THE RELATIVE SIGNIFICANCE OF CO2-FIXING ENZYMES IN THE METABOLISM OF RAT BRAIN

To evaluate the relative significance of CO₂-fixing enzymes in the metabolism of rat brain, the subcellular distribution of pyruvate carboxylase, phosphoenolpyruvate carboxykinase, NADP-isocitrate dehydrogenase and NADP-malate dehydrogenase, as well as the fixation of H¹⁴CO₃^? by the cytosol and the mitochondria was investigated. Pyruvate carboxylase and phosphoenol-pyruvate carboxykinase are mainly localized in the mitochondria whereas NADP-isocitrate dehydrogenase and NADP-malate dehydrogenase are present in both the cytosol and the mitochondria. In the presence of pyruvate rat brain mitochondria fixed H¹⁴CO₃^? at a rate of about 170 nmol/g of tissue/min whereas these organelles fixed negligible amounts of H¹⁴CO₃^? in the presence of α-ketoglutarate or phosphoenolpyruvate. Rat brain cortex slices fixed H¹⁴CO₃^? at a rate of about 7 nmol/g of tissue/min and it was increased by two-fold when pyruvate was added to the incubation medium. The carboxylation of α-ketoglutarate and pyruvate by the reversal of the cytosolic NADP-isocitrate dehydrogenase and NADP-malate dehydrogenase respectively was very low as compared to that by pyruvate carboxylase. The rate of carboxylation reaction of both NADP-isocitrate dehydrogenase and NADP-malate dehydrogenase was only about 1/10th of that of decarboxylation reaction of the same enzyme. It is suggested that under physiological conditions these two enzymes do not play a significant role in CO₂-fixation in the brain. In rat brain cytosol, citrate is largely metabolized to α-ketoglutarate by a sequential action of aconitate hydratase and NADP-isocitrate dehydrogenase. The operation of the citrate-cleavage pathway in rat brain cytosol is demonstrated. The data show that among four CO₂-fixing enzymes, pyruvate carboxylase, an anaplerotic enzyme, plays the major role in CO₂-fixation in the brain.     

C4 characteristics of photosynthesis in the C3 alga Hydrodictyon africanum

Abstract Results obtained with Hydrodictyon africanum, and data from the literature, show that most green algae of the chlorophyte type (e.g. Chlorella, Chlamydomonas, Hydrodictyon) differ in their photosynthetic C fixation characteristics from most green algae of the charophyte type (e.g. Spirogyra, Chara) and from C₃ higher plants. The chlorophyte algae fix inorganic carbon by the photosynthetic carbon reduction cycle pathway, but have a low CO₂ compensation point in 250 μM O₂, a low inhibition of CO₂ fixation from 10 μM CO₂/250 μM O₂ when compared with 10 μM CO₂/zero O₂, and a low half-saturation constant for CO₂. These three characteristics are different from those of charophytes and C₃ higher plants, and resemble those of C₄ higher plants. It is suggested that these characteristics of chlorophyte algae are the result of a ‘CO₂ concentrating mechanism’ which increases the CO₂/O₂ ratio at the site of ribulose bisphosphate carboxylase-oxygenase action in a similar way to that achieved by the C₄?C₃ acid cycle in C₄ plants. In the chlorophyte algae, however, CO₂ concentration probably involves active HCO₃^? transport at the inner membrane of the chloroplast envelope. Active HCO₃^? transport can occur at the plasmalemma of charophyte algae and submerged aquatic higher plants as well as chlorophyte algae, so it is unlikely to explain the differences between the two groups of aquatic green plants. Differences in the properties of ribulose bisphosphate carboxylase-oxygenase, and differences in CO₂ production in the light, also seem inadequate to account for the different photosynthetic characteristics. The chlorophyte type of ‘C0₂ concentrating mechanism’ appears to be common in other classes of eukaryotic algae, and in cyanophytes. Some of the ‘advanced’ members of these eukaryotic algal classes (including the chlorophytes) may lack the mechanism, while some ‘primitive’ charophytes may retain the mechanism which their ancestors presumably possessed.     

Products of photosynthetic 14CO2 fixation and related enzyme activities in fruiting structures of chickpea

Fruiting structures of a number of legumes including chickpea are known to carry out photosynthetic CO₂ assimilation, but the pathway of CO₂ fixation and particularly the role of phosphoenolpyruvate carboxylase (EC 4.1.1.31) in these tissues is not clear. Activities of some key enzymes of the Calvin cycle and C₄ metabolism, rates of ¹⁴CO₂ fixation in light and dark, and initial products of photosynthetic ¹⁴CO₂ fixation were determined in podwall and seedcoat (fruiting structures) and their subtending leaf in chickpea (Cicer arietinum L.). Compared to activities of ribulose-1,5-bisphosphate carboxylase (EC 4.1.1.39) and other Calvin cycle enzyme, viz. NADP⁺-glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.13), NAD⁺-glyceraldehyde-3-phosphate dehydrogenase (EC 1.2.1.12) and ribulose-5-phosphate kinase (EC 2.7.1.19), the levels of phosphoenolpyruvate carboxylase and other enzymes of C₄ metabolism viz. NADP⁺-malate dehydrogenase (EC 1.1.1.82), NAD⁺-malate dehydrogenase (EC 1.1.1.37), NADP⁺ malic enzyme (EC 1.1.1.40), NAD⁺-malic enzyme (EC 1.1.1.39), glutamate oxaloacetate transaminase (EC 2.6.1.1) and glutamate pyruvate transaminase (EC 2.6.1.2), were generally much higher in podwall and seedcoat than in the leaf. Podwall and seedcoat fixed ¹⁴CO₂ in light and dark at much higher rates than the leaf. Short-term assimilation of ¹⁴CO₂ by illuminated fruiting structures produced malate as the major labelled product with less labelling in 3-phosphoglycerate, whereas the leaf showed a major incorporation into 3-phosphoglycerate. It seems likely that the fruiting structures of chickpea utilize phosphoenolpyruvate carboxylase for recapturing the respired carbon dioxide.     

14CO2 Fixation and Compartmentation of Carbon Metabolism in a Recombined Chloroplast- 'Cytoplasm' System

The Compartmentation of ¹⁴CO₂ fixation and concomitant metabolism of ^l4C-iabelled products in a recombined system, composed of isolated intact spinach (Spinacia oleracea) chloroplasls and a ‘cytoplasm’ fraction, has been studied. Addition of ‘cytoplasm’ to chloroplasts fixing ¹⁴CO₂ increased the label in hexoae monophosphates outside the chloroplasts at the expense of excreted dihydroxyacetone phosphate. The label in ammo acids was increased both inside and outside the chloroplasts. The results support the view that chloroplasls are not able to make 2-oxoacids for amino acid synthesis directly from fixed CO₂, but have to co-operate with the cytoplasm and other organelles. The results also show that recombined systems can be useful for studies on the compartmen tation of carbon metabolism in pholosynthesizing plant tissues.     

Photosynthesis, immediate export and carbon partitioning in source leaves of C3, C3-C4 intermediate, and C4Panicum and Flaveria species at ambient and elevated CO2 levels

Immediate export in leaves of C₃‐C₄ intermediates were compared with their C₃ and C₄ relatives within the Panicum and Flaveria genera. At 35 Pa CO₂, photosynthesis and export were highest in C₄ species in each genera. Within the Panicum, photosynthesis and export in ‘type I’ C₃‐C₄ intermediates were greater than those in C₃ species. However, ‘type I’ C₃‐C₄ intermediates exported a similar proportion of newly fixed ¹⁴C as did C₄ species. Within the Flaveria, ‘type II’ C₃‐C₄ intermediate species had the lowest export rather than the C₃ species. At ambient CO₂, immediate export was strongly correlated with photosynthesis. However, at 90 Pa CO₂, when photosynthesis and immediate export increased in all C₃ and C₃‐C₄ intermediate species, proportionally less C was exported in all photosynthetic types than that at ambient CO₂. All species accumulated starch and sugars at both CO₂ levels. There was no correlation between immediate export and the pattern of ¹⁴C‐labelling into sugars and starch among the photosynthetic types within each genus. However, during CO₂ enrichment, C₄Panicum species accumulated sugars above the level of sugars and starch normally made at ambient CO₂, whereas the C₄Flaveria species accumulated only additional starch.     

Poa annua shows inter-generational differences in response to elevated CO2

Inter-generational effects on the growth of Poa annua (L.) in ambient and elevated atmospheric CO₂ conditions (350 and 550 μl l^–1, respectively) were studied in two different experiments. Both experiments showed similar results. In a greenhouse experiment growth, measured as the numbers of tillers produced per week, was compared for plants grown from first and second generation seeds. Second generation seeds were obtained from plants grown for one whole generation in either ambient or elevated atmospheric CO₂ (‘ambient’ and ‘elevated’ seeds, respectively). First generation plants and second generation ‘ambient’ plants did not respond to elevated CO₂. Second generation ‘elevated’ plants produced significantly more tillers in elevated CO₂. In the second experiment model terrestrial ecosystems growing in the Ecotron and which included Poa annua were used. Above-ground biomass after one and two generations of growth were compared. At the end of Generation 1 no difference was found in biomass production while at the end of Generation 2 biomass increased in elevated CO₂ by 50%. The implications for climate change research are discussed.     

The Effect of Sodium Chloride on the Balance between the C3- and C4-Carbon Fixation Pathways

Young leaves of salt-depleted Aeluropus litoralis Parl. plants show CO₂ fixation by the C₃-carbon fixation pathway. No detectable activity of phosphoenol pyruvate (PEP) carboxylase was found. When A. litoralis plants were exposed to a NaCl solution, the leaves showed a high activity of PEP carboxylase as well as a significant CO₂ fixation by the C₄-pathway. — Also in Zea mays L. and Chloris gayana Kunth., the presence of NaCl in the medium influences the balance between phosphoenol pyruvate carboxylase and ribulose-1,5-diphosphate carboxylase.     

On the significance of C3—C4 intermediate photosynthesis to the evolution of C4 photosynthesis

Abstract Evidence is drawn from previous studies to argue that C₃—C₄ intermediate plants are evolutionary intermediates, evolving from fully-expressed C₃ plants towards fully-expressed C₄ plants. On the basis of this conclusion, C₃—C₄ intermediates are examined to elucidate possible patterns that have been followed during the evolution of C₄ photosynthesis. An hypothesis is proposed that the initial step in C₄-evolution was the development of bundle-sheath metabolism that reduced apparent photorespiration by an efficient recycling of CO₂ using RuBP carboxylase. The CO₂-recycling mechanism appears to involve the differential compartmentation of glycine decarboxylase between mesophyll and bundle-sheath cells, such that most of the activity is in the bundlesheath cells. Subsequently, elevated phosphoenolpyruvate (PEP) carboxylase activities are proposed to have evolved as a means of enhancing the recycling of photorespired CO₂. As the activity of PEP carboxylase increased to higher values, other enzymes in the C₄-pathway are proposed to have increased in activity to facilitate the processing of the products of C₄-assimilation and provide PEP substrate to PEP carboxylase with greater efficiency. Initially, such a ‘C₄-cycle’ would not have been differentially compartmentalized between mesophyll and bundlesheath cells as is typical of fully-expressed C₄ plants. Such metabolism would have limited benefit in terms of concentrating CO₂ at RuBP carboxylase and, therefore, also be of little benefit for improving water- and nitrogen-use efficiencies. However, the development of such a limited C₄-cycle would have represented a preadaptation capable of evolving into the leaf biochemistry typical of fully-expressed C₄ plants. Thus, during the initial stages of C₄-evolution it is proposed that improvements in photorespiratory CO₂-loss and their influence on increasing the rate of net CO₂ assimilation per unit leaf area represented the evolutionary ‘driving-force’. Improved resourceuse efficiency resulting from an efficient CO₂-concentrating mechanism is proposed as the driving force during the later stages.     

Metabolism of Propane, n-Propylamine, and Propionate by Hydrocarbon-Utilizing Bacteria

Studies were conducted on the oxidation and assimilation of various three-carbon compounds by a gram-positive rod isolated from soil and designated strain R-22. This organism can utilize propane, propionate, or n-propylamine as sole source of carbon and energy. Respiration rates, enzyme assays, and ¹⁴CO₂ incorporation experiments suggest that propane is metabolized via methyl ketone formation; propionate and n-propylamine are metabolized via the methylmalonyl-succinate pathway. Isocitrate lyase activity was found in cells grown on acetate and was not present in cells grown on propionate or n-propylamine. ¹⁴CO₂ was incorporated into pyruvate when propionate and n-propylamine were oxidized in the presence of NaAsO₂, but insignificant radioactivity was found in pyruvate produced during the oxidation of propane and acetone. The n-propylamine dissimilatory mechanism was inducible in strain R-22, and amine dehydrogenase activity was detected in cells grown on n-propylamine. Radiorespirometer and ¹⁴CO₂ incorporation studies with several propane-utilizing organisms indicate that the methylmalonyl-succinate pathway is the predominant one for the metabolism of propionate.     

N-poor ecosystems may respond more to elevated [CO2] than N-rich ones in the long term. A model analysis of grassland.

The Hurley Pasture Model was used to examine the short and long-term responses of grazed grasslands in the British uplands to a step increase from 350 to 700 μmol mol^–1 CO₂ concentration ([CO₂]) with inputs of 5 or 100 kg N ha^–1 y^–1. In N-rich grassland, [CO₂] doubling quickly increased net primary productivity (NPP), total carbon (C_sys) and plant biomass by about 30%. By contrast, the N-poor grassland underwent a prolonged ‘transient’, when there was little response, but eventually NPP, C_sys and plant biomass more than doubled. The ‘transient’ was due to N immobilization and severe depletion of the soil mineral N pool. The large long-term response was due to slow N accumulation, as a result of decreased leaching, decreased gaseous N losses and increased N₂-fixation, which amplified the CO₂ response much more in the N-poor than in the N-rich grassland. It was concluded that (i) ecosystems use extra carbon fixed at high [CO₂] to acquire and retain nutrients, supporting the contention of Gifford et al. (1996 ), (ii) in the long term, and perhaps on the real timescale of increasing [CO₂], the response (in NPP, C_sys and plant biomass) of nutrient-poor ecosystems may be proportionately greater than that of nutrient-rich ones, (iii) short-term experiments on nutrient-poor ecosystems may observe only the transient responses, (iv) the speed of ecosystem responses may be limited by the rate of nutrient accumulation rather than by internal rate constants, and (v) ecosystem models must represent processes affecting nutrient acquisition and retention to be able to simulate likely real-world CO₂ responses.     

Rapid appearance of 13C in biogenic isoprene when 13CO2 is fed to intact leaves

Biogenic isoprene substantially affects atmospheric chemistry, but it is not known how or why many plants, especially trees, make isoprene. We fed ¹³CO₂ to leaves of Quercus rubra and monitored the incorporation of ¹³C into isoprene by mass spectrometry. After feeding ¹³CO₂ for 9 min we found all possible labelling patterns from completely unlabelled to fully labelled isoprene. By 18 min, 84% of the carbon atoms in isoprene were ¹³C. Labelling of the last 20% of the carbon atoms was much slower than labelling of the first 80%. The rate of labelling of isoprene was similar to that reported for phosphoglyceric acid indicating that there is a close linkage between the carbon source for isoprene synthesis and the photosynthetic carbon reduction pathway.     

14C – a tool for separation of autotrophic and heterotrophic soil respiration

We assessed the potential of using ¹⁴C contents of soil respired CO₂ to calculate the contributions of heterotrophic and autotrophic respiration to total soil respiration. The partitioning of these fluxes is of utmost importance to evaluate implications of environmental change on soil carbon cycling and sequestration. At three girdled forest stands in Sweden and Germany, where the tree root (autotrophic) respiration had been eliminated, we measured both flux rates and ¹⁴C contents of soil respired CO₂ in girdled and control plots in the summers of 2001 or 2002. At all stands, CO₂ flux rates were slightly higher in the control plots, whereas the ¹⁴C contents of respired CO₂ tended to be higher in the girdled plots. This was expected and confirmed that heterotrophically respired CO₂ cycles more slowly through the forest ecosystem than autotrophically respired CO₂. On the basis of these data, the contributions of hetero‐ and autotrophic respiration to total soil respiration were calculated using two separate approaches (i.e. based on flux rates or ¹⁴C). Fractions of heterotrophic respiration ranged from 53% to 87%. Values calculated by both approaches did not differ significantly from each other. Finally, we compared the ¹⁴C contents of soil respired CO₂ in the girdled plots with the ¹⁴C contents of heterotrophically respired CO₂ calculated by three different ¹⁴C models. None of the models matched the measured data sufficiently. In addition, we suspect that inherent effects of girdling may cause the ¹⁴C content of CO₂ respired in the girdled plots to be lower than ‘true’ heterotrophically respired CO₂ in an undisturbed plot. Nevertheless, we argue that measurements and modeling of ¹⁴C can be developed into a valuable tool for separating heterotrophic and autotrophic soil respiration (e.g. when girdling cannot be performed).     

The sensitivity of C3 photosynthesis to increasing CO2 concentration: a theoretical analysis of its dependence on temperature and background CO2 concentration

The atmospheric CO₂ concentration has increased from the pre-industrial concentration of about 280 μmol mol⁻¹ to its present concentration of over 350 μmol mol⁻¹, and continues to increase. As the rate of photosynthesis in C₃ plants is strongly dependent on CO₂ concentration, this should have a marked effect on photosynthesis, and hence on plant growth and productivity. The magnitude of photo-synthetic responses can be calculated based on the well-developed theory of photosynthetic response to intercellular CO₂ concentration. A simple biochemically based model of photosynthesis was coupled to a model of stomatal conductance to calculate photosynthetic responses to ambient CO₂ concentration. In the combined model, photosynthesis was much more responsive to CO₂ at high than at low temperatures. At 350 μmol mol⁻¹, photosynthesis at 35°C reached 51% of the rate that would have been possible with non-limiting CO₂, whereas at 5°C, 77% of the CO₂ non-limited rate was attained. Relative CO₂ sensitivity also became smaller at elevated CO₂, as CO₂ concentration increased towards saturation. As photosynthesis was far from being saturated at the current ambient CO₂ concentration, considerable further gains in photosynthesis were predicted through continuing increases in CO₂ concentration. The strong interaction with temperature also leads to photosynthesis in different global regions experiencing very different sensitivities to increasing CO₂ concentrations.     

An inexpensive system for exposing plants in the field to elevated concentrations of CO2

An inexpensive, potentially mobile field exposure system is described which may be easily constructed by a small workshop. It may be operated as an open-top with a frustrum or covered with a polycarbonate ‘lid’. The system is cost-effective for CO₂ exposure work because the small size allows provision of CO₂-enriched atmospheres over prolonged periods at relatively low cost. A preliminary assessment of the chambers has been made and concentrations can be maintained at ±6% for a target atmosphere of 680 cm³ m^?3 CO₂ under normal operating conditions. Other chamber environmental conditions are reported.     

Carbonic anhydrase and C4 photosynthesis: a transgenic analysis

Carbonic anhydrase (CA, EC 4.2.1.1) catalyses the first reaction in the C₄ photosynthetic pathway, the conversion of atmospheric CO₂ to bicarbonate in the mesophyll cytosol. To examine the importance of the enzyme to the functioning of the C₄ photosynthetic pathway, Flaveria bidentis (L.) Kuntze, a C₄ dicot, was genetically transformed with an antisense construct in which the cDNA encoding a putative cytosolic CA (CA3) was placed under the control of a constitutive promoter. Some of the primary transformants had impaired CO₂ assimilation rates and required high CO₂ for growth. The T₁ progeny of four primary transformants were used to examine the quantitative relationship between leaf CA activity and CO₂ assimilation rate. CA activity was determined in leaf extracts with a mass spectrometric technique that measured the rate of ¹⁸O exchange from doubly labelled ¹³C¹⁸O₂. Steady‐state CO₂ assimilation rates were unaffected by a decrease in CA activity until CA activity was less than 20% of wild type when they decreased steeply. Transformants with less than 10% of wild‐type CA activity had very low CO₂ assimilation rates and grew poorly at ambient CO₂ partial pressure. Reduction in CA activity also increased the CO₂ partial pressure required to saturate CO₂ assimilation rates. The present data show that CA activity is essential for the functioning of the C₄ photosynthetic pathway.     

Photoassimilation of CO2 by Isolated Bundle-Sheath Strands of Zea mays

The role of malate decarboxylation as a source of CO₂ and NADPH for the evolution of photosynthesis of isolated maize bundle-sheath strands has been investigated. The bundle-sheath cells were supplied with malate plus NADP, in the presence of intermediates of the Calvin cycle to increase the rate of CO₂ fixation. The effects of malate addition on the rate of 3 phospoglycerate synthesis, with non-saturating concentrations of bicarbonate, can be explained by an increase of the cellular pool of CO₂ in the cells due to malate decarboxylation. The CO₂ reacting with RuDP to give phosphoglycerate corresponds effectively to carbon atom 4 of malate. Malate addition produces an enhancement of the rate of CO₂ incorporation which is much more important when the reducing power is the limiting process for the evolution of the Calvin cycle (with phosphoglycerate as added substrate and/or in the presence of DCMU. These results demonstrate the utilization of NADPH produced by malate decarboxylation for the regeneration of RuDP. NADPH can also reverse the reaction of malate decarboxylation and gives rise to a synthesis of malate by carboxylation of pyruvate. In contrast, the pattern of ¹⁴C distribution among compounds is not strongly modified by malate addition. This result suggests that PGA reduction in the whole leaf must occur also in mesophyll cells to allow correctregeneration of the reduced compounds of the photosynthetic cycle.     

Photosynthetic energy supply for NO3− assimilation in Scenedesmus

NO₃^?-dependent O₂ in synchronous Scenedesmus obtusiusculus Chod. in the absence of CO₂ is stoichiometric with NH₄⁺ excretion, indicating a close coupling of NO₃^? reduction to non-cyclic electron flow. Also in the presence of CO₂, NO₃^? stimulates O₂ evolution as manifested by an increase in the O₂/CO₂ ratio from 0.96 to 1.11. This quotient was increased to 1.36 by addition of NO₂^?, without competitive interaction with CO₂ fixation, indicating that the capacity for non-cyclic electron transport at saturating light is non-limiting for simultaneous reduction of NO₃^? and CO₂ at high rates. During incubation with NO₃^?+ CO₂, no NH₄⁺ is released to the outer medium, whereas during incubation with NO₂^?+ CO₂, excess NH₄⁺ is formed and excreted. NO₃^? uptake is stimulated by CO₂, and this stimulation is also significant when the cellular energy metabolism is restricted by moderate concentrations of carbonyl cyanide-p-trifluoromethoxyphenylhydrazone, whereas NO₃^? uptake in the absence of CO₂ is severely inhibited by the uncoupler. Also under energy-restricted conditions NO₃^? uptake is not competitive with CO₂ fixation. Antimycin A is inhibitory for NO₃^? uptake in the absence of CO₂, and there is no enhancement of NO₃^? uptake by CO₂ in the presence of antimycin A. It is assumed that the energy demand for NO₃^? uptake is met by energy fixed as triosephosphates in the Calvin cycle. Antimycin A possibly affects the transfer of reduced triose phosphates from the chloroplast to the cytoplasm. Active carbon metabolism also seems to exert a control effect on NO₃^? assimilation, inducing complete incorporation of all NO₃^? taken up into amino acids. This control effect is not functional when NO₂^? is the nitrogen source. Active carbon metabolism thus seems to be essential both for provision of energy for NO₃^? uptake and for regulation of the process.     

Indirect effects of soil moisture reverse soil C sequestration responses of a spring wheat agroecosystem to elevated CO2

Increased plant productivity under elevated atmospheric CO₂ concentrations might increase soil carbon (C) inputs and storage, which would constitute an important negative feedback on the ongoing atmospheric CO₂ rise. However, elevated CO₂ often also leads to increased soil moisture, which could accelerate the decomposition of soil organic matter, thus counteracting the positive effects via C cycling. We investigated soil C sequestration responses to 5 years of elevated CO₂ treatment in a temperate spring wheat agroecosystem. The application of ¹³C‐depleted CO₂ to the elevated CO₂ plots enabled us to partition soil C into recently fixed C (C_new) and pre‐experimental C (C_old) by ¹³C/¹²C mass balance. Gross C inputs to soils associated with C_new accumulation and the decomposition of C_old were then simulated using the Rothamsted C model ‘RothC.’ We also ran simulations with a modified RothC version that was driven directly by measured soil moisture and temperature data instead of the original water balance equation that required potential evaporation and precipitation as input. The model accurately reproduced the measured C_new in bulk soil and microbial biomass C. Assuming equal soil moisture in both ambient and elevated CO₂, simulation results indicated that elevated CO₂ soils accumulated an extra ～40–50 g C m^?2 relative to ambient CO₂ soils over the 5 year treatment period. However, when accounting for the increased soil moisture under elevated CO₂ that we observed, a faster decomposition of C_old resulted; this extra C loss under elevated CO₂ resulted in a negative net effect on total soil C of ～30 g C m^?2 relative to ambient conditions. The present study therefore demonstrates that positive effects of elevated CO₂ on soil C due to extra soil C inputs can be more than compensated by negative effects of elevated CO₂ via the hydrological cycle.     

Lack of C4 photosynthetic metabolism in ears of C3 cereals

There is continuing controversy over whether a degree of C₄ photosynthetic metabolism exists in ears of C₃ cereals. In this context, CO₂ exchange and the initial products of photosynthesis were examined in flag leaf blades and various ear parts of two durum wheat (Triticum durum Desf.) and two six-rowed barley (Hordeum vulgare L.) cultivars. Three weeks after anthesis, the CO₂ compensation concentration at 210 mmol mol^?1 O₂ in durum wheat and barley ear parts was similar to or greater than that in flag leaves. The O₂ dependence of the CO₂ compensation concentration in durum wheat ear parts, as well as in the flag leaf blade, was linear, as expected for C₃ photosynthesis. In a complementary experiment, intact and attached ears and flag leaf blades of barley and durum wheat were radio-labelled with ¹⁴CO₂ during a 10s pulse, and the initial products of fixation were studied in various parts of the ears (awns, glumes, inner bracts and grains) and in the flag leaf blade. All tissues assimilated CO₂ mainly by the Calvin (C₃) cycle, with little fixation of ¹⁴CO₂ into the C₄ acids malate and aspartate (about 10% or less). These collective data support the conclusion that in the ear parts of these C₃ cereals C₄ photosynthetic metabolism is nil.     

Uptake of CO2 by aquatic vegetation

Abstract Photosynthesis by aquatic plants based on the supply of CO₂ from air-equilibrated solutions may be limited by the low diffusion coefficient of CO₂ in water. For plants in which the transport of CO₂ from the bulk medium is by diffusion, and the initial carboxylation uses RUBISCO, CO₂ supply can be increased by growth in habitats with fast water flow over the surface (reducing unstirred layer thickness), or with heterotrophically-augmented CO₂ levels, including the direct use of sediment CO₂. Many aquatic plants using RUBISCO as their initial carboxylase counter the limitations on CO₂ supply via the operation of biophysical CO₂ concentrating mechanisms which are based on active transport of HCO^?₃, CO₂ or H⁺ at the plasmalemma, and use bulk-phase HCO^?₃ or CO₂ as the C source. A final group of aquatic plants use biochemical CO₂ concentrating mechanisms based on auxiliary carboxylation by PEPc: C₄-like and Crassulacean Acid Metabolism–like processes are involved. These various mechanisms for increasing CO₂ supply to RUBISCO also help to offset the low specific reaction rate of aquatic plant RUBISCOs at low [CO₂] and low [CO₂]: [CO₂]. In addition to overcoming restrictions on CO₂ supply, the various methods of increasing inorganic C availability may also be important in alleviating shortages of nitrogen or photons.