The acquisition and accumulation of inorganic carbon by the unicellular green alga Chlorella ellipsoidea

Abstract. The uptake and accumulation of inorganic carbon has been investigated in Chlorella ellipsoidea cells grown at acid or alkaline pH. Carbonic anhydrase (CA) was detected in ceil extracts but not in intact cells and CA activity in acid-grown cells was considerably less than that in alkali-grown cells. Both cell types demonstrates low K_1/2 (CO₂) values in the range pH 7.0–8.0 and these were unaffected by O₂ concentration. The CO₂ compensation concentrations of acid- and alkali-grown cells suspended in aqueous media were not significantly different in the range of pH 6.0–8.0, but at pH 5.0, the CO₂ compensation concentrations of acid-grown cells (57.4cm³ m⁻³) were lower than those of alkali-grown cells (79.2cm³ m⁻³). The rate of photo-synthetic O₂ evolution in the range pH 7.5–8.0 exceeded the calculated rate of CO₂ supply two- to three-fold, in both acid- and alkali-grown cells, indicating that HCO₃⁻ was taken up by the cells. Accumulation of inorganic carbon was measured at pH 7.5 by silicone-oil centri-fugation, and the concentration of unfixed inorganic carbon was found to be 5.1 mol m⁻³ in acid-grown and 6.4mol m⁻³ in alkali-grown cells. These concentrations were 4.6- and 5.9-fold greater than in the external medium. These results indicate that photorespiration is suppressed in both acid- and alkali-grown cells by an intracellular accumulation of inorganic carbon due, in part, to an active uptake of bicarbonate.     

Catabolite inactivation of the maltose transporter in nitrogen-starved yeast could be due to the stimulation of general protein turnover

Addition of glucose to Saccharomyces cerevisiae inactivates the maltose transporter. The general consensus is that this inactivation, called catabolite inactivation, is one of the control mechanisms developed by this organism to use glucose preferentially whenever it is available. Using nitrogen-starved cells (resting cells), it has been shown that glucose triggers endocytosis and degradation of the transporter in the vacuole. We now show that maltose itself triggers inactivation and degradation of its own transporter as efficiently as glucose. This fact, and the observation that glucose inactivates a variety of plasma membrane proteins including glucose transporters themselves, suggests that catabolite inactivation of the maltose transporter in nitrogen-starved cells is not a control mechanism specifically directed to ensure a preferential use of glucose. It is proposed that, in this metabolic condition, inactivation of the maltose transporter might be due to the stimulation of the general protein turnover that follows nitrogen starvation.     

Enhancement of rice canopy carbon gain by elevated CO(2) is sensitive to growth stage and leaf nitrogen concentration

Increasing our understanding of the factors regulating seasonal changes in rice canopy carbon gain (C(gain): daily net photosynthesis -- night respiration) under elevated CO(2) concentrations ([CO(2)]) will reduce our uncertainty in predicting future rice yields and assist in the development of adaptation strategies. In this study we measured CO(2) exchange from rice (Oryza sativa) canopies grown at c. 360 and 690 micromol mol(-1)[CO(2)] in growth chambers continuously over three growing seasons. Stimulation of C(gain) by elevated [CO(2)] was 22-79% during vegetative growth, but decreased to between -12 and 5% after the grain-filling stage, resulting in a 7-22% net enhancement for the whole season. The decreased stimulation of C(gain) resulted mainly from decreased canopy net photosynthesis and partially from increased respiration. A decrease in canopy photosynthetic capacity was noted where leaf nitrogen (N) decreased. The effect of elevated [CO(2)] on leaf area was generally small, but most dramatic under ample N conditions; this increased the stimulation of whole-season C(gain). These results suggest that a decrease in C(gain) enhancement following elevated CO(2) levels is difficult to avoid, but that careful management of nitrogen levels can alter the whole-season C(gain) enhancement.     

Competitive dominance of the cyanobacterium Microcystis aeruginosa in nutrient-rich culture conditions with special reference to dissolved inorganic carbon uptake

The aim of the present study was to determine what factors contribute to the competitive advantage of cyanobacteria in eutrophic conditions. Mixed species batch culture experiments were conducted at three pHs (8.2, 8.8, 10.2) and irradiances (30, 90, 180 μmol photons m^?2 s^_1) between Microcystis aeruginosa Kützing and Staurastrum dorsidentiferum W. et West or Synedra ulna (Nitzsch) Ehrenberg, which always resulted in the dominance of M. aeruginosa. The final yields of competitors were often significantly lower than when cultured singly. The dominance of the surface‐loving M. aeruginosa appears to be related to its advantageous C0₂ uptake. To clarify the importance of dissolved inorganic carbon (DIC) as a selective factor, the effect of aeration was examined with M. aeruginosa. The growth of M. aeruginosa quickly stopped in nonaerated conditions, while it continued growing throughout the culture when aerated. This result indicates that DIC limitation easily occurs in static conditions and the viewpoint of C0₂ exchange efficiency would be helpful in discussing the competitive advantage of M. aeruginosa. When the three species were cultured at various ratios of surface area to volume (s/v), postulating it as an index of the gas exchange efficiency, the increases in population densities strongly correlated with s/v, and the relationships between the specific growth rate and s/v corresponded well to a Monod type saturation function. We found that M. aeruginosa had the lowest half‐saturation constant among the three, reflecting its high affinity for DIC.     

Regulation of 5-phosphoribosyl-1-pyrophosphate synthesis in human fibroblasts by the concentration of inorganic phosphate

During the growth cycle of normal fibroblasts and of fibroblasts deficient in glucose-6-phosphate dehydrogenase activity, the concentration of 5-phosphoribosyl-1-pyrophosphate and of P_i, as well as the activity of 5-phosphoribosyl-1-pyrophosphate synthetase, decreased to stable values in confluent cultures. A high degree of correlation (0.89 and 0.91 for two normal and 0.69 for one glucose-6-phosphate dehydrogenase-deficient cell strain, respectively) was shown between intracellular P_i and 5-phosphoribosyl-1-pyrophosphate concentrations under varying culture and incubation conditions. 5-phosphoribosyl-1-pyrophosphate concentrations were elevated in normal fibroblasts incubated with methylene blue only if intracellular P_i levels were high. Neither methylene blue nor 6-aminonicotinamide, singly, affected intracellular P_i concentrations. However, when normal cells were pretreated with 6-aminonicotinamide and then with methylene blue, intracellular P_i decreased, 5-phosphoribosyl-1-pyrophosphate was depleted, and its rate of generation decreased. Under similar conditions, glucose-6-phosphate dehydrogenase-deficient fibroblasts maintained unaltered P_i levels, and 5-phosphoribosyl-1-pyrophosphate concentration and generation were slightly increased. The decrease in intracellular P_i in normal cells after the combined treatment was commensurate with an accumulation of 6-phosphogluconate, which did not take place in mutant cells. The changes in 5-phosphoribosyl-1-pyrophosphate synthesis, whether due to the stage of growth or various experimental manipulations, were always concordant with changes in intracellular P_i level. The regulatory role of P_i is consistent with the known enzymic properties of 5-phosphoribosyl-1-pyrophosphate synthetase.     

The requirement for external carbonic anhydrase in diatoms is influenced by the supply and demand for dissolved inorganic carbon

Photosynthesis by marine diatoms contributes significantly to the global carbon cycle. Due to the low concentration of CO₂ in seawater, many diatoms use extracellular carbonic anhydrase (eCA) to enhance the supply of CO₂ to the cell surface. While much research has investigated how the requirement for eCA is influenced by changes in CO₂ availability, little is known about how eCA contributes to CO₂ supply following changes in the demand for carbon. We therefore examined how changes in photosynthetic rate influence the requirement for eCA in three centric diatoms. Modeling of cell surface carbonate chemistry indicated that diffusive CO₂ supply to the cell surface was greatly reduced in large diatoms at higher photosynthetic rates. Laboratory experiments demonstrated a trend of an increasing requirement for eCA with increasing photosynthetic rate that was most pronounced in the larger species, supporting the findings of the cellular modeling. Microelectrode measurements of cell surface pH and O₂ demonstrated that individual cells exhibited an increased contribution of eCA to photosynthesis at higher irradiances. Our data demonstrate that changes in carbon demand strongly influence the requirement for eCA in diatoms. Cell size and photosynthetic rate will therefore be key determinants of the mode of dissolved inorganic carbon uptake.     

Exogenous inorganic carbon sources for photosynthesis in seawater by members of the Fucales and the Laminariales (Phaeophyta): ecological and taxonomic implications

Summary Characteristics of inorganic carbon assimilation by photosynthesis in seawater were investigated in six species of the Fucales (five Fucaceae, one Cystoseiraceae) and four species of the Laminariales (three Laminariaceae, one Alariaceae) from Arbroath, Scotland. All of the algae tested could photosynthesise faster at high external pH values than the uncatalysed conversion of HCO ₃ ^- to CO₂ can occur, i.e. can use external HCO ₃ ^- . They all had detectable extracellular carbonic anhydrase activity, suggesting that HCO ₃ ^- use could involve catalysis of external CO₂ production, a view supported to some extent by experiments with an inhibitor of carbonic anhydrase. All of the algae tested had CO₂ compensation concentrations at pH 8 which were lower than would be expected from diffusive entry of CO₂ supplying RUBISCO as the initial carboxylase, consistent with the operation of energized entry of HCO ₃ ^- and / or CO₂ acting as a CO₂ concentrating mechanism. Quantitative differences among the algae examined were noted with respect to characteristics of inorganic C assimilation. The most obvious distinction was between the eulittoral Fucaceae, which are emersed for part of, or most of, the tidal cycle, and the other three families (Cystoseiraceae, Laminariaceae, Alariaceae) whose representatives are essentially continually submersed. The Fucaceae examined are able to photosynthesise at high pH values, and have lower CO₂ compensation concentrations, and lower K_1/2 values for inorganic C use in photosynthesis, at pH 8, than the other algae tested. Furthermore, the Fucaceae are essentially saturated with inorganic C for photosynthesis at the normal seawater concentration at pH 8 and 10°C. These characteristics are consistent with the dominant role of a CO₂ concentrating mechanism in CO₂ acquisition by these plants. Other species tested have characteristcs which suggest a less effective HCO ₃ ^- use and CO₂ concentrating mechanism, with the Laminariaceae being the least effective; unlike the Fucaceae, photosynthesis by these algae is not saturated with inorganic C in normal seawater. Taxonomic and ecological implications of these results are considered in relation to related data in the literature.     

Quantifying the impact of daily and seasonal variation in sap pH on xylem dissolved inorganic carbon estimates in plum trees

In studies on internal CO₂ transport, average xylem sap pH (pH_x) is one of the factors used for calculation of the concentration of dissolved inorganic carbon in the xylem sap ([]). Lack of detailed pH_x measurements at high temporal resolution could be a potential source of error when evaluating [] dynamics. In this experiment, we performed continuous measurements of CO₂ concentration ([CO₂]) and stem temperature (T_stem), complemented with pH_x measurements at 30‐min intervals during the day at various stages of the growing season (Day of the Year (DOY): 86 (late winter), 128 (mid‐spring) and 155 (early summer)) on a plum tree (Prunus domestica L. cv. Reine Claude d'Oullins). We used the recorded pH_x to calculate [] based on T_stem and the corresponding measured [CO₂]. No statistically significant difference was found between mean [] calculated with instantaneous pH_x and daily average pH_x. However, using an average pH_x value from a different part of the growing season than the measurements of [CO₂] and T_stem to estimate [] led to a statistically significant error. The error varied between 3.25 ± 0.01% under‐estimation and 3.97 ± 0.01% over‐estimation, relative to the true [] data. Measured pH_x did not show a significant daily variation, unlike [CO₂], which increased during the day and declined at night. As the growing season progressed, daily average [CO₂] (3.4%, 5.3%, 7.4%) increased and average pH_x (5.43, 5.29, 5.20) decreased. Increase in [CO₂] will increase its solubility in xylem sap according to Henry's law, and the dissociation of [] will negatively affect pH_x. Our results are the first quantifying the error in [] due to the interaction between [CO₂] and pH_x on a seasonal time scale. We found significant changes in pH_x across the growing season, but overall the effect on the calculation of [] remained within an error range of 4%. However, it is possible that the error could be more substantial for other tree species, particularly if pH_x is in the more sensitive range (pH_x > 6.5).     

Matrix approach to land carbon cycle modeling: A case study with the Community Land Model

The terrestrial carbon (C) cycle has been commonly represented by a series of C balance equations to track C influxes into and effluxes out of individual pools in earth system models (ESMs). This representation matches our understanding of C cycle processes well but makes it difficult to track model behaviors. It is also computationally expensive, limiting the ability to conduct comprehensive parametric sensitivity analyses. To overcome these challenges, we have developed a matrix approach, which reorganizes the C balance equations in the original ESM into one matrix equation without changing any modeled C cycle processes and mechanisms. We applied the matrix approach to the Community Land Model (CLM4.5) with vertically‐resolved biogeochemistry. The matrix equation exactly reproduces litter and soil organic carbon (SOC) dynamics of the standard CLM4.5 across different spatial‐temporal scales. The matrix approach enables effective diagnosis of system properties such as C residence time and attribution of global change impacts to relevant processes. We illustrated, for example, the impacts of CO₂ fertilization on litter and SOC dynamics can be easily decomposed into the relative contributions from C input, allocation of external C into different C pools, nitrogen regulation, altered soil environmental conditions, and vertical mixing along the soil profile. In addition, the matrix tool can accelerate model spin‐up, permit thorough parametric sensitivity tests, enable pool‐based data assimilation, and facilitate tracking and benchmarking of model behaviors. Overall, the matrix approach can make a broad range of future modeling activities more efficient and effective.     

Terrestrial higher-plant response to increasing atmospheric [CO2] in relation to the global carbon cycle

Terrestrial higher plants exchange large amounts of CO₂ with the atmosphere each year; c. 15% of the atmospheric pool of C is assimilated in terrestrial-plant photosynthesis each year, with an about equal amount returned to the atmosphere as CO₂ in plant respiration and the decomposition of soil organic matter and plant litter. Any global change in plant C metabolism can potentially affect atmospheric CO₂ content during the course of years to decades. In particular, plant responses to the presently increasing atmospheric CO₂ concentration might influence the rate of atmospheric CO₂ increase through various biotic feedbacks. Climatic changes caused by increasing atmospheric CO₂ concentration may modulate plant and ecosystem responses to CO₂ concentration. Climatic changes and increases in pollution associated with increasing atmospheric CO₂ concentration may be as significant to plant and ecosystem C balance as CO₂ concentration itself. Moreover, human activities such as deforestation and livestock grazing can have impacts on the C balance and structure of individual terrestrial ecosystems that far outweigh effects of increasing CO₂ concentration and climatic change. In short-term experiments, which in this case means on the order of 10 years or less, elevated atmospheric CO₂ concentration affects terrestrial higher plants in several ways. Elevated CO₂ can stimulate photosynthesis, but plants may acclimate and (or) adapt to a change in atmospheric CO₂ concentration. Acclimation and adaptation of photosynthesis to increasing CO₂ concentration is unlikely to be complete, however. Plant water use efficiency is positively related to CO₂ concentration, implying the potential for more plant growth per unit of precipitation or soil moisture with increasing atmospheric CO₂ concentration. Plant respiration may be inhibited by elevated CO₂ concentration, and although a naive C balance perspective would count this as a benefit to a plant, because respiration is essential for plant growth and health, an inhibition of respiration can be detrimental. The net effect on terrestrial plants of elevated atmospheric CO₂ concentration is generally an increase in growth and C accumulation in phytomass. Published estimations, and speculations about, the magnitude of global terrestrial-plant growth responses to increasing atmospheric CO₂ concentration range from negligible to fantastic. Well-reasoned analyses point to moderate global plant responses to CO₂ concentration. Transfer of C from plants to soils is likely to increase with elevated CO₂ concentrations because of greater plant growth, but quantitative effects of those increased inputs to soils on soil C pool sizes are unknown. Whether increases in leaf-level photosynthesis and short-term plant growth stimulations caused by elevated atmospheric CO₂ concentration will have, by themselves, significant long-term (tens to hundreds of years) effects on ecosystem C storage and atmospheric CO₂ concentration is a matter for speculation, not firm conclusion. Long-term field studies of plant responses to elevated atmospheric CO₂ are needed. These will be expensive, difficult, and by definition, results will not be forthcoming for at least decades. Analyses of plants and ecosystems surrounding natural geological CO₂ degassing vents may provide the best surrogates for long-term controlled experiments, and therefore the most relevant information pertaining to long-term terrestrial-plant responses to elevated CO₂ concentration, but pollutants associated with the vents are a concern in some cases, and quantitative knowledge of the history of atmospheric CO₂ concentrations near vents is limited. On the whole, terrestrial higher-plant responses to increasing atmospheric CO₂ concentration probably act as negative feedbacks on atmospheric CO₂ concentration increases, but they cannot by themselves stop the fossil-fuel-oxidation-driven increase in atmospheric CO₂ concentration. And, in the very long-term, atmospheric CO₂ concentration is controlled by atmosphere-ocean C equilibrium rather than by terrestrial plant and ecosystem responses to atmospheric CO₂ concentration.     

Ecophysiology matters: linking inorganic carbon acquisition to ecological preference in four species of microalgae (Chlorophyceae)

The effect of CO₂ supply is likely to play an important role in algal ecology. Since inorganic carbon (C_i) acquisition strategies are very diverse among microalgae and C_i availability varies greatly within and among habitats, we hypothesized that C_i acquisition depends on the pH of their preferred natural environment (adaptation) and that the efficiency of C_i uptake is affected by CO₂ availability (acclimation). To test this, four species of green algae originating from different habitats were studied. The pH‐drift and C_i uptake kinetic experiments were used to characterize C_i acquisition strategies and their ability to acclimate to high and low CO₂ conditions and high and low pH was evaluated. Results from pH drift experiments revealed that the acidophile and acidotolerant Chlamydomonas species were mainly restricted to CO₂, whereas the two neutrophiles were efficient bicarbonate users. CO₂ compensation points in low CO₂‐acclimated cultures ranged between 0.6 and 1.4 μM CO₂ and acclimation to different culture pH and CO₂ conditions suggested that CO₂ concentrating mechanisms were present in most species. High CO₂ acclimated cultures adapted rapidly to low CO₂ condition during pH‐drifts. C_i uptake kinetics at different pH values showed that the affinity for C_i was largely influenced by external pH, being highest under conditions where CO₂ dominated the C_i pool. In conclusion, C_i acquisition was highly variable among four species of green algae and linked to growth pH preference, suggesting that there is a connection between C_i acquisition and ecological distribution.     

Leaf and canopy scale drivers of genotypic variation in soybean response to elevated carbon dioxide concentration

The atmospheric [CO₂] in which crops grow today is greater than at any point in their domestication history and represents an opportunity for positive effects on seed yield that can counteract the negative effects of greater heat and drought this century. In order to maximize yields under future atmospheric [CO₂], we need to identify and study crop cultivars that respond most favorably to elevated [CO₂] and understand the mechanisms contributing to their responsiveness. Soybean (Glycine max Merr.) is a widely grown oilseed crop and shows genetic variation in response to elevated [CO₂]. However, few studies have studied the physiological basis for this variation. Here, we examined canopy light interception, photosynthesis, respiration and radiation use efficiency along with yield and yield parameters in two cultivars of soybean (Loda and HS93‐4118) previously reported to have similar seed yield at ambient [CO₂], but contrasting responses to elevated [CO₂]. Seed yield increased by 26% at elevated [CO₂] (600 μmol/mol) in the responsive cultivar Loda, but only by 11% in HS93‐4118. Canopy light interception and leaf area index were greater in HS93‐4118 in ambient [CO₂], but increased more in response to elevated [CO₂] in Loda. Radiation use efficiency and harvest index were also greater in Loda than HS93‐4118 at both ambient and elevated [CO₂]. Daily C assimilation was greater at elevated [CO₂] in both cultivars, while stomatal conductance was lower. Electron transport capacity was also greater in Loda than HS93‐4118, but there was no difference in the response of photosynthetic traits to elevated [CO₂] in the two cultivars. Overall, this greater understanding of leaf‐ and canopy‐level photosynthetic traits provides a strong conceptual basis for modeling genotypic variation in response to elevated [CO₂].     

Regulation of the induction of bicarbonate uptake by dissolved CO2 in the marine diatom, Phaeodactylum tricornutum

Physiological properties of photosynthesis were determined in the marine diatom, Phaeodactylum tricornutum UTEX640, during acclimation from 5% CO₂ to air and related to H₂CO₃ dissociation kinetics and equilibria in artificial seawater. The concentration of dissolved inorganic carbon at half maximum rate of photosynthesis (K_0·5[DIC]) value in high CO₂‐grown cells was 1009 mmol m ^{? 3} but was reduced three‐fold by the addition of bovine carbonic anhydrase (CA), whereas in air‐grown cells K_0·5[DIC] was 71 mmol m ^{? 3}, irrespective of the presence of CA. The maximum rate of photosynthesis (P_max) values varied between 300 and 500 μ mol O₂ mg Chl ^{? 1} h ^{? 1} regardless of growth pCO₂. Bicarbonate dehydration kinetics in artificial seawater were re‐examined to evaluate the direct HCO₃ ^? uptake as a substrate for photosynthesis. The uncatalysed CO₂ formation rate in artificial seawater of 31·65°/_oo of salinity at pH 8·2 and 25 °C was found to be 0·6 mmol m ^{? 3} min ^{? 1} at 100 mmol m ^{? 3} DIC, which is 53·5 and 7·3 times slower than the rates of photosynthesis exhibited in air‐ and high CO₂‐grown cells, respectively. These data indicate that even high CO₂‐grown cells of P. tricornutum can take up both CO₂ and HCO₃ ^? as substrates for photosynthesis and HCO₃ ^? use improves dramatically when the cells are grown in air. Detailed time courses were obtained of changes in affinity for DIC during the acclimation of high CO₂‐grown cells to air. The development of high‐affinity photosynthesis started after a 2–5 h lag period, followed by a steady increase over the next 15 h. This acclimation time course is the slowest to be described so far. High CO₂‐grown cells were transferred to controlled DIC conditions, at which the concentrations of each DIC species could be defined, and were allowed to acclimate for more than 36 h. The K_0·5[DIC] values in acclimated cells appeared to be correlated only with [CO_2(aq)] in the medium but not to HCO₃ ^? , CO₃^{2 ?} , total [DIC] or the pH of the medium and indicate that the critical signal regulating the affinity of cells for DIC in the marine diatom, P. tricornutum, is [CO_2(aq)] in the medium.     

Seasonal changes in nitrate use by three woody species: the importance of the leaf-expansion period

Seasonal changes in plant NO₃ ⁻-N use were investigated by measuring leaf nitrate reductase activity (NRA), leaf N concentration, and leaf expansion in one evergreen woody species (Quercus glauca Thunb.) and two deciduous woody species [Acer palmatum Thunb. and Zelkova serrata (Thunb.) Makino]. Leaf N concentration was highest at the beginning of leaf expansion and decreased during the expansion process to a steady state at the point of full leaf expansion in all species. The leaf NRA of all species was very low at the beginning of leaf expansion, followed by a rapid increase and subsequent decrease. The highest leaf NRA was observed in the middle of the leaf-expansion period, and the lowest leaf NRA occurred in summer for all species. Significant positive correlations were detected between leaf NRA and leaf expansion rates, while leaf N concentrations were negatively correlated with leaf area. In the evergreen Q. glauca, the N concentration in current buds increased before leaves opened; concurrently, the N concentration in 1-year-old leaves decreased by 25%. Our results show that the leaf-expansion period is the most important period for NO₃ ⁻-N assimilation by broadleaf tree species, and that decreases in leaf N concentration through the leaf-expansion period are at least partly compensated for by newly assimilated NO₃ ⁻-N in current leaves.