Strategies for carbon acquisition in the red marine macroalga Coccotylus truncatus from the Baltic Sea

Marine macroalgae need carbon-concentrating mechanisms because they have only limited access to CO₂ in their natural environment. Previous studies have shown that one important strategy common to many algae is the activity of periplasmic carbonic anhydrases that catalyse the dehydration of HCO₃^- into CO₂. The latter can then cross the plasma membrane by passive diffusion. We hypothesised that an active (energy-consuming) mechanism might also be involved in the membrane transport of CO₂, as is the case in a number of microalgae. Coccotylus truncatus was chosen as a model organism for this study because it belongs to a group of algae that usually lack direct HCO₃^- uptake: sublittoral red algae. The method used to study carbon uptake was pH drift of the seawater medium surrounding the algae in a closed vessel, with and without the addition of specific inhibitors or proton buffers. Measured parameters included pH, total inorganic carbon and alkalinity of the seawater medium. Our results suggest that, in C. truncatus, periplasmic carbonic anhydrase as well as H⁺ extrusion, probably driven by a vanadate-sensitive P-type H⁺-ATPase (proton pump), are involved in CO₂ uptake. No direct uptake of HCO₃^- was discovered. This paper also presents data on the buffer capacity of several proton buffers and the carbon-uptake inhibitors acetazolamide, 4,4'-diisothiocyano-stilbene-2,2'-disulfonate (DIDS) and orthovanadate in Baltic Sea water with a salinity of 6.5 psu.     

Carbon isotope effects during precipitation of barium carbonate: implications for environmental studies

Here we demonstrate that precipitation of dissolved inorganic carbon (DIC) to at pH 10.26, 11.38 and 12.11 does not cause measurable carbon isotope fractionation. However, NaOH solution prepared under standard laboratory procedure and used to increase pH value for precipitation of BaCO₃ can be contaminated by atmospheric CO₂. In our experiment, δ¹³C value in the contaminant DIC was −24.30‰, which resulted in decrease in the δ¹³C(DIC) value by 0.03, 0.07 and 0.22 at pH 10.26, 11.38 and 12.11, respectively.     

Growth rate and carbon affinity ofUlva lactuca under controlled levels of carbon,pH and oxygen

We examined the growth rate (µ) ofUlva lactuca L. (collected from Roskilde Fjord, Denmark in 1987) at different levels of dissolved inorganic carbon (DIC), pH and oxygen in two culture facilities. Growth was faster in Facility A (µ _max ca 0.3 d^–1) than in B (µ _max ca 0.2 d^–1), probably because of more efficient stirring and higher light intensity. The growth-DIC response curve exhibited low half-saturation constant (K _1/2) values (0.35 mM DIC in A, 0.55 mM in B) and growth rates close toµ _max at natural seawater concentration of 2 mM DIC. Growth rate showed a low sensitivity to oxygen over a wide range of DIC and oxygen concentrations. Collectively, the results demonstrated an efficient mechanism for DIC use, unaffected by acclimatization to DIC concentrations between 0.2 and 3 mM. The growth rate decreased little between pH 7.5 and 9 at 2 mM DIC, but steeply above pH 9 approaching zero just above pH 10. The decline of growth at high pH may result from direct pH effects on cell pH, reduced HCO ₃ ^- availability and impaired operation of the carbon uptake process. The growth responses ofU. lactuca to DIC, pH and oxygen resembled those observed in previous short-term photosynthetic experiments. This similarity is probably due to the fast growth ofU. lactuca which means that photosynthetic products are rapidly converted into cell growth. Based on the culture experiments we argue that field plants ofU. lactuca not exposed to stagnant water and DIC depletion are likely to be limited in growth by environmental factors other than DIC (e.g. light and nutrients). Dense mats ofU. lactuca, however, may show reduced growth as a result of DIC depletion, high pH and self-shading.     

Effects of O2, pH and DIC on photosynthetic net-O2 evolution by marine macroalgae

Net photosynthetic O₂ evolution by five marine macroalgae:Ulva lactuca L.,Enteromorpha sp.,Ceramium strictum Harvey,Fucus serratus L., andF. vesiculosus L., collected from Danish waters in the summer of 1983 was followed at increasing O₂ and with pH either fixed close to pH 7, 8 or 9, or drifting upwards during photosynthesis in a closed chamber to determine the effects of changing O₂, pH and DIC (dissolved inorganic carbon) on photosynthesis. Increasing O₂, increasing pH and decreasing DIC together limited O₂ evolution. Raising the O₂ concentration with pH and DIC held constant resulted in less inhibition of net-O₂ evolution than when all three factors acted together. The O₂ inhibition of photosynthesis was similar to the reported O₂ inhibition of ribulose 1,5-bisphosphate carboxylase isolated from lower and higher plants. Net-O₂ evolution as a function of the molar ratio of O₂ to HCO^– ₃ + CO₂ in solution provided a general, linear relationship (r ² = 0.72 to 0.84), predicting inhibition of photosynthesis based on O₂ pH and DIC changing together. Slopes of this relationship, representing competition between O₂ and carbon based on external concentrations, were similar for the five taxonomically different algae, suggesting that similar processes act to reduce net-O₂ evolution.     

Photosynthetic utilization of CO2(aq) and HCO 3 - in Thalassia testudinum (Hydrocharitaceae)

The effects of total dissolved inorganic carbon (DIC), free carbon dioxide [CO_2(aq)], and bicarbonate (HCO ₃ ^- ) concentrations on net photosynthetic oxygen evolution of the marine angiosperm Thalassia testudinum Banks ex König collected from Biscayne Bay (1988) and from Tampa Bay (1990), Florida, USA, were examined. Rates of photosynthesis declined by 85% from pH 7.25 to 8.75 in buffered seawater media with constant DIC concentration (2.20 mM), suggesting a strong influence of CO_2(aq) concentration. A plateau in the pH-response curve between pH 7.75 and 8.50 indicated possible utilization of HCO ₃ ^- . Responses of photosynthesis measured in buffered seawater media of varying DIC concentrations (0.75 to 13.17 mM) and pH (7.8 to 8.61) demonstrated that photosynthesis is rate-limited at ambient DIC levels. Photosynthesis increased in media with increasing HCO ₃ ^- concentrations but near-constant CO_2(aq) levels, confirming HCO ₃ ^- assimilation. Calculated half-saturation constants (K _s )for CO_2(aq) and HCO ₃ ^- indicated a high affinity for the former [K _s (CO₂)=3 to 18 M] and a much lower affinity for the latter [K _s (HCO ₃ ^- )=1.22 to 8.88 mM]. Calculated V _max values for HCO ₃ ^- were generally higher than those for CO_2(aq), suggesting relatively efficient HCO ₃ ^- utilization, despite the apparent low affinity for this carbon form.     

Diffusive boundary layers and photosynthesis of the epilithic algal community of coral reefs

The effects of mass transfer resistance due to the presence of a diffusive boundary layer on the photosynthesis of the epilithic algal community (EAC) of a coral reef were studied. Photosynthesis and respiration of the EAC of dead coral surfaces were investigated for samples from two locations: the Gulf of Aqaba, Eilat (Israel), and One Tree Reef on the Great Barrier Reef (Australia). Microsensors were used to measure O₂ and pH at the EAC surface and above. Oxygen profiles in the light and dark indicated a diffusive boundary layer (DBL) thickness of 180–590 μm under moderate flow (~0.08 m s^?1) and >2,000 μm under quasi-stagnant conditions. Under light saturation the oxygen concentration at the EAC surface rose within a few minutes to 200–550% air saturation levels under moderate flow and to 600–700% under quasi-stagnant conditions. High maximal rates of net photosynthesis of 8–25 mmol O₂ m^?2 h^?1 were calculated from measured O₂ concentration gradients, and dark respiration was 1.3–3.3 mmol O₂ m^?2 h^?1. From light–dark shifts, the maximal rates of gross photosynthesis at the EAC surface were calculated to be 16.5 nmol O₂ cm^?3 s^?1. Irradiance at the onset of saturation of photosynthesis, E _k, was <100 µmol photons m^?2 s^?1, indicating that the EAC is a shade-adapted community. The pH increased from 8.2 in the bulk seawater to 8.9 at the EAC surface, suggesting that very little carbon in the form of CO₂ occurs at the EAC surface. Thus the major source of dissolved inorganic carbon (DIC) must be in the form of HCO₃ ^?. Estimates of DIC fluxes across the DBL indicate that, throughout most of the daytime under in situ conditions, DIC is likely to be a major limiting factor for photosynthesis and therefore also for primary production and growth of the EAC.     

Diurnal changes in seawater carbonate chemistry speciation at increasing atmospheric carbon dioxide

Natural variability in seawater pH and associated carbonate chemistry parameters is in part driven by biological activities such as photosynthesis and respiration. The amplitude of these variations is expected to increase with increasing seawater carbon dioxide (CO₂) concentrations in the future, because of simultaneously decreasing buffer capacity. Here, we address this experimentally during a diurnal cycle in a mesocosm CO₂ perturbation study. We show that for about the same amount of dissolved inorganic carbon (DIC) utilized in net community production diel variability in proton (H⁺) and CO₂ concentrations was almost three times higher at CO₂ levels of about 675 ± 65 in comparison with levels of 310 ± 30 μatm. With a simple model, adequately simulating our measurements, we visualize carbonate chemistry variability expected for different oceanic regions with relatively low or high net community production. Since enhanced diurnal variability in CO₂ and proton concentration may require stronger cellular regulation in phytoplankton to maintain respective gradients, the ability to adjust may differ between communities adapted to low in comparison with high natural variability.     

Effects of host-tissue homogenate of the scleractinian coral Plesiastrea versipora on glycerol metabolism in isolated symbiotic dinoflagellates

Symbiotic dinoflagellate algae (Symbiodinium sp.) isolated from the scleractinian coral Plesiastrea versipora and incubated in homogenized host tissue released 4 to 7 times as much glycerol (14 to 46 nmol glycerol/10⁶ algae) as those incubated in seawater (3 to 6 nmol glycerol/10⁶ algae) after 4 h incubation in the light. During this period, no release of triglycerides was detected. Intracellular glycerol increased 2- to 3-fold in algae incubated in host homogenate, but remained unchanged in algae incubated in seawater at a concentration of 0.82 ± 0.47 nmol glycerol/10⁶ algae. In each incubation condition, intracellular triglyceride levels increased. However, in algae incubated in host homogenate, the intracellular levels of triglycerides reached only about 75% of the amount reached in algae incubated in seawater (max. 18.55 ± 2.40 nmol glycerol/10⁶ cells). Host homogenate did not stimulate the release of glycerol from algae during dark incubation. These data show that the glycerol released by algae incubated in host-tissue homogenate was derived from increased synthesis of glycerol or from diversion of some glycerol or other photosynthetic intermediates from incorporation into algal triglyceride stores, and did not come from existing stores. Received: 20 December 1996 / Accepted: 9 January 1997     

Changes in pH as a measure of photosynthesis by marine macroalgae

An automatically operated method for high precision measurements of steady-state photosynthesis by macroalgae was developed. Changes in pH and oxygen content of seawater passing the algae in a flowthrough system, could be measured with extremely high accuracy over very long periods of time. The method is especially suitable for measurements on flowthrough systems with high rates of water exchanges (i.e. short retention time), and can be used to study exchange processes for marine plants, animals and small ecosystems. Since the same measuring unit is used for several flowthrough chambers, the method is very suitable for comparisons between different species, or between differently pretreated specimens of the same species (e.g. in toxicological studies). The method was used to study the ratio: [oxygen production] to [CO₂+H⁺ uptake] at different light intensities for several macroalgae belonging to different systematic groups and from different habitats. At lower photosynthetic rates this ratio was similar for all of the algae studied (1.17±0.02). For brown algae of the fucacean family, the ratio increased by 0.08 units at higher photosynthetic rates. This increase was thought to be related to the crassulacean acid metabolism (CAM)-like strategies connected to these algae. For all other algae studied, the ratio remained constant or decreased slightly (at most by 0.04 units) at higher photosynthetic rates. The relations between the abovementioned ratio and the photosynthetic quotient are discussed on a theoretical basis.     

Carbon metabolism and strobilation in Cassiopea andromedea (Cnidaria: Scyphozoa): Significance of endosymbiotic dinoflagellates

Scyphopolyps and scyphomedusae of Cassiopea andromeda Forskål (Cnidaria, Scyphozoa) containing dinoflagellate endosymbionts (zooxanthellae) were investigated for rates and pathways of carbon fixation. Photosynthesis by the algae, accounting for 80 and 15 mol C h^-1 on a dry weight basis in medusae and polyps, respectively, by far exceeds dark incorporation of inorganic carbon by the intact association. Photosynthetic carbon fixation is operated via C₃ pathway of carbon reduction. DCMU-treatment (1×10^-6 M and 1×10^-5 M) completely inhibits light-dependent carbon assimilation. Major photosynthates presumably involved in a metabolite flow from algal symbionts to animal tissue are glycerol and glucose. A total of 5–10% net algal photosynthate appears to be seleased in vivo to the host. This is probably less than the energy supply ultimately required for the nutrition of the polyps and medusae. The presence of zooxanthellae proved to be indispensable for strobilation in the scyphopolyps. However, photosynthesis by algal symbionts as well as photosynthate release is obviously not essential for the initiation of ephyrae as is shown by DCMU-treatment, culture in continous darkness, and aposymbiotic controls. It is therefore concluded that strobilation is supported, but not triggered by algal photosynthetic activity. The induction of strobilation thus seems to depend on a more complex system of regulation.     

The mechanism of calcification and its relation to photosynthesis and respiration in the scleractinian coral <Emphasis Type="Italic"> Galaxea fascicularis</Emphasis>

The mechanism of calcification and its relation to photosynthesis and respiration were studied with Ca²⁺, pH and O₂ microsensors using the scleractinian coral Galaxea fascicularis. Gross photosynthesis (Pg), net photosynthesis (Pn) and dark respiration (DR) were measured on the surface of the coral. Light respiration (LR) was calculated from the difference between Pg and Pn. Pg was about seven times higher than Pn; thus, respiration consumes most of the O₂ produced by the algal symbiont's photosynthesis. The respiration rate in light was ca. 12 times higher than in the dark. The coupled Pg and LR caused an intense internal carbon and O₂ cycling. The resultant product of this cycle is metabolic energy (ATP). The measured ATP content was about 35% higher in light-incubated colonies than in dark-incubated ones. Direct measurements of Ca²⁺ and pH were made on the outer surface of the polyp, inside its coelenteron and under the calicoblastic layer. The effects on Ca²⁺ and pH dynamics of switching on and off the light were followed in these three compartments. Ca²⁺ concentrations decreased in light on the surface of the polyp and in the coelenteron. They increased when the light was switched off. The opposite effect was observed under the calicoblastic layer. In light, the level of Ca²⁺ was lower on the polyp surface than in the surrounding seawater, and even lower inside the coelenteron. The concentration of calcium under the calicoblastic layer was about 0.6 mM higher than in the surrounding seawater. Thus Ca²⁺ can diffuse from seawater to the coelenteron, but metabolic energy is needed for its transport across the calicoblastic layer to the skeleton. The pH under the calicoblastic layer was more alkaline compared with the polyp surface and inside the coelenteron. This rise in pH increased the supersaturation of aragonite from 3.2 in the dark to 25 in the light, and brought about more rapid precipitation of CaCO₃. When ruthenium red was added, Ca²⁺ and pH dynamics were inhibited under the calicoblastic layer. Ruthenium red is a specific inhibitor of Ca-ATPase. The results indicated that Ca-ATPase transports Ca²⁺ across the calicoblastic layer to the skeleton in exchange for H⁺. Addition of dichlorophenyldimethylurea completely inhibited photosynthesis. The calcium dynamics under the calicoblastic layer continued; however, the process was less regular. Initial rates were maintained. We conclude that light and not energy generation triggers calcium uptake; however, energy is also needed.     

δ<Superscript>13</Superscript>C and δ<Superscript>15</Superscript>N values in reef corals <Emphasis Type="Italic">Porites lutea</Emphasis> and <Emphasis Type="Italic">P. cylindrica</Emphasis> and in their epilithic and endolithic algae

In summer 1998, shallow water corals at Sesoko Island, Japan (26°38′N, 127°52′E) were damaged by bleaching. In August 2003, partially damaged colonies of the massive Porites lutea and the branching P. cylindrica were collected at depths of 1.0–2.5 m. The species composition of epilithic algal communities on dead skeletal surfaces of the colonies (‘red turfs’, ‘green turfs’, ‘red crusts’) and the endolithic algae (living in coral skeletons) growing close to and away from living coral polyps was determined. Carbon and nitrogen stable isotope values of organic matter (δ¹³C and δ¹⁵N) from all six of these biological entities were determined. There were no significant differences in the isotope composition of coral tissues of the two corals, with P. lutea having δ¹³C of −15.3 to −9.6‰ and δ¹⁵N of 4.7–6.1‰ and P. cylindrica having similar values. Polyps in both species living close to an interface with epilithic algae had similar isotope values to polyps distant from such an interface. Despite differences in the relative abundance of the algal species in red turfs and crusts, their δ¹³C and δ¹⁵N values were not significantly different from each other (−18.2 to −13.9, −20.6 to −16.2, 1.1–4.3, and 3.3 to 4.9‰, respectively). The green algal turf had significantly higher δ¹³C values (−14.9 to −9.3‰) than that of red turfs and crusts but similar δ¹⁵N (1.2–4.1‰) to the red algae. The data do not suggest that adjoining associations of epilithic algae and coral polyps exchange carbon- and nitrogen-containing metabolites to a significant extent. The endolithic algae in the coral skeletons had δ¹³C values of −14.8 to −12.3‰ and δ¹⁵N of 4.0–5.4‰. Thus they did not differ significantly from the coral polyps in their carbon and nitrogen isotope values. The similarity in carbon isotope values between the coral polyps and endolithic algae may be attributed to a common source of CO₂ for zooxanthellae and endolithic algae, namely, from respiration by the coral host. While it is difficult to fully interpret similarity in the nitrogen isotope composition of coral tissue and of green endolithic algae and the difference in δ¹⁵N between green epilithic and endolithic algae, the data are consistent with nitrogen-containing metabolites from the scleractinian coral serving as a significant source of nitrogen for the endolithic algae.     

Estimating marine biogeochemical rates of the carbonate pH system—A Kalman filter tested

Oxygen (O₂), nitrate (NO₃), dissolved inorganic carbon (DIC) or pCO₂, and pH or total alkalinity (TA), are useful indices of marine chemical, physical and biological processes operating on varying time-scales. Although these properties are increasingly being monitored at high frequency, they have not been extensively used for studying ecosystem dynamics. We test whether we can estimate time-evolving biogeochemical rates (e.g. primary production, respiration, calcification and carbonate dissolution, and nitrification) from synthetic high frequency time-series of O₂, NO₃, DIC, pCO₂, TA or pH. More specifically, a Kalman filter has been implemented in a very simplified biogeochemical model describing the dynamics of O₂, NO₃, DIC and TA and linking the concentration data to biogeochemical fluxes. Different sets of concentration data are assimilated and biogeochemical rates are estimated. The frequency of assimilation required to get acceptable results is investigated and is compared with the frequency of sampling in the field or in controlled experimental settings.Smoothing of the data to remove data noise before assimilation improves the estimation of the biogeochemical rates. The best estimated rates are obtained when assimilating O₂, NO₃ and TA although the assimilation of DIC instead of TA also gives satisfactory results. In case pH or pCO₂ is assimilated rather than DIC or TA, the linearization of the (now nonlinear) observation equation introduces perturbations and the Kalman filter behaves suboptimal. We conclude that, given the resolution of data required, the tool has potential to estimate biogeochemical rates of the carbonate system under controlled settings.     

Relative growth of different species of marine algae in wasterwater-seawater mixtures

In recent studies, we developed a combined nutrient removal-marine aquaculture process for the tertiary treatment of wastewater and the production of commercially important shellfish. Part of this process consists of an outdoor mass cultivation system for marine algae. During our previous experiments we observed that marine diatoms almost exclusively are the dominant algal species in our outdoor cultures. To better understand this phenomenon of diatom dominance we grew 16 species of marine algae in continuous monoculture under laboratory conditions simulating to some degree the conditions prevailing in our outdoor experiments. Species such as Skeletonema costatum, Monochrysis lutheri and Tetraselmis sp., which were never dominant in our outdoor cultures, grew as well in monoculture, as Phaeodactylum tricornutum, frequently, the prevalent species outdoors. However, when monocultures of Dunaliella tertiolecta and Thalassiosira pseudonana (3H) were purposely contaminated with P. tricornutum, the latter species quickly became dominant. It is suggested that a complex interaction of environmental factors is usually responsible for the dominance of a particular species; one such factor may be the nitrogen source in the growth media. Under conditions of virtually, complete nitrogen assimilation, the carbon: nitrogen ratio in the algae was high (7 to 8) when NO ₃ ^- –N was the source of nitrogen, and low (4 to 6) when NH ₄ ⁺ –N was the prime form of nitrogen. When algal growth was low, resulting in a large inorganic nitrogen residue, the carbon:nitrogen ratio was low regardless of whether NO ₃ ^- –N or NH ₄ ⁺ –N was the main nitrogen source.Contribution No. 3297 from the Woods Hole Oceanographic Institution.     

Zooxanthellae providing assimilatory power for the incorporation of exogenous acetate in Heteroxenia fuscescens (Cnidaria: Alcyonaria)

The soft coral Heteroxenia fuscescens (Ehrb.) and its isolated zooxanthellae (endosymbiotic dinoflagellates) were investigated with particular regard to uptake and utilization of exogenously supplied ¹⁴C-acetate in the light and in the dark. The incorporation of ¹⁴C from ¹⁴C-acetate into the host tissue and into the zooxanthellae was consistently much higher in the light than in the dark. The incorporated ¹⁴C-acetate was rapidly metabolized by the host and algae and was recovered from different assimilate fractions. The major proportion of radiocarbon from metabolized ¹⁴C-acetate was located in host tissue. The CHCl₃-soluble fraction composed of diverse lipids showed the strongest ¹⁴C-labelling. Zooxanthellae isolated prior to incubation accounted for about 80% of the acetate incorporation recorded for zooxanthellae in situ (in vivo). It is concluded from a comparison of acetate incorporation and conversion under light and dark conditions that most of the lipid reserve of the host tissue originates from fatty acids, which are synthesized within the algal symbionts and are then translocated to the heterotrophic partner via extrusion. The acetate units needed for lipid synthesis are obtained by absorption of free acetate from dissolved organic matter (DOM) in the seawater as well as by photosynthetic assimilation of inorganic carbon. Thus, in H. fuscescens, lipogenesis is operated as a light-driven process to which the zooxanthellae considerably contribute assimilatory power by performing fatty acid synthesis and translocation of lipid compounds to their intracellular environment (host cell). A metabolic scheme is proposed to account for the different pathways of carbon conversion observed in H. fuscescens. The incubations took place in August 1980 and the analytical part from October 1980 to January 1984.     

The size and function of the internal inorganic carbon pool of the foraminifer Amphistegina lobifera

The existence of an internal inorganic carbon pool in the perforate foraminifer Amphistegina lobifera, as suggested recently (ter Kuile and Erez 1987), has been established by direct measurements using a new ¹⁴C tracer method. The imperforate species Amphisorus hemprichii does not contain such a pool. The size of the pool in A. lobifera is proportional to its calcification rate and approximately equals the amount of carbon incorporated into the skeleton during 24 h. Time course experiments show that inorganic carbon (Ci) is photoassimilated at constant rates by the algal symbionts, that the pool is filled to maximum capacity in ca. 24 h, and that Ci incorporation into the skeleton starts only after the pool is filled up. During the chase phase of pulsechase experiments, all ¹⁴C initially residing in the pool is transferred to the skeleton, indicating that the pool serves for calcification and not for photosynthesis. Uptake of Ci into the pool occurs only in the light, indicating that energy may be required for this process. Furthermore, calculations of the Ci concentration inside the pool suggest that it is higher by 2 to 3 orders of magnitude compared to seawater concentration, suggesting that its accumulation is an energy dependent process.     

Interactive effects of nitrogen and dissolved inorganic carbon on photosynthesis,growth, and ammonium uptake of the macroalgae Cladophora vagabunda and Gracilaria tikvahiae

Dissolved inorganic carbon (DIC) is rarely considered limiting for macroalgae, but some research suggests that under conditions of N sufficiency, photosynthetic capacity is enhanced with DIC enrichment. During spring (April–May) and summer (July–August) 1993, we investigated the interactive effects of nitrogen (N) and DIC on photosynthetic capacity, growth, and nutrient uptake rates of the macroalgae, Cladophora vagabunda (L.) van den Hoek and Gracilaria tikvahiae (McLachlan), dominant species in a temperate eutrophic estuary (Cape Cod, Massachusetts, USA). Water-column CO₂ concentrations showed significant diurnal fluctuations, ranging from a morning CO₂ peak (21 M) to an afternoon low (13 M) during summer, probably associated with metabolic activities in a thick algal mat. Results from instantaneous photosynthesis measurements and microcosm experiments indicate that DIC limits photosynthetic capacity and growth rates of C. vagabunda during summer, perhaps related to tissue N sufficiency and low water-column CO₂ concentrations. For example, this species showed enhanced growth (F=8.69, P<0.02) under DIC but not N enrichment. G. tikvahiae showed marginal DIC enhancement of maximum photosynthetic rate, while growth was significantly stimulated by addition of N. Reduced thallus N of this species during the summer further identifies N as the primary factor limiting growth. In addition, G. tikvahiae has the ability to use DIC in its several forms, while C. vagabunda primarily uses dissolved CO₂. DIC enrichment resulted in a depression of NH₄ ⁺ uptake rates for both species, particularly during summer at saturating (60 M) ammonium levels, suggesting competition between NH₄ ⁺ uptake and DIC acquisition under conditions of N sufficiency. Dominance of C. vagabunda and G. tikvahiae in areas undergoing eutrophication has been attributed to their successful procurement and storage of N as well as to high growth rates. The present study revealed that under conditions of N sufficiency during summer, DIC may control rates of production of these opportunistic macroalgae.     

Host feeding and nutrient sufficiency for zooxanthellae in the sea anemone Aiptasia pallida

Nutrient sufficiency of zooxanthellae in the sea anemone Aiptasia pallida cultured in low nutrient seawater depends on the availability of particulate food to the host. Zooxanthellae in anemones unfed for 20 to 30 d exhibited the following characteristics of nutrient deficiency: cell division rates decreased; chlorophyll a content gradually decreased from 2 to <1 pg cell^–1; and C:N ratios increased from 7.5 to 16. Over a 3-mo period, algal populations in unfed anemones gradually decreased, indicating that zooxanthellae were lost faster than they were replaced by division. The mitotic index of zooxanthellae in unfed anemones was stimulated either by feeding the host or by the addition of inorganic N and P to the medium. Whether algae are nutrient-limited in hosts under field conditions has not been examined fully; however, C:N ratios in zooxanthellae from field-collected hosts are slightly higher (9.4 vs 7.5) than in hosts fed to repletion in laboratory cultures. This observation might indicate N limitation in the field.     

Mass transfer limitation of photosynthesis of coral reef algal turfs

Algal turfs are the major primary producing component on many coral reefs and this production supports higher levels in the complex reef trophic web. Rates of metabolism of algal turfs are related positively to water motion, consistent with limitation by the diffusion of a substance through a boundary layer. Based on engineering mass transfer theory, we hypothesized that photosynthesis of algal turfs is controlled by rates of mass transfer and responses of photosynthesis to increasing flow speed should be predicted by engineering correlations. This hypothesis was tested in ten experiments where photosynthesis was estimated in a flume/respirometer from changes in dissolved oxygen at eight flow speeds between 0.08 and 0.52 m/s. Flow in the flume and over the reef at Kaneohe Bay, Oahu, Hawaii was estimated using hot-film thermistor and electromagnetic current meters. Rates of photosynthesis were related positively to flow in all experiments and plots of the log of the average Sherwood number (Sh _meas) versus log Reynolds number (Re _D) for each experiment are lower than predicted for mass transfer through a turbulent boundary layer. Algal turf-covered plates are characterized as hydrodynamically transitional to fully rough surfaces and the lower than predicted slopes suggest that roughness reduces rates of mass transfer. A negative correlation between algal turf biomass and slopes of the log Sh _meas−log Re _D plots suggests that mass transfer to algal turfs is affected significantly by the physical structure of the algal community. Patterns of photosynthesis based on changes in dissolved oxygen and dissolved inorganic carbon concentrations (DIC) indicate that the flow speed effect is not the result of increased flux of oxygen from the algal turfs, and combined with the short response time to flow speed, suggest that DIC may limit rates of photosynthesis. Although there are differences between flow in the flume and flow over algal turfs on the reef, these results suggest that photosynthesis is controlled, at least in part, by mass transfer. The chemical engineering approach provides a framework to pose further testable hypotheses about how algal canopy height, flow oscillation, turbulence, and substratum roughness may modulate rates of metabolism of coral reef algal turfs.     

Photosynthetic carbon acquisition in the red algaGracilaria conferta

Photosynthetic properties of the common red algaGracilaria conferta, collected from the eastern Mediterranean Sea were investigated in 1989, in order to begin evaluating its adaptative strategies with regard to the inorganic carbon composition of seawater, and to test whether the alleged C₄ photosynthesis of anotherGracilaria species is common within the genus. Net photosynthetic rates ofG. conferta were, under ambient conditions of inorganic carbon (ca. 10µM, CO₂ and 2.2 mM HCO ₃ ^- ), not sensitive to O₂ over the range 10 to 300µM, and the CO₂ compensation point was low (ca. 0.005µM). Ribulose-1,5-bisphosphate carboxylase/oxygenase was the major carboxylating enzyme, with a crude extract activity of 175µmol CO₂ g^–1 fresh wt h^–1 while phosphoenolpyruvate carboxylase and phosphoenolpyruvate carboxykinase were present at 70 and 20%, respectively, of that activity. No activities of the decarboxylases NAD-and NADP-malic enzyme could be detected. The¹⁴C pulse-chase incorporation pattern showed thatG. conferta fixes inorganic carbon via the photosynthetic carbon reduction cycle only, with no evidence for photosynthetic C₄ acid metabolism. Photosynthesis at the natural seawater pH of 8.2 was, at 25°C and saturating light, saturated at the ambient inorganic carbon concentration of 2.5 mM. It is proposed that, under ambient inorganic carbon conditions, a CO₂ concentrating system other than C₄ metabolism provides an internal CO₂ concentration sufficient to suppress the O₂ effect on ribulose-1,5-bisphosphate carboxylase/oxygenase and, thus, on photorespiration, in a medium where the external free CO₂ concentration is lower than theK _m(CO₂) of the carboxylating enzyme. Since inorganic carbon, under natural saturating light conditions, seems not to be a limiting factor for photosynthesis ofG. conferta, it likely follows that other nutrients limit the growth of this alga in nature.