Kinetics and Apparent K(m) of Oxygen Cycle under Conditions of Limiting Carbon Dioxide Fixation

A mass spectrometer with a membrane inlet was used to monitor light-driven O₂ evolution, O₂ uptake, and CO₂ uptake in suspensions of algae (Scenedesmus obliquus). We observed the following. (a) The rate of O₂ uptake, which, in the presence of iodoacetamide, replaces the uptake of CO₂, showed a distinct plateau (V_max) beyond ~30% O₂ and was half-maximal at ~8% O₂. We concluded that this light-driven O₂ uptake process, which does not involve carbon compounds, is saturated at lower O₂ concentrations than are photorespiration and glycolate formation. (b) In the absence of inhibitor, O₂ evolution was relatively unaffected by the presence or absence of CO₂. During the course of CO₂ depletion, electron flow to CO₂ was replaced by an equivalent flow to O₂. (c) There was a distinct delay between the cessation of CO₂ uptake and the increase in O₂ uptake. We ascribe this delay to the transient utilization of another electron acceptor—possibly bicarbonate or another bound form of CO₂.     

Photorespiratory phenomena in maize: oxygen uptake, isotope discrimination, and carbon dioxide efflux

Concurrent O₂ evolution, O₂ uptake, and CO₂ uptake by illuminated maize (Zea mays) leaves were measured using ¹³CO₂ and ¹⁸O₂. Considerable O₂ uptake occurred during active photosynthesis. At CO₂ compensation, O₂ uptake increased. Associated with this increase was a decrease in O₂ release such that a stoichiometric exchange of O₂ occurred. The rate of O₂ exchange at CO₂ compensation was directly related to O₂ concentration in the atmosphere at least up to 8% (v/v).     

Photosynthetic o(2) exchange kinetics in isolated soybean cells

Light-dependent O₂ exchange was measured in intact, isolated soybean (Glycine max. var. Williams) cells using isotopically labeled O₂ and a mass spectrometer. The dependence of O₂ exchange on O₂ and CO₂ was investigated at high light in coupled and uncoupled cells. With coupled cells at high O₂, O₂ evolution followed similar kinetics at high and low CO₂. Steady-state rates of O₂ uptake were insignificant at high CO₂, but progressively increased with decreasing CO₂. At low CO₂, steady-state rates of O₂ uptake were 50% to 70% of the maximum CO₂-supported rates of O₂ evolution. These high rates of O₂ uptake exceeded the maximum rate of O₂ reduction determined in uncoupled cells, suggesting the occurrence of another light-induced O₂-uptake process (i.e. photorespiration).

Rates of O₂ exchange in uncoupled cells were half-saturated at 7% to 8% O₂. Initial rates (during induction) of O₂ exchange in uninhibited cells were also half-saturated at 7% to 8% O₂. In contrast, steady-state rates of O₂ evolution and O₂ uptake (at low CO₂) were half-saturated at 18% to 20% O₂. O₂ uptake was significantly suppressed in the presence of nitrate, suggesting that nitrate and/or nitrite can compete with O₂ for photoreductant.

These results suggest that two mechanisms (O₂ reduction and photorespiration) are responsible for the light-dependent O₂ uptake observed in uninhibited cells under CO₂-limiting conditions. The relative contribution of each process to the rate of O₂ uptake appears to be dependent on the O₂ level. At high O₂ concentrations (≥40%), photorespiration is the major O₂-consuming process. At lower (ambient) O₂ concentrations (≤20%), O₂ reduction accounts for a significant portion of the total light-dependent O₂ uptake.

     

On the nature of the oxygen uptake in the light by Chondrus crispus. Effects of inhibitors,temperature and light intensity

The nature of the different processes of O₂ uptake involved in the light in the red macroalga Chondrus crispus Stackhouse (Rhodophyta, Gigartinales) was investigated. At limiting CO₂, INH (2.5 mM) did not alter the O₂ uptake rate. Glycolate was not excreted and did not accumulate within the cells. KCN reduced the rate of O₂ uptake in the light by 76% at limiting CO₂ and by 43% at saturating CO₂, but caused > 95% inhibition of O₂ evolution. DCMU (5 μM) totally blocked the photosynthetic electron transport chain, but allowed a residual O₂ uptake of 3.0±0.6 μmol O₂ .h^?1.g^?1 FW, irrespective of the CO₂ concentration. In saturating CO₂, a high light intensity pretreatment significantly stimulated the rate of O₂ uptake compared to net O₂ evolution, suggesting the persistence, in the light, of mitochondrial respiration. Irrespective of the CO₂ concentration, the optimum temperature for O₂ evolution was 17°C whereas dark O₂ uptake increased linearly with temperature. In contrast, O₂ uptake in the light showed an optimum at 17°C in limiting CO₂, and 21–25° C in saturating CO₂; its Q₁₀ was 2.4 at limiting CO₂, a value close to that of RuBP oxygenase, and 3.1 at saturating CO₂, a value close to that of dark respiration. It is concluded that: 1) mitochondrial respiration and Mehler reaction are both involved at all CO₂ concentrations, 2) RuBP oxygenase activity cannot account for more than 45%, and Mehler reaction for less than 20%, of the total O₂ uptake observed in the light at limiting CO₂.     

Photoreduction of O(2) Primes and Replaces CO(2) Assimilation

A mass spectrometer with a membrane inlet system was used to monitor directly gaseous components in a suspension of algae. Using labeled oxygen, we observed that during the first 20 seconds of illumination after a dark period, when no net O₂ evolution or CO₂ uptake was observed, O₂ evolution was normal but completely compensated by O₂ uptake. Similarly, when CO₂ uptake was totally or partially inhibited, O₂ evolution proceeded at a high (near maximal) rate. Under all conditions, O₂ uptake balanced that fraction of the O₂ evolution which could not be accounted for by CO₂ uptake.     

Relationship between Respiration and Photosynthesis in Guard Cell and Mesophyll Cell Protoplasts of Commelina communis L

A mass spectrometric method combining ¹⁶O/¹⁸O and ¹²C/¹³C isotopes was used to quantify the unidirectional fluxes of O₂ and CO₂ during a dark to light transition for guard cell protoplasts and mesophyll cell protoplasts of Commelina communis L. In darkness, O₂ uptake and CO₂ evolution were similar on a protein basis. Under light, guard cell protoplasts evolved O₂ (61 micromoles of O₂ per milligram of chlorophyll per hour) almost at the same rate as mesophyll cell protoplasts (73 micromoles of O₂ per milligram of chlorophyll per hour). However, carbon assimilation was totally different. In contrast with mesophyll cell protoplasts, guard cell protoplasts were able to fix CO₂ in darkness at a rate of 27 micromoles of CO₂ per milligram of chlorophyll per hour, which was increased by 50% in light. At the onset of light, a delay observed for guard cell protoplasts between O₂ evolution and CO₂ fixation and a time lag before the rate of saturation suggested a carbon metabolism based on phosphoenolpyruvate carboxylase activity. Under light, CO₂ evolution by guard cell protoplasts was sharply decreased (37%), while O₂ uptake was slowly inhibited (14%). A control of mitochondrial activity by guard cell chloroplasts under light via redox equivalents and ATP transfer in the cytosol is discussed. From this study on protoplasts, we conclude that the energy produced at the chloroplast level under light is not totally used for CO₂ assimilation and may be dissipated for other purposes such as ion uptake.     

The Sequence of Change within the Photosynthetic Apparatus of Wheat following Short-Term Exposure to Ozone

The basis of inhibition of photosynthesis by single acute O₃ exposures was investigated in vivo using analyses based on leaf gas exchange measurements. The fully expanded second leaves of wheat plants (Triticum aestivum L. cv Avalon) were fumigated with either 200 or 400 nanomoles per mole O₃ for between 4 and 16 hours. This reduced significantly the light-saturated rate of CO₂ uptake and was accompanied by a parallel decrease in stomatal conductance. However, the stomatal limitation, estimated from the relationship between CO₂ uptake and the internal CO₂ concentration, only increased significantly during the first 8 hours of exposure to 400 nanomoles per mole O₃; no significant increase occurred for any of the other treatments. Analysis of the response of CO₂ uptake to the internal CO₂ concentration implied that the predominant factor responsible for the reduction in light-saturated CO₂ uptake was a decrease in the efficiency of carboxylation. This was 58 and 21% of the control value after 16 hours at 200 and 400 nanomoles per mole O₃, respectively. At saturating concentrations of CO₂, photosynthesis was inhibited by no more than 22% after 16 hours, indicating that the capacity for regeneration of ribulose bisphosphate was less susceptible to O₃. Ozone fumigations also had a less pronounced effect on light-limited photosynthesis. The maximum quantum yield of CO₂ uptake and the quantum yield of oxygen evolution showed no significant decline after 16 hours with 200 nanomoles per mole O₃, requiring 8 hours at 400 nanomoles per mole O₃ before a significant reduction occurred. The photochemical efficiency of photosystem II estimated from the ratio of variable to maximum chlorophyll fluorescence and the atrazine-binding capacity of isolated thylakoids demonstrated that photochemical reactions were not responsible for the initial inhibition of CO₂ uptake. The results suggest that the apparent carboxylation efficiency appears to be the initial cause of decline in photosynthesis in vivo following acute O₃ fumigation.     

Simultaneous measurement of oscillations in oxygen evolution and chlorophyll a fluorescence in leaf pieces

In spinach (Spinacia oleracea) and barley (Hordeum vulgare) leaves, chlorophyll a fluorescence and O₂ evolution have been measured simultaneously following re-illumination after a dark interval or when steady state photosynthesis has been perturbed by changes in the gas phase. In high CO₂ concentrations, both O₂ and fluorescence can display marked dampening oscillations that are antiparallel but slightly out of phase (a rise or fall in fluorescence anticipating a corresponding fall or rise in O₂ by about 10 to 15 seconds). Infrared gas analysis measurements showed that CO₂ uptake behaved like O₂ evolution both in the period of oscillation (about 1 minute) and in its relation to fluorescence. In the steady state, oscillations were initiated by increases in CO₂ or by increases or decreases in O₂. Oscillations in O₂ or CO₂ did not occur without associated oscillations in fluorescence and the latter were a sensitive indicator of the former. The relationship between such oscillations in photosynthetic carbon assimilation and chlorophyl a fluorescence is discussed in the context of the effect of ATP or NADPH consumption on known quenching mechanisms.     

Photosynthesis and photorespiration in whole plants of wheat

Wheat was cultivated in a small phytotronic chamber. ¹⁸O₂ was used to measure the O₂ uptake by the plant, which was recorded simultaneously with the O₂ evolution, net CO₂ uptake, and transpiration. At normal atmospheric CO₂ concentration, photorespiration, measured as O₂ uptake, was as important as the net photosynthesis. The level of true O₂ evolution was independent of CO₂ concentration and stayed nearly equal to the sum of net CO₂ photosynthesis and O₂ uptake. We conclude that at a given light intensity, O₂ and CO₂ compete for the reducing power produced at constant rate by the light reactions of photosynthesis.     

Oxygen exchange in leaves in the light

Photosynthetic O₂ production and photorespiratory O₂ uptake were measured using isotopic techniques, in the C₃ species Hirschfeldia incana Lowe., Helianthus annuus L., and Phaseolus vulgaris L. At high CO₂ and normal O₂, O₂ production increased linearly with light intensity. At low O₂ or low CO₂, O₂ production was suppressed, indicating that increased concentrations of both O₂ and CO₂ can stimulate O₂ production. At the CO₂ compensation point, O₂ uptake equaled O₂ production over a wide range of O₂ concentrations. O₂ uptake increased with light intensity and O₂ concentration. At low light intensities, O₂ uptake was suppressed by increased CO₂ concentrations so that O₂ uptake at 1,000 microliters per liter CO₂ was 28 to 35% of the uptake at the CO₂ compensation point. At high light intensities, O₂ uptake was stimulated by low concentrations of CO₂ and suppressed by higher concentrations of CO₂. O₂ uptake at high light intensity and 1000 microliters per liter CO₂ was 75% or more of the rate of O₂ uptake at the compensation point. The response of O₂ uptake to light intensity extrapolated to zero in darkness, suggesting that O₂ uptake via dark respiration may be suppressed in the light. The response of O₂ uptake to O₂ concentration saturated at about 30% O₂ in high light and at a lower O₂ concentration in low light. O₂ uptake was also observed with the C₄ plant Amaranthus edulis; the rate of uptake at the CO₂ compensation point was 20% of that observed at the same light intensity with the C₃ species, and this rate was not influenced by the CO₂ concentration. The results are discussed and interpreted in terms of the ribulose-1,5-bisphosphate oxygenase reaction, the associated metabolism of the photorespiratory pathway, and direct photosynthetic reduction of O₂.     

Active CO(2) Transport by the Green Alga Chlamydomonas reinhardtii

Mass spectrometric measurements of dissolved free ¹³CO₂ were used to monitor CO₂ uptake by air grown (low CO₂) cells and protoplasts from the green alga Chlamydomonas reinhardtii. In the presence of 50 micromolar dissolved inorganic carbon and light, protoplasts which had been washed free of external carbonic anhydrase reduced the ¹³CO₂ concentration in the medium to close to zero. Similar results were obtained with low CO₂ cells treated with 50 micromolar acetazolamide. Addition of carbonic anhydrase to protoplasts after the period of rapid CO₂ uptake revealed that the removal of CO₂ from the medium in the light was due to selective and active CO₂ transport rather than uptake of total dissolved inorganic carbon. In the light, low CO₂ cells and protoplasts incubated with carbonic anhydrase took up CO₂ at an apparently low rate which reflected the uptake of total dissolved inorganic carbon. No net CO₂ uptake occurred in the dark. Measurement of chlorophyll a fluorescence yield with low CO₂ cells and washed protoplasts showed that variable fluorescence was mainly influenced by energy quenching which was reciprocally related to photosynthetic activity with its highest value at the CO₂ compensation point. During the linear uptake of CO₂, low CO₂ cells and protoplasts incubated with carbonic anhydrase showed similar rates of net O₂ evolution (102 and 108 micromoles per milligram of chlorophyll per hour, respectively). The rate of net O₂ evolution (83 micromoles per milligram of chlorophyll per hour) with washed protoplasts was 20 to 30% lower during the period of rapid CO₂ uptake and decreased to a still lower value of 46 micromoles per milligram of chlorophyll per hour when most of the free CO₂ had been removed from the medium. The addition of carbonic anhydrase at this point resulted in more than a doubling of the rate of O₂ evolution. These results show low CO₂ cells of Chlamydomonas are able to transport both CO₂ and HCO₃⁻ but CO₂ is preferentially removed from the medium. The external carbonic anhydrase is important in the supply to the cells of free CO₂ from the dehydration of HCO₃⁻.     

Photorespiration in Air and High CO(2)-Grown Chlorella pyrenoidosa

Oxygen inhibition of photosynthesis and CO₂ evolution during photorespiration were compared in high CO₂-grown and air-grown Chlorella pyrenoidosa, using the artificial leaf technique at pH 5.0. High CO₂ cells, in contrast to air-grown cells, exhibited a marked inhibition of photosynthesis by O₂, which appeared to be competitive and similar in magnitude to that in higher C₃ plants. With increasing time after transfer to air, the photosynthetic rate in high CO₂ cells increased while the O₂ effect declined. Photorespiration, measured as the difference between ¹⁴CO₂ and ¹²CO₂ uptake, was much greater and sensitive to O₂ in high CO₂ cells. Some CO₂ evolution was also present in air-grown algae; however, it did not appear to be sensitive to O₂. True photosynthesis was not affected by O₂ in either case. The data indicate that the difference between high CO₂ and air-grown algae could be attributed to the magnitude of CO₂ evolution. This conclusion is discussed with reference to the oxygenase reaction and the control of photorespiration in algae.     

Gas Exchange Characteristics of the Submerged Aquatic Crassulacean Acid Metabolism Plant, Isoetes howellii

The submerged aquatic plant Isoetes howellii Engelmann possesses Crassulacean acid metabolism (CAM) comparable to that known from terrestrial CAM plants. Infrared gas analysis of submerged leaves showed Isoetes was capable of net CO₂ uptake in both light and dark. CO₂ uptake rates were a function of CO₂ levels in the medium. At 2,500 microliters CO₂ per liter (gas phase, equivalent to 1.79 milligrams per liter aqueous phase), Isoetes leaves showed continuous uptake in both the light and dark. At this CO₂ level, photosynthetic rates were light saturated at about 10% full sunlight and were about 3-fold greater than dark CO₂ uptake rates. In the dark, CO₂ uptake rates were also a function of length of time in the night period. Measurements of dark CO₂ uptake showed that, at both 2,500 and 500 microliters CO₂ per liter, rates declined during the night period. At the higher CO₂ level, dark CO₂ uptake rates at 0600 h were 75% less than at 1800 h. At 500 microliters CO₂ per liter, net CO₂ uptake in the dark at 1800 h was replaced by net CO₂ evolution in the dark at 0600 h. At both CO₂ levels, the overnight decline in net CO₂ uptake was marked by periodic bursts of accelerated CO₂ uptake. CO₂ uptake in the light was similar at 1% and 21% O₂, and this held for leaves intact as well as leaves split longitudinally. Estimating the contribution of light versus dark CO₂ uptake to the total carbon gain is complicated by the diurnal flux in CO₂ availability under field conditions.     

Photosynthetic oxygen exchange in C4 grasses: the role of oxygen as electron acceptor

C₄ grasses of the NAD‐ME type (Astrebla lappacea, Eleusine coracana, Eragrostis superba, Leptochloa dubia, Panicum coloratum, Panicum decompositum) and the NADP‐ME type (Bothriochloa bladhii, Cenchrus ciliaris, Dichanthium sericeum, Panicum antidotale, Paspalum notatum, Pennisetum alopecuroides, Sorghum bicolor) were used to investigate the role of O₂ as an electron acceptor during C₄ photosynthesis. Mass spectrometric measurements of gross O₂ evolution and uptake were made concurrently with measurements of net CO₂ uptake and chlorophyll fluorescence at different irradiances and leaf temperatures of 30 and 40 °C. In all C₄ grasses gross O₂ uptake increased with increasing irradiance at very high CO₂ partial pressures (pCO₂) and was on average 18% of gross O₂ evolution. Gross O₂ uptake at high irradiance and high pCO₂ was on average 3.8 times greater than gross O₂ uptake in the dark. Furthermore, gross O₂ uptake in the light increased with O₂ concentration at both high CO₂ and the compensation point, whereas gross O₂ uptake in the dark was insensitive to O₂ concentration. This suggests that a significant amount of O₂ uptake may be associated with the Mehler reaction, and that the Mehler reaction varies with irradiance and O₂ concentration. O₂ exchange characteristics at high pCO₂ were similar for NAD‐ME and NADP‐ME species. NAD‐ME species had significantly greater O₂ uptake and evolution at the compensation point particularly at low irradiance compared to NADP‐ME species, which could be related to different rates of photorespiratory O₂ uptake. There was a good correlation between electron transport rates estimated from chlorophyll fluorescence and gross O₂ evolution at high light and high pCO₂.     

Effect of CO(2), O(2), and Light on Photosynthesis and Photorespiration in Wheat

Unidirectional O₂ fluxes were measured with ¹⁸O₂ in a whole plant of wheat cultivated in a controlled environment. At 2 or 21% O₂, O₂ uptake was maximum at 60 microliters per liter CO₂. At lower CO₂ concentrations, it was strongly inhibited, as was photosynthetic O₂ evolution. At 2% O₂, there remained a substantial O₂ uptake, even at high CO₂ level; the O₂ evolution was inhibited at CO₂ concentrations under 330 microliters per liter. The O₂ uptake increased linearly with light intensity, starting from the level of dark respiration. No saturation was observed at high light intensities. No significant change in the gas-exchange patterns occurred during a long period of the plant life. An adaptation to low light intensities was observed after 3 hours illumination. These results are interpreted in relation to the functioning of the photosynthetic apparatus and point to a regulation by the electron acceptors and a specific action of CO₂. The behavior of the O₂ uptake and the study of the CO₂ compensation point seem to indicate the persistence of mitochondrial respiration during photosynthesis.     

CO(2) and O(2) Exchange in Two Mosses, Hypnum cupressiforme and Dicranum scoparium

Photosynthetic CO₂ and O₂ exchange was studied in two moss species, Hypnum cupressiforme Hedw. and Dicranum scoparium Hedw. Most experiments were made during steady state of photosynthesis, using ¹⁸O₂ to trace O₂ uptake. In standard experimental conditions (photoperiod 12 h, 135 micromoles photons per square meter per second, 18°C, 330 microliters per liter CO₂, 21% O₂) the net photosynthetic rate was around 40 micromoles CO₂ per gram dry weight per hour in H. cupressiforme and 50 micromoles CO₂ per gram dry weight per hour in D. scoparium. The CO₂ compensation point lay between 45 and 55 microliters per liter CO₂ and the enhancement of net photosynthesis by 3% O₂versus 21% O₂ was 40 to 45%. The ratio of O₂ uptake to net photosynthesis was 0.8 to 0.9 irrespective of the light intensity. The response of net photosynthesis to CO₂ showed a high apparent K_m (CO₂) even in nonsaturating light. On the other hand, O₂ uptake in standard conditions was not far from saturation. It could be enhanced by only 25% by increasing the O₂ concentration (saturating level as low as 30% O₂), and by 65% by decreasing the CO₂ concentration to the compensation point. Although O₂ is a competitive inhibitor of CO₂ uptake it could not replace CO₂ completely as an electron acceptor, and electron flow, expressed as gross O₂ production, was inhibited by both high O₂ and low CO₂ levels. At high CO₂, O₂ uptake was 70% lower than the maximum at the CO₂ compensation point. The remaining activity (30%) can be attributed to dark respiration and the Mehler reaction.     

Analysis of the Relative Increase in Photosynthetic O2 Uptake When Photosynthesis in Grapevine Leaves Is Inhibited following Low Night Temperatures and/or Water Stress

We found similarities between the effects of low night temperatures (5°C–10°C) and slowly imposed water stress on photosynthesis in grapevine (Vitis vinifera L.) leaves. Exposure of plants growing outdoors to successive chilling nights caused light- and CO₂-saturated photosynthetic O₂ evolution to decline to zero within 5 d. Plants recovered after four warm nights. These photosynthetic responses were confirmed in potted plants, even when roots were heated. The inhibitory effects of chilling were greater after a period of illumination, probably because transpiration induced higher water deficit. Stomatal closure only accounted for part of the inhibition of photosynthesis. Fluorescence measurements showed no evidence of photoinhibition, but nonphotochemical quenching increased in stressed plants. The most characteristic response to both stresses was an increase in the ratio of electron transport to net O₂ evolution, even at high external CO₂ concentrations. Oxygen isotope exchange revealed that this imbalance was due to increased O₂ uptake, which probably has two components: photorespiration and the Mehler reaction. Chilling- and drought-induced water stress enhanced both O₂ uptake processes, and both processes maintained relatively high rates of electron flow as CO₂ exchange approached zero in stressed leaves. Presumably, high electron transport associated with O₂ uptake processes also maintained a high ΔpH, thus affording photoprotection.     

Evidence for Cyclic Photophosphorylation during CO(2) Fixation in Intact Chloroplasts: Studies with Antimycin A, Nitrite, and Oxaloacetate

This study examines the effect of antimycin A and nitrite on ¹⁴CO₂ fixation in intact chloroplasts isolated from spinach (Spinacia oleracea L.) leaves. Antimycin A (2 micromolar) strongly inhibited CO₂ fixation but did not appear to inhibit or uncouple linear electron transport in intact chloroplasts. The addition of small quantities (40-100 micromolar) of nitrite or oxaloacetate, but not NH₄Cl, in the presence of antimycin A restored photosynthesis. Antimycin A inhibition, and the subsequent restoration of photosynthetic activities by nitrite or oxaloacetate, was observed over a wide range of CO₂ concentration, light intensity, and temperature. High O₂ concentration (up to 240 micromolar) did not appear to influence the extent of the inhibition by antimycin A, nor the subsequent restoration of photosynthetic activity by nitrite or oxaloacetate. Studies of O₂ exchanges during photosynthesis in cells and chloroplasts indicated that 2 micromolar antimycin A stimulated O₂ uptake by about 25% while net O₂ evolution was inhibited by 76%. O₂ uptake in chloroplasts in the presence of 2 micromolar antimycin A was 67% of total O₂ evolution. These results suggest that only a small proportion of the O₂ uptake measured was directly linked to ATP generation. The above evidence indicates that cyclic photophosphorylation is the predominant energy-balancing reaction during photosynthesis in intact chloroplasts. On the other hand, pseudocyclic O₂ uptake appears to play only a minimal role.     

Effect of Photon Fluence Rate on Oxygen Evolution and Uptake by Chlamydomonas reinhardtii Suspensions Grown in Ambient and CO(2)-Enriched Air

A closed system consisting of an assimilation chamber furnished with a membrane inlet from the liquid phase connected to a mass spectrometer was used to measure O₂ evolution and uptake by Chlamydomonas reinhardtii cells grown in ambient (0.034% CO₂) or CO₂-enriched (5% CO₂) air. At pH = 6.9, 28°C and concentrations of dissolved inorganic carbon (DIC) saturating for photosynthesis, O₂ uptake in the light (U_o) equaled O₂ production (E_o) at the light compensation point (15 micromoles photons per square meter per second). E_o and U_o increased with increasing photon fluence rate (PFR) but were not rate saturated at 600 micromoles photons per square meter per second, while net O₂ exchange reached a saturation level near 500 micromoles photons per square meter per second which was nearly the same for both, CO₂-grown and air-grown cells. Comparison of the U_o/E_o ratios between air-grown and CO₂-grown C. reinhardtii showed higher values for air-grown cells at light intensities higher than light compensation. For both, air-grown and CO₂-grown algae the rates of mitochondrial O₂ uptake in the dark measured immediately before and 5 minutes after illumination were much lower than U_o at PFR saturating for net photosynthesis. We conclude that noncyclic electron flow from water to NADP⁺ and pseudocyclic electron flow via photosystem I to O₂ both significantly contribute to O₂ exchange in the light. In contrast, mitochondrial respiration and photosynthetic carbon oxidation cycle are regarded as minor O₂ consuming reactions in the light in both, air-grown and CO₂-grown cells. It is suggested that the “extra” O₂ uptake by air-grown algae provides ATP required for the energy dependent CO₂/HCO₃⁻ concentrating mechanism known to be present in these cells.     

Light-Dependent Oxygen Uptake, Glycolate, and Ammonia Release in l-Methionine Sulfoximine-Treated Chlamydomonas

Glycolate and ammonia excretion plus oxygen exchanges were measured in the light in l-methionine-dl-sulfoximine treated air-grown Chlamydomonas reinhardii. At saturating CO₂ (between 600 and 700 microliters per liter CO₂) neither glycolate nor ammonia were excreted, whereas at the CO₂ compensation concentration (<10 microliters per liter CO₂) treated algae excreted both glycolate and ammonia at rates of 37 and 59 nanomoles per minute per milligram chlorophyll, respectively. From the excretion values we calculate the amount of O₂ consumed through the glycolate pathway. The calculated value was not significantly different from the component of O₂ uptake sensitive to CO₂ obtained from the difference between O₂ uptake of the CO₂ compensation point and at saturating CO₂. This component was about 40% of stationary O₂ uptake measured at the CO₂ compensation point. From these data we conclude that glyoxylate decarboxylation in air-grown Chlamydomonas represents a minor pathway of metabolism even in conditions where amino donors are deficient and that processes other than glycolate pathway are responsible for the O₂ uptake insensitive to CO₂.