Photosynthetic inorganic carbon uptake and accumulation in two marine diatoms

Some physiological characteristics of photosynthetic inorganic carbon uptake have been examined in the marine diatoms Phaeodactylum tricornutum and Cyclotella sp. Both species demonstrated a high affinity for inorganic carbon in photosynthesis at pH7.5, having K_1/2(CO₂) in the range 1.0 to 4.0mmol m^?3 and O_2? and temperature-insensitive CO₂ compensation concentrations in the range 10.8 to 17.6 cm³ m^?3. Intracellular accumulation of inorganic carbon was found to occur in the light; at an external pH of 7.5 the concentration in P. tricornutum was twice, and that in Cyclotella 3.5 times, the concentration in the suspending medium. Carbonic anhydrase (CA) was detected in intact Cyclotella cells but not in P. tricornutum, although internal CA was detected in both species. The rates of photosynthesis at pH 8.0 of P. tricornutum cells and Cyclotella cells treated with 0.1 mol m^?3 acetazolamide, a CA inhibitor, were 1.5- to 5-fold the rate of CO₂ supply, indicating that both species have the capacity to take up HCO₃^? as a source of substrate for photosynthesis. No Na⁺ dependence for HCO₃^? could be detected in either species. These results indicate that these two marine diatoms have the capacity to accumulate inorganic carbon in the light as a consequence, in part, of the active uptake of bicarbonate.     

PHYSIOLOGICAL CHARACTERISTICS OF PHOTOSYNTHESIS IN PORPHYRIDIUM CRUENTUM: EVIDENCE FOR BICARBONATE TRANSPORT IN A UNICELLULAR RED ALGA1

Various physiological characteristics of photosynthesis in the unicellular red alga Porphyridium cruentum Naegeli have been investigated. The rate of photosynthesis was optimal at 25° C and pH 7.5 and was not inhibited by 21% oxygen over a temperature range of 5 to 35° C. Kinetics of whole cell photosynthesis as a function of substrate concentration gave a K_1/2, (CO₂) of 0.3 μM. CO₂ compensation point, measured in a closed system at pH 7.5, was a constant 6.7 m?L · L^?1 over the temperature range 15 to 30° C and was unaffected by O₂ concentration. Whole cell photosynthesis, measured in a closed system at alkaline pH, showed that the rates of oxygen evolution were greatly in excess of the rate of CO₂ supply from the spontaneous dehydration of HCO₃^? in the medium. This indicates that bicarbonate is utilized by the cell to support this photosynthetic rate. These physiological characteristics of Porphyridium cruentum are consistent with the hypothesis that this alga transports bicarbonate across the plasmalemma.     

The effect of temperature and oxygen on the CO2 compensation point of the marine alga Ulva lactuca

Abstract The CO₂ compensation point of Ulva lactuca frond sections has been measured in artificial seawater using a sensitive gas-chromatographic method. Under nitrogen the compensation point remained relatively constant at 3–6 cm³ m⁻³ at temperatures from 10 to 30°C while in air-saturated medium (0.3 kg m⁻³ O₂) the compensation point rose from 5 cm³ m⁻³ at 10°C to 11 cm³ m⁻³ at 30°C. These responses of the compensation point to temperature and oxygen concentration indicate that there is little photorespiratory CO₂ loss in this marine macroalga, and the low values of these compensation points indicate that inorganic carbon is actively accumulated by the plant.     

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.     

Measurement of the photorespiratory activity of the submerged aquatic plant Myriophyllum spicatum L.

Abstract The CO₂ compensation concentrations (points) of leaves of the submerged vascular aquatic plant Myriophyllum spicatum L. were determined in a closed aqueous system at pH 7.0 by a gas chromatographic technique and over the range 10–30deg;C were found to range from 36 to 46 cm³m^?3 in medium equilibrated with 21% O₂ (0.03 kgm^?3), and 25 to 35 cm³m^?3 in medium equilibrated with 2% O₂ (0.03 kgm^?3). The rates of true (TPS) and apparent (APS) photosynthesis of leaves were measured in medium equilibrated with 21% O₂ and buffered at pH 7.0, at subsaturating concentrations (12.8–18.8 mmol m^?3) of dissolved inorganic carbor. (DIC) containing H¹⁴CO₃, by determining the initial rates of uptake by the leaves of DIC and ¹⁴C-activity from the medium. The rate of photorespiration, the difference between TPS and APS, was 7.0–13.3% of TPS over the range of 10–25°C and rose to 29% of TPS at 35°C. The magnitude of the compensation point of this plant is therefore similar to, but is much less O₂-sensitive than, those of C₃ plants, and the photorespiratory rate, at DIC concentrations near the CO₂ compensation point, is very low compared to that of C₃ plants.     

Rising CO2 levels and their potential significance for carbon flow in photosynthetic cells

Abstract. In the first part of this review, I discuss how we can predict the direct short-term effect of enhanced CO₂ on photosynthetic rate in C₃ terrestrial plants. To do this, I consider: (1) to what extent enhanced CO₂ will stimulate or relieve demand on partial processes like carboxylation, light harvesting and electron transport, the Calvin cycle, and end-product synthesis; and (2) the extent to which these various processes actually control the rate of photosynthesis. I conclude that control is usually shared between Rubisco (which responds sensitively to CO₂) and other components (which respond less sensitively), and that photosynthesis will be stimulated by 25–75% when the CO₂ concentration is doubled from 35 to 70 Pa. This is in good agreement with the published responses. In the next part of the review, I discuss the evidence that most plants undergo a gradual inhibition of photosynthesis during acclimation to enhanced CO₂. I argue that this is related to an inadequate demand for carbohydrate in the remainder of the plant. Differences in the long-term response to CO₂ may be explained by differences in the sink-source status of plants, depending upon the species, the developmental stage, and the developmental conditions. In the third part of the review, I consider the biochemical mechanisms which are involved in ‘sink’ regulation of photosynthesis. Accumulating carbohydrate could lead to a direct inhibition of photosynthesis, involving mechanical damage by large starch grains or Pi-limitation due to inhibition of sucrose synthesis. I argue that Pi is important in the short-term regulation of partitioning to sucrose and starch, but that its contribution to ‘sink’ regulation has not yet been conclusively demonstrated. Indirect or ‘adaptive’ regulation of photosynthesis is probably more important, involving decreases in amounts of key photosynthetic enzymes, including Rubisco. This decreases the rate of photosynthesis, and potentially would allow resources (e.g. amino acids) to be remobilized from the leaves and reinvested in sink growth to readjust the sink-source balance. In the final part of the review, I argue that similar changes of Rubisco and, possibly, other proteins are probably also involved during acclimation to high CO₂.     

Estimation of the whole-plant CO2 compensation point of tobacco (Nicotiana tabacum L.)

The whole-plant CO₂ compensation point (Γ_plant) is the minimum atmospheric CO₂ level required for sustained growth. The minimum CO₂ requirement for growth is critical to understanding biosphere feedbacks on the carbon cycle during low CO₂ episodes; however, actual values of Γ_plant remain difficult to calculate. Here, we have estimated Γ_plant in tobacco by measuring the relative leaf expansion rate at several low levels of atmospheric CO₂, and then extrapolating the leaf growth vs. CO₂ response to estimate CO₂ levels where no growth occurs. Plants were grown under three temperature treatments, 19/15, 25/20 and 30/25°C day/night, and at CO₂ levels of 100, 150, 190 and 270 μmol CO₂ mol⁻¹ air. Biomass declined with growth CO₂ such that Γ_plant was estimated to be approximately 65 μmol mol⁻¹ for plants grown at 19/15 and 30/25°C. In the first 19 days after germination, plants grown at 100 μmol mol⁻¹ had low growth rates, such that most remained as tiny seedlings (canopy size <1 cm²). Most seedlings grown at 150 μmol mol⁻¹ and 30/25°C also failed to grow beyond the small seedling size by day 19. Plants in all other treatments grew beyond the small seedling size within 3 weeks of planting. Given sufficient time (16 weeks after planting) plants at 100 μmol mol⁻¹ eventually reached a robust size and produced an abundance of viable seed. Photosynthetic acclimation did not increase Rubisco content at low CO₂. Instead, Rubisco levels were unchanged except at the 100 and 150 μmol mol⁻¹ where they declined. Chlorophyll content and leaf weight per area declined in the same proportion as Rubisco, indicating that leaves became less expensive to produce. From these results, we conclude that the effects of very low CO₂ are most severe during seedling establishment, in large part because CO₂ deficiency slows the emergence and expansion of new leaves. Once sufficient leaf area is produced, plants enter the exponential growth phase and acquire sufficient carbon to complete their life cycle, even under warm conditions (30/25°C) and CO₂ levels as low as 100 μmol mol⁻¹.     

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.     

Effects of solar radiation on the endemic Mediterranean red alga Rissoella verruculosa: photosynthetic performance, pigment content and the activities of enzymes related to nutrient uptake

The effects of full-spectrum solar radiation and of solar radiation deprived of total u.v. radiation (λ<395 nm) on the endemic Mediterranean red alga Rissoella verruculosa (Bertoloni) J. Agardh were studied in situ in early summer, from sunrise to sunset. Photosynthetic performance, pigment content and the activities of enzymes related to nutrient uptake, were monitored under both radiation conditions throughout a daylight period.
The doses of solar radiation, measured on the day during which the experiments were carried out (24 June 1997), were 9228·25, 1109·70 and 13·03 kJ m⁻² for PAR (λ=400–700 nm), u.v.-A (λ=315–400 nm) and u.v.-B (λ=280–315 nm), respectively. Under these conditions, a clear daily variation in photosynthetic performance was found. However, no significant differences were detected between the two radiation conditions. At noon, strong sunlight impaired O₂ evolution by 75%, but complete recovery occurred during the afternoon, reaching similar values to those measured in the early morning. By contrast, photoinhibition of optimal quantum yield ( F _v/ F _m), and the relative electron transport rate was followed by only a slight recovery during the afternoon. The rate of photosynthesis in air (simulating emersion conditions), estimated by CO₂ exchange, showed a negative balance at noon, which was accentuated in plants exposed to the full-solar spectrum.
Significant changes in the activities of nitrate reductase and carbonic anhydrase were found throughout the day under both radiation conditions. In thalli receiving solar radiation deprived of u.v., total CA activity decreased throughout the day. However, in thalli exposed to full-spectrum solar radiation, the activity of the enzyme tended to increase in the afternoon, correlating with an increase in NR activity.     

Generation of 10,154 expressed sequence tags from a leafy gametophyte of a marine red alga, Porphyra yezoensis.

A total of 10,154 5'-end expressed sequence tags (EST) were established from the normalized and size-selected cDNA libraries of a marine red alga, Porphyra yezoensis. Among the ESTs, 2140 were unique species, and the remaining 8014 were grouped into 1127 species. Database search of the 3267 non-redundant ESTs by BLAST algorithm showed that the sequences of 1080 species (33.1%) have similarity to those of registered genes from various organisms including higher plants, mammals, yeasts, and cyanobacteria, while 2187 (66.9%) are novel. Codon usage analysis in the coding regions of 101 non-redundant EST groups showing significant similarity to known genes indicated the higher GC contents at the third position of codons (79.4%) than the first (62.2%) and the second position (45.0%), suggesting that the genome has been exposed to high GC pressure during evolution. The sequence data of individual ESTs are available at the web site http://www.kazusa.or.jp/en/plant/porphyra/EST/.     

Photosynthetic acclimation of maize to growth under elevated levels of carbon dioxide

The effects of elevated CO₂ concentrations on the photochemistry, biochemistry and physiology of C₄ photosynthesis were studied in maize (Zea mays L.). Plants were grown at ambient (350 μL L⁻¹) or ca. 3 times ambient (1100 μL L⁻¹) CO₂ levels under high light conditions in a greenhouse for 30 d. Relative to plants grown at ambient CO₂ levels, plants grown under elevated CO₂ accumulated ca. 20% more biomass and 23% more leaf area. When measured at the CO₂ concentration of growth, mature leaves of high-CO₂-grown plants had higher light-saturated rates of photosynthesis (ca. 15%), lower stomatal conductance (71%), higher water-use efficiency (225%) and higher dark respiration rates (100%). High-CO₂-grown plants had lower carboxylation efficiencies (23%), measured under limiting CO₂, and lower leaf protein contents (22%). Activities of a number of C₃ and C₄ cycle enzymes decreased on a leaf-area basis in the high-CO₂-grown plants by 5–30%, with NADP-malate dehydrogenase exhibiting the greatest decrease. In contrast, activities of fructose 1,6-bisphosphatase and ADP-glucose pyrophosphorylase increased significantly under elevated CO₂ condition (8% and 36%, respectively). These data show that the C₄ plant maize may benefit from elevated CO₂ through acclimation in the capacities of certain photosynthetic enzymes. The increased capacity to synthesize sucrose and starch, and to utilize these end-products of photosynthesis to produce extra energy by respiration, may contribute to the enhanced growth of maize under elevated CO₂. Received: 30 April 1999 / Accepted: 17 June 1999     

Characteristics of photosynthetic carbon metabolism of spikelets in rice

In lemmas and paleae of rice, the amount of pyruvate, Pi dikinase (PPDK) protein increased dramatically 6 d after anthesis and this change was consistent with that in the activity of PPDK. Since lemmas and paleae at this stage also showed high activities of the other marker enzymes of C4 pathway including phosphot enolpyruvate carboxylase (Imaizumi et al. (1990) Plant Cell Physiol 31: 835–843), photosynthetic carbon metabolism with lemmas at this stage were characterized. In a 14C pulse-12C chase study by photosynthetic CO2 fixation, about 35% and 25% of 14C fixed in lemmas were incorporated initially into 3-phosphoglycerate (3-PGA) and C4 acids, respectively. This suggests that lemmas participate mainly in C3-type photosynthetic metabolism, but that lemmas may also participate in the metabolism of C4 acids to some extent. To clarify this possibility, large amounts of 14C-labeled C4 acids were synthesized in vivo by a light-enhanced dark CO2 fixation (LED) method and the fate of 14C in C4 acids in the light was investigated. The percentage distribution of 14C in C-4 position of malate was about 90% and 83% after 10 s of photosynthetic 14CO2 fixation and 110 s of LED, respectively. Some of the 14C incorporated into C4 acids was transferred into 3-PGA and sugar phosphates. The possibility of direct fixation of CO2 by phosphot enolpyruvate carboxylase and metabolic pathway of CO2 released by decarboxylation of malate produced were discussed.     

The freshwater red alga Batrachospermum turfosum (Florideophyceae) can acclimate to a wide range of light and temperature conditions

Batrachospermum turfosum Bory is one of the generalists among the few red algae that have adapted to freshwater habitats, occurring in a variety of primarily shaded, nutrient-poor micro-habitats with lotic (running) or lentic (standing) waters. Seasonal variations in water level and canopy cover can expose this sessile alga to widely fluctuating temperatures, solar irradiation and nutrient availability. Here we report on the ecophysiology of B. turfosum collected from an ultra-oligotrophic bog pool in the Austrian Alps. Photosynthesis as a function of photon fluence density (PFD) and temperature was studied by measuring oxygen evolution in combination with chlorophyll fluorescence. In addition, the effects of ultraviolet radiation (UVR) on photosynthetic pigments were analysed using HPLC and spectrophotometric methods, and cellular ultrastructure was studied using transmission electron microscopy. We found that B. turfosum is adapted to low light, with a light compensation point (I_c) and a light saturation point (I_k) of 8.4 and 29.7 µmol photons m^–2 s^–1, respectively, but also tolerates higher PFDs of ~1000 µmol photons m^–2 s^–1, and is capable of net photosynthesis at temperatures between 5°C and 35°C. Exposure to either UV-A or UV-AB for 102 h led to a strong transient drop in effective quantum yield (ΔF/F_M’), followed by an acclimation to about 70% of initial ΔF/F_M’ values. Ultrastructural changes included the accumulation of plastoglobules and dilated membranes after UVR treatment. Although all photosynthetic pigments strongly decreased upon UVR exposure and no UV-photoprotectants (e.g. mycosporine-like amino acids) could be detected, the alga was capable of recovering ΔF/F_M’ and phycobiliproteins after UVR treatment. In summary, B. turfosum tolerates a wide range of irradiation and temperature regimes, and these traits may be the basis for its successful adaptation to challenging environments.     

Carbonic anhydrase activity and inorganic carbon fluxes in low- and high-C1 cells of Chlamydomonas reinhardtü and Scenedesmus obliquus

Carbonic anhydrase (CA) activity associated with high- and low-dissolved inorganic carbon (C₁) grown cells was examined in whole cells by measuring ¹⁸O exchange from doubly labeled CO₂ (¹³C¹⁸O¹⁸O). Both algal species showed the presence of extracellular (periplasmic) as well as intracellular CA activity, which were both greatly increased in low-C₁ cells. The periplasmic CA activity was at least 40-fold higher in lowcompared to high-C₁ cells in both C. reinhardtii and S. obliquus. while low-C₁ cells of S. obliquus showed the highest activity of internal CA. The CA inhibitor ethoxyzolamide showed a strong inhibition of the C₁ uptake process in both C. reinhardtii and S. obliquus as in cyanobacteria. which may indicate that the nature of the primary uptake process is similar in both green algae and cyanobacteria. By using a mass spectrometnc disequilibrium technique it was possible to separate the C₁ fluxes of net HCO^?₃-uptake and net CO₂-uptake during steady-state photosynthesis in high- and Sow-C₁ grown cells of Chlamydomonas reinhardtii (WT. 2137+) and Scenedesmus obliquus (WT. D3). It was found that both high- and low-C₁ cells of the two algae can utilize both CO₂ and HCO^?₃ for photosynthesis, although low-C₁ cells have a higher affinity for the uptake of both C₁ species. Induction at low-C₁ causes an increase in the affinity of both species for HCO^?₃ and CO₂; changes in net CO₂-uptake were, however, significantly greater.     

The regulation of photosynthetic rate and activation of extracellular carbonic anhydrase under CO2-limiting conditions in the marine diatom Skeletonema costatum

In the marine diatom Skeletonema costatum , carbonic anhydrase activity exterior to the plasma membrane (CA_ext) was detected only when the available CO₂ concentration was less than 5·0 mmol m^–3, this activity being unaffected by the total dissolved inorganic carbon concentration. The inhibition of CA_ext by dextran bound sulphonamide (DBS) demonstrated the key role of this enzyme in maintaining photosynthetic rate under CO₂-limited conditions. Treatment with trypsin followed by affinity chromatography on p-aminomethylbenzene-sulphamide agarose and subsequent SDS-PAGE analysis revealed a polypeptide from carbon-replete cells of identical molecular mass to the CA_ext released by trypsin from CO₂-limited cells. Redox activity in the plasma membrane of intact cells was measured by following the light-dependent reduction of ferricyanide or NADP, the greatest activity being shown by CO₂-limited cells. Overall the results suggest that high rates of redox activity under conditions of CO₂-limitation were required for the activation of CA_ext.     

Bicarbonate uptake in the marine macroalga Ulva sp. is inhibited by classical probes of anion exchange by red blood cells

We demonstrate in this work that HCO _inf3 ^sup– uptake in the marine macroalga Ulva sp. features functional resemblances to anion transport mediated by anion exchangers of mammalian cell membranes. The evidence is based on (i) competitive inhibition of photosynthesis by the classical red-blood-cell anion-exchange blockers 4,4-dinitrostilbene-2,2-disulfonate and 4-nitro-4-isothiocyanostilbene-2,2-disulfonate under conditions where HCO _inf3 ^sup– , but not CO₂, was the inorganic carbon form taken up; (ii) inhibition of HCO _inf3 ^– uptake by pyridoxal phospate, indicating the involvement of lysine residues in the binding/translocation of HCO _inf3 ^sup– ; and (iii) inhibition of HCO _inf3 ^sup– (but not of CO₂) uptake by exofacial trypsin treatments, indicating the functional involvement of a plasmalemma protein. It is suggested that HCO _inf3 ^sup– uptake mediated by such a putative anion transporter can be a fundamental step in providing inorganic carbon for the CO₂-concentrating system of marine marcoalgae in an environment where the HCO _inf3 ^sup– concentration is high, but the CO₂ concentration and rates of uncatalyzed HCO _inf3 ^sup– dehydration are low.Abbreviations CI ionorganic carbon - DIDS 4,4-diisothiocyanostilbene-2,2-disulfonate - DNDS 4,4-dinitrostilbene-2,2-disulfonate - NIDS 4-nitro-4-isothiocyanostilbene-2,2-disulfonate - PLP pyridoxal phosphate - Rubisco ribulose-1,5-bisphosphate carboxylase-oxygenase     

The interactive effects of light and inorganic carbon on aquatic plant growth

Submerged aquatic macrophytes grow across a wide, often coupled, range of light and inorganic carbon availabilities, and each single factor influences photosynthesis and acclimation. Here we examine the interactive effects of light and inorganic carbon on the growth of Elodea canadensis and Callitriche cophocarpa. The plants were grown in the laboratory at a range of light intensities (0–108 μmol m⁻²s⁻¹) and four inorganic carbon regimes in a crossed factorial design. Plant growth rates, measured over 3–4 weeks of incubation, increased in response to increasing light intensity and inorganic carbon availability, and significant interactive effects were observed. The light-use efficiency for growth at low light increased 2-fold for Callitriche and 6-fold for Elodea between the lowest and highest inorganic carbon concentrations applied. Also, the growth rate at the highest light intensity increased with inorganic carbon availability, but the relative increase was smaller than at low light. Both species acclimated to the light and carbon regime such that the chlorophyll content declined at low and high light intensities and the initial slopes of the photosynthetic CO₂ and HCO₃⁻ response curves declined at high levels of CO₂. Callitriche responded less markedly than Elodea to changing inorganic carbon availability during growth, and the initial slope of the photosynthetic HCO₃⁻ response curve, in particular, was greatly reduced (>90%) in Elodea by high CO₂. It is suggested that the coupled responses of aquatic macrophytes to light and inorganic carbon influence their ability to develop dense stands at high light in shallow water and to extend to greater depths in waters rich in inorganic carbon.