Efficient CO2-Reducing Activity of NAD-Dependent Formate Dehydrogenase from Thiobacillus sp. KNK65MA for Formate Production from CO2 Gas

NAD-dependent formate dehydrogenase (FDH) from Candida boidinii (CbFDH) has been widely used in various CO₂-reduction systems but its practical applications are often impeded due to low CO₂-reducing activity. In this study, we demonstrated superior CO₂-reducing properties of FDH from Thiobacillus sp. KNK65MA (TsFDH) for production of formate from CO₂ gas. To discover more efficient CO₂-reducing FDHs than a reference enzyme, i.e. CbFDH, five FDHs were selected with biochemical properties and then, their CO₂-reducing activities were evaluated. All FDHs including CbFDH showed better CO₂-reducing activities at acidic pHs than at neutral pHs and four FDHs were more active than CbFDH in the CO₂ reduction reaction. In particular, the FDH from Thiobacillus sp. KNK65MA (TsFDH) exhibited the highest CO₂-reducing activity and had a dramatic preference for the reduction reaction, i.e., a 84.2-fold higher ratio of CO₂ reduction to formate oxidation in catalytic efficiency (k _cat/K _B) compared to CbFDH. Formate was produced from CO₂ gas using TsFDH and CbFDH, and TsFDH showed a 5.8-fold higher formate production rate than CbFDH. A sequence and structural comparison showed that FDHs with relatively high CO₂-reducing activities had elongated N- and C-terminal loops. The experimental results demonstrate that TsFDH can be an alternative to CbFDH as a biocatalyst in CO₂ reduction systems.     

Gradients of Intercellular CO(2) Levels Across the Leaf Mesophyll

Most current photosynthesis models, and interpretations of many wholeleaf CO₂ gas exchange measurements, are based on the often unstated assumption that the partial pressure of CO₂ is nearly uniform throughout the airspaces of the leaf mesophyll. Here we present measurements of CO₂ gradients across amphistomatous leaves allowed to assimilate CO₂ through only one surface, thus simulating hypostomatous leaves. We studied five species: Eucalyptus pauciflora Sieb. ex Spreng., Brassica chinensis L., Gossypium hirsutum L., Phaseolus vulgaris L., and Spinacia oleracea L. For Eucalyptus, maximum CO₂ pressure differences across the leaf mesophyll were 73 and 160 microbar when the pressures outside the lower leaf surface were 310 and 590 microbar, respectively. Using an approximate theoretical calculation, we infer that if the CO₂ had been supplied equally at both surfaces then the respective mean intercellular CO₂ pressures would have been roughly 12 and 27 microbar less than the pressures in the substomatal cavities in these cases. For ambient CO₂ pressures near 320 microbar, the average and minimum pressure differences across the mesophyll were 45 and 13 microbar. The corresponding mean intercellular CO₂ pressures would then be roughly 8 and 2 microbar less than those in the substomatal cavities. Pressure differences were generally smaller for the four agricultural species than for Eucalyptus, but they were nevertheless larger than previously reported values.     

Acclimation of Two Tomato Species to High Atmospheric CO(2): I. Sugar and Starch Concentrations

Lycopersicon esculentum Mill. cv Vedettos and Lycopersicon chmielewskii Rick, LA 1028, were exposed to two CO₂ concentrations (330 or 900 microliters per liter) for 10 weeks. Tomato plants grown at 900 microliters per liter contained more starch and more sugars than the control. However, we found no significant accumulation of starch and sugars in the young leaves of L. esculentum exposed to high CO₂. Carbon exchange rates were significantly higher in CO₂-enriched plants for the first few weeks of treatment but thereafter decreased as tomato plants acclimated to high atmospheric CO₂. This indicates that the long-term decline of photosynthetic efficiency of leaf 5 cannot be attributed to an accumulation of sugar and/or starch. The average concentration of starch in leaves 5 and 9 was always higher in L. esculentum than in L. chmielewskii (151.7% higher). A higher proportion of photosynthates was directed into starch for L. esculentum than for L. chmielewskii. However, these characteristics did not improve the long-term photosynthetic efficiency of L. chmielewskii grown at high CO₂ when compared with L. esculentum. The chloroplasts of tomato plants exposed to the higher CO₂ concentration exhibited a marked accumulation of starch. The results reported here suggest that starch and/or sugar accumulation under high CO₂ cannot entirely explain the loss of photosynthetic efficiency of high CO₂-grown plants.     

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.     

Isolation and Characterization of a Mutant of Chlamydomonas reinhardtii Deficient in the CO(2) Concentrating Mechanism

A Chlamydomonas reinhardtii mutant has been isolated that cannot grow photoautotrophically on low CO₂ concentrations but can grow on elevated CO₂. In a test cross, the high CO₂-requirement for growth showed a 2:2 segregation. This mutant, designated CIA-5, had a phenotype similar to previously identified mutants that were defective in some aspect of CO₂ accumulation. Unlike previously isolated mutants, CIA-5 did not have detectable levels of the periplasmic carbonic anhydrase, an inducible protein that participates in the acquisition of CO₂ by C. reinhardtii. CIA-5 also did not accumulate inorganic carbon to levels higher than could be accounted for by diffusion. This mutant strain did not synthesize any of the four polypeptides preferentially made by wild type C. reinhardtii when switched from an environment containing elevated CO₂ levels to an environment low in CO₂. It is concluded that this mutant fails to induce the CO₂ concentrating system and is incapable of adapting to low CO₂ conditions.     

Photosynthesis of Grass Species Differing in Carbon Dioxide Fixation Pathways: IV. ANALYSIS OF REDUCED OXYGEN RESPONSE IN PANICUM MILIOIDES AND PANICUM SCHENCKII

Reduced photorespiration has been reported in Panicum milioides on the basis of lower CO₂ compensation concentrations than in C₃ species, lower CO₂ evolution in the light, and less response of apparent photosynthesis to O₂ concentration. The lower response to O₂ in P. milioides could be due to reduced O₂ competition with CO₂ for reaction with ribulose 1,5-bisphosphate, to a reduced loss of CO₂, or to an initial fixation of CO₂ by phosphoenolpyruvate carboxylase. Experiments were carried out with Panicum maximum Jacq., a C₄ species having no apparent photorespiration; tall fescue (Festuca arundinacea Schreb.), a C₃ species; P. milioides Nees ex Trin.; and Panicum schenckii Hack. The latter two species are closely related and have low photorespiration rates. CO₂ exchange was measured at five CO₂ concentrations ranging from 0 to 260 microliters per liter at both 2 and 21% O₂. Mesophyll conductance or carboxylation efficiency was estimated by plotting substomatal CO₂ concentrations against apparent photosynthesis. In the C₄ species P. maximum, mesophyll conductance was 0.96 centimeters per second and was unaffected by O₂ concentration. At 21% O₂ mesophyll conductance of tall fescue was decreased 32% below the value at 2% O₂. Decreases in mesophyll conductance at 21% O₂ for P. milioides and P. schenckii were similar to that for tall fescue. On the other hand, loss of CO₂ in CO₂-free air, estimated by extrapolating the CO₂ response curve to zero CO₂, was increased from 1.8 to 6.5 milligrams per square decimeter per hour in tall fescue as O₂ was raised from 2-21%. Loss of CO₂ was less than 1 milligram per square decimeter per hour for P. milioides and P. schenckii and was unaffected by O₂. The results suggest that the reduced O₂ response in P. milioides and P. schenckii is due to a lower loss of CO₂ in the light rather than less inhibition of carboxylation by O₂, since the decrease in carboxylation efficiency at 21% O₂ was similar for P. milioides, P. schenckii, and tall fescue. The inhibition of apparent photosynthesis by 21% O₂ in these three species at low light intensities was similar at 31 to 36% which also indicates similar O₂ effects on carboxylation. Apparent photosynthesis at high light intensity was inhibited less by 21% O₂ in P. milioides (16.8%) and P. schenckii (23.8%) than in tall fescue (28.4%). This lower inhibition in the Panicum species may have been due to a higher degree of recycling of photorespired CO₂ in these species than in tall fescue.     

Effects of increased CO2 levels on growth, photosynthesis, ammonium uptake and cell composition in the macroalga Hypnea spinella (Gigartinales, Rhodophyta)

The red seaweed Hypnea spinella (Gigartinales, Rhodophyta), was cultured at laboratory scale under three different CO₂ conditions, non-enriched air (360?ppm CO₂) and CO₂-enriched air at two final concentrations (750 and 1,600?ppm CO₂), in order to evaluate the influence of increased CO₂ concentrations on growth, photosynthetic capacity, nitrogen removal efficiency, and chemical cellular composition. Average specific growth rates of H. spinella treated with 750 and 1,600?ppm CO₂-enriched air increased by 85.6% and 63.2% compared with non-enriched air cultures. CO₂ reduction percentages close to 12% were measured at 750?ppm CO₂ with respect to 5% and 7% for cultures treated with air and 1,600?ppm CO₂, respectively. Maximum photosynthetic rates were enhanced significantly for high CO₂ treatments, showing P _max values 1.5-fold higher than that for air-treated cultures. N–NH ₄ ⁺ consumption rates were also faster for algae growing at 750 and 1,600?ppm CO₂ than that for non-enriched air cultures. As a consequence of these experimental conditions, soluble carbohydrates increased and soluble protein contents decreased in algae treated with CO₂-enriched air. However, internal C and N contents remained constant at the different CO₂ concentrations. No significant differences in data obtained with both elevated CO₂ treatments, under the assayed conditions, indicate that H. spinella is saturated at dissolved inorganic carbon concentrations close by twice the actual atmospheric levels. The results show that increased CO₂ concentrations might be considered a key factor in order to improve intensively cultured H. spinella production yields and carbon and nitrogen bioremediation efficiencies.     

Why the free floating macrophyte Stratiotes aloides mainly grows in highly CO2-supersaturated waters

We examined how the freely floating macrophyte, Stratiotes aloides L., sampled from a CO₂-supersaturated pond, changes leaf morphology, photosynthesis and inorganic carbon acquisition during its different submerged and emerged life stages in order to evaluate whether S. aloides requires consistently supersaturated CO₂ conditions to grow and complete its life cycle. Submerged rosettes formed from over-wintering turions had typical traits of submerged plants with high specific leaf area and low chlorophyll a concentrations. Emergent leaf parts of mature, floating specimens had typical terrestrial traits with stomata, low specific leaf area and high chlorophyll a content, while offsets formed vegetatively and basal, submerged parts of mature plants showed traits in between. All submerged leaf types exhibited some ability to use HCO₃⁻ but only rosettes formed from turions had efficient HCO₃⁻ use. Rosettes also had the highest CO₂ affinity and maximum CO₂-saturated photosynthesis in water. Half-saturation constants for CO₂ (21–74 μM CO₂) were for all submerged leaf parts 5–140 times lower than the concentrations of free CO₂ in the pond (350–2800 μM CO₂). Emergent leaves were less efficient in water but had significantly higher photosynthesis than submerged, mature leaf parts in air, and rates of photosynthesis of emergent leaves in air were three to five times higher than rates of CO₂-saturated photosynthesis of the three submerged leaf types in water. Underwater photosynthetic rates estimated at CO₂ concentrations corresponding to air equilibrium were not sufficiently high to support any noticeable growth except for rosettes, in which bicarbonate utilization combined with high CO₂ affinity resulted in photosynthetic rates corresponding to almost 34% of maximum rates at high free CO₂. We conclude that S. aloides requires consistently high CO₂-supersaturation to support high growth and to complete its life cycle, and we infer that this requirement explains why S. aloides mainly grows in ponds, ditches and reed zones that are characterized by strong CO₂-supersaturation.     

Carbon Dioxide in Boreal Surface Waters: A Comparison of Lakes and Streams

The quantity of carbon dioxide (CO₂) emissions from inland waters into the atmosphere varies, depending on spatial and temporal variations in the partial pressure of CO₂ (pCO₂) in waters. Using 22,664 water samples from 851 boreal lakes and 64 boreal streams, taken from different water depths and during different months we found large spatial and temporal variations in pCO₂, ranging from below atmospheric equilibrium to values greater than 20,000???atm with a median value of 1048???atm for lakes (n?=?11,538 samples) and 1176???atm for streams (n?=?11,126). During the spring water mixing period in April/May, distributions of pCO₂ were not significantly different between stream and lake ecosystems (P?>?0.05), suggesting that pCO₂ in spring is determined by processes that are common to lakes and streams. During other seasons of the year, however, pCO₂ differed significantly between lake and stream ecosystems (P?<?0.0001). The variable that best explained the differences in seasonal pCO₂ variations between lakes and streams was the temperature difference between bottom and surface waters. Even small temperature differences resulted in a decline of pCO₂ in lake surface waters. Minimum pCO₂ values in lake surface waters were reached in July. Towards autumn pCO₂ strongly increased again in lake surface waters reaching values close to the ones found in stream surface waters. Although pCO₂ strongly increased in the upper water column towards autumn, pCO₂ in lake bottom waters still exceeded the pCO₂ in surface waters of lakes and streams. We conclude that throughout the year CO₂ is concentrated in bottom waters of boreal lakes, although these lakes are typically shallow with short water retention times. Highly varying amounts of this CO₂ reaches surface waters and evades to the atmosphere. Our findings have important implications for up-scaling CO₂ fluxes from single lake and stream measurements to regional and global annual fluxes.     

Nematode Genera in Forest Soil Respond Differentially to Elevated CO2

Previous reports suggest that fungivorous nematodes are the only trophic group in forest soils affected by elevated CO₂. However, there can be ambiguity within trophic groups, and we examined data at a genus level to determine whether the conclusion remains similar. Nematodes were extracted from roots and soil of loblolly pine (Pinus taeda) and sweet gum (Liquidambar styraciflua) forests fumigated with either ambient air or CO₂-enriched air. Root length and nematode biomass were estimated using video image analysis. Most common genera included Acrobeloides, Aphelenchoides, Cephalobus, Ditylenchus, Ecphyadorphora, Filenchus, Plectus, Prismatolaimus, and Tylencholaimus. Maturity Index values and diversity increased with elevated CO₂ in loblolly pine but decreased with elevated CO₂ in sweet gum forests. Elevated CO₂ treatment affected the occurrence of more nematode genera in sweet gum than loblolly pine forests. Numbers were similar but size of Xiphinema decreased in elevated CO₂. Abundance, but not biomass, of Aphelenchoides was reduced by elevated CO₂. Treatment effects were apparent at the genus levels that were masked at the trophic level. For example, bacterivores were unaffected by elevated CO₂, but abundance of Cephalobus was affected by CO₂ treatment in both forests.     

Does elevated CO2 protect photosynthesis from damage by high temperature via modifying leaf water status in maize seedlings?

We hypothesized that decreased stomatal conductance (g _s) at elevated CO₂ might decrease transpiration (E), increase leaf water potential (Ψ_W), and thereby protect net photosynthesis rate (P _N) from heat damage in maize (Zea mays L) seedlings. To separate long-term effects of elevated CO₂, plants grew at either ambient CO₂ or elevated CO₂. During high-temperature treatment (HT) at 45°C for 15 min, leaves were exposed either to ambient CO₂ (380 μmol mol^?1) or to elevated CO₂ (560 μmol mol^?1). HT reduced P _N by 25 to 38% across four CO₂ combinations. However, the g _s and E did not differ among all CO₂ treatments during HT. After returning the leaf temperature to 35°C within 30 min, g _s and E were the same or higher than the initial values. Leaf water potential (Ψ_W) was slightly lower at ambient CO₂, but not at elevated CO₂. This study highlighted that elevated CO₂ failed in protecting P _N from 45°C via decreasing g _s and Ψ_W.     

Why the free floating macrophyte Stratiotes aloides mainly grows in highly CO2-supersaturated waters

We examined how the freely floating macrophyte, Stratiotes aloides L., sampled from a CO₂-supersaturated pond, changes leaf morphology, photosynthesis and inorganic carbon acquisition during its different submerged and emerged life stages in order to evaluate whether S. aloides requires consistently supersaturated CO₂ conditions to grow and complete its life cycle. Submerged rosettes formed from over-wintering turions had typical traits of submerged plants with high specific leaf area and low chlorophyll a concentrations. Emergent leaf parts of mature, floating specimens had typical terrestrial traits with stomata, low specific leaf area and high chlorophyll a content, while offsets formed vegetatively and basal, submerged parts of mature plants showed traits in between. All submerged leaf types exhibited some ability to use HCO₃⁻ but only rosettes formed from turions had efficient HCO₃⁻ use. Rosettes also had the highest CO₂ affinity and maximum CO₂-saturated photosynthesis in water. Half-saturation constants for CO₂ (21–74 μM CO₂) were for all submerged leaf parts 5–140 times lower than the concentrations of free CO₂ in the pond (350–2800 μM CO₂). Emergent leaves were less efficient in water but had significantly higher photosynthesis than submerged, mature leaf parts in air, and rates of photosynthesis of emergent leaves in air were three to five times higher than rates of CO₂-saturated photosynthesis of the three submerged leaf types in water. Underwater photosynthetic rates estimated at CO₂ concentrations corresponding to air equilibrium were not sufficiently high to support any noticeable growth except for rosettes, in which bicarbonate utilization combined with high CO₂ affinity resulted in photosynthetic rates corresponding to almost 34% of maximum rates at high free CO₂. We conclude that S. aloides requires consistently high CO₂-supersaturation to support high growth and to complete its life cycle, and we infer that this requirement explains why S. aloides mainly grows in ponds, ditches and reed zones that are characterized by strong CO₂-supersaturation.     

Salt-Induced Metabolic Changes in Dunaliella salina

An increase of medium NaCl concentration induces Dunaliella cells to evolve O₂ photosynthetically even in the absence of CO₂. This NaCl-induced O₂ evolution may reflect the induced conversion of reserve carbohydrate to glycerol. The quantum yield for the NaCl-induced O₂ evolution, in the absence of CO₂, is 1.5-fold higher than that obtained for CO₂ fixation. Since the synthesis of glycerol from reserve carbohydrate in the absence of CO₂ requires only 0.5 ATP/NADPH, whereas photosynthesis requires at least 1.3 ATP/NADPH, it is concluded that the ATP/2e⁻ ratio coupled to NADP reduction in Dunaliella is lower than required for CO₂ fixation.     

Effects of elevated CO2 on the nutrient compositions and enzymes activities of Nilaparvata lugens nymphs fed on rice plants

Elevated CO 2 may reduce the tolerance of Nilaparvata lugen (N. lugens) to adverse environmental factors through the biological and physiological degeneration of N. lugens. In an artificial climate box, under 375 and 750 μL L 1 CO 2 levels, the rice stems nutrient content, the nutrient content and enzyme activities of N. lugens nymph fed on rice seedlings exposed to ambient and elevated CO 2 were studied. The results showed that rice stems had significantly higher protein and total amino acid levels under ambient than elevated CO 2 levels. Nymphs had significantly higher protein levels in the ambient CO 2 treatment, while their glucose levels were significantly lower under ambient CO 2 conditions. Significantly higher trypsin activity was observed in nymphs grown in elevated CO 2 . Significantly lower activities of the protective enzymes total superoxide dismutase and catalase were observed in the nymphs under ambient CO 2 . Meanwhile, the activity of the detoxification enzyme glutathione S-transferase was significantly higher in the ambient CO 2 treatment. Measuring how energy and resources were allocated to enzymes in N. lugens nymphs under elevated CO 2 conditions can provide a more meaningful evaluation of their metabolic tolerances to adverse climatic conditions.     

Impact of Different Carbon Dioxide Concentrations in the Olfactory Response of Sipha flava (Hemiptera: Aphididae) and its Predators

The increasing level CO₂ may altered host plant physiology and hence affect the foraging behavior of herbivore insects and predator. Hence, the aim of this study was provides evidence that host plants grown at different levels of CO₂ can alter the choice behavior of aphid, Sipha flava and their natural enemies, Cycloneda sanguinea and Diomus seminulus. The plant used was Pennisetum purpureum, cultivar Cameron Piracicaba growing in greenhouse (mean value of CO₂?=?440 ppm), climatic chamber with constant value of CO₂?=?500 ppm and climatic chamber with fluctuating CO₂ (mean value?=?368 ppm). A glass Y-shape olfactometer was used to verify the insects responses towards elephant grass plants cultivated under different conditions. The aphids were statistically more attracted by plants grown with constant CO₂ level (500 ppm) than by plants grown with fluctuating CO₂ level or plants grown in greenhouse. There was no difference in S. flava preference to non-infested versus infested plants by conspecifics. The predator C. sanguinea did not show difference between plants grown with constant CO₂ level and infested or not with S. flava. However, the predator D. seminulus showed higher preference to plants grown with constant CO₂ level and infested with S. flava. This study showed that the response of S. flava and its predators were affected by plants grown under different levels of CO₂.     

Adaptation of Chlamydomonas reinhardtii High-CO(2)-Requiring Mutants to Limiting CO(2)

Photosynthetic characteristics of four high-CO₂-requiring mutants of Chlamydomonas reinhardtii were compared to those of wild type before and after a 24-hour exposure to limiting CO₂ concentrations. The four mutants represent two loci involved in the CO₂-concentrating system of this unicellular alga. All mutants had a lower photosynthetic affinity for inorganic carbon than did the wild type when grown at an elevated CO₂ concentration, indicating that the genetic lesion in each is expressed even at elevated CO₂ concentrations. Wild type and all four mutants exhibited adaptive responses to limiting CO₂ characteristic of the induction of the CO₂-concentrating system, resulting in an increased affinity for inorganic carbon only in wild type. Although other components of the CO₂-concentrating system were induced in these mutants, the defective component in each was sufficient to prevent any increase in the affinity for inorganic carbon. It was concluded that the genes corresponding to the ca-1 and pmp-1 loci exhibit at least partially constitutive expression and that all components of the CO₂-concentrating system may be required to significantly affect the photosynthetic affinity for inorganic carbon.     

Metabolic effects of CO2 anaesthesia in Drosophila melanogaster

Immobilization of insects is necessary for various experimental purposes, and CO₂ exposure remains the most popular anaesthetic method in entomological research. A number of negative side effects of CO₂ anaesthesia have been reported, but CO₂ probably brings about metabolic modifications that are poorly known. In this work, we used GC/MS-based metabolic fingerprinting to assess the effect of CO₂ anaesthesia in Drosophila melanogaster adults. We analysed metabolic variation of flies submitted to acute CO₂ exposure and assessed the temporal metabolic changes during short- and long-term recovery. We found that D. melanogaster metabotypes were significantly affected by the anaesthetic treatment. Metabolic changes caused by acute CO₂ exposure were still manifested after 14 h of recovery. However, we found no evidence of metabolic alterations when a long recovery period was allowed (more than 24 h). This study points to some metabolic pathways altered during CO₂ anaesthesia (e.g. energetic metabolism). Evidence of short-term metabolic changes indicates that CO₂ anaesthesia should be used with utmost caution in physiological studies when a short recovery is allowed. In spite of this, CO₂ treatment seems to be an acceptable anaesthetic method provided that a long recovery period is allowed (more than 24 h).     

Some observations on the carbon dioxide burst in chlorella and chlamydomonas

In the course of mass spectrometer studies with the algae Chlorella and Chlamydomonas, data were obtained which indicate that the CO₂ burst and gulp are sensitive to oxygen. Furthermore, the CO₂ burst was found to be strongly suppressed when wave lengths shorter than 460 nanometers were blocked at intensities adequate to saturate photosynthesis. Under appropriate conditions at 30°, the CO₂ burst was interrupted by a brief CO₂ gulp and the post illumination gulp by a brief burst of CO₂. The post illumination gulp of CO₂ could be induced during illumination by interposition of a filter blocking wave lengths shorter than 460 nanometers. These data are discussed in relation to earlier reports of the phenomenon and briefly as they affect the detection of photorespiration.