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)     

Effect of Tribenzylphosphate on the Phosphate Translocator of Isolated Pea Chloroplasts

The effect of tribenzylphosphate on the activity of the phosphatetranslocator of intact pea chloroplasts was tested. The translocatoractivity was followed by O₂ evolution, ¹⁴CO₂ fixation and ³²Pback-exchange. The reagent inhibited 3-phosphoglycerate dependent-photosyntheticactivities probably through an interaction with the PGA translocator. (Received September 11, 1985; Accepted November 21, 1985)     

DNA Synthesis in Chloroplasts: III. THE DNA GYRASE INHIBITORS NALIDIXIC ACID AND NOVOBIOCIN INHIBIT BOTH THYMIDINE INCORPORATION INTO DNA AND PHOTOSYNTHETIC OXYGEN EVOLUTION BY ISOLATED CHLOROPLASTS

The influence of two DNA gyrase inhibitors, nalidixic acid andnovobiocin, on DNA synthesis in isolated pea chloroplasts wasexamined. Novobiocin at 1–5 mol m^–3 markedly lowered[³H]thymidine incorporation into DNA (30–95% inhibition);while less effective, nalidixic acid at similar concentrationsalso diminished incorporation (25–35% inhibition). Theinhibition of chloroplast DNA (ctDNA) biosynthesis by nalidixicacid and novobiocin was confirmed by autoradiography and densitometry.These data are consistent with the view that chloroplasts containa DNA gyrase-like enzyme which is necessary for DNA replication.Despite this, interpretation of the results is not straightforward,as both nalidixic acid and novobiocin also inhibited photosyntheticactivity. Each substance (at millimolar levels) reduced ferricyanide-dependentO₂ evolution in isolated chloroplasts. However, at lower concentrations(0.05–0.3 mol m^–3) they slightly enhanced photosyntheticelectron flow; thus, these compounds may act as uncouplers ofphotophosphorylation as well as inhibitors of electron transport.Nalidixic acid and novobiocin at relatively low (0.1 mol m^–3)concentrations also strongly reduced CO₂-dependent O₂ evolution(an index of CO₂ photo-assimilation) in isolated plastids. Thus,caution must be exercised in assessing results from studiesin which nalidixic acid and novobiocin are used with whole plants,cells, protoplasts or isolated chloroplasts. Key words: Chloroplast, DNA replication, novobiocin, nalidixic acid, DNA gyrase     

青海省三江源区人工草地生态系统CO2通量

 了解三江源人工草地净生态系统CO₂交换(Net ecosystem CO₂ exchange, NEE)的季节变化规律和主要生物因子及环境因子对这些过程的影响将有助于认识青藏高原人工草地生态系统碳循环、生态价值、功能,以及对三江源区的生态安全的重要意义。该研究利用涡度相关技术,于2005年9月1日至2006年8月31日对位于青海腹地的垂穗披碱草（Elymus nutans）人工草地的NEE及生物和环境因子进行观测, 阐明NEE及其组分的动态变化特征和影响因子。三江源区人工草地生态系统的日最大吸收量为2.38 g C·m^－2·d^－1,出 现在7月30日。日间最大吸收率和最大排放率都出现在8月,分别为-6.82和2.95μmol CO₂·m^－2·s^－1。在生长季, 白天的NEE主要受光合有效辐射(Photosynthe tically active rad iation, PAR)变化控制,同时又与叶面积指数和群落多样性交互作用,共同调节光合速率和光合效率的强度。最大光合同化速率为2.46～10.39μmol CO₂·m^－2·s^－1,表观初始光能利用率为0.013～0.070μmol CO₂·μmol^－1 PAR。 在碳交换日过程中,NEE并不完全随着 PAR的增加而增大,当PAR超过某一值（>1 200μmol ·m^－2·s^－1）时,NEE随PAR的增加而降低。受温度的影响,生长季的生态系统的呼吸商Q₁₀（1.8）小于非生长季节的 2.6）。 生态系统呼吸主要受温度的控制,同时也受到叶面积指数的显著影响。生长季昼夜温差大并不利于生态系统的碳获取。 三江源区人工草地生态系统是一个较强的碳汇,为-49.35 g C·m^－2·a^－1。     

Light-enhanced carbon dioxide fixation in isolated chloroplasts

Preillumination of leaves of spinach, soybean and maize in theabsence of CO₂ greatly enhanced the capacity for fixing CO₂in an immediately following dark period. Lightenhanced darkCO₂-fixation was further observed in isolated chloroplasts ofspinach and soybean. When isolated chloroplasts were illuminated,CO₂-fixing capacity in the subsequent dark period increasedrapidly at first and later more slowly attaining a stationaryvalue in about 20 min. When the light was turned off at thisstage, the capacity decreased very rapidly becoming zero inabout 10 min. The magnitude of the enhanced dark fixation andits decay in the dark were not influenced by the presence orabsence of atmospheric oxygen. In both leaves and isolated chloroplasts,no significant change in oxygen (21%) occurred in distributionpatterns of radioactivity in products fixed by photosynthetic,or light-enhanced, dark, ¹⁴CO₂-fixation. In preilluminated leaves¹⁴C was incorporated into sucrose in the subsequent dark period,indicating that the photosynthetic carbon reduction cycle isoperating in light-enhanced dark fixation in higher plants. ¹Present address: Noda Institute for Scientific Research, Noda,Chiba Prefecture (Received August 10, 1970; )     

Studies in the Respiratory and Carbohydrate Metabolism of Plant Tissues1: XVII. THE EFFECT OF IODOACETATE ON THE CO2 OUTPUT, PATHWAYS OF GLUCOSE RESPIRATION AND CONTENTS OF PHOSPHATE ESTERS IN STRAWBERRY LEAVES

When specifically labelled glucose was fed to strawberry leaves,the C₆/C₁, quotient (rate of release of ¹⁴CO₂ from glucose-6-¹⁴C/rateof release of ¹⁴CO₂ from glucose-1¹⁴C ranged from 0.27 to 0.35in leaves in water and from 0.46 to 0.96 in leaves fed withiodoacetate. These quotients indicate that both the glycolyticand the pentose phosphate pathways participate in the respirationof strawberry leaves, with a greater contribution from the formerin the iodoacetate increased CO₂ output. Concurrently with the increase of CO² output in iodoacetate,the contents of glucose-6-phosphate (G6P), fructose-6 (F6P)and fructose-1,6-diphosphate (FDP) increased greatly; therewas a smaller increase of phosphoenol-pyruvate (PEP). The increasein the CO₂ output in iodoacetate may be explained solely onthe basis that the increases of G6P and FDP accelerate the ratesrespectively of the pentose phosphate pathway and of glycolysisand traffic into the tricarboxylic acid cycle. The increasein content of G6P and FDP is attributed to an increase in theaccessibility of enzymes and substrates caused by iodoacetate.Alternatively the increased CO₂ output in iodoacetate may bepartly due to uncoupling of oxidative phosphorylation.     

STUDIES ON THE RE-ASSIMILATION OF RESPIRATORY CO2 IN ILLUMINATED LEAVES

Experiments were performed, using rice, barley and Hydrangealeaves, to examine the re-assimilation of respiratory ¹⁴CO₂while photosynthesis is going on in an open air flow system. It was found that the leaves which had assimilated ¹⁴CO₂ beforehandevolved, when kept under photosynthesizing conditions, threeto four tenths (variable according to plant species and externalconditions) of the amount of ¹⁴CO₂ to be produced in the dark.Such an incomplete re-utilization of ¹⁴CO₂ was observed alsoin spinach leaf homogenate as well as in the leaves which hadpreviously absorbed ¹⁴C-glucose. The ¹⁴CO₂ output in rice leaves was found to be acceleratedby the light of high intensity. A possibility of light stimulationon the respiration was suggested. (Received October 7, 1961; )     

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; )     

EFFECT OF {alpha}-HYDROXYSULFONATES ON PHOTOCHEMICAL REACTIONS OF SPINACH CHLOROPLASTS AND PARTICIPATION OF GLYOXYLATE IN PHOTOPHOSPHORYLATION

1. The effect of -hydroxy sulfonates and sulfite, inhibitors ofglycolate oxidase, on the photochemical reactions of spinachchloroplasts was studied. The photo reduction of ferricyanideand NADP was not affected by the poisons, whereas the photophosphorylationand ¹⁴CO₂ fixation were inhibited.
2. Glyoxylate was photoreducedby the chloroplasts in the presenceof PPNR and glyoxylate reductase,and this reduction was acceleratedby the addition of NADP.ATP formation accompanied with thereduction of glyoxylate bythe illuminated chloroplasts wasobserved. It is supposed thatglyoxylate oxidizes the photoreducedNADPH₂ or PPNR and thusthe photophosphorylation is stimulated.

¹A part of this paper was presented at the annual meeting ofAgricultural Society of Japan, in August, 1964. ²Present address: Radiation Center of Osaka Prefecture, Sakai,Osaka.     

The Effect of Subjecting Peas to Air Enriched with Carbon Dioxide: II. RESPIRATION AND THE METABOLISM OF THE MAJOR ACIDS

Whole peas at about 75 per cent of their maximum fresh weightwere subjected to 5–30 per cent CO₂ in air for periodsof from 1–6 d, then returned to air for a further 1 d.Samples were withdrawn at intervals and organic acids, ^TCO₂and ethanol estimated as well as the rate of respiration. Slicesof cotyledons suspended in water were also subjected to highconcentrations of CO₂ in air for 3 h. The rate of respiration was inhibited progressively by increasein CO₂ content of the tissue. The high internal CO₂ contentof the intact pea causes an inhibition of its rate of respirationby about 25 per cent. Alcohol production commenced at between10 and 15 per cent CO₂ in the ambient gas and slowly increasedin rate up to 37 per cent. The CO₂-air mixtures reduced the content of malate, pyruvateand -oxoglutarate, increased that of succinate and left citrateunaffected. On return to air malate rose rapidly and succinatefell slowly to their original concentrations. During the sameperiod the concentration of PEP fell sharply and after about1 h rose again, whereas oxalacetate showed a reverse response.It is argued that the rapid re-synthesis of malate was by carboxylationof PEP to oxalacetate and that this reaction was stimulatedby a change in pH rather than by the direct effect of the changein concentration of CO₂. In one experiment ¹⁴CO₂ was supplied for 2 h before return toair and the movement of ¹⁴C followed for 6 h. The results supportthe method of re-synthesis of malate proposed.     

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     

Effect of linolenate on photosynthesis by intact spinach chloroplasts II. Influence of preillumination of leaves on the inhibition of photosynthesis by linolenate

The inhibitory effect of linolenate on intact spinach chloroplastsdepends on the level of the internal pool of metabolites. Chloroplastsfrom preilluminated leaves or chloroplasts artificially loadedwith 3-phosphoglyceric acid required higher concentrations oforthophosphate for maximal rates of CO₂ dependent O₂ evolutionthan untreated chloroplasts. The loaded chloroplasts were moresensitive to linolenate, and in the presence of linolenate theoptimal phosphate concentration was shifted toward lower values.We propose that the inhibition of photosynthesis by linolenateis due to inhibition of the "phosphate translocator". ¹ Part of this work has been published in the Book of Abstracts,4th International Congress on Photosynthesis, Reading, U.K.,1977, p. 265–266. ² This work is part of a doctoral programme carried out by L.Mv6 Akamba in this laboratory. ³ To whom reprint requests should be adressed. (Received October 14, 1978; )     

Bicarbonate stimulation of oxygen evolution, ferricyanide reduction and photoinactivation using isolated chloroplasts

Dependence of Hill reaction (ferricyanide reduction) by isolated(broken) chloroplasts on bicarbonate ion increases with timeof illumination (upto 4 min) in HCO₃^- free reaction mixture.The stimulation caused by HCO₃^- is independent of light intensitydown to very low intensities indicating an involvement of thision in early photochemical events of photosystem II. Oxygenevolution was found to be more dependent than ferricyanide reductionon HCO₃^-. The existence of an endogenous non-oxygen evolvingelectron donor in chloroplasts is thus suggested. HCO₃^- is alsoshown to greatly increase the rate of photoinactivation duringHill reaction. (Received March 2, 1974; )     

准噶尔荒漠早春短命植物的光合特性及生物量分配特点

准噶尔荒漠分布的早春短命植物不仅具有十分独特的生物学特点,而且在荒漠植物群落演替、物种多样性维持及土壤改良与防治水土流失等方面具有重要的生态学价值。该文运用Li-6400开放式气体交换光合作用测定系统,对分布于准噶尔荒漠的16种早春短命植物生长盛期的净光合速率(P_n)、蒸腾速率(T_r)、水分利用效率(WUE)等特征进行了测定,并对其中7种植物与生长相关的生物量分配特征进行了分析。结果表明:1)16种植物的最大P_n、 最大T_r及WUE分别为8.07~35.96 μmol CO₂·m^-2·s^-1、3.16~29.64 mmol H₂O·m^-2·s^-1、0.54~4.26 μmol CO₂·mmol^-1H₂O;种间最大P_n与最大气孔导度(Stomatal conductance, G_s)之间存在正相关关系,其相关系数为0.77(p<0.05),线性回归斜率为26.36 μmol·mmol^-1;从光合速率对胞间CO₂浓度及光量子通量密度的响应曲线来看,这类植物的表观CO₂补偿点均在4~5 Pa之间(28~30 ℃),表观羧化效率为0.64~1.86 μmol CO₂·m^-2·s^-1·Pa^-1,表观量子效率为0.05~0.06。2)从生物量分配来看,所测植物的个体生物量为0.05~0.39 g;单株总叶面积为 3.24~51.40 cm²;单位叶面积干重为0.40~0.77 g·m^-2,根在总生物量中所占比例为5.72%~19.43%,单株叶面积比在2.92~9.00 m²·kg^-1之间。种间根所占生物量的比与对应的WUE之间的比较分析结果表明,二者之间存在显著的正相关关系,其相关系数r为0.93(p<0.01)。这些结果表明,所观测的早春短命植物具有典型的C₃植物特征,相比其它类型的荒漠植物具有较高的单位叶面积P_n、高T_r及低WUE,并且在生长发育过程中表现出很低的根/地上生物量比、较高的叶面积比和单位叶面积干重,说明它们具有相对高的生长速率,这与其生长发育节律相一致,反映了它们与准噶尔荒漠环境相适应的特点。     

The Transport and Metabolism of Urea in Chara australis: II. UREASE AND THE DISTRIBUTION OF TRANSPORTED UREA

The ureolytic enzyme in Chara was investigated. This enzymewas shown to be a urease with an unusually high affinity forurea(K_m = 158 mmol m^-3). Little inhibition of urease activitywas found when intact Chara cells were exposed to the ureaseinhibitors hydroxyurea, acetohydroxamic acid and N-ethylmaleimide,although there was some inhibition of urea uptake. The distribution of radioactivity amongst the amino acid, organicacid and sugar/neutral fractions, determined by ion-exchangechromatography, was very similar whether the Chara internodeswere exposed to ¹⁴C-urea or to H¹⁴CO₃. This suggests that thefraction of the urea-carbon liberated by the urease as CO₂ andretained by the cell is used in photosynthetic carbon-fixation.During the initial 15 min of ¹⁴C-urea uptake, label appearsin the vacuole only in the form of unmetabolized urea. Afterthis time a variety of labelled compounds appear in the vacuole,presumably reflecting the gradual movement of carbon-fixationproducts from the chloroplasts to the cytoplasm and thence intothe vacuole. Key words: Urea transport, metabolism, Chara, urease     

Photosynthetic Properties of Green Barley Leaves after Treatment with Pyridazinone Herbicides--Comparison with the Effects of Diuron

Photosynthetic characteristics of detached green barley leavesafter 72 h of treatment with 0·2 mol m^–3 of thepyridazinone herbicides SAN 6706, SAN 9785 and SAN 9789 werestudied. For comparison, the effects of 0·01 mol m^–3diuron were also investigated. Pyridazinone herbicides causedonly a slight reduction of the total carotene content of thebarley leaves. The total chlorophyll content, as well as thelinolenic to linoleic acid ratio of chloroplast glycerolipids,however, remained unchanged. Diuron treatment caused total inhibitionof electron transport, as revealed by fast fluorescence inductionof leaves and the Hill reaction activity of chloroplasts. The¹⁴CO₂-nxation by the leaves and the light-induced fluorescencequenching were also completely inhibited in vivo by diuron.Pyridazinone herbicides left 20–40% of the ¹⁴CO₂-fixationfound in the control, in spite of the fact that their fast fluorescenceinduction tracings showed inhibition in the electron transport.Chloroplasts isolated from the leaves treated with pyridazinoneswere found to be highly active in mediating the ferricyanide-dependentHill reaction. In order to test the ability of pyridazinonesto inhibit photosynthetic electron transport in vivo, their‘prompt’ effect on fluorescence was also investigated.It is concluded that pyridazinone herbicides can readily andrapidly enter the chloroplasts and inhibit the photosyntheticelectron transport in vivo. The differences between the long-termeffects of pyridazinones and those of diuron suggest differencesin the inhibitory effectiveness on the various photosyntheticparameters between the two herbicide groups. It is suggestedthat pyridazinones can leave the chloroplasts during isolationowing to the loose binding onto the thylakoid membranes. Key words: Pyridazinone herbicides, electron transport, fluorescence induction     

Effect of abscisic acid on differentiation and ribulose diphosphate carboxylase of chloroplasts

Daucus carota tissues were grown on Murashige-Skoog medium (MS)at different concentrations with abscisic acid (ABA). Sevenbands of chloroplast fractions were obtained on a sucrose gradient.At 10^–5M, ABA highly increased chlorophyll and proteinnitrogen content of medium density chloroplasts. With increasingage of the tissues, the most active chloroplasts according totheir ¹⁴CO₂ fixation were found in smaller numbers. When treatedwith 10^–5 M ABA, 34 day-old tissues cultivated in vitroshowed the chloroplast pattern of 110 day-old tissues. The effectof ABA—given to the tissues during a short pretreatmentor continuously present in the culture medium—on the ribulosediphosphate carboxylase activity was analysed. It was foundthat ABA at 10^–5 M strongly inhibited ¹⁴CO₂ fixation. (Received December 20, 1977; )     

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; )     

Glycine as a substrate for photorespiration

Substrates for photorespiration were examined by feeding ¹⁴Clabeled compounds to tobacco and corn leaf segments and by measuring¹⁴CO₂ evolution in light and darkness. CO₂ release in the darkwas rapid, but in light CO₂ release was slow due to refixationby photosynthesis. Carboxyl labeled glycine was more rapidlydecarboxylated than were glyoxylate, glycolate or serine. Hydroxypyridinemethanesulfonate, an inhibitor of glycolate oxidase, blocked CO₂ releasefrom glycolate but not from glycine. Isonicotynyl hydrazideblocked CO₂ release from both glycine and glycolate. DCMU blockedphotosynthetic refixation of the released CO₂, consequentlythe rates of CO₂ release in light and dark were about equal.It was concluded that CO₂ release during photo-respiration camefrom the conversion of 2 molecules of glycine to one serineand one CO₂. ¹⁴CO₂ release from glycine-l-¹⁴C in the dark or with DCMU inlight can be used as an assay for photorespiration ability. CO₂ release from glycine and glycolate by corn leaf segmentsin the dark proceeded at the rate of that in normal tobaccoleaf. This result, together with other work on O₂ exchange andenzymatic analysis, indicates that corn and other plants docarry on photorespiration, but it is not manifested by CO₂ releasein light. A yellow tobacco mutant, Consolation 402, had high rates ofphotorespiration by the ¹⁴CO₂ assay, nearly half (or more) asmany peroxisomes as chloroplasts, and high rates of CO₂ releasefrom glycine-l-¹⁴C or glycolate-l-¹⁴C. A common tobacco, BrightYellow, had lower rates of photorespiration, fewer visible peroxisomes,and slower decarboxylation of glycine and glycolate. The amount of ¹⁴CO₂ release from glycine-l-¹⁴C or glycolate-l-¹⁴Cincreased only slightly when the temperature was raised from25 to 35°C. ¹Parts of this work were abstracted at the Annual Meeting (April,1969) of Japanese Society of Plant Physiologists, Kanazawa ²Department of Biochemistry, Michigan State University, EastLansing, Michigan, U.S.A. (Received September 3, 1969; )     

Photosynthesis in Cell Suspension Culture of a C4 Plant, Gisekia pharnaceoides L.

Photomixotrophic cell suspension culture was established fromthe leaf derived callus cells of Gisekia pharnaceoides L., aC₄ dicotyledonous weed. The late log phase cells possessed shade-typecharacters such as low chlorophyll a/b ratio, less pronouncedO₂ evolution and CO₂ fixation, saturation of photosyntheticCO₂ fixation at low intensity. The chloroplasts from these cellscontained granal stacking with high degree of a very few granawhich are characterized by their wide and high degree of stackings. The predominant labelling of 3-phosphoglyceric acid and sugarphosphates (40% of the total ¹⁴C incorporated) during 5 s exposureto ¹⁴CO₂ in light and subsequent decrease in percentage of ¹⁴Cin these compounds with increase in exposure time indicatedthe operation of the C₃ pathway in these cells. The simultaneoussynthesis of malate (23% of the total ¹⁴C incorporated) is relatedto the much pronounced glycolytic and tricarboxylic acid cycleactivities in these cells. The initial proliferation of callimainly from the zones of vascular supplies in the leaf, highstarch content of the cells, presence of large starch grainsin all the chloroplasts, activities of Calvin cycle enzymes,heavy labelling of C₃ type intermediates and less labellingof aspartate as early photosynthates and rapid accumulationof radioactivity into starch during ¹⁴CO₂ assimilation indicatedthat most of the cells in photomixotrophic culture were derivedfrom bundle sheath cells or the leaf cells of Gisekia changetheir function under culture conditions. ¹Present address: Tropical Botanic Garden and Research Institute,Navaranga Road, Trivandrum 695 011, India. (Received January 29, 1982; Accepted June 4, 1983)