EFFECT OF CHROMATIC LIGHTS ON PHYCOBILIN FORMATION IN A BLUE-GREEN ALGA, TOLYPOTHRIX TENUIS

1. The formation of phycobilin pigments in a blue-green alga Tolypothrixtenuis was investigated with special reference to the effectsof preillumination with colored lights.
2. It was discoveredthat the algal cells are capable of formingphycobilin pigmentsin the dark, if they have been previouslyilluminated for severalhours in the presence of CO₂.
3. The color of light applied inthe later period of preillumination(chromatic illumination)was found to affect the ratio of phycoerythrinto phycocyaninformed in the subsequent dark period. A greenlight acceleratesthe dark-formation of phycoerythrin, a redlight that of phycocyanin,and the two lights counteractingwith each other in their effects.
4. These directive effects of the "chromatic illumination" canbe accomplished within a very short period, for instance, in3 minutes if it is preceded by sufficient "preillumination"with an incandescent or day light fluorescent light. The reactionsoccurring during the period of chromatic illumination does notrequire the presence of CO₂ and the aerobic condition.
5. Thealga can be grown heterotrophically when supplied with casaminoacids and glucose. Under such a condition the alga forms phycocyanintogether with chlorophyll and carotenoids, but not phycoerythrin.
6. On the basis of the results obtained, a tentative scheme forthe biosynthesis of phycobilin pigments in the alga was proposed,assuming the light-induced formation of unknown precursors whichare converted into phycocyanin and phycoerythrin in the subsequentdark period.

(Received July 4, 1960; )     

CHANGES IN EMISSION SPECTRA OF PHOTOSYNTHETIC PIGMENTS IN VIVO

1. The effects of 3-(4'-chlorophenyl)-1, 1-dimethylurea (CMU)onthe fluorescence of photosynthetic pigments in vivo wereinvestigatedin blue-green, red and brown algae and in isolatedspinach chloroplasts.CMU caused an increase in steady statelevel of fluorescenceof chlorophyll a, but did not influencethe fluorescence ofphycobilins. The spectrum of the fluorescenceincrement hada peak at 685 m/µ and a shoulder at 730–740mµ.These two bands probably arise from chlorophyll a(C_f684) belongingto pigment system II.
2. On excitation of chlorophylla in a red alga, Porphyra yezoensis,a fluorescence band witha peak at 720 mµ was observedbesides a shoulder at 685mµ. The 720 m band is inferredto arise from chlorophylla (probably, C_f-1) in pigment systemI.
3. On addition of CMUto the algal cells, the induction of fluorescencewas modifiedto take a simple time course. The induction wasobserved onlywith respect to the fluorescence of chlorophylla, but not inthe fluorescence of phycobilins. The spectrumof the "transient"fluorescence showed two emission bands ofchlorophyll a at 685mµ and 740 mµ, and was quitesimilar in form tothe spectrum of the CMU-caused increase insteady state fluorescence.
4. These facts were interpreted in terms of the correlation offluorescence of chlorophyll a and the photochemical reactionsof photosynthesis

(Received July 20, 1967; )     

COMPONENTS OF THE ELECTRON-TRANSFERRING SYSTEM IN ACETOBACTER SUBOXYDANS AND RECONSTRUCTION OF THE LACTATE OXIDATION SYSTEM

1. Cytochromes a₁⁵⁹⁰, b⁵⁶⁰, c₁⁵⁵⁴ and c₁⁵⁵² were isolated andpurifiedfrom a strain of Acetobacter suboxydans. The proceduresusedwere described in detail.
2. The main cytochrome band at550-560 mµ in intact cellssplitted at liquid air temperatureinto two bands, 551 mµ(strong) and 559 mµ (weak).
3. Optical and physiological properties of the four cytochromeswere investigated. Lactic dehydrogenase activity was found tobe associated with cytochrome c₁⁵⁵⁴. The two c₁-type cytochromes,especially cytochrome c₁⁵⁵⁴, persisted in their reduced formafter the purification through many steps.
4. By some combinationsof isolated components reconstruction ofthe oxygen uptake systemcould be realized.
5. The oxygen-consuming activity of purifiedoxidase preparationswas accelerated by a-tocopherol but notby Emasoll 4130 andTween 80.
6. Some discussions were made onthe nature of terminal oxidase,the role of cytochrome c₁⁵⁵²in the electron-transport system,and persistence of reducedstate of c₁-type cytochromes.
7. A possible scheme of the electron-transferringsystem of Acetobactersuboxydans was presented.

(Received May 16, 1960; )     

CHANGES IN RESTING POTENTIAL AND ION ABSORPTION INDUCED BY LIGHT IN A SINGLE PLANT CELL

1. Effect of light on ion absorption and resting potential of theinternodal cell of Nitella flexilis was investigated under variousconditions.
2. On illumination, the resting potential increasedby about 30mVin 10^–4 M KCl and by about 60 mV in 10^–4M NaClsolution. A similar photoelectric response was also observedin 10^–3 M KCl, 10^–2 M CaCl₂ and 5 x 10^–2 MCaCl₂ solutions, but not at all in 10^–2 M KCl solution.
3. Absorption of ions by the cell took place in parallel withthelight-induced change in resting potential.
4. Red and bluelights were very effective in increasing the restingpotential,while green light was almost ineffective. These differenteffectsof color lights were in good agreement with their effectsinincreasing the osmotic value of the cell.
5. The photoelectricresponse was not affected by phenylurethane,which, on the otherhand, strongly inhibited the light-inducedion absorption.
6. Theuptake of ions by the cell from the external medium intothevacuole is assumed to proceed in two different steps: thefirstis the process involving the ion movements across theoutermostplasmalemma, and the second is that involved in thetransportof ions through the cytoplasmic layer and tonoplast.The formerprocess is considered to be influenced by the increasein restingpotential probably caused by the light absorbed bychlorophyll.The process was, however, suggested to be independentof photosynthesis.On the other hand, the latter process issupposed to be relatedto photosynthesis. A discussion was madealong this line.

(Received July 26, 1962; )     

Are Mistletoes Shade Plants? CO2Assimilation and Chlorophyll Fluorescence of Temperate Mistletoes and their Hosts

Mistletoes usually have slower rates of photosynthesis thantheir hosts. This study examines CO₂assimilation, chlorophyllfluorescence and the chlorophyll content of temperate host–parasitepairs (nine hosts parasitized by Ileostylus micranthus and Carpodetusserratus parasitized by Tupeia antarctica). The hosts of I.micranthus had higher mean annual CO₂assimilation (3.59 ±0.41 µmol m^-2 s^-1) than I. micranthus(2.42 ± 0.20µmol m^-2 s^-1), and C. serratus(2.41 ± 0.43 µmolm^-2 s^-1) showed higher CO₂assimilation than T. antarctica(0.67± 0.64 µmol m^-2 s^-1). Hosts saturated at significantlyhigher electron transport rates (ETR) and light levels thanmistletoes. The positive relationship between CO₂assimilationand electron transport suggests that the lower CO₂assimilationrates in mistletoes are a consequence of lower electron transportrates. When photosynthetic rates, ETR and chlorophyll a /b ratioswere adjusted for photosynthetically active radiation, hostsdid not have significantly higher CO₂assimilation (3.21 ±0.37 µmol m^-2 s^-1) than mistletoes (2.54 ± 0.41µmol m^-2 s^-1), but still had significantly higher ETRand chlorophyll a / b ratios. The electron transport rates,saturating light and chlorophyll a / b ratios of sun leavesfrom mistletoes were similar to host shade leaves. These responsesindicate that in comparison with their hosts, mistletoe leaveshave the photosynthetic characteristics of the leaves of shadeplants. Copyright 2000 Annals of Botany Company CO₂assimilation, photosynthetic active radiation (PAR), chlorophyll fluorescence, electron transport rate (ETR), photochemical quenching (q_p), non-photochemical quenching (q_n), sun and shade leaves, chlorophyll content, Ileostylus micranthus, Tupeia antarctica, New Zealand     

LIGHT-INDUCED REDUCTION OF NITRATE, NITRITE AND HYDROXYLAMINE IN A BLUE-GREEN ALGA, ANABAENA CYLINDRICA

1. Reduction of nitrate, nitrite and hydroxylamine by intact cellsof Anabaena cylindrica was investigated with special referenceto the stimulating effect of light on these processes.
2. Itwas found that in light and under anaerobic condition thesecompounds are reduced to ammonia, with the production of extraoxygen. The stoichiometry of the reactions under these conditionscan be represented as follows: HNO₂+H₂O=NH₂+2O₂ HNO₂+H₂O=NH₂+1O₂ NH₂OH+H₂O=NH₂+O₂+H₂O
3. Reduction of nitrite and hydroxylaminewas markedly suppressedby CMU in the light but not in the dark.KCN inhibited reductionto the same extent both in the lightand in the dark. Reductionin the light was much less sensitiveto the uncoupling agent,DNP, than was that in the dark.
4. Atlow light intensities, CO_2– was suppressed by 20–30per cent by the simultaneous provision of nitrite, but the nitritereduction was not affected at all by CO₂. At high light intensities,reduction of nitrate and nitrite was considerably acceleratedby CO₂
5. On the basis of these findings, a possible mechanismfor thelight stimulation of the reactions in question was brieflydiscussed.

(Received August 22, 1962; )     

LACTATE OXIDATION SYSTEM IN ACETOBACTER SUBOXYDANS, WITH SPECIAL REFERENCE TO CARBON MONOXIDE-BINDING PIGMENT

1. Lactate oxidation system was investigated in Acetobacter suboxydans,which was found to have cytochromes a and b, but no cytochromec. Haemoprotein 558 was also found to exist.
2. Carbon monoxideinhibited the lactate oxidation in the darkbut not in the light.WARBURG'S partition constant was estimatedto be 7.
3. Additionof haemoprotein 558 noticeably enhanced the oxygenuptake bythe LDH preparation, which was obtained from bacterialcellsand partially purified.
4. Haemoprotein 558 has protohaem asits prosthetic group. Notonly its absorption spectrum is reminiscentof that of peroxidase,but it also shows peroxidase-like reactivitywith some ligandswith a few exceptions.
5. Ferrohaemoprotein558 reacts with oxygen, forming, at first,a complex, whichhas its SORET absorption peak at 423 mµ.The absorptionmaximum then shifts to 417 mµ, indicatinga transformationto another compound. One of these two productsis likely tobe the oxygenated ferro-haemoprotein 558.
6. The mutual transformationbetween cytochrome B and haemoprotein558 is discussed.

(Received October 8, 1965; )     

Biological activity of phyllosinol, a phytotoxic compound isolated from a culture filtrate of Phyllosticta sp.

1. Phyllosinol is a phytotoxic metabolite of Phyllosticta sp. Thissubstance at 100 µg/ml produced dark grey necrotic lesionson the leaf of red clover. Sensitivities of various plant speciesto phyllosinol differed both quantitatively and qualitatively.
2. Phyllosinol reduced root growth in rice seedlings by 60% at10^–4 M, whereas stimulation of root elongation occurredat a concentration range from 10^–9 to 10^–5 M.
3. Phyllosinolat 2.5x10^–4M promoted adventitious root formationin epicotylsof Azukia cuttings by about 100%. Promotion waspartly reducedby simultaneous application of cysteine.
4. IAA-induced elongationof isolated Avena coleoptile sectionswas inhibited by phyllosinolat a concentration range from 10^–5to 10^–3M.
5. Sulfhydrylcompounds, i.e. cysteine and glutathione relievedinhibitioncaused by phyllosinol in IAA-induced elongation ofAvena coleoptilesections.
6. GA₃-induced elongation of wheat leaf sections wasslightly inhibitedby phyllosinol at 10^–4M.
7. Phyllosinolalso has antibiotic activity. Among the organismstested, Phycomycetesand Gram-negative bacteria appeared mostsusceptible to phyllosinol.

(Received April 21, 1970; )     

Photosynthesis of Stands of Tomato, Cucumber and Sweet Pepper Measured in Greenhouses under Various CO2-concentrations

The rate of net photosynthesis (P) of whole plant stands oftomato (Lycopersicon esculentum Mill.), cucumber (Cucumis sativusL.) and sweet pepper (Capsicum annuum L.) was measured in sixlong-term experiments in large greenhouses under normal operatingconditions and CO₂-concentrations between 200 and 1200 µmolmol^-1. The objective was to quantify the responses to lightand carbon dioxide and to obtain data sets for testing simulationmodels. The method of measuring canopy photosynthesis involvedan accurate estimation of the greenhouse CO₂ balance, usingnitrous oxide (N₂O) as tracer gas to determine, on-line, theexchange rate between greenhouse and outside air. The estimatedrelative error in the observed P was about ± 10%, exceptthat higher relative errors could occur under particular conditions. A regression equation relating P to the photosynthetically activeradiation, the CO₂ concentration and the leaf area index explained83-91% of the variance. The main canopy photosynthesis characteristicscalculated with the fitted regression equations were: canopyP_max 5-9 g m^-2 h^-1 CO₂ uptake; ratio P_max/LAI 1·5-3 gm^-2 h^-1; light compensation point 32-86 µmol s^-1 m^-2;light use efficiency (quantum yield) at low light 0·06-0·10µmol µmol^-1 and CO₂ compensation point 18-54 µmolmol^-1. The results were related to the prevailing conditions.Copyright1994, 1999 Academic Press Canopy photosynthesis, Capsicum annuum L., carbon dioxide, CO₂, CO₂ balance, CO₂ use efficiency, cucumber, _{Cucumis sativus} L., glasshouse, greenhouse, light use efficiency, Lycopersicon esculentum Mill., sweet pepper, tomato, tracer gas     

Reactions of birch leaves to changes in light during early ontogeny: comparison between in vivo and in vitro techniques to measure carbon uptake

Trends in several photosynthetic parameters and their responseto changed growth light were followed for 15 d in leaves ofyoung birch saplings using a rapid-response gas exchange measuringequipment. These in vivo measurements were compared to biochemicalassays that were made from the same leaves after the gas exchangestudies. The measurements were made on leaves that were selectedprior to the study and were at that time of similar age. Forthe first 7 d the photosynthetic parameters were followed fromthe growth conditions of moderate light (200 µmol m^–2s^–1; referred to as controls later in the text). On day7 some of the saplings were transferred to grow either underhigh (450 µmol m^–2 s^–1; referred to as highlight plants) or low (75 µmol m^–2 s^–1; referredto as low light plants) light and the capability of the preselectedleaves for acclimation was followed for 6 d. For comparison,at the end of the experiment the measurements were made on bothcontrols and on young leaves that had developed under high andlow light. Generally the in vivo measured rate of CO₂ uptake (gross photosynthesis)both at 310 ppm CO₂ and 2000 ppm CO₂ corresponded very wellto the biochemically determined CO₂ fixation capacity in vitroafter rapid extraction (measured as the initial and total activityof Rubisco, respectively). However, if the flux of CO₂ intothe chloroplasts was limited by the closure of the stomata,as was the case of the high light plants, then the in vitromeasured Rubisco activity was greater than the in vivo measuredCO₂ uptake. V_max, calculated from the mesophyll conductanceat 1% O₂, exceeded the initial activity of Rubisco (assayedat saturating RuBP and CO₂) constantly by 60%. The catalyticactivity of Rubisco in birch leaves was overall very low, evenwhen calculated from the total activity of Rubisco (K_cat 0.63–1.18 s^–1), when compared to herbaceous C₃ species. Signs of light acclimation were not observed in most of thephotosynthetic parameters and in chloroplast structure whenmature birch leaves were subjected to changes in growth lightfor 6 d. However, the change of the growth light either to highor low light caused day-to-day fluctuations in most of the measuredphotosynthetic parameters and in the case of the high lightplants signs of photoinhibition and photodestruction were alsoobserved (decrease in the amount of chlorophyll and increasein chlorophyll a/b ratio). As a result of these fluctuationsthese plants achieved a new and lower steady-state conditionbetween the light and dark reactions, as judged from the molarratio of RuBP to Rubisco binding site. Key words: Acclimation, photosynthesis, light, Rubisco, birch     

Enhancement of Peroxidase-Dependent Oxidation of Sinapyl Alcohol by an Apoplastic Component, 4-Coumaric Acid Ester Isolated from Epicotyls of Vigna angularis L.

A water-soluble component that enhanced the peroxidase-dependent(POX-dependent) oxidation of sinapyl alcohol was isolated fromepicotyls of Vigna angularis. This compound was an ester of4-coumaric acid and a hexose, and it was found in both the apoplastand the symplast. The ester was oxidized by a basic POX isozyme(K_m, about 20 µM) and by an acidic POX isozyme (K_m, about40 µM) that had been partially purified from the apoplasticfraction of epicotyls of V. angularis. These POX isozymes oxidizedsinapyl alcohol at only a very low rate, but a 15-fold enhancementwas observed upon addition of the ester. The concentrationsof the ester required for the half-maximal enhancement weresimilar to the K_m values of the ester for its oxidation by therespective isozymes. The apoplastic concentration of the esterwas higher than 130 µM, suggesting that this ester mightact as a donor of electrons to the apoplastic POX isozymes insitu. Coniferyl alcohol also enhanced the POX-catalyzed oxidationof sinapyl alcohol. The concentrations of coniferyl alcoholrequired for half-maximal enhancement of the oxidation of sinapylalcohol were about 23 and 250 µM when reactions were catalyzedby the basic and acidic POXs, respectively. These values weresimilar to the K_m values of coniferyl alcohol for its oxidationby the respective isozymes. These results suggest that 4-coumaricacid ester and coniferyl alcohol, if it is present in the apoplast,can enhance the POX-dependent oxidation of sinapyl alcohol inthe apoplast of epicotyls of V. angularis. (Received July 1, 1996; Accepted February 5, 1997)     

Acclimation of Lolium temulentum to enhanced carbon dioxide concentration

Acclimation of single plants of Lolium temulentum to changing[CO₂] was studied on plants grown in controlled environmentsat 20°C with an 8 h photoperiod. In the first experimentplants were grown at 135 µ;mol m^–2 s^–1 photosyntheticphoton flux density (PPFD) at 415µl l^–1 or 550µll^–1 [CO₂] with some plants transferred from the lowerto the higher [CO₂] at emergence of leaf 4. In the second experimentplants were grown at 135 and 500 µmol m^–2 s^–1PPFD at 345 and 575 µl l^–1 [CO₂]. High [CO₂] during growth had little effect on stomatal density,total soluble proteins, chlorophyll a content, amount of Rubiscoor cytochrome f. However, increasing [CO₂] during measurementincreased photosynthetic rates, particularly in high light.Plants grown in the higher [CO₂] had greater leaf extension,leaf and plant growth rates in low but not in high light. Theresults are discussed in relation to the limitation of growthby sink capacity and the modifications in the plant which allowthe storage of extra assimilates at high [CO₂]. Key words: Lolium, carbon dioxide, photosynthesis, growth, stomatal density     

LIGHT-INDUCED OXYGEN EVOLUTION CAUSED BY HYDROGEN PEROXIDE IN CATALASE-INHIBITED CHLORELLA CELLS

1. Chlorella cells and spinach chioroplasts, whose catalase activityhad been more than 90% inhibited by 10^–5 M azide, werefound to decompose H₂O₂ photochemically to liberate oxygen,indicating that H₂O₂ was used as an oxidant of the HILL reaction.
2. That, however, the observed phenomena cannot be fully accountedfor in terms of the HILL reaction with H₂O₂ was revealed bythe observation that an extract of Chiorella cells, which hadbeen completely freed from chlorophyll, also showed a light-acceleratedO2 evolution from H₂O₂ in the presence of 10^–5 M azide.This extract contained a large quantity of catalase, which seemedto have been, in some way, involved in the reaction in question.
3. The catalatic H₂O₂ decomposition caused by crystalline catalaseof mammalian liver (in the presence of 10^–5 M azide) wasnot accelerated by the effect of light.

¹ Present address: Department of Biology, Faculty of Science,Niigata University, Niigata. (Received June 4, 1961; )     

Simultaneous changes in cell quotas of microcystin, chlorophyll a, protein and carbohydrate during different growth phases of a batch culture experiment with Microcystis aeruginosa

The cell quotas of microcystin (Q_mcyst), protein (Q_prot), chlorophylla (Q_chloro) and carbohydrate (Q_carbo), as well as the net productionrates of these parameters, were determined during the exponentialand stationary phases in nine batch cultures of Microcystisaeruginosa (CYA 228) at light regimes from 33 to 53 µmolphotons m^–2 s^–1. The following results were obtained.(i) A parallel pattern was found in the changes of Q_mcyst, Q_prot,Q_chloro and Q_carbo during the entire growth cycle and significantcorrelations were recorded between Q_mcyst and Q_prot, Q_chloroand Q_carbo. (ii) The net microcystin production rate (µ_mcyst)was positively correlated with the specific cell division rate(µ_c), the chlorophyll production rate (µ_chloro)and the protein production rate. (iii) A significant inverselinear relationship was found between µ_c and Q_mcyst, i.e.cultures with a positive µ_c had a Q_mcyst between 110 and400 fg microcystin cell^–1, while declining cultures hadQ_mcyst values >400 fg microcystin cell^–1. Maximum variationin Q_mcyst within cultures was 3.5-fold. Collectively, the resultsshow that cells produced microcystin at rates approximatingthose needed to replace losses to daughter cells during divisionand that microcystin was produced in a similar way to proteinand chlorophyll, indicating a constitutive microcystin production.     

Long Term Effects of High CO2 Concentration on Photosynthesis of Water Hyacinth (Eichhornia crassipes (Mart.) Solms)

The photosynthetic response to CO₂ concentration, light intensityand temperature was investigated in water hyacinth plants (Eichhorniacrassipes (Mart.) Solms) grown in summer at ambient CO₂ or at10000 µmol(CO₂) mol^–1 and in winter at 6000 µmol(CO₂)mol^–1 Plants grown and measured at ambient CO₂ had highphotosynthetic rate (35 µmo1(CO₂) m^–2 s^–1),high saturating photon flux density (1500–2000) µmolm^–2 s^–1 and low sensitivity to temperature in therange 20–40 °C. Maximum photosynthetic rate (63 µmol(CO₂)m^–2 s^–1) was reached at an internal CO₂ concentrationof 800 µmol mol^–1. Plants grown at high CO₂ in summerhad photosynthetic capacities at ambient CO₂ which were 15%less than for plants grown at ambient CO₂, but maximum photosyntheticrates were similar. Photosynthesis by plants grown at high CO₂and high light intensity had typical response curves to internalCO₂ concentration with saturation at high CO₂, but for plantsgrown under high CO₂ and low light and plants grown under lowCO₂ and high light intensity photosynthetic rates decreasedsharply at internal CO₂ concentrations above 1000 µmol^–1. Key words: Photosynthesis, CO₂, enrichment, Eichhornia crassipes     

EFFECT OF ULTRAVIOLET LIGHT UPON VARIOUS PHYSIOLOGICAL ACTIVITIES OF CHLORELLA CELLS AT DIFFERENT STAGES IN THEIR LIFE CYCLE

1. Using Chlorella ellipsoidea as material, investigations weremade of the effects of ultraviolet irradiation upon variousactivities of cells at different developmental stages in theirlife cycle. Cell activities investigated were photosynthesis,respira tion, over-all growth, modes of synchronous growth andcell division as well as the formation of nucleic acids. Theu. v.- light applied was 30 µµW/cm²in intensityand 2537 Å in wavelength.
2. The most u. v.-sensitive wasthe over-all growth activity, andin this respect the irradiationapplied at the L₂-stage wasmore inhibitive than that givenat the D-stage. The next mostvulnerable was the photosyntheticactivity, the sensitivitybeing the same in the D- and L-cells.The most resistant towardu.v. was the endogenous respirationof D-cells followed by theirrespiration using exogenous glucoseas substrate. The L₂-cellsappeared to be unable to use exogenousglucose as substrateof respiration, but their endogenous respirationwas considerablystronger than that of D-cells, and its u. v.-sensitivitywasthe same as that of glucose respiration of D-cells.
3. WhenD-cells were u. v. irradiated immediately before the startofsynchronous culture, growth and cell division as well astheformation of DNA and RNA were retarded in proportion totheu. v.-dose applied. The division number (n) was normal (around4) at lower u.v.-doses (1-2 minute irradiation), but was reducedto a half (about 2) at a higher dose.
4. When, during the synchronousculture, 1-minute u.v.- irradiationwas applied at various stagesof the ripening phase, the divisionwas retarded, but the cells,after attaining an abnormally largesize, divided into about8. If the irradiation was given atthe L₄-stage, the divisionnumber was practically unmodified(n=4.5), although the divisionwas somewhat retarded comparedwith that of the control culture.When a 1-minute irradiationwas given at the L₂-stage, thereoccurred an apparent stimulationof DNA- and RNA-formation,a phenomenon which corresponds tothe production of a largernumber of daughter cells than itwas the case in control cultures.
5. Thus the cells which were moderately u.v.-irradiated at differentstages of synchronous culture were able to complete their lifecycle, but later a certain portion of irradiated cells becameunable to grow normally.

¹Present address: Department of Biochemistry, Dartmouth MedicalSchool, Hanover, New Hampshire, U.S.A. (Received March 6, 1961; )     

Properties of Phosphoenolpyruvate Carboxylase and Carbohydrate Accumulation in the C3-CAM Intermediate Sedum telephium L. Grown Under Different Light and Watering Regimes

A comparison of the activity and properties of the enzyme phosphoenolpyruvatecarboxylase (PEPC) was made for plants of Sedum telephium L.grown under low (70 µmol m^–2 s^–1) or high(500µmol m^–2 s^–1) PPFD and subjected to varyingdegrees of water stress. Under well-watered conditions onlyplants grown under high PPFD accumulated titratable acidityovernight and the extractable activity of PEPC was almost 2-foldhigher in these plants than in plants grown under low PPFD.Increasing drought stress resulted in a substantial increasein the activity of PEPC extracted both during the light anddark periods and a decrease in the sensitivity to inhibitionby malic acid. The magnitude of these changes was determinedby the severity and duration of drought and by light intensity.A comparison of the kinetic properties of PEPC from severelydroughted plants revealed that plants droughted under high PPFDhad a lower K_m for PEP than plants under low PPFD. Additionof 2·0 mol m^–3 malate resulted in an increase inthe K_m for PEP, with plants draughted under low PPFD havinga significantly higher K_m in the presence of malic acid comparedto those under high _PPFD. Response to the activator glc-6-P,which lowered the K_m for PEP, also varied between plants grownunder the two light regimes. Under well-watered conditions PEPCextracted from plants under high PPFD was more sensitive toactivation by glc-6-P than those under low PPFD. After the severedrought treatment, however, the K_m for PEP in the presence ofglc-6-P was similar for enzyme extracted from plants grown underboth light regimes. Soluble sugars and starch were depletedovernight and were both possible sources of substrate for PEPC.With increasing drought, however, the depletion of starch relativeto soluble sugars increased under both light regimes. The propertiesof PEPC and the characteristics of carbohydrate accumulation/depletionare discussed in relation to the regulation of CAM in S. telephiumgrown under different light and watering regimes. Key words: PEP carboxylase, CAM, carbohydrates, Sedum telephium     

Effect of elevated CO2 on the utilization of light energy in Nothofagus fusca and Pinus radiata

Red beech (Nothofagus fusca (Hook. F.) Oerst.; Fagaceae) andradiata pine (Pinus radiata D. Don; Pinaceae) were grown for16 months in large open-top chambers at ambient (37 Pa) andelevated (66 Pa) atmospheric partial pressure of CO₂, and incontrol plots (no chamber). Summer-time measurements showedthat photosynthetic capacity was similar at elevated CO₂ (lightand CO₂-saturated value of 17.2 µmol m^–2 s^–1for beech, 13.5 µmol m^–2 s^–1 for pine), plantsgrown at ambient CO₂ (beech 21.0 µmol^–2 s^–1,pine 14.9 µmol m^–2s^–1) or control plants grownwithout chambers (beech 23.2 µmol m^–2 s^–1,pine 12.9 µmol m^–2 s^–1). However, the higherCO₂ partial pressure had a direct effect on photosynthetic rate,such that under their respective growth conditions, photosynthesisfor the elevated CO₂ treatment (measured at 70 Pa CO₂ partialpressure: beech 14.1 µmol m^–2 s^–1 pine 10.3)was greater than in ambient (measured at 35 Pa CO₂: beech 9.7µmol m^–2 s^–1, pine 7.0 µmol m^–2s^–1) or control plants (beech 10.8 µmol m^–2s^–1, pine 7.2 µmol m^–2 s^–1). Measurementsof chlorophyll fluorescence revealed no evidence of photodamagein any treatment for either species. The quantity of the photoprotectivexanthophyll cycle pigments and their degree of de-epoxidationat midday did not differ among treatments for either species.The photochemical efficiency of photosystem II (yield) was lowerin control plants than in chamber-grown plants, and was higherin chamber plants at ambient than at elevated CO₂. These resultssuggest that at lower (ambient) CO₂ partial pressure, beechplants may have dissipated excess energy by a mechanism thatdoes not involve the xanthophyll cycle pigments. Key words: Carotenoids, chlorophyll fluorescence, photosynthesis, photoinhibition, photoprotection, xanthophyll cycle     

Acclimation in Seedlings of a Tropical Tree, Bischofia javanica, Following a Stepwise Reduction in Light

Acclimation of fully developed leaves of Bischofia javanicaBlume to shadelight was examined. Seedlings were grown undersimulated daylight (1000 µmol m^–2 s^–1), thentransferred to a simulated shadelight (40 µmol m^–2s^–1). When a high-light leaf was transferred to low light, large negativenet photosynthetic rates (P_m) were recorded. This decrease wasrapid, but within 7 d the rate increased and became equal tothe low-light control leaf. These changes in photosynthesisdid not follow the pattern of changes in stomatal conductance(g_s). Transfer to the low light resulted in a dramatic decreasein leaf weight per unit area (L_w), and most of the decreasesin L_w occurred within 3 d of transfer when the P_m of the transferredleaf was well below that of the low-light control leaf. There was a significant decrease in chlorophyll a in the transferredleaf without an appreciable change in chlorophyll b resultingin a large decrease in the chlorophyll a to chlorophyll b ratio.Leaf chlorophylls per unit area were higher in the transferredleaf than the low-light control leaf. Maximum photosyntheticrate in the transferred leaf was decreased by 40% compared tothat for the high-control leaf, but was almost at the same extenthigher than the low-light control leaf The results are discussedin the context of carbon gain capacity of its seedlings underlight-limiting forest understorey habitats. Bischofia, chlorophylls, light, photosynthesis, shade acclimation, tree seedlings, tropical tree     

SYNCHRONOUS CULTURE OF CHLORELLA: I. KINETIC ANALYSIS OF THE LIFE CYCLE OF CHLORELLA ELLIPSOIDEA AS AFFECTED BY CHANGES OF TEMPERATURE AND LIGHT INTENSITY

1. Using the technique of synchronous culture, investigationsweremade of the effects of temperature and light-intensityon cellularlife cycle of Chlorella ellipsoidea. Some improvementsin theculture technique for obtaining a good synchrony of algalgrowthwere described.
2. By following the changes of averagecell volume and cell numberoccurring during culturing, therates of the following processesof life cycle were determined:(i) "growth" (or the increasein cell mass) occurring from thestage of smaller cells (D_a)to the stage of ripened cell (L₃),(ii) "ripening" (or processofformation of "nuclear substances"as estimated from the averagenumber of daughter cells formedfrom single mother cell), and(iii) " maturing and division" which leads to the full maturationof mother cells (L-cells)and their division into separate daughtercells (D-cells).
3. "Growth"and "ripening" were found to be dependent in light,"maturingand division" light-independent. The time requiredfor "growth"and "ripening" (C) is dependent on temperaturebut independentof light intensity, the onset of "maturing anddivision" occurringat the same time (D) of culturing undervaried light intensities.The average cell volume at this stage(L₃),however, was foundto be markedly modified by light intensity;larger with highertemperatures (see Fig. 4).
4. Changes in incubation temperature(under the condition of saturatinglight intensities) were foundto affect the life cycle in thefollowing way: (i) The timeof onset of "maturing and division"(D), varies markedly withculturing temperature; earlier athigher temperatures, (ii)The average cell volume at this stagealso depends on temperature; smaller at higher temperatures.
5. The average number of daughtercells (n) emerging from singlemother cells, was found to beuninfluenced by culturing temperature;(4.0–4.1 underthe conditions of the present study). Itwas found that thedivision number n is remarkably varied bychanging the lightintensity in the "growth" and "ripening"phases; 2.0 at 1 kilolux,3.7 at 5 kilolux, 4.2 at saturatinglight intensities (10 and25 kilolux). This finding was explainedby assuming a light-dependentformation of "nuclear substances"during the "growth" and "ripening"phases, the quantity of thesubstances in the cell at L₃ stagedeterminig the division number.
6. The experimental data wereanalyzed reaction kinetically, therate constants and othercharacteristics of the reactions constitutingthe processesof life cycle were determined, and values forthe apparent activationenergy for each reaction were computed.The reactions were discussedwith special reference to theirrelationship with photosyntheticprocess was discussed.

(Received November 7, 1959; )