Growth limitation of submerged aquatic macrophytes by inorganic carbon

1. This study determined the effects of CO₂ and HCO₃- enrichment on in situ growth of two submerged macrophytes, Elodea canadensis and Callitriche cophocarpa, in two Danish lakes: Lake Hampen and Lake Væng. Lake Hampen is an oligotrophic low-alkaline lake (0.4 meq ^?1) and Lake Væng is mesotrophic with an alkalinity of 1.1 meq 1-^?1. In Lake Hampen experiments were carried out throughout the growth season, whereas experiments in Lake Væng were restricted to late summer. The CO₂ and HCO₃-enrichment procedures used increased the concentration of free-CO₂ by 500–1000 μM and the concentration of HCO₃- by about 80 μM. 2. The concentration of free-CO₂ in Lake Hampen was about five times atmospheric equilibrium concentration (55 μM) in early summer declining to virtually zero at the end of summer. 3. Under ambient conditions Callitriche, which is restricted to CO₂ use, was unable to grow and survive in both lakes. In contrast, Elodea, which has the potential to use HCO₃- in photosynthesis, grew at rates varying from 0.046 to 0.080 day^?1 over the season. 4. Under CO₂ enrichment the growth rate of Callitriche varied from 0.089 to 0.124 day^?1 and for Elodea from 0.076 to 0.117 day^?1 over the season. Enrichment with HCO₃-affected Elodea only and only to a limited extent. This may be a result of insufficient increase in [HCO₃-] upon enrichment or to a limited capacity of the plants to take up HCO₃-. 5. The substantial stimulation of in situ growth of Elodea and Callitriche by enhanced concentrations of free-CO₂ shows that inorganic carbon is an important determinant of growth of submerged macrophytes and that inorganic carbon limitation of in situ growth may be a common phenomenon in nature, even in lakes with an alkalinity as high a 1 meq 1-^?1. Inorganic carbon, however, is only one of many parameters important for growth, and the growth rates of Elodea at both ambient and high free-CO₂ were closely coupled to day length and photon irradiance, indicating that light had an ultimate control on growth.     

The metabolic cost of bicarbonate use in the submerged plant Elodea nuttallii

The aquatic plant Elodea nuttallii (Planch.) St. John has been shown to express plasticity in the source of inorganic carbon it uses for photosynthesis. An investigation was undertaken to determine what effect the switch from CO₂ to HCO₃⁻ use had on the growth of E. nuttallii. Plants were grown under reduced CO₂ availability that favoured the switch, together with control plants (CO₂ at equilibrium with air) that continued to use CO₂ only. The extent to which both sets of plants could utilise HCO₃⁻ was determined (as the ratio of oxygen evolution at pH 9 and 6.5), and several measures of growth were made. Although reduced CO₂ availability produced an increase in HCO₃⁻ utilisation, no differences were found in the measured growth of the plants. Therefore, it was possible to estimate, from the difference between the estimated rate of photosynthesis of the plants utilising HCO₃⁻ and those using CO₂ only, the approximate cost of constructing, maintaining and running the bicarbonate utilisation mechanism in this species as 69 μmol photons m⁻² s⁻¹. This value can be used to estimate an irradiance of circa 80 μmol m⁻² s⁻¹ below which HCO₃⁻ use would not be expected in this species, an irradiance commonly experienced by submerged macrophytes in the field.     

A role for mitochondrial carbonic anhydrase in limiting CO2 leakage from low CO2-grown cells of Chlamydomonas reinhardtii

A model is presented which quantifies a possible role for the carbonic anhydrase in the mitochondrial matrix of Chlamydomonas reinhardtii which incorporates the observation that the expression of this enzyme is increased under growth conditions in which the expression of the carbon dioxide-concentrating mechanism is increased. It is assumed that the inorganic carbon enters the cytosol from the medium, and leaves the cytosol to the plastids, as HCO₃⁻ and that there is negligible carbonic anhydrase activity in the cytosol. The role of the mitochondrial carbonic anhydrase is suggested to be the conversion to HCO₃^– of the CO₂ produced in the mitochondria in the light from tricarboxylic acid cycle activity and from decarboxylation of glycine in any photorespiratory carbon oxidation cycle activity which is not suppressed by the carbon concentrating mechanism. If there is a HCO₃⁻ channel in the inner mitochondrial membrane then almost all of the inorganic carbon leaves the mitochondria as HCO₃⁻, thus limiting the potential for CO₂ leakage through the plasmalemma. This mechanism could increase inorganic C supply to ribulose bisphosphate carboxylase-oxygenase by some 10% at the energetic expense of less than 1% of the total ATP generation by plastids plus mitochondria.     

Photosynthetic response to globally increasing CO2 of co‐occurring temperate seagrass species

Photosynthesis of most seagrass species seems to be limited by present concentrations of dissolved inorganic carbon (DIC). Therefore, the ongoing increase in atmospheric CO₂ could enhance seagrass photosynthesis and internal O₂ supply, and potentially change species competition through differential responses to increasing CO₂ availability among species. We used short‐term photosynthetic responses of nine seagrass species from the south‐west of Australia to test species‐specific responses to enhanced CO₂ and changes in HCO₃^?. Net photosynthesis of all species except Zostera polychlamys were limited at pre‐industrial compared to saturating CO₂ levels at light saturation, suggesting that enhanced CO₂ availability will enhance seagrass performance. Seven out of the nine species were efficient HCO₃^? users through acidification of diffusive boundary layers, production of extracellular carbonic anhydrase, or uptake and internal conversion of HCO₃^?. Species responded differently to near saturating CO₂ implying that increasing atmospheric CO₂ may change competition among seagrass species if co‐occurring in mixed beds. Increasing CO₂ availability also enhanced internal aeration in the one species assessed. We expect that future increases in atmospheric CO₂ will have the strongest impact on seagrass recruits and sparsely vegetated beds, because densely vegetated seagrass beds are most often limited by light and not by inorganic carbon.     

Effects of phytoplankton on the distribution of submerged macrophytes in a small canal

Submerged macrophytes have been disappearing from the Kanto Plain, Japan since the 1960s. This disappearance is usually attributable to the interaction between macrophytes and phytoplankton. Phytoplankton contributes to shading of the available light and changes the availability of inorganic carbon from free CO₂ to HCO ₃ ^? for use in photosynthesis. However, limited information is available about the interaction between carbon fraction and submerged macrophytes through phytoplankton abundance. In this short note, we observe the distribution of submerged macrophytes and phytoplankton in a small canal. We found that, despite high photosynthetically active radiation (PAR) in the downstream region, low free CO₂ concentration through phytoplankton abundance can deplete free CO₂ for submerged macrophytes. In contrast, the upstream region exhibited macrophytes in an environment with high free CO₂ concentration. The stable carbon isotope ratio of submerged macrophytes follows this pattern, with more positive values occurring in the downstream region and more negative values in the upstream region. It has been reported that phytoplankton limits light availability for submerged macrophytes, but carbon availability could also be a factor in the distribution of submerged macrophytes. Because the source of water for submerged macrophytes is groundwater, its preservation possibly plays a key role for the restoration of submerged macrophytes.     

Growth and Photosynthesis of the Cyanobacterium Synechococcus leopoliensis in HCO(3)-Limited Chemostats

Synechococcus leopoliensis was grown in HCO₃⁻-limited chemostats. Growth at 50% the maximum rate occurred when the inorganic carbon concentration was 10 to 15 micromolar (or 5.6 to 8.4 nanomolar CO₂). The O₂ to CO₂ ratios during growth were as high as 192,000 to 1. At growth rates below 80% the maximum rate, essentially all the supplied inorganic carbon was converted to organic carbon, and the cells were carbon limited. Carbon-limited cells used HCO₃⁻ rather than CO₂ for growth. They also exhibited a very high photosynthetic affinity for inorganic carbon in short-term experiments. Cells growing at greater than 80% maximum growth rate, in the presence of high dissolved inorganic carbon, were termed carbon sufficient. These cells had photosynthetic affinities that were about 1000-fold lower than HCO₃⁻-limited cells and also had a reduced capacity for HCO₃⁻ transport. HCO₃⁻-limited cells are reminiscent of the air-grown cells of batch culture studies while the carbon sufficient cells are reminiscent of high-CO₂ grown cells. However, the low affinity cells of the present study were growing at CO₂ concentrations less than air saturation. This suggests that supranormal levels of CO₂ not required to induce the physiological changes usually ascribed to high CO₂ cells.     

CO2 is the main inorganic C species entering photosynthetically active leaf protoplasts of the freshwater macrophyte Ranunculus penicillatus ssp. pseudofluitans

Submerged aquatic macrophytes growing in water where free CO₂ is unavailable (above pH 8·2) must use mechanisms to supply external dissolved inorganic carbon in a form available to chloroplasts (CO₂). Active transport of HCO₃^– across the plasmalemma has not been proven to be widespread in aquatic macrophytes and catalytic conversion of HCO₃^– to CO₂ is the usual supply mechanism in submerged macrophytes. The interaction of leaf form and function in this respect was investigated in the linear, submerged leaves of Ranunculus penicillatus (Dumort.) Bab ssp. pseudofluitans (Syme) S.Webster. Viable protoplasts were isolated using a mixture of cell wall degrading enzymes optimized for this species. Protoplast viabilities greater than 80% after 5 h of isolation were achieved. Photosynthetic rates of isolated protoplasts were comparable with that of intact plant tissue. Results of carbon isotopic disequilibrium experiments showed that CO₂ was the preferred species of dissolved inorganic carbon for photosynthesis by protoplasts and that HCO₃^– which predominates in the plant’s natural environment mainly contributes by supplying CO₂ outside the cells.     

Diurnal variation in light and carbon limitation of photosynthesis by two species of submerged freshwater macrophyte with a differential ability to use bicarbonate

SUMMARY.

• 1 Rates of photosynthetic oxygen evolution by Callitriche cophocarpa and Ranunculus peltatus in stream were measured on live occasions during the light period on 2 days at ambient light and ambient inorganic carbon, ambient light and saturating inorganic carbon, saturating light and ambient inorganic carbon, saturating light and saturating inorganic carbon and air-equilibrium inorganic carbon and ambient light.
• 2 Despite an ambient CO₂ concentration of about 220 μm , which is about ten times air-equilibrium, the concentration of inorganic carbon was more limiting than light on all the occasions that rates were measured. On average, rates of photosynthesis at ambient concentrations of CO₂ were about 130 and 425 μmol O₂ g^?1 DW h^?1 for C. cophocarpa and R. peltatus, respectively. These rates as a percentage of carbon saturated rates were only about 35% for C. cophocarpa and about 60% for R. peltatus. Ambient rates as a percentage of light saturated rates were about 80% for C. cophocarpa and about 95% for R. peltatus. Only in early morning and late evening where the photon irradiance was below 160 μmol m^?2 s^?1 was there evidence for slight light limitation.
• 3 Based on results from pH-drift experiments and from rates of photosynthesis as a function of CO₂ concentration in the presence and absence of HCO₃^?, C. cophocarpa was unable, but R. peltatus able to use HCO₃^? at an ambient HCO₃^? concentration of about 0·84 mm . The greater rates of photosynthesis at ambient CO₂ concentration and the lesser limitation by inorganic carbon shown by R. peltatus compared to C. cophocarpa was the result of HCO₃^?-use as laboratory experiments showed that R. peltatus performed similarly to C. cophocarpa if the HCO₃^? concentration was reduced to 60 μm .

     

Acclimation of photosynthesis to supersaturated CO2 in aquatic plant bicarbonate users

相似文献   

The effect of pH on the inorganic carbon source for photosynthesis in the seagrass Zostera muelleri irmisch ex aschers

The ability of the seagrass Zostera muelleri Irmisch ex Aschers. to use HCO₃⁻ as well as CO₂ for photosynthesis was investigated by measuring photosynthetic O₂ evolution over a range of pH values. It was found that the apparent K_m CO₂ fell from 0.128 mM at pH 7.9 to 0.016 mM at pH 9.1 indicating that HCO₃⁻ as well as CO₂ may act as a substrate for photosynthesis.The true K_m CO₂ could not be determined due to inhibition of photosynthesis at pHs less than 7.8 K_m CO₂ must be at least 0.128 mM, the apparent K_m at pH 7.9, and is probably of the order of 0.200 mM CO₂, the same as that reported for other marine plants. K_m HCO₃⁻¹ is about 20 mM when CO₂-dependent photosynthesis is minimal. Such a high K_m HCO₃⁻ resembles values reported for freshwater, rather than marine plants.Photosynthetic O₂ evolution is not saturated with respect to total inorganic carbon in natural seawater (pH 8.2). It is suggested that the distinctive shoulder from pH 8.1 to 8.5 in the pH profile of photosynthetic O₂ evolution at a constant concentration of inorganic carbon is caused by an effect of pH on HCO₃⁻ uptake. The effect of pH on HCO₃⁻ uptake was determined by constructing a pH profile of photosynthesis at constant HCO₃⁻ concentration, and subtracting the estimated contribution of CO₂ to photosynthesis from this rate. The resultant curve has a maximum at pH 8.4 and declines sharply at pHs less than 8.     

Scaling of photosynthetic production of aquatic macrophytes – a review

Most studies on photosynthetic production of aquatic macrophytes have been made on detached leaves and algal thalli. This may have given the false impression that production is often saturated by light and that inorganic carbon and nutrients are more important limiting factors. However, studies on the more relevant ecological scale of macrophyte communities lead to a completely different perception because community production is light limited due to intense self‐shading. Relatively high irradiances are needed for photosynthesis to balance respiratory costs and not even maximum irradiances at noon during summer saturate photosynthetic production. The fundamental importance of light is confirmed by the close coupling to community light absorptance of both maximum production in high‐light environments and efficiency of light use in low‐light environments. The upper boundaries of light‐limited and light‐saturated production are distinctly and linearly related to community absorptance. Moreover, higher diversity in the community has a positive and stabilizing influence on light absorptance and production because different species supplement each other temporally and spatially. Close predictions of actual production rates in macroalgal communities throughout the year are possible solely from determinations of incident and absorbed irradiance. Along with the increasing regulating role of light from leaves or thalli to entire communities the importance of temperature, inorganic carbon and other resources decreases. Thus, ten‐fold rise of CO₂ relative to atmospheric saturation does not enhance maximum production in dense communities of efficient HCO₃^?‐users and only doubles production of pure CO₂‐users. A challenge therefore exists establishing the importance of light for photosynthetic production of macrophytes from individuals to communities and re‐evaluating the postulated main importance of inorganic carbon and temperature in the scenarios of globally rising CO₂ and temperature.     

SOURCES OF INORGANIC CARBON FOR PHOTOSYNTHESIS BY THREE SPECIES OF MARINE DIATOM1

The utilization of inorganic carbon by three species of marine diatom, Skeletonema costatum (Grev.) Cleve. Ditylum brightwellii (West) Grun., and Chaetoceros calcitrans Paulsen was investigated using an inorganic carbon isotopic disequilibnum technique and inorganic carbon dose-response curves. Stable carbon isotope data of the diatoms are also presented. Observed rates of photosynthetic oxygen evolution were greater than could be accounted for by the theoretical rate of CO₂ supply from the uncatalyzed dehydration of HCO₃^? in the external medium, suggesting use of HCO₃^? as an inorganic carbon source. Data from the isotopic disequilibrium experiment demonstrate the use of both HCO₃^? and CO₂ for photosynthesis. Carbon isotope discrimination values support the use of HCO₃^? by the diatoms.     

EFFECTS OF CO2 ENRICHMENT ON THE BLOOM‐FORMING CYANOBACTERIUM MICROCYSTIS AERUGINOSA (CYANOPHYCEAE): PHYSIOLOGICAL RESPONSES AND RELATIONSHIPS WITH THE AVAILABILITY OF DISSOLVED INORGANIC CARBON1

Microcystis aeruginosa Kütz. 7820 was cultured at 350 and 700 μL·L ^{? 1} CO₂ to assess the impacts of doubled atmospheric CO₂ concentration on this bloom‐forming cyanobacterium. Doubling of CO₂ concentration in the airflow enhanced its growth by 52%–77%, with pH values decreased and dissolved inorganic carbon (DIC) increased in the medium. Photosynthetic efficiencies and dark respiratory rates expressed per unit chl a tended to increase with the doubling of CO₂. However, saturating irradiances for photosynthesis and light‐saturated photosynthetic rates normalized to cell number tended to decrease with the increase of DIC in the medium. Doubling of CO₂ concentration in the airflow had less effect on DIC‐saturated photosynthetic rates and apparent photosynthetic affinities for DIC. In the exponential phase, CO₂ and HCO₃ ^? levels in the medium were higher than those required to saturate photosynthesis. Cultures with surface aeration were DIC limited in the stationary phase. The rate of CO₂ dissolution into the liquid increased proportionally when CO₂ in air was raised from 350 to 700 μL·L ^{? 1}, thus increasing the availability of DIC in the medium and enhancing the rate of photosynthesis. Doubled CO₂ could enhance CO₂ dissolution, lower pH values, and influence the ionization fractions of various DIC species even when the photosynthesis was not DIC limited. Consequently, HCO₃ ^? concentrations in cultures were significantly higher than in controls, and the photosynthetic energy cost for the operation of CO₂ concentrating mechanism might decrease.     

The Underwater Life of Secondarily Aquatic Plants: Some Problems and Solutions

The freshwater secondarily aquatic plants, most of which are higher plants, are those returned to the water environment after spending a period of time living on land. The readaptation to living underwater has made it necessary for these plants to put in place morphological and functional strategies to cope with some major problems due to features of the aquatic environment, but also deriving from the specialized organization of their “terrestrial” bodies. The poor O₂ availability underwater accounted for the evolution of wide aerenchyma tissues throughout the plant organs to improve the photosynthetic O₂ flux from the shoot to the roots buried in anoxic sediments and to the neighboring rhizosphere. This favors sediment oxygenation, sustains the aerobic metabolism of roots, and improves the availability and uptake of mineral nutrients, whose delivery to the entire plants, without a transpirational flux, is ensured by an acropetal mass transport depending on root pressure, guttation from hydathodes and channeling by apoplast closure around the vascular tissues. A great expansion of leaf surfaces and an enhanced surface:volume ratio of chloroplast-rich photosynthetic cells help to contact the water medium and to increase the cell/environment exchanges to gain inorganic carbon. Furthermore, different physiological mechanisms operate to cope with the scarce availability of CO₂ and the prevalence of HCO₃ ^? as inorganic carbon form in water. Some of them, like cell wall acidification through H⁺ extrusion by a light-dependent APTase or activation of an apoplastic carbonic anhydrase, operate outside the cells, leading to a conversion of HCO₃ ^? to CO₂, which then diffuses into the cells. Others, on the contrary, act inside the cells to load the active site of Rubisco with CO₂, thus favoring photosynthesis and lowering photorespiration. Aquatic macrophytes with isoetid life form, moreover, can obtain most ot the fixed CO₂ from sediments. In submerged species, in additin to the C₃ cycle, the C₄ and CAM-like photosynthetic metabolisms can also operate, and are modulated by the environmental inorganic carbon availability and the plant photosynthetic demand. Interestingly, in the aquatic plants the C₄ pathway, which can be concomitant with the C₃ one, does not depend on the Kranz anatomy of leaves, but relies on the intracellular compartmentation of carboxylative and decarboxylative enzymes. The CAM-like pathway, defined AAM, which also coexists with the C₃, allows the submerged plants to fix CO₂ in the dark, thus exploiting the higher CO₂ availability in the water medium during the night, and extending to 24?h the period of inorganic carbon assimilation. In almost all the aquatic macrophytes the AAM is only expressed in the submersion state, whereas it is quickly inactivated in emerging leaves in a cell by cell way.     

Two Systems for Concentrating CO(2) and Bicarbonate during Photosynthesis by Scenedesmus

Scenedesmus cells grown on high CO₂, when adapted to air levels of CO₂ for 4 to 6 hours in the light, formed two concentrating processes for dissolved inorganic carbon: one for utilizing CO₂ from medium of pH 5 to 8 and one for bicarbonate accumulation from medium of pH 7 to 11. Similar results were obtained with assays by photosynthetic O₂ evolution or by accumulation of dissolved inorganic carbon inside the cells. The CO₂ pump with K_0.5 for O₂ evolution of less than 5 micromolar CO₂ was similar to that previously studied with other green algae such as Chlamydomonas and was accompanied by plasmalemma carbonic anhydrase formation. The HCO₃⁻ concentrating process between pH 8 to 10 lowered the K_0.5 (DIC) from 7300 micromolar HCO₃⁻ in high CO₂ grown Scenedesmus to 10 micromolar in air-adapted cells. The HCO₃⁻ pump was inhibited by vanadate (K_i of 150 micromolar), as if it involved an ATPase linked HCO₃⁻ transporter. The CO₂ pump was formed on low CO₂ by high-CO₂ grown cells in growth medium within 4 to 6 hours in the light. The alkaline HCO₃⁻ pump was partially activated on low CO₂ within 2 hours in the light or after 8 hours in the dark. Full activation of the HCO₃⁻ pump at pH 9 had requirements similar to the activation of the CO₂ pump. Air-grown or air-adapted cells at pH 7.2 or 9 accumulated in one minute 1 to 2 millimolar inorganic carbon in the light or 0.44 millimolar in the dark from 150 micromolar in the media, whereas CO₂-grown cells did not accumulate inorganic carbon. A general scheme for concentrating dissolved inorganic carbon by unicellular green algae utilizes a vanadate-sensitive transporter at the chloroplast envelope for the CO₂ pump and in some algae an additional vanadate-sensitive plasmalemma HCO₃⁻ transporter for a HCO₃⁻ pump.     

贵州喀斯特森林三种植物对不同坡位环境的光合生理响应

该研究以贵州普定喀斯特森林中、下坡位生长的构树( Broussonetia papyrifera)、朴树( Celtis sinensis)和光滑悬钩子( Rubus tsangii)为材料,通过对碳酸酐酶( CA)活性、光合作用日变化、净光合速率对CO2与光的响应曲线、叶绿素荧光特性以及稳定碳同位素组成等指标的测定,进而对比分析三种植物不同的光合生理响应特性。结果表明：构树光合作用过程的无机碳源既可来自大气中的CO2,也可以在气孔部分闭合的情况下利用细胞内的HCO3－,下坡位的构树较高的CA活性使其利用HCO3－的效率会更高,并能在较低光强下具有较高的光能利用效率。这可能与下坡位的构树具有较高的CA活性有关,对下坡位具有更好的适应性。朴树光合无机碳的同化能力最低,且光合无机碳源较单一,主要利用大气CO2,其较慢的生长速率使其对无机碳的需求最低,且能保持较稳定的无机碳同化速率。相对来说,中坡位的朴树具有相对较高的净光合速率和光能利用效率,对中坡位表现出较好的适应性。光滑悬钩子主要利用大气中的CO2进行光合作用。中坡位的光滑悬钩子具有较强的光能利用效率,并表现出较高的净光合速率,光滑悬钩子对中坡位同样表现出较好的适应性。该研究结果为喀斯特生态脆弱区植被重建过程中树种的选择及合理配置提供了科学依据。     

Studies of Elodea nuttallii grown under photorespiratory conditions. I. Photosynthetic characteristics

Abstract. The photosynthetic characteristics of Elodea nuttallii grown in wastewater in continuous flow reactors in a greenhouse were investigated. The diurnal changes in dissolved inorganic carbon (DIC), dissolved oxygen (DO) and pH were monitored. Photosynthesis removed both CO₂(aq) and HCO₃^? from the reactors. A stoichiometry of 1.19:1 was observed between HCO₃^? removal during photosynthesis and OH^? production during photosynthesis, consistent with theories regarding direct bicarbonate utilization. In laboratory experiments, the light compensation points (г_PPFD) were similar (31–35μmol m^?2 s^?1) to reported values for other macrophytes; however, the light saturation level was high (1100μmol m^?2 s^?1) and similar to values reported for aerial portions Of heterophyllous macrophytes. The kinetics of photosynthetic oxygen evolution (K_m (CO₂) = 96mmol m^?3; V_max= 133mmol g^?1 Chl h^?1) and the CO₂ compensation point (г= 44cm³ m^?3) suggested an adaptive, low photorespiratory state in response to low carbon concentrations. Photosynthetic V_max values were slightly, but significantly higher (P 0.001) at pH 8.0 compared to pH 4.5. While CO₂ utilization at pH 8 could account for most of the observed phototsynthetic rates, an HCO₃^? component was present, suggesting two separate transport systems for HCO₃^? and CO₂(aq) in E. nuttallii. The activity of RUBISCO (160.3 mmol g^?1 Chl h^?1 was one of the highest reported values for aquatic macrophytes. Compared to RUBISCO, we observed lower activities of the β-carboxylating enzymes phopho enolpyruvate carboyxlase (PEPcase), 24.1 mmol g^?1 Chl h^?1; phosphor enol pyruvate carboxykinase (PEPCKase), 14 mmol g^?1 Chl h^?1. This suggests that the potential light-independent fixation of carbon in E. nuttallii was much less than RUBISCO-dependent fixation. The RUBISCO/PEPcase ratio was 6.6, indicating that E. nuttallii was similar to Myriophyllum sp. in possessing a physiological adaptation to low CO₂ levels which is hypothesized to include carbonic anhydrase (CA) and an active transport system for HCO₃^?. CA levels were surprisingly low in E. nuttallii (14.2 EUmg Chl^?).     

CO2 uptake and transport in leaf mesophyll cells

Abstract The acquisition of inorganic carbon for photosynthetic assimilation by leaf mesophyll cells and chloroplasts is discussed with particular reference to membrane permeation of CO₂ and HCO^?₃. Experimental evidence indicates that at the apoplast pH normally experienced by leaf mesophyll cells (pH 6–7) CO₂ is the principal species of inorganic carbon taken up. Uptake of HCO^?₃ may also occur under certain circumstances (i.e. pH 8.5), but its contribution to the net flux of inorganic carbon is small and HCO^?₃ uptake does not function as a CO₂-concentrating mechanism. Similarly, CO₂ rather than HCO^?₃ appears to be the species of inorganic carbon which permeates the chloroplast envelope. In contrast to many C₃ aquatic plants and C₄ plants, C₃ terrestrial plants lack specialized mechanisms for the acquisition and transport of inorganic carbon from the intercellular environment to the site of photosynthetic carboxylation, but rely upon the diffusive uptake of CO₂.     

Effect of photon flux density on inorganic carbon accumulation and net CO2 exchange in a high-CO2-requiring mutant of Chlamydomonas reinhardtii

The effect of photon flux density on inorganic carbon accumulation and photosynthetic CO₂ assimilation was determined by CO₂ exchange studies at three, limiting CO₂ concentrations with a ca-1 mutant of Chlamydomonas reinhardiii. This mutant accumulates a large internal inorganic carbon pool in the light which apparently is unavailable for photosynthetic assimilation. Although steady-state photosynthetic CO₂ assimilation did not respond to the varying photon flux densities because of CO₂ limitation, components of inorganic-carbon accumulation were not clearly light saturated even at 1100 mol photons m^-2 s^-1, indicating a substantial energy requirement for inorganic carbon transport and accumulation. Steady-state photosynthetic CO₂ assimilation responded to external CO₂ concentrations but not to changing internal inorganic carbon concentrations, confirming that diffusion of CO₂ into the cells supplies most of the CO₂ for photosynthetic assimilation and that the internal inorganic carbon pool is essentially unavailable for photosynthetic assimilation. The estimated concentration of the internal inorganic carbon pool was found to be relatively insensitive to the external CO₂ concentration over the small range tested, as would be expected if the concentration of this pool is limited by the internal to external inorganic carbon gradient. An attempt to use this CO₂ exchange method to determine whether inorganic carbon accumulation and photosynthetic CO₂ assimilation compete for energy at low photon flux densities proved inconclusive.     

HCO3− and CO2 leakage from Synechococcus UTEX 625

The leakage of various inorganic carbon species from air-grown cells of Synechococcus UTEX 625 was investigated after a light to dark transition or during a light period using a mass spectrometer under a wide variety of experimental conditions. Total inorganic carbon efflux and CO₂ efflux during the initial period of darkness were measured with or without carbonic anhydrase in the reaction medium respectively. The HCO₃^? efflux after a light to dark transition was estimated by difference. Carbon dioxide efflux in the light was measured by inhibiting CO₂ transport with either Na₂S or COS₃ or quenching the ¹³C inorganic carbon transport by the addition of ¹²C inorganic carbon in excess. In cells in which CO₂ fixation was inhibited, when only the HCO₃^? transport system was fully operative, CO₂ effluxed continuously during the light period at a rate equal to about 25% of that in darkness. When only the CO₂ transport system was operative, HCO₃^? effluxed during the light period. The difference between the light and dark efflux rates was consistent with a 0.6 unit decrease in the intracellular pH upon darkening the cells. The permeabilities of the cell for CO₂ (2.94 ± 0.14 ± 10^?8ms^?1; mean ± SE, n=137) and HCO₃^? (1.4–1.7 ± 10^?9 ms^?1) were calculated.