber production was induced by decreasingthe photoperiod, increasing the sucrose concentration and includingcytokinin in the medium, partial sealing promoted conspicuoushypertrophy of the lenticels. These responses of potato wereprevented if the ethylene absorbant mercuric perchlorate wasenclosed together with the cultures. Plant tissue culture, poor aeration, ethylene, leaf expansion, Ficus lyrata Warb., Gerbera jamesonii Bolus, Solanum tuberosum L. cv. Red Craig's Royal     

Changes in the Morphology of Roots and Leaves of Carob Seedlings Induced by Nitrogen Source and Atmospheric Carbon Dioxide

Carob seedlings were grown hydroponically for 9 weeks under360 and 800 µl l^-1CO₂. One of two nitrogen sources, nitrateor ammonium, was added to the nutrient medium at concentrationsof 3 mol m^-3. Root systems of the developing plants suppliedwith nitrate compared to those supplied with ammonium were characterizedby:(a)more biomass on the lower part of the root;(b)fewer lateralroots of first and second order;(c)longer roots;(d)higher specificroot length;(e)a smaller root diameter. The morphology of theroot systems of nitrate-fed plants changed in the presence ofelevated carbon dioxide concentrations, resembling, more closely,that of ammonium-fed plants. Total leaf area was higher in ammonium-than in nitrate-fed plants. Nitrate-fed plants had greater totalleaf area in the presence of high carbon dioxide than in normalCO₂, due to an increase in epidermal cell size that led to developmentof larger leaflets with lower stomatal frequency. The observedchanges in the morphology of roots and shoots agreed with theresults observed for total biomass production. Nitrate-fed plantsincreased their biomass production by 100% in the presence ofelevated CO₂compared to 15% in ammonium-fed plants, indicatingthat the response of carob to high CO₂concentrations is verydependent on the nitrogen source. Under elevated CO₂, nitrate-grownplants had a larger content of sucrose in both roots and shoots,while no significant difference was observed in the contentof sucrose in ammonium-grown plants, whether in ambient or enrichedcarbon dioxide. Hence, the differences in soluble carbohydratecontents can, at least partly, account for differences in rootand shoot morphology.Copyright 1997 Annals of Botany Company Ceratonia siliquaL.; carob; ammonium; carbohydrate; carbon dioxide; nitrate; morphology; sucrose     

The Influence of Volatile Substances from Incubated Litter of Pinus radiata on Seed Germination

Vapour from incubated Pinus radiata D. Don litter caused a depressionin the germination of Lolium perenne L. seeds and responsesvarying from stimulation to depression in germination of Trifoliumrepens L. seeds. These effects could be attributed to ethyleneand carbon dioxide. Experiments using ethylene and carbon dioxidesupported this conclusion. Pinus radiata, litter, Lolium perenne, Trifolium repens, seed germination, ethylene, carbon dioxide     

The Kinetics of Isocitrate Lyase Formation in Chlorella: Evidence for the Promotion of Enzyme Synthesis by Photophosphorylation

The addition of acetate to aerobic Chlorella pyrenoidosa indarkness was followed by the formations of isocitrate lyaseactiity. After a lag period of 40 minutes the formation proceededat a constant rate. By use of actylamide gel electrophoresisit was shown that the increase in enzyme activity was accompaniedby the formation of a new protein which, after separation byelectrophoresis, contained isocitrate lyase activity. The formationof isocitrate lyase was repressed by glucose; it was repressedby light in the presence of carbon dioxide, but not when DCMUwas added. In light, plus DCMU, isocitrate lyase was formedanaerobically and the capacity for photo-formation of isocitratelyase was saturated at 500 ergs/cm²/sec. In this respect theprocess resembled the photo-conversion of glucose to polysaccharidebut differed from the photo-assimilation of carbon dioxide whichbecame saturated at a heigher light intensity. Monochromaticlight of 706 mµ wavelength supported both isocitrate layseformation and the conversion of glucose to polysaccharide butnot carbon dioxide fixation. It is concluded that ATP generatedby cyclic photophosphorylatin can provide the energy for isocitratelyase synthesis in Chlorella.     

Heterocyst Formation in the Blue-green Alga Anabaens doliolum--A Study of Some Aspects of Photoregulation

Of the different wavelengths of visible spectrum, red light(630–680 nm) supports maximum heterocyst production andits effect depends upon incident energy and the exposure period.The action spectrum of heterocyst formation corresponds withthe absorption spectrum of major photosynthetic pigments. Absenaof carbon dioxide is inhibitory to heterocyst formation, butsugars can partially substitute for carbon dioxide in the light,not in the dark. The inductive effect of red light is not reversedby low or high energy green or far-red. Such results are consistentwith a photosynthetic role of light in heterocyst formation,although a direct activation of some enzymes by light may alsobe of importance. Anabaena dollolum, blue-green alga, heterocyst formation, light     

The Effects of Oxygen, Carbon Dioxide and Ethylene on Ethylene Biosynthesis in Relation to Shoot Extension in Seedlings of Rice (Oryza sativa) and Barnyard Grass (Echinochloa oryzoides)

Exposing dark-grown seedlings for 3 d to oxygen deficiency (0or 5 kPa) or to additions of carbon dioxide (10 kPa) or ethylene(0·1 Pa) slowed shoot extension in Echinochloa oryzoides,while in rice it was promoted by these treatments, except that5 kPa oxygen was without effect. In E. oryzoides this was dueto reduced growth of the mesocotyl, and in rice to enhancedgrowth of the coleoptile. These responses to carbon dioxideand oxygen deficiency were not consequences of increased ethyleneproduction, since this remained unchanged by carbon dioxideand depressed by oxygen shortage in both species. Furthermore,exogenous ethylene and the ethylene action inhibitor 2,5-norbornadieneeach failed to influence extension in anoxic seedlings, indicatingno regulatory role for ethylene in the absence of oxygen. However,concentrations of the ethylene precursor 1 -aminocyclopropane-1-carboxylic acid (ACC) were increased by carbon dioxide and0 kPa or 5 kPa oxygen, although after 72 h without oxygen totalACC production (i.e. changes in ethylene + ACC + MACC) was suppressedin both species. There was little effect on bound ACC [putativemalonyl-ACC (MACC)] formation. Transferring anaerobic (0 kPa)seedlings to oxygenated conditions (21 kPa) resulted in abnormallyfast rates of ethylene formation, possibly due to the accumulationof ACC under anoxia. This post-anoxic ethylene may have contributedto the faster extension by rice coleoptiles and slower extensionby mesocotyls of E. oryzoides compared with those of seedlingsmaintained continuously in air. Echinochloa oryzoides [Ard.] Fritsch, barnyard grass, Oryza sativa L, rice, oxygen shortage, carbon dioxide, ethylene biosynthesis, shoot extension, 1-aminocyclopropane-1-carboxylic acid (ACC), malonyl-ACC, GC-MS     

Metabolic Systems in the 'Root' of Brassica napus L.

This is a study of the effects of possible intermediary metaboliteson the respiration of root tissue from Brassica napus usingthe Warburg micro-manometric technique. It is concluded thatascorbic acid is oxidized by two systems, one of which appearsto be a direct oxidase and the other a dehydrogenase. No evidenceof peroxidase activity was secured. A substantial fraction ofthe total respiratory activity was insensitive to cyanide andazide. The biologically important organic acids were oxidizedwith the production of carbon dioxide. Glutamic and asparticacids were metabolized with great rapidity, glycine and alaninemuch more slowly. A scheme integrating these results is outlinedand compared with the respiratory systems existing in potato.     

Studies on the Growth in Culture of Plant Cells: XI. THE INFLUENCE OF SHAKING RATE ON THE GROWTH OF SUSPENSION CULTUEES

The influence of shaking rates (expressed as revolutions permin) on orbital shaking platforms (1 in (2.54 cm) diam. rotarymotion) on the growth of cell suspension cultures of Acer pseudoplatanusL. and Atropa belladonna cultivar lutea Döll are described.By following cell growth and respiration and the levels of oxygenand carbon dioxide in the media during the progress of incubationit is concluded that the reduction of growth at sub-optimalshaking rates is not due to oxygen deficiency or toxic accumulationof carbon dioxide. The growth of the Atropa cell suspensionin ‘closed systems’ has been studied by the developmentof modified culture vessels and evidence obtained that the reducedgrowth in the systems is due to the formation by the culturesof an unidentified volatile growth inhibitor and not to eitheroxygen depletion or toxic accumulation of either carbon dioxideor ethylene. It is suggested that the reduced growth in ‘opensystems’ cultures at sub-optimal shaking speeds is eitherdue to retention of this volatile inhibitor or to restrictionof nutrient uptake by the existence of a stationary liquid-phaseboundary to the cells.     

In-Phase Cycling of Photosynthesis and Conductance at Saturating Carbon Dioxide Pressure Induced by Increases in Water Vapour Pressure Deficit

Bunce, J. A. 1987. In-phase cycling of photosynthesis and conductanceat saturating carbon dioxide pressure induced by increases inwater vapour pressure deficit.—J. exp. Bot. 38: 1413–1420. The leaf to air water vapour deficit was increased suddenlyfrom about 1·0 to 2·5 IcPa for single leaves ofsoybean (Glycine max L. Merr.) plants held at 30 °C, 2·0mmol m ^–2 s^–1 photosynthetic photon flux density(PPFD) and carbon dioxide pressures saturating to photosynthesis.After a lag of about 10 min, photosynthetic rate and stomatalconductance to water vapour began to decrease, and then cycledin phase with each other. The period of the cydes was about20 min. During these cycles the substomatal carbon dioxide pressurewas constant in the majority of leaves examined, and was alwaysabove saturation for photosynthesis. Epidermal impressions showedthat most stomata changed in aperture during the cycles, andthat very few were ever fully closed. Water potential measuredon excised discs changed by at most 0·1 MPa from theminima to the maxima in transpiration rate. In contrast, forleaves of sunflower (Helianthus animus L.) grown at low PPFD,the increase in VPD led to leaf wilting and decreased photosynthesis,followed by recovery of turgor and photosynthesis as stomatalconductance began to decrease. In these leaves photosynthesisand conductance then cycled approximately 180° out of phase.It is suggested that in soybeans decreased leaf conductanceinduced by high VPD provided a signal which decreased the rateof photosynthesis at carbon dioxide saturation by a mechanismthat was not related to a water deficit in the mesophyll. Key words: Photosynthesis, stomatal conductance, cycling, vapour pressure deficit     

Effects of Metabolic Inhibitors on the Calcification of a Freshwater Green Alga, Gloeotaenium loitlesbergarianum Hansgirg. 1. Effects of Some Photosynthetic and Respiratory Inhibitors

The effect of a number of metabolic inhibitors on the calcificationof Gloeotaenium loitlesbergarianum Hansgirg, a freshwater greenalga, was studied. The inhibitors used were methylamine, trimethylamine,mercuric chloride, imidazole, fluoride, arsenate, atrazine,DCMU and dinitrophenol. The effects of these inhibitors showthat transport, or stimulation of respiratory carbon dioxideevolution inhibits calcification. Calcification in Gloeotaeniumis, at least partly, due to a local rise in pH as a result ofphotosynthetic carbon dioxide assimilation. There is also someevidence that, apart from its role in carbon dioxide assimilation,photosynthesis supplies the additional energy needed for calcification. Calcification, Gloeotaenium loitlesbergarianum Hansgirg, green algae, Chlorophyceae, metabolic inhibitors, photosynthesis, respiration     

The Kinetics of Extracellular Glycollate Production by Chlorella pyrenoidosa

Extracellular glycollate is liberated by Chlorella pyrenoidosaduring growth in medium bubbled with air or 3 per cent carbondioxide in air. With air the rate of release of glycollate percell decreases, with 3 per cent carbon dioxide it increases,with increase in cell number. Glycollate is released duringshort-term experiments when C. pyrenoidasa, grown under lowlight and high carbon dioxide, is transferred suddenly to highlight and low carbon dioxide. No other combination of thesefactors produces a comparable release of glycollate. The quantityof glycollate released in short-term experiments increases exponentiallywith the relative growth-rate of the culture from which thecells are derived. A crucial condition for maximum glycollaterelease is that growth of the culture prior to the experimentshould not be limited by carbon-dixoide concentration. The effectof pH is related to its effect on growth-rate; i.e. C. pyrenoidosahas a lower relative growth-rate at pH 8.3 and produces correspondinglyless glycollate than faster growing cultures at pH 6.4. Duringshort-term experiments under high light and low carbon dioxidethe rate of glycollate release drops after 50–100 minutessuggesting exhaustion of the glycollate precursor.     

The Oxidation of 14C-labelled Glucose by Chlorella vulgaris: 1. The Time Course of 14CO2 Production

Glucose, either uniformly labelled with¹⁴C, or specificallylabelled in the I, 2, or 6 position, was added to C. vulgaris.Radio-active carbon dioxide was produced initially ten timesfaster from glucose-I-¹⁴C than from glucose-6-¹⁴C. This differencewas found with carbohydrate-starved cultures, exponentiallygrowing cultures, and cultures assimilating ammonia or nitraterapidly. A similar difference was also found with C. pyrenoidosaand Ankistrodesmus. 37 per cent. of the ¹⁴C added as glucose-¹-¹⁴Cto exponentially growing cells was recovered as carbon dioxidebut generally the recovery was less than this. Only 5 per cent.of ¹⁴C added as glucose-6-¹⁴C was recovered as carbon dioxide.The specific activity of the carbon dioxide produced was considerablylower than that of the carbon in the added glucose.