Oxygen inhibition of dissolved inorganic carbon uptake in unicellular green algae

Unicellular green algae have a dissolved inorganic carbon (DIC) concentrating mechanism, commonly known as the DIC pump, to concentrate inorganic carbon into cells and chloroplasts. The DIC pump activity is normally measured as the K_0.5(DIC) that equals the CO₂ plus HCO₃‐ concentration at a cited pH at which the rate of DIC‐dependent photosynthetic O₂ evolution is half‐maximal, or by the amount of intra‐cellular DIC accumulation in 15–60 s, using a limited amount of NaH¹⁴CO₃, measured by the silicone oil cen‐trifugation technique. The dissolved oxygen in the assay inhibits or reduces the DIC uptake by the cells of unicellular green algae Chlamydomonas reinhardtii Dangeard, strain 137 and in a cell wall‐less marine algae Dunaliella tertiolecta Butcher. The algal cells concentrated the highest amount of DIC when little or no oxygen was present in the assay medium. The results suggest that the amount of O₂ and DIC must be carefully monitored before DIC‐pump assay.     

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.     

Interactions of blue light and inorganic carbon supply in the control of light-saturated photosynthesis in brown algae

Photosynthetic capacities of five species of brown algae in red light were found to be strongly limited by the inorganic carbon supply of natural sea water. Under these conditions, pH 8·2 and dissolved inorganic carbon concentration (DIG) of 2·1 mol m^?3, a short pulse of blue light was found to increase the subsequent rate of photosynthesis in saturating red light. The degree of blue light stimulation varied between species, ranging from an increase of over 200% of the original rate in Colpomenia peregrins to only 10% in Dictyota dichotoma. Increasing the DIG concentration of sea water by bicarbonate addition resulted in carbon saturation of photosynthesis in all five species. Blue light stimulation was greatly reduced at these higher DIG concentrations. The response in Laminaria digitata was examined in more detail by manipulation of pH and DIG to produce solutions with different concentrations of dissolved CO₂. At a CO₂ concentration typical of normal sea water (12·4 mmol m^?3), blue light treatment increased photosynthetic rate by approximately 50%. Blue light stimulation was increased to over 150% at CO₂ concentrations below that of sea water, whereas at concentrations above that of sea water, the effect was diminished. Therefore, the effect of blue light on photosynthetic capacity appears to involve an increase in the rate of supply of carbon dioxide to the plant.     

UPTAKE,EFFLUX, AND PHOTOSYNTHETIC UTILIZATION OF INORGANIC CARBON BY THE MARINE EUSTIGMATOPHYTE NANNOCHLOROPSIS SP.1

Uptake, efflux and utilization of inorganic carbon were investigated in the marine eustigmatophyte Nannochloropsis sp. grown under an air level of CO₂. Maximal photosynthetic rate was hardly affected by raising the pH porn 5.0 to 9.0. The apparent photosynthetic affinity for dissolved inorganic carbon (DIC) was 35 μM DIC between pH 6.5 to 9.0, but increased approximately threefold at pH 5.0 suggesting that HCO₃- was the main DIC species used from the medium. No external carbonic anhydrase (CA) activity could be detected by the pH drift method. However, application of ethoxyzolamide (an inhibitor of CA) resulted an a significant inhibition of photosynthetic O₂ evolution and carbon utilization, suggesting involvement of internal CA or CA-like activity in DIC utilization. Under high light conditions, the rate of HCO₃^? uptake and its internal conversion to CO₂ apparently exceeded the rate of carbon fixation, resulting in a large leak of CO₂ from the cells to the external medium. When the cells were exposed to low DIC concentrations, the ratio of internal to external DIC concentration was about eight. On the other hand, in the presence of 2 mM DIC, conditions prevailing in the marine environment, the internal concentration of DIC was only 50% higher than the external one.     

Two Polypeptides in the Inner Chloroplast Envelope of Dunaliella tertiolecta Induced by Low CO(2)

Unicellular green algae have a mechanism for concentrating dissolved inorganic carbon (DIC) only when grown in low CO₂. To find proposed transporter protein(s) for DIC, we isolated intact chloroplasts from Dunaliella tertiolecta cells, separated the chloroplast envelopes by isopyknic centrifugation, and separated their polypeptides by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Two peptides of apparent molecular masses of 45 and 47 kD were constituents of the inner chloroplast envelope only if the cells had been adapted to low CO₂ in the light or grown in low CO₂. These two low CO₂-induced peptides appear to be part of the algal DIC pump.     

Carbon acquisition characteristics of six microalgal species isolated from a subtropical reservoir: potential implications for species succession

CO ₂ levels in freshwater systems can fluctuate widely, potentially influencing photosynthetic rates and growth of phytoplankton. Given the right conditions, this can lead to bloom formation and affect water quality. This study investigated the acquisition of dissolved inorganic carbon (DIC ) by six species of microalgae, a cyanobacterium Cylindrospermopsis raciborskii , the diatoms Cyclotella sp., Nitzschia sp., and the green algae Stichococcus sp., Staurastrum sp., and Monoraphidium sp., all isolated from a subtropical reservoir in Australia. Carbon acquisition characteristics, specifically the affinity for DIC , internal pH , and internal DIC concentrations were measured. Affinities for CO ₂ () ranged between 0.7 and 6 μM CO ₂. This was considerably lower than air‐equilibrated surface water CO ₂ concentrations, and below reported affinities for CO ₂ of RuBisCO suggesting operation of active carbon dioxide concentrating mechanisms (CCM s) in all species. Internal pH was lowest for Cyclotella sp. at 7.19, and highest for Staurastrum sp., at 7.71. At 180 μM external DIC , ratios of internal:external CO ₂ ranged from 2.5 for Nitzschia sp. to 14 in C. raciborskii . Internal HCO ₃^? concentration showed a linear relationship with surface area to biovolume ratio (SA :Vol). We hypothesized that species with a higher SA :Vol suffer more from diffusive escape of CO ₂, thus storage of DIC as bicarbonate is favored in these strains. For C. raciborskii , under stratified summer conditions, its strong CCM , and resilient photosynthetic characteristics may contribute to its bloom forming capacity.     

CO2‐CONCENTRATING MECHANISMS OF THE POTENTIALLY TOXIC DINOFLAGELLATE PROTOCERATIUM RETICULATUM (DINOPHYCEAE,GONYAULACALES)1

The low CO₂ concentration in seawater poses severe restrictions on photosynthesis, especially on those species with form II RUBISCO. We found that the potentially toxic dinoflagellate Protoceratium reticulatum Clap. et J. Lachm. possesses a form II RUBISCO. To cast some light on the mechanisms this organism undergoes to cope with low CO₂ availability, we compared cells grown at atmospheric (370 ppm) and high (5000 ppm) CO₂ concentrations, with respect to a number of physiological parameters related to dissolved inorganic carbon (DIC) acquisition and assimilation. The photosynthetic affinity for DIC was about one order of magnitude lower in cells cultivated at high [CO₂]. End‐point pH‐drift experiments suggest that P. reticulatum was not able to efficiently use HCO₃^? under our growth conditions. Only internal carbonic anhydrase (CA) activity was detected, and its activity decreased by about 60% in cells cultured at high [CO₂]. Antibodies raised against a variety of algal CAs were used for Western blot analysis: P. reticulatum extracts only cross‐reacted with anti‐ß‐CA sera, and the amount of immunoreactive protein decreased in cells grown at high [CO₂]. No pyrenoids were observed under all growth conditions. Our data indicate that P. reticulatum has an inducible carbon‐concentrating mechanism (CCM) that operates in the absence of pyrenoids and with little intracellular CO₂ accumulation. Calculations on the impact of the CA activity to photosynthetic growth [CO₂] suggest that it is an essential component of the CCM of P. reticulatum and is necessary to sustain the photosynthetic rates observed at ambient CO₂.     

Seasonal and tidal variability of environmental carbon related physico-chemical variables and inorganic C acquisition in Gracilariopsis longissima and Enteromorpha intestinalis from Los Toruños salt marsh (Cádiz Bay, Spain)

The seasonal and tidal variability of inorganic C acquisition mechanisms, photosynthesis, internal composition and growth were studied in two co-occurring macroalgae in Los Toruños salt marsh (Cádiz Bay), Gracilariopsis longissima and Enteromorpha intestinalis. This variability was monitored together with physico-chemical variables affecting carbon availability, photosynthesis, and growth. The environmental variables, such as light, temperature, pH, salinity, oxygen, alkalinity, dissolved inorganic carbon (DIC) and CO₂, displayed not only an expected seasonal cycle but also a daily (tidal) variability, with abrupt and rapid changes influenced by biological activities, physical variables, tidal state and tidal timing. In contrast to environmental variables, photosynthesis, pigments and C:N composition were affected by seasonal changes but not by tidal regimes, as organisms integrated these short-term fluctuations in physico-chemical variables. Photosynthesis, pigments and internal N composition were maximal in autumn and minimal in summer for both species. Growth showed a seasonal trend, displaying a summer drop with negative values. This response can be the result of extreme values of environmental variables (temperature, light, pH, nutrients, and the shortage of DIC) in summer, in comparison with higher growth rates in September onwards. The use of inhibitors of carbon acquisition in situ at normal DIC concentrations (2.2. mM) revealed species-dependent differences. While the external carbonic anhydrase (CA) activity showed a constitutive character in G. longissima, it showed little effect in E. intestinalis, which relies on internal CA activity. The 4, 4′-diisothiocyanatostilbene-2,2′-disulfonate (DIDS)-sensitive bicarbonate transport in G. longissima was effective in winter. In contrast, DIDS stimulated photosynthesis in summer, and relieved AZ inhibition. This response could suggest a stimulation of a H⁺ extrusion mediated-CO₂ transport in periods of low CO₂ availability.     

The role of external carbonic anhydrase in the photosynthetic use of inorganic carbon in the deep-water alga Phyllariopsis purpurascens (Laminariales, Phaeophyta)

Mechanisms of inorganic carbon assimilation were investigated in the deep-water alga Phyllariopsis purpurascens (C. Agardh) Henry et South (Laminariales, Phaeophyta). The gross photosynthetic rate as a function of external pH, at a constant concentration of 2 mM dissolved inorganic carbon (DIC), decreased sharply from pH 7.0 to 9.0, and was not substantially different from 0 above pH 9.0. These data indicate that P. purpurascens is inefficient in the use of external HCO₃ ⁻ as a carbon source in photosynthesis. Moreover, the photosynthetic rate as a function of external DIC and the highest pH (9.01 ± 0.07) that this species can achieve in a closed system were consistent with a low capacity to use HCO₃ ⁻, in comparison to many other species of seaweeds. The role of external carbonic anhydrase (CA; EC 4.2.1.1) on carbon uptake was investigated by measuring both the HCO₃ ⁻-dependent O₂ evolution and the CO₂ uptake, at pH 5.5 and 8.0, and the rate of pH change in the external medium, in the presence of selected inhibitors of extra- and intracellular CA. Photosynthetic DIC-dependent O₂ evolution was higher at pH 5.5 (where CO₂ is the predominant form of DIC) than at pH 8.0 (where the predominant chemical species is HCO₃ ⁻). Both intra- and extracellular CA activity was detected. Dextran-bound sulfonamide (DBS; a specific inhibitor of extracellular CA) reduced the photosynthetic O₂ evolution and CO₂ uptake at pH 8.0, but there was no effect at pH 5.5. The pH-change rate of the medium, under saturating irradiance, was reduced by DBS. Phyllariopsis purpurascens has a low efficiency in the use of HCO₃ ⁻ as carbon source in photosynthesis; nevertheless, the ion can be used after dehydration, in the external medium, catalyzed by extracellular CA. This mechanism could explain why the photosynthetic rate in situ was higher than that supported solely by the diffusion of CO₂ from seawater. Received: 6 March 1998 / Accepted: 22 June 1998     

Different mechanisms of inorganic carbon acquisition in red macroalgae (Rhodophyta) revealed by the use of TRIS buffer

The effects on photosynthesis of acetazolamide (AZ, an inhibitor of the external carbonic anhydrase) and TRIS buffer at pH 8.7 were assessed in 24 species of red macroalgae. Only Palmaria palmata was unaffected by both substances. The rest of species were classified into three groups according to their sensitivity to TRIS and AZ. Photosynthesis of fourteen species was significantly inhibited by both TRIS and AZ. Inhibition by TRIS varied from almost 100% to 25% while AZ produced similar effects. Inhibition by TRIS was completely reverted by increasing the dissolved inorganic carbon concentration (DIC). This species group had half-saturation constants for photosynthesis (K_m(DIC)) ranging from 0.5 to 1.1 mM of DIC. TRIS produced a significant increase of K_m(DIC). Altogether, these results indicate that the algae sensitive to TRIS are capable of using HCO₃⁻ efficiently at pH 8.7. Furthermore, the buffering capacity of TRIS was responsible for its inhibitory effect on photosynthesis suggesting that HCO₃⁻ use was facilitated by excretion of protons outside the plasma membrane, which creates regions of low pH resulting in a higher-than-ambient CO₂ concentration. In contrast, photosynthesis by two Porphyra species analysed was slightly stimulated by TRIS and completely inhibited by AZ, suggesting that the mechanism was different. In a third group of seaweeds, photosynthesis was insensitive to TRIS but it was significantly inhibited by AZ. These species had relatively high values of K_m(DIC) indicating that they relied on purely diffusive entry of CO₂ generated by external carbonic anhydrase activity. Consequently, the results demonstrate that external carbonic anhydrase is widespread among red macroalgae since only P. palmata was insensitive to AZ. The functional significance of this enzyme was quite variable among the tested species.     

Detection of a variable intracellular acid‐labile carbon pool in Thalassiosira weissflogii (Heterokontophyta) and Emiliania huxleyi (Haptophyta) in response to changes in the seawater carbon system

Accumulation of an intracellular pool of carbon (C_i pool) is one strategy by which marine algae overcome the low abundance of dissolved CO₂ (CO_2(aq)) in modern seawater. To identify the environmental conditions under which algae accumulate an acid‐labile C_i pool, we applied a ¹⁴C pulse‐chase method, used originally in dinoflagellates, to two new classes of algae, coccolithophorids and diatoms. This method measures the carbon accumulation inside the cells without altering the medium carbon chemistry or culture cell density. We found that the diatom Thalassiosira weissflogii [(Grunow) G. Fryxell & Hasle] and a calcifying strain of the coccolithophorid Emiliania huxleyi [(Lohmann) W. W. Hay & H. P. Mohler] develop significant acid‐labile C_i pools. Ci pools are measureable in cells cultured in media with 2–30 µmol l^?1 CO_2(aq), corresponding to a medium pH of 8.6–7.9. The absolute C_i pool was greater for the larger celled diatoms. For both algal classes, the C_i pool became a negligible contributor to photosynthesis once CO_2(aq) exceeded 30 µmol l^?1. Combining the ¹⁴C pulse‐chase method and ¹⁴C disequilibrium method enabled us to assess whether E. huxleyi and T. weissflogii exhibited thresholds for foregoing accumulation of DIC or reduced the reliance on bicarbonate uptake with increasing CO_2(aq). We showed that the C_i pool decreases with higher CO₂:HCO₃^? uptake rates.     

Acclimation of photosynthesis to supersaturated CO2 in aquatic plant bicarbonate users

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Historical perspective on microalgal and cyanobacterial acclimation to low- and extremely high-CO2 conditions

Reports in the 1970s from several laboratories revealed that the affinity of photosynthetic machinery for dissolved inorganic carbon (DIC) was greatly increased when unicellular green microalgae were transferred from high to low-CO₂ conditions. This increase was due to the induction of carbonic anhydrase (CA) and the active transport of CO₂ and/or HCO₃ ⁻ which increased the internal DIC concentration. The feature is referred to as the `CO₂-concentrating mechanism (CCM)'. It was revealed that CA facilitates the supply of DIC from outside to inside the algal cells. It was also found that the active species of DIC absorbed by the algal cells and chloroplasts were CO₂ and/or HCO₃ ⁻, depending on the species. In the 1990s, gene technology started to throw light on the molecular aspects of CCM and identified the genes involved. The identification of the active HCO₃ ⁻ transporter, of the molecules functioning for the energization of cyanobacteria and of CAs with different cellular localizations in eukaryotes are examples of such successes. The first X-ray structural analysis of CA in a photosynthetic organism was carried out with a red alga. The results showed that the red alga possessed a homodimeric β-type of CA composed of two internally repeating structures. An increase in the CO₂ concentration to several percent results in the loss of CCM and any further increase is often disadvantageous to cellular growth. It has recently been found that some microalgae and cyanobacteria can grow rapidly even under CO₂ concentrations higher than 40%. Studies on the mechanism underlying the resistance to extremely high CO₂ concentrations have indicated that only algae that can adopt the state transition in favor of PS I could adapt to and survive under such conditions. It was concluded that extra ATP produced by enhanced PS I cyclic electron flow is used as an energy source of H⁺-transport in extremely high-CO₂ conditions. This same state transition has also been observed when high-CO₂ cells were transferred to low CO₂ conditions, indicating that ATP produced by cyclic electron transfer was necessary to accumulate DIC in low-CO₂ conditions. This revised version was published online in August 2006 with corrections to the Cover Date.     

Coexistence,size overlap and population regulation in tidal vs. non-tidal Palaemon prawns

Summary The effect of increased habitat heterogeneity in tidal areas on coexistence between Palaemon prawns was studied at eight sites along the European Atlantic coast. Two species which are sympatric in non-tidal areas, Palaemon adspersus Rathke and P. squilla (L.) are largely allopatric in tidal areas, and the sympatric size difference decreases in allopatry. In tidal areas the smaller species, P. squilla, is restricted to brown algal belts and rockpools. A third species, P. serratus (Pennant), larger than the others, occurs under oceanic salinities in subtidal brown algal belts and there has forced P. squilla to restrict its habitat distribution to adjacent intertidal rockpools. At estuarine salinities, however, P. squilla also inhabits the brown algal belts. A larger diel variation in stomach fullness index in P. squilla than in P. adspersus persists in non-tidal areas.Abiotic factors probably restrict the two largest species, viz. P. adspersus and P. serratus, to subtidal environments; these species are sensitive to the extremes in salinity, temperature or O₂ levels characteristic of the intertidal zone. Competition and/or predation probably relegates P. squilla to the intertidal zone. In non-tidal areas, where this zone is reduced, P. squilla increases its niche width and coexists with P. adspersus, and the size differentiation associated with sympatry may reduce interspecific competition.     

GROWTH RATE DEPENDENT OPTIMUM RATIOS IN SELENASTRUM MINUTUM (CHLOROPHYTA): IMPLICATIONS FOR COMPETITION,COEXISTENCE AND STABILITY IN PHYTOPLANKTON COMMUNITIES 12

Steady-State growth equations predict that the optimum C:P ratio (R) of Selenastrum minutum (Naeq.) Collins should change by a factor of 20 over the growth range of this organism. Chemostat cultures were established at fixed inflow C:P ratios chosen such that a transition from carbon to phosphorus limitation should occur solely as a result of increasing the steady-state growth rate. Measurements of residual dissolved inorganic carbon (DIC), cellular C:P, the kinetics of photosynthesis with respect to [DIC] and the response of culture biomass lo DIC or K₂HPO₄ additions were obtained. These results show that optimum ratios are growth rate dependent and that this dependency can be predicted based on steady-stale algal growth equations. A theoretical analysis was undertaken evaluating the range of growth rate dependent changes in the optimum ratio which could be expected for different nutrient pairs. Further analysis showed that, under certain conditions, the growth rate dependence of the optimum ratio may alter the breadth of zones of stable coexistence between species and allow for either the formation or complete elimination of such zones.     

潮汐作用和干湿交替对盐沼湿地碳交换的影响机制研究进展

潮汐盐沼湿地具有高的碳积累速率和低的CH_4排放量,是地球上最密集的碳汇之一。同时,气候变暖和海平面上升可能使得盐沼湿地更迅速的捕获和埋藏大气中的CO_2,因此盐沼湿地的"蓝碳"在减缓气候变化方面扮演着重要角色。潮汐盐沼湿地与其他湿地类型最大的区别和最显著的特征是在周期性潮汐作用下出现淹没和暴露,同时伴随盐分表聚与淋洗的干湿交替,可能是控制盐沼湿地碳交换过程和碳收支平衡的关键因素。但是,当前潮汐水动力过程及其周期性干湿交替对盐沼湿地碳交换关键过程和碳汇形成机制的影响尚不十分清楚。另外,以往相关研究通常孤立地考虑垂直方向上CO_2或CH_4交换或横向方向上的可溶性有机碳(DOC)、可溶性无机碳(DIC)、颗粒有机碳(POC)交换通量对盐沼湿地碳平衡进行评估,显然不够准确。因此,为了精确评估和预测盐沼湿地蓝碳的吸存能力,必须系统研究潮汐不同阶段对盐沼湿地碳交换过程的影响;深入分析潮汐作用下盐沼湿地碳交换的微生物机制;关注潮汐水动力作用对盐沼湿地DOC、DIC和POC产生、释放以及向邻近水体输出的影响;阐明潮汐作用对盐沼湿地碳汇形成机制的影响;纳入潮汐水动力过程作为变量,建立盐沼湿地碳循环模型。     

Effects of herbivory and the season of disturbance on algal succession in a tropical intertidal shore,Phuket, Thailand

The effects of herbivory and the season of disturbance on species composition and algal succession were experimentally tested at a tropical intertidal shore, Phuket Island, Thailand. Dead coral patches were cleared of all organisms during both the dry and rainy seasons in order to study the effects of season on algal succession and cages were set up to exclude fish herbivory. Algal succession in this intertidal habitat showed a simple pattern and took a year from the early Ulva paradoxa C. Agardh stage to the late Polysiphonia sphaerocarpa Børgesen stage. The abundance of algae during succession was under the influence of seasonal change. U. paradoxa reproduced and recruited throughout the year. Caging effects did not apparently influence algal abundance, perhaps because resident herbivorous damselfishes excluded other herbivores from their territories and maintained their algal “farms”. Unexpectedly, the percent cover of Ulva in the caged plots was lower than in uncaged plots. This pattern may indicate that caging excluded damselfishes only, but allowed small herbivores that consumed substantial amounts of soft filamentous algae in the cages.     

Photosynthetic performance of freshwater Rhodophyta in response to temperature, irradiance, pH and diurnal rhythm

Responses of net photosynthetic rates to temperature, irradiance, pH/inorganic carbon and diurnal rhythm were analyzed in 15 populations of eight freshwater red algal species in culture and natural conditions. Photosynthetic rates were determined by oxygen concentration using the light and dark bottles technique. Parameters derived from the photosynthesis–irradiance curves indicated adaptation to low irradiance for all freshwater red algae tested, confirming that they tend to occur under low light regimes. Some degree of photo‐inhibition (β= ‐0.33–0.01 mg O₂ g^?1 DW h^?1 (μmol photons m^?2 s^?1)^?1) was found for all species/populations analyzed, whereas light compensation points (Ic) were very low (≤ 2 μmol photons m‐ photons s^?1) for most algae tested. Saturation points were low for all algae tested (Ik = 6–54 μmol photons m^?2 s^?1; Is = 20–170 umol photons m^?2 s^?1). Rates of net photosynthesis and dark respiration responded to the variation in temperature. Optimum temperature values for net photosynthesis were variable among species and populations so that best performances were observed under distinct temperature conditions (10, 15, 20 or 25°C). Rates of dark respiration exhibited an increasing trend with temperature, with highest values under 20–25°C. Results from pH experiments showed best photosynthetic performances under pH 8.5 or 6.5 for all but one species, indicating higher affinity for inorganic carbon as bicarbonate or indistinct use of bicarbonate and free carbon dioxide. Diurnal changes in photosynthetic rates revealed a general pattern for all algae tested, which was characterized by two relatively clear peaks, with some variations around it: a first (higher) during the morning (07.00–11.00 hours.) and a second (lower) in the afternoon (14.00–18.00 hours). Comparative data between the ‘Chantransia’ stage and the respective gametophyte for one Batrachospermum population revealed higher values (ca 2‐times) in the latter, much lower than previously reported. The physiological role of the ‘Chantransia’ stage needs to be better analyzed.     

Influence of carbon supply on the circadian rhythmicity of photosynthesis and its stimulation by blue light in Ectocarpus siliculosus: clues to the mechanism of inorganic carbon acquisition in lower brown algae

Stimulation or light-saturated rates of photosynthesis in Ectocarpus siliculosus (Dillwyn) Lyngb. by blue light was eliminated by increasing dissolved inorganic carbon (DIC) or by lowering pH in natural seawater. The amplitude of the circadian rhythm of photosynthesis was also diminished under these conditions, and the pH compensation points in a closed system were higher in the presence of blue light and during the circadian day. These observations suggest that blue light and the circadian clock regulate the activity of a carbon acquisition system in these plants. The inhibitor of external carbonic anhydrase, acetazolamide, reduced overall rates of photosynthesis by only about 30%, but ethoxyzolamide suppressed the circadian rhythm of photosynthesis almost completely and markedly reduced the duration of responses to blue light pulses. Similar patterns were obtained when photosynthesis was measured in strongly limiting DIC concentrations (0–0.5 mol m^?3). Since blue light stimulated photosynthesis under these conditions of strong carbon limitation, we suggest that blue light activates the release of CO₂ from an internal CO₂ store. We propose a metabolic pathway with similarities to that of CAM plants. Non-photosynthetic fixation leads to the accumulation of a storage metabolite. The circadian clock and blue light control the mobilization of CO₂ at the site of decarboxylation of this metabolite. In the presence of continuous blue light the pathway is proposed to cycle and act as a pump for CO₂ into the chloroplasts. This hypothesis helps to explain a number of previously reported peculiarities of brown algal photosynthesis.     

ENHANCED TRANSPORT OF INORGANIC CARBON INTO ALGAL CELLS AND ITS IMPLICATIONS FOR THE BIOLOGICAL FIXATION OF CARBON1,2

Kinetics of uptake of inorganic carbon by the freshwater green alga Chlamydomonas reinhardtii Dang. suggest that rates of fixation may be enhanced at low tensions of CO₂ by transport of bicarbonate from the cell surface to the chloroplast. Results are evaluated in the context of models that treat diffusion and reaction of dissolved inorganic carbon across a 3 dimensional finite boundary layer, and they are consistent with the claim that CO₂ alone is the substrate used during carbon fixation. An alternative hypothesis, which presumes that both CO₂ and bicarbonate are used as substrates, yields predictions which are inconsistent with the data. Instead, bicarbonate seems to act only as a vehicle for the transport of inorganic carbon into the cell, thereby adding its flux to that of CO₂, and enhancing rates of synthesis that would otherwise be restricted by uptake of CO₂ alone.