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.     

Some properties of calcium-activated adenosine triphosphatase from the hermatypic coralGalaxea fascicularis

Samples of the hermatypic coralGalaxea fascicularis were collected between April 1987 and April 1990 from coral reefs off Singapore (103 °45E; 1 °13N). Ca²⁺-activated adenosine triphosphatase (ATPase) activity was detected in the plasma-membrane-enriched heavy microsomal fraction ofG. fascicularis. The high affinity component hadKm andVmax values of 0.0021 mM and 0.050 µmol Pi mg^–1 protein min^–1, respectively; corresponding values for the low affinity component were 0.15 mM and 0.85 µmol mg^–1 protein min^–1. The activity of the high affinity component was inhibited 80 and 50%, respectively, by the anticalmodulin drugs calmidazolium and chlorpromazine. The low affinity component of the Ca²⁺-ATPase may represent activities of alkaline phosphatase, Ca²⁺-ATPase from membranes of mitochondria and endoplasmic reticulum, or calmodulin-dissociated plasma membrane Ca²⁺-ATPase resulting from the removal of Ca²⁺ by EDTA during the isolation process. The high affinity Ca²⁺-ATPase is probably the enzyme responsible for Ca²⁺ extrusion from the cells ofG. fascicularis. The high and low affinity components of this Ca²⁺-ATPase could use ATP and ADP as substrates. Maximum activities of both components were registered at pH 7 and at 45°C. Ruthenium red, a specific inhibitor of Ca²⁺-ATPase, inhibited the activities of the high and low affinity Ca²⁺-ATPase by 100 and 60%, respectively. Inhibition of the activities of both components was also observed with sulphydryl reagents (PCMB and mersalyl). However, DCMU, diamox, dinitrophenol, iodoacetate, fluoride, cyanide, ouabain, oligomycin B and L-phenylalanine had no effect on the enzyme activities.     

Role of glutamine synthetase in ammonia assimilation by symbiotic marine dinoflagellates (zooxanthellae)

The dinoflagellate symbionts (zooxanthellae) present in many reef corals aid in the survival of the symbiotic unit in nitrogen deficient tropical waters by providing additional routes of nitrogen uptake and metabolism. The enzymatic pathway of ammonia assimilation from seawater and the re-assimilation of coral ammonium waste by zooxanthellae was studied by examining the affinity of glutamine synthetase for one of its substrates, ammonia. Glutamine synthetase activity was measured in dinoflagellates of the species Symbiodinium microadriaticum found in symbiotic association with various marine coelenterates. Michaelis-Menten kinetics for the substrate ammonia were determined for freshly isolated dinoflagellates from Condylactis gigantea (apparent NH₃ K_m=33 M) and for cultured dinoflagellates from Zoanthus sociatus (apparent NH₃ K_m=60 M). On the basis of the low apparent K_ms for NH₃, it appears that ammonia assimilation by these symbiotic dinoflagellates occurs via the glutamine synthetase/glutamate synthase pathway. Additionally, the uptake of exogenous ammonium by an intact coelenterate-dinoflagellate symbiosis was strongly inhibited by 0.5 mM methionine sulfoximine, and inhibitor of glutamine synthetase.     

Characteristics of the uptake system for L-lysine and L-arginine inPhaeodactylum tricornutum

Cells ofPhaeodactylum tricornutum Bohlin develop the ability to take up L-lysine when they are deprived of nitrogen (illuminated in nitrogen-free medium), carbon (incubated in darkness) or both. Cells with a developed uptake system take up and accumulate lysine in an unchanged form. Uptake occurs under either aerobic or anaerobic conditions and is dependent on the presence of sodium⁺ ions (K _s ^{Na +}=,ca. 10 mM). Some potassium⁺ ions are necessary for uptake, presumably within the cells, but with potassium⁺-replete cells, increasing K⁺ concentration depresses lysine uptake. The lysine-uptake porter also transports L-arginine.K _s values are about 1.5 M for lysine and 0.5 M for arginine. It is, however, possible that the uptake system developed by incubating cells in darkness differs from that produced in light; it shows a pronounced pH optimum at pH 8.5, whereas the activity of the light-developed system declines from pH 6.5 to pH 9.0 and correlates well with the concentration of lysine⁺. The uptake system developed in darkness may also have a higher affinity for lysine. Lysine uptake is not inhibited by 1 mM concentrations of nitrate, nitrate, ammonium, or urea nor by similar concentrations of amphoteric or acidic amino acids.     

Purification and characterisation of octopine dehydrogenase from the marine nemertean Cerebratulus lacteus (Anopla: Heteronemerta): comparison with scallop octopine dehydrogenase

Octopine dehydrogenase from the nemertean Cerebratulus lacteus was purified over 1000-fold to almost homogeneity. The enzyme does not bind to arginine Sepharose 4B. It has a monomeric structure with a relative molecular mass of 40000. Two isoenzymes were identified with isoelectric points of 5.6 and 5.4, whereas the purified isoenzymes of Pecten jacobaeus adductor mucles (which bind to arginine Sepharose 4B) had lower IEP's of 4.9 and 4.7. Apparent K_m's of the nemertean ODH for arginine and pyruvate are dependent on the respective co-substrate concentration. This phenomenon may result in activation of ODH and, thus, production of octopine in locomotory highly active individuals while attacking food, especially when this takes place in a hypoxic habitat, such as decaying mud near the high-water mark. The apparent K_m's for octopine (0.22 mM) and NAD⁺ (14 M) are low. Octopine is a substrate inhibitor for the reverse reaction above 2 mM, and a product inhibitor of the forward reaction by 50% at 1.2 mM. Therefore, only small amounts of octopine are likely to accumulate in vivo. Amino acid substrate specificity is limited to guanidino amino acids. We believe that the amino acid substrate specificity is not an evolutionary modification, but rather that it is narrowed to guanidino amino acids (or even specificity to arginine) in those species where ODH has a physiological function in maintaining redox balance during exercise. The specificity for keto acids is dependent on chain length, (-ketobutyrate>-ketocapronate); a second carboxyl group inactivates the enzyme.     

Gene flow among population of a teleost (painted greenling,Oxylebius pictus) from Puget Sound to southern California

Net photosynthetic oxygen evolution in Amphiroa anceps (Lamarck) Decaisne is inhibited at high oxygen concentrations. Photosynthesis is highest between pH 6.5 and 7.5. At pH 9 to 10 there is still a significant photosynthetic rate, suggesting that this alga can use HCO _- ³ as a substrate for photosynthesis. At pH 7.0 to 8.5, the photosynthetic rate saturates at a total inorganic carbon concentration (C_i) greater than 3 mM. At pH 8.5 and 8.8, calcification rate continues to increase with increasing concentration of C_i. Between pH 7 and 9, the calcification rate in the light in A. foliacea Lamouroux is proportional to the photosynthetic rate, whereas at higher pH where the photosynthetic rate is very low, the calcification rate is stimulated by the higher concentration of CO _2- ³ ion. At all pH values examined, the calcification rate of living plants in the dark and of dead plants is directly proportional to the CO _2- ³ ion concentration, suggesting little metabolic involvement in calcification processes in the dark, whereas calcification in live A. foliacea in the light is influenced both by the photosynthetic rate and the CO _2- ³ ion concentration in the medium.     

NADP+-dependent glutamate dehydrogenase from Acropora formosa: purification and properties

As an initial step in our study of nitrogen metabolism in the coral/algal symbiosis we have purified glutamate dehydrogenase (EC 1.4.1.4) to homogeneity from polyp tissue of the staghorn coral Acropora formosa collected from Magnetic Island (North Queensland) in 1985–1986. The purified enzyme had a specific activity of 78 U mg^-1. The native enzyme had a relative molecular weight, M _r, of 360 000 (±20 000), and appears to be a hexamer with subunits of M _r=56000 (±3 000). Like the enzyme from other coelenterates, the coral glutamate dehydrogenase (GDH) was absolutely specific with respect to the coenzyme substrate (NADP⁺/NADPH), and was insensitive to allosteric regulation by nucleotides; unlike other coelenterate GDHs, the coral enzyme was absorlutely specific for ammonium as amino group donor in the reductive amination reaction, and major differences in kinetic properties were apparent. Linear Michaelis-Menten kinetics were observed for the substrates a-ketoglutarate, NADPH and NADP⁺, the K _m values being 0.93, 0.11 and 0.03 mM, respectively. However glutamate dehydrogenase displayed biphasic kinetics with respect to l-glutamate and ammonium, indicating two apparent K _m values (18 and 81 mM for l-glutamate and 9.2 and 416 mM for ammonium). The enzyme also exhibits Scatchard plots, Hill coefficients and cooperativity indices characteristic of enzymes displaying negative cooperativity.     

Na<Superscript>+</Superscript>/K<Superscript>+</Superscript>–ATPase activity and gill ultrastructure in the hyper-hypo-regulating crab<Emphasis Type="Italic"> Chasmagnathus granulatus</Emphasis> acclimated to dilute,normal, and concentrated seawater

We studied Na⁺/K⁺ –ATPase activity and ultrastructure in gills of the hyper-hypo-regulating crab Chasmagnathus granulatus Dana, 1851 acclimated to different salinities: 10, 30 and 45, known to be hypo-, iso-, and hyper-osmotic to the hemolymph, respectively. After centrifugation of homogenates at 11,000 g, Na⁺/K⁺–ATPase activity was almost entirely found in the pellets from the posterior (6–8) and anterior (3–5) gills, whereas very little was detected in the supernatant liquid. Specific activity of gill 6 was 41.3, 30.2, and 28.2 µmol Pi h^–1 mg prot^–1 for crabs acclimated to 10, 30, and 45, respectively, the result for 10 being significantly higher than those at 30 and 45. Although the concentration of sodium at which the reaction rate is half-maximal (K _M) was similar in the three acclimation salinities, only the enzyme from crabs acclimated to 10 was inhibited by high sodium concentration. Specific activity of gill 5 increased with the increment in external salinity (10.1, 15, and 18.1 µmol Pi h^–1 mg prot^–1 for 10, 30, and 45, respectively), the only significant difference being that between the extreme salinities. The epithelium thickness of the dorsal portion of gill 6 showed a variation among salinities: 21.7, 15.8 and 17.2 µm for 10, 30 and 45, respectively. There were significant differences in epithelium thickness between the 10 and the other salinities. In all three salinities, the ultrastructure of gill 6 epithelium showed a high density of mitochondria, estimated by their volume fraction (Vv _m=0.307–0.355). These mitochondria were packed between extensive basolateral membrane interdigitations in ionocytes and pillar cells. Gill 5 showed three cell types: pillars which possess mitochondria packed between membrane folds only in their interdigitations with neighbouring cells; type-I cells 8.0 µm thick with low density of mitochondria (Vv _m=0.088), and type-II cells, 9.9 µm thick and rich in mitochondria (Vv _m=0.423), but lacking basolateral interdigitations. Vv _m of type-I cells of gill 5 was significantly lower than those of type-II cells of the same gill and the ionocytes of gill 6. No significant difference in Vv _m was detected between the latter cell types.Communicated by P.W. Sammarco, Chauvin     

Photosynthetic carbon acquisition in the red alga Gracilaria conferta

In this continuing study on photosynthesis of the marine red alga Gracilaria conferta, it was found that ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) in crude extracts had a K _m (CO₂) of 85 M. Since seawater contains only ca. 10 M CO₂, it appears that this alga must possess a CO₂ concetrating system in order to supply sufficient CO₂ to the vicinity of the enzyme. Because this species is a C₃ plant (and thus lacks the C₄ system for concentrating CO₂), but can utilize HCO₃ ^- as an exogenous carbon source, we examined whether HCO₃ ^- uptake could be the initial step of such a CO₂ concetrating system. The surface pH of G. conferta thalli was 9.4 during photosynthesis. At this pH, estimated maximal uncatalyzed HCO₃ ^- dehydration (CO₂ formation) within the unstirred layer was too slow to account for measured phostosynthetic rates, even in the presence of an external carbonic anhydrase inhibitor. This observation, and the marked pH increase in the unstirred layer following the onset of light, suggests that a HCO₃ ^- transport system (probably coupled to transmembrane H⁺/OH^- fluxes) operates at the plasmalemma level. The involvement of surface-bound carbonic anhydrase in such a system remains, however, obscure. The apparent need of marine macroalgae such as G. conferta for CO₂ concentrating mechanisms is discussed with regard to their low affinity of Rubisco to CO₂ and the low rate of CO₂ supply in water. The close similarity between rates of Rubisco carboxylation and measured photosynthesis further suggests that the carboxylase activity, rather than inorganic carbon transport and intercoversion events, could be an internal limiting factor for photosynthetic rates of G. conferta.     

Biosynthetically active glutamine synthetase in the marine diatom Phaeodactylum tricornutum: optimization of the forward-reaction assay

The activity of glutamine synthetase (GS) was measured in the marine diatom Phaeodactylum tricornutum Bohlin (Strain SME) by a biosynthetic assay, based on -glutamyl hydroxamate synthesis, and referred to as the forwardreaction assay. The effects of pH, temperature and different homogenizing buffer preparations on enzyme activity, linearity of reactions, and substrate-saturation kinetics were investigated. The resultant data provide the basis for establishing optimum experimental conditions for a standard assay. Affinities of P. tricornutum GS for glutamate, ATP and Mg²⁺ were similar to those recorded elsewhere for a variety of other phytoplankton species using true biosynthetic assays based on release of inorganic phosphate, whereas the affinity for hydroxylamine was two orders of magnitude lower than that for ammonium, with an apparent K _m value in the millimolar range. This, together with negative results obtained during earlier attempts to detect GS activity in P. tricornutum using the true biosynthetic assay, indicates that the GS of this alga has a lower affinity for ammonium than that of other phytoplankton species. Dual substrate kinetics demonstrated that apparent K _m and V _m values for glutamate were directly proportional to the concentration of ATP, thus giving indirect evidence of a correlation between GS activity and the adenylate energy charge. Comparisons between synthetase activities obtained with the optimized forward-reaction assay and transferase activities reported from other studies on various phytoplankton species revealed discrepancies which, to a great extent, probably arise from differences in the growth conditions of the organisms.     

Nitrate reductase activity in Zostera marina

Eelgrass (Zostera marina L.) has access to nutrient pools in both the water column and sediments. We investigated the potential for eelgrass to utilize nitrate nitrogen by measuring nitrate reductase (NR) activity with an in vivo tissue assay. Optimal incubation media contained 60 mM nitrate, 100 mM phosphate, and 0.5% 1-propanol at pH 7.0. Leaves had significantly higher NR activity than roots (350 vs 50 nmoles NO ₂ ^– produced g FW^–1 h^–1). The effects of growing depth (0.8 m MLW, 1.2 m, 3.0 m, 5.0 m) and location within the eelgrass meadow (patch edge vs middle) on NR activity were examined using plants collected from three locations in the Woods Hole area, Massachusetts, USA, in July 1987. Neither depth nor position within the meadow appear to affect NR activity. Nitrate enrichment experiments (200 M NO ₃ ^– for 6 d) were conducted in the laboratory to determine if NR activity could be induced. Certain plants from shallow depth (1.2 m) showed a significant response to enrichment, with NR activity increasing from >100 up to 950 nmoles NO ₂ ^– g FW^–1 h^–1 over 6 d. It appears that Z. marina growing in very shallow water (0.8 m) near a shoreline may be affected by ground water or surface run-off enrichments, since plants from this area exhibited rates up to 1 600 nmol NO ₂ ^– g FW^–1 h^–1. Water samples from this location consistently had slightly higher NO ₃ ^– concentrations (1.4 M) than all other collection sites (0.7 M). Thus, it is possible that chronic run-off or localized groundwater inputs can create sufficient NO ₃ ^– enrichment in the water column to induce nitrate reductase activity in Zostera leaves.     

Regulation of fructose-2,6-bisphosphate content in mantle tissue of the sea musselMytilus galloprovincialis

6-phosphofructo-2-kinase (PFK-2) from the mantle of the sea musselMytilus galloprovincialis Lmk, collected from the Ría de Arosa (NW Spain) in 1990, was purified 550-fold by extraction and sequential affinity chromatography on Affi-gel Blue and ATP-agarose columns. The enzyme was a dimer with a native molecular weight of 100 kilodaltons (KDa) and a subunitM _r of 53 KDa. PFK-2 activity is dependent on the presence of P_i. At physiological P_i concentrations, the enzyme exhibits hyperbolic kinetics with both ATP and Fru-6-P, withK _m values of 0.62 and 0.37 mM respectively. In vivo, PFK-2 activity is limited by the concentration of Fru-6-P which is low in comparison with theK _m for this substrate. Citrate and PEP inhibited PFK-2 activity.     

A comparative study of the glutamate dehydrogenase activity in several species of marine phytoplankton

Kinetic studies on the NADP-linked glutamate dehydrogenase (GDH) activity in 8 species of marine phytoplankton have been carried out. The pH optima, linearity of reactions, apparent energies of activation, and apparent K _m values for NADPH, NH ₄ ^>+ and -ketoglutarate were determined. Based on these data, an accurate, sensitive enzyme assay procedure is presented which was successfully tested on shipboard during a recent cruise in an upwelling region. Preliminary studies on inhibition and activation of the NADP-linked GDH activity have also been carried out. In addition to NADP-linked GDH activity, NAD-linked GDH activity was discovered in certain species of phytoplankton. The possible physiological roles of NAD-and NADP-linked GDH enzymes are discussed.Contribution No. 911 from the Department of Oceanography, University of Washington. This research was supported by the National Science Foundation Grants GA-34165 and GX-33502.     

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.     

Purification and partial characterisation of phenoloxidase from the colonial ascidian Botryllus schlosseri

Phenoloxidase (PO) from the colonial ascidian Botryllus schlosseri was purified using two different chromatographic strategies. A three-step purification was developed in order to maintain enzyme activity, whereas an easier purification procedure was adopted to obtain enough PO for the production of specific polyclonal antibodies. The enzyme showed optimal pH and temperature values of 7.0 to 7.5 and 35 °C, respectively, and a K _m value of 4.62 ± 0.76 mM was estimated using l-DOPA as substrate. A molecular weight of 160 kDa was determined after SDS-PAGE under non-reducing conditions. The addition of the reducing agent β-mercaptoethanol caused the disappearance of the 160 kDa band and the appearance of a new band at 80 kDa, suggesting that active PO is a dimer and the two subunits are linked by disulphide bridges. Received: 14 December 1998 / Accepted: 24 August 1999     

Inorganic carbon uptake by photosynthetically active protoplasts of the red macroalga Chondrus crispus

Photosynthetically active protoplasts were isolated from Chondrus crispus Stackh. by treating thalli with -carrageenase produced from batch culture of Pseudomonas carrageenovora. Using the silicone oil centrifugation technique, it was found that the protoplasts: (1) did not generally accumulate inorganic carbon (C_i) above the concentration in their incubation medium; (2) were saturated at C_i concentrations of 3 to 4 mM; (3) had an intracellular pH of 7.50 when incubated at pH 7.5; and (4) their initial carbon fixation rate was reduced by carbonic anhydrase inhibitors. Although the carbon fixation rate of the protoplasts was about 30% that of thallus fragments, presumably due to the relatively harsh protoplast isolation treatment, the behavior of the protoplasts was similar to that of fragments. This similarity indicates that the protoplasts are photosynthetically active and behave as thallus fragments. Further, the data are consistent with the hypothesis that C. crispus acquires C_i for photosynthesis by the diffusion of CO₂ across the plasma membrane.     

Hepatopancreas mitochondria from Mytilus edulis:

A method for the isolation of intact coupled mitochondria from the hepatopancreas of Mytilus edulis has been developed. These mitochondria prefer lipid substrates and accept a range of fatty acid chain lengths. Acetyl carnitine is also oxidized at high rates. Evidence for the presence of the tricarboxylate and the dicarboxylate transporters is presented. The absence of the pyruvate transporter or inhibition of pyruvate dehydrogenase is indicated by the absence of pyruvate oxidation. Glutamate is oxidized at high rates with a pH optimum of 7.5 based on the RCR. Cysteine oxidation may indicate the presence of the enzymes involved in the synthesis of taurine. Ornithine transport is also indicated, implying the existence of the urea cycle. Alanine, glycine, proline, asparagine, arginine, aspartate and citrulline were not oxidized at detectable levels. The osmotic optimum for the oxidation of glutamate extends from 600 to 1000 mOsm l^-1. The state 3 rate is more profoundly affected by the osmolarity than the state 4 rate. The role of osmolarity in the control of oxidation of amino acids during volume regulation of osmotically stressed M. edulis is suggested. Mitochondrial volume changes correlate with the optimal range of osmolarity for glutamate oxidation based on the RCR.     

The ammonium/methylammonium uptake system of Symbiodinium microadriaticum

The substrate analogue [¹⁴C]-methylammonium was used to study ammonium/methylammonium uptake by Symbiodinium microadriaticum (zooxanthellae). The value of the Michaelis constant (K _m) for the uptake system was approximately 35 M with methylammonium as substrate; ammonium was a competitive inhibitor of methylammonium uptake, and the K _m for ammonium uptake (determined as the inhibition constant, K _i, for methylammonium) was 6.6 M. Methylammonium uptake by zooxanthellae was light-dependent. Methylammonium uptake rates of zooxanthellae which had been freshly isolated from the hermatypic coral Acropora formosa (0.85±0.05x10^-10 mol min^-1 cell^-1) were lower than those of axenic cultures of the zooxanthellae from Montipora verrucosa (Acroporidae) grown under various nitrogen regimes (1.6 to 12x10^-10 mol min^-1 cell^-1). Maximum uptake rates were found for ammonium-starved cultured M. verrucosa zooxanthellae (10.2 to 12x10^-10 mol min^-1 cell^-1); M. verrucosa zooxanthellae growing with ammonium as nitrogen source and zooxanthellae which had been freshly isolated from A. formosa gave similar and considerably lower uptake rates (0.85 to 1.6x10^-1 mol min^-1 cell^-1). These results suggest that either coral tissue contains sufficient ammonium to repress synthesis of the uptake system of the algal symbionts or, alternatively, there are additional barriers to ammonium transport for zooxanthellae in vivo.     

Photosynthetic utilisation of inorganic carbon by seagrasses from Zanzibar, East Africa

Photosynthetic rates of eight seagrass species from Zanzibar were limited by the inorganic carbon composition of natural seawater (2.1 mM, mostly in the form of HCO₃ ⁻), and they exhibited more than three time higher rates at inorganic carbon saturation (>6 mM). The intertidal species that grew most shallowly, Halophila ovalis, Halodule wrightii and Cymodocea rotundata, showed the highest affinity for inorganic carbon (K _1/2 = ca. 2.5 mM), followed by the subtidal species (K _1/2 > 5 mM). Photosynthesis of H. wrightii, C. rotundata, Cymodocea serrulata and Enhalus acoroides was >50% inhibited by acetazolamide, a membrane-impermeable inhibitor of carbonic anhydrase, indicating that extracellular HCO₃ ⁻ dehydration is an important part of their inorganic carbon uptake. Photosynthetic rates of H. wrightii, Thalassia hemprichii, Thalassodendron ciliatum, C. serrulata and E. acoroides were strongly reduced by changing the seawater pH from 8.2 to 8.6 in a closed system. In H. ovalis, C. rotundata and Syringodiumisoetifolium, photosynthesis at pH 8.6 was maintained at a higher level than could be caused by the ca. 30% CO₂ concentration which remained in the closed experimental systems at that pH, pointing toward HCO₃ ⁻ uptake in those species. It is suggested that the ability of H. ovalis and C. rotundata to grow in the high, frequently air-exposed, intertidal zone may be related to a capability to take up HCO₃ ⁻ directly, since this is a more efficient way of HCO₃ ⁻ utilisation than extracellular HCO₃ ⁻ dehydration under such conditions. The inability of all species to attain maximal photosynthetic rates under natural conditions of inorganic carbon supports the notion that seagrasses may respond favourably to any future increases in marine CO₂ levels. Received: 19 March 1997 / Accepted: 31 March 1997