Nutrient uptake and alkaline phosphatase (ec 3:1:3:1) activity of emiliania huxleyi (PRYMNESIOPHYCEAE) during growth under n and p limitation in continuous cultures

Emiliania huxleyi (strain L) expressed an exceptional P assimilation capability. Under P limitation, the minimum cell P content was 2.6 fmol P·cell^?1, and cell N remained constant at all growth rates at 100 fmol N·cell^?1. Both, calcification of cells and the induction of the phosphate uptake system were inversely correlated with growth rate. The highest (cellular P based) maximum phosphate uptake rate (V_max^P) was 1400 times (i.e. 8.9 h^?1) higher than the actual uptake rate. The affinity of the P‐uptake system (dV/dS) was 19.8 L·μmol^?1·h^?1 at μ = 0.14 d^?1. This is the highest value ever reported for a phytoplankton species. V_max and dV/dS for phosphate uptake were 48% and 15% lower in the dark than in the light at the lowest growth rates. The half‐saturation constant for growth was 1.1 nM. The coefficient for luxury phosphate uptake (Q_maxt/Q_min) was 31. Under P limitation, E. huxleyi expressed two different types of alkaline phosphatase (APase) enzyme kinetics. One type was synthesized constitutively and possessed a V_max and half‐saturation constant of 43 fmol MFP·cell^?1·h^?1 and 1.9 μM, respectively. The other, inducible type of APase expressed its highest activity at the lowest growth rates, with a V_max and half‐saturation constant of 190 fmol MFP·cell^?1·h^?1 and 12.2 μM, respectively. Both APase systems were located in a lipid membrane close to the cell wall. Under N‐limiting growth conditions, the minimum N quotum was 43 fmol N·cell^?1. The highest value for the cell N‐specific maximum nitrate uptake rate (V_max^N) was 0.075 h^?1; for the affinity of nitrate uptake, 0.37 L·μmol^?1·h^?1. The uptake rate of nitrate in the dark was 70% lower than in the light. N‐limited cells were smaller than P‐limited cells and contained 50% less organic and inorganic carbon. In comparison with other algae, E. huxleyi is a poor competitor for nitrate under N limitation. As a consequence of its high affinity for inorganic phosphate, and the presence of two different types of APase in terms of kinetics, E. huxleyi is expected to perform well in P‐controlled ecosystems.     

PHOTOSYNTHESIS AND CALCIFICATION BY EMILIANIA HUXLEYI (PRYMNESIOPHYCEAE) AS A FUNCTION OF INORGANIC CARBON SPECIES

To test the possibility of inorganic carbon limitation of the marine unicellular alga Emiliania huxleyi (Lohmann) Hay and Mohler, its carbon acquisition was measured as a function of the different chemical species of inorganic carbon present in the medium. Because these different species are interdependent and covary in any experiment in which the speciation is changed, a set of experiments was performed to produce a multidimensional carbon uptake scheme for photosynthesis and calcification. This scheme shows that CO₂ that is used for photosynthesis comes from two sources. The CO₂ in seawater supports a modest rate of photosynthesis. The HCO is the major substrate for photosynthesis by intracellular production of CO₂ (HCO+ H⁺→ CO₂+ H₂O → CH₂O + O₂). This use of HCO is possible because of the simultaneous calcification using a second HCO, which provides the required proton (HCO+ Ca²⁺→ CaCO₃+ H⁺). The HCO is the only substrate for calcification. By distinguishing the two sources of CO₂ used in photosynthesis, it was shown that E. huxleyi has a K_½ for external CO₂ of “only” 1.9 ± 0.5 μM (and a V_max of 2.4 ± 0.1 pmol·cell⁻¹·d⁻¹). Thus, in seawater that is in equilibrium with the atmosphere ([CO₂]= 14 μM, [HCO]= 1920 μM, at fCO₂= 360 μatm, pH = 8, T = 15° C), photosynthesis is 90% saturated with external CO₂. Under the same conditions, the rate of photosynthesis is doubled by the calcification route of CO₂ supply (from 2.1 to 4.5 pmol·cell⁻¹·d⁻¹). However, photosynthesis is not fully saturated, as calcification has a K_½ for HCO of 3256 ± 1402 μM and a V_max of 6.4 ± 1.8 pmol·cell⁻¹·d⁻¹. The H⁺ that is produced during calcification is used with an efficiency of 0.97 ± 0.08, leading to the conclusion that it is used intracellularly. A maximum efficiency of 0.88 can be expected, as NO uptake generates a H⁺ sink (OH⁻ source) for the cell. The success of E. huxleyi as a coccolithophorid may be related to the efficient coupling between H⁺ generation in calcification and CO₂ fixation in photosynthesis.     

SEASONAL EFFECT ON THE RELATIONSHIP BETWEEN THE PHOTOSYNTHETIC CAPACITY OF INTERTIDAL MICROPHYTOBENTHOS AND TEMPERATURE1

The response of the photosynthetic capacity (P_max) of microphytobenthos to short-term variations of temperature (in the range 5–35° C) was assessed on a seasonal basis. The relationship is described mathematically, and relevant physiological parameters are identified: P_MAX, the maximum value of P_max achieved at T_opl, the optimum temperature. Estimated values of T_opt do not change significantly throughout the year and remain close to 25° C. It is thus concluded that T_opt is not influenced by seasonal variations in the daily range of mud surface temperature. Identical conclusions hold for T_max (ca. 38° C), the thermal threshold beyond which no photosynthesis occurs. Conversely, P_MA estimates exhibit substantial variability: P_MAX (mean ± root mean square error) is highest in April (11.18 ± 0.42 [μg C · [μg Chl a]^?1· h^?1) during the beginning of the annual increase in temperature, photoperiod, and maximum irradiance and is lowest in December (3.04 ± 0.16 μg C · [μg Chl a]^?1· h^?l). From an ecological point of view, the short-term and seasonal variations of P_MAX suggest that the microphytobenthic community takes advantage of the abiotic spring environmental conditions, allowing the onset of the bloom. Nevertheless, no “acclimation strategy” (i.e. shifts in T_opt and T_max that prevent temperature inhibition in summer or improve photosynthetic rates in winter) is apparent from our results.     

Temperature response of photosynthetic light‐ and carbon‐use characteristics in the red seaweed Gracilariopsis lemaneiformis (Gracilariales,Rhodophyta)

The red seaweed Gracilariopsis is an important crop extensively cultivated in China for high‐quality raw agar. In the cultivation site at Nanao Island, Shantou, China, G. lemaneiformis experiences high variability in environmental conditions like seawater temperature. In this study, G. lemaneiformis was cultured at 12, 19, or 26°C for 3 weeks, to examine its photosynthetic acclimation to changing temperature. Growth rates were highest in G. lemaneiformis thalli grown at 19°C, and were reduced with either decreased or increased temperature. The irradiance‐saturated rate of photosynthesis (P_max) decreased with decreasing temperature, but increased significantly with prolonged cultivation at lower temperatures, indicating the potential for photosynthesis acclimation to lower temperature. Moreover, P_max increased with increasing temperature (~30 μmol O₂ · g^?1FW · h^?1 at 12°C to 70 μmol O₂ · g^?1FW · h^?1 at 26°C). The irradiance compensation point for photosynthesis (I_c) decreased significantly with increasing temperature (28 μmol photons · m^?2 · s^?1 at high temperature vs. 38 μmol photons · m^?2 · s^?1 at low temperature). Both the photosynthetic light‐ and carbon‐use efficiencies increased with increasing growth or temperatures (from 12°C to 26°C). The results suggested that the thermal acclimation of photosynthetic performance of G. lemaneiformis would have important ecophysiological implications in sea cultivation for improving photosynthesis at low temperature and maintaining high standing biomass during summer. Ongoing climate change (increasing atmospheric CO₂ and global warming) may enhance biomass production in G. lemaneiformis mariculture through the improved photosynthetic performances in response to increasing temperature.     

PHOSPHORUS AND NITROGEN NUTRITION IN CHONDRUS CRISPUS (RHODOPHYTA): EFFECTS ON TOTAL PHOSPHORUS AND NITROGEN CONTENT,CARRAGEENAN PRODUCTION,AND PHOTOSYNTHETIC PIGMENTS AND METABOLISM1

The existence of a phenomenon in phosphorus (P) nutrition comparable to the “Neish effect” in nitrogen (N) nutrition (an inverse relation between seawater N enrichment and carrageenan content) was investigated in the temperate red alga Chondrus crispus Stackhouse. Plants were preconditioned for 17 d and then cultured under varying enrichments of P (0, 3, 6, 10, 15 μM P·wk^?1) and a constant N enrichment (53.5 μM N·wk^?1) for 5 wk. Tissue total P, tissue total N, and carrageenan contents were then determined. Identical experiments were performed using C. crispus collected during the fall, winter, spring, and summer seasons. The procedure was repeated using material collected during the following fall season and cultured under constant P (6 μM P·wk^?1) and varying N enrichments (0, 3, 6, 10, 25 μM N·wk^?1). In the fall (P) experiment, carrageenan content was the highest [53.1 ± 0.3% DW (dry weight)], and tissue total P content was the lowest (1.71 ± 0.27 mg P·g DW^?1) in plants that received no P enrichment. Carrageenan content was stable (46.1 ± 1.8% DW) for plants given enrichments of 3 μM P·wk^?1 and greater. Thus, a decrease in carrageenan content, concomitant with an increase in tissue total P content, was observed, but only at tissue total P levels below 2 mg P·g DW^?1. As these levels were always higher than 2 mg P·g DW^?1 in the winter, spring, and summer experiments, carrageenan content remained constant within each season at 46.2 ± 1.3, 43.1 m 0.7, and 44.5 ± 0.6% DW, respectively. Nitrogen enrichment of plants collected in the fall did not affect carrageenan content, which was stable at 49.3 ± 0.9% DW. When these plants were compared with those of the previous fall experiment (6 μM P·wk^?1 and 53.5 μM N·wk^?1), a slight increase in carrageenan content was noted. Thus, at sufficiently high concentration, N also decreased carrageenan content in C. crispus. Phosphorus nutrition had no significant effect on photosynthesis versus irradiance parameters (P_max, α, R_d, I_c, and I_k), the contents of the photosynthetic pigments chlorophyll-a, phycoerythrin (PE), phycocyanin (PC), and allophycocyanin (APC), and the ratios PE:APC and PC:APC. In contrast, N nutrition affected both P_maxand the photosynthetic pigment contents. The data indicate that N limitation reduces the number of phycobilisomes but not their size. The greater reduction in phycobiliprotein than chlorophyll-acontent corroborates the natural bleaching phenomenon regularly observed in C. crispus populations during summer when N levels are generally low in seawater. These results suggest that C. crispus in the temperate waters of the Bay of Fundy may experience N limitation, but P limitation is unlikely.     

Uptake kinetics and storage capacity of dissolved inorganic phosphorus and corresponding dissolved inorganic nitrate uptake in Saccharina latissima and Laminaria digitata (Phaeophyceae)

Uptake rates of dissolved inorganic phosphorus and dissolved inorganic nitrogen under unsaturated and saturated conditions were studied in young sporophytes of the seaweeds Saccharina latissima and Laminaria digitata (Phaeophyceae) using a “pulse‐and‐chase” assay under fully controlled laboratory conditions. In a subsequent second “pulse‐and‐chase” assay, internal storage capacity (ISC) was calculated based on V_M and the parameter for photosynthetic efficiency F_v/F_m. Sporophytes of S. latissima showed a V_S of 0.80 ± 0.03 μmol · cm^?2 · d^?1 and a V_M of 0.30 ± 0.09 μmol · cm^?2 · d^?1 for dissolved inorganic phosphate (DIP), whereas V_S for DIN was 11.26 ± 0.56 μmol · cm^?2 · d^?1 and V_M was 3.94 ± 0.67 μmol · cm^?2 · d^?1. In L. digitata, uptake kinetics for DIP and DIN were substantially lower: V_S for DIP did not exceed 0.38 ± 0.03 μmol · cm^?2 · d^?1 while V_M for DIP was 0.22 ± 0.01 μmol · cm^?2 · d^?1. V_S for DIN was 3.92 ± 0.08 μmol · cm^?2 · d^?1 and the V_M for DIN was 1.81 ± 0.38 μmol · cm^?2 · d^?1. Accordingly, S. latissima exhibited a larger ISC for DIP (27 μmol · cm^?2) than L. digitata (10 μmol · cm^?2), and was able to maintain high growth rates for a longer period under limiting DIP conditions. Our standardized data add to the physiological understanding of S. latissima and L. digitata, thus helping to identify potential locations for their cultivation. This could further contribute to the development and modification of applications in a bio‐based economy, for example, in evaluating the potential for bioremediation in integrated multitrophic aquacultures that produce biomass simultaneously for use in the food, feed, and energy industries.     

PHOTOSYNTHESIS AND RESPIRATION OF THE CONCHOCELIS STAGE OF ALASKAN PORPHYRA (BANGIALES,RHODOPHYTA) SPECIES IN RESPONSE TO ENVIRONMENTAL VARIABLES1

Photosynthesis and respiration of three Alaskan Porphyra species, P. abbottiae V. Krishnam., P. pseudolinearis Ueda species complex (identified as P. “pseudolinearis” below), and P. torta V. Krishnam., were investigated under a range of environmental parameters. Photosynthesis versus irradiance (P–I) curves revealed that maximal photosynthesis (P_max), irradiance at maximal photosynthesis (I_max), and compensation irradiance (I_c) varied with salinity, temperature, and species. The P_max of Porphyra abbottiae conchocelis varied between 83 and 240 μmol O₂ · g dwt^?1 · h^?1 (where dwt indicates dry weight) at 30–140 μmol photons · m^?2 · s^?1 (I_max) depending on temperature. Higher irradiances resulted in photoinhibition. Maximal photosynthesis of the conchocelis of P. abbottiae occurred at 11°C, 60 μmol photons · m^?2·s^?1, and 30 psu (practical salinity units). The conchocelis of P. “pseudolinearis” and P. torta had similar P_max values but higher I_max values than those of P. abbottiae. The P_max of P. “pseudolinearis” conchocelis was 200–240 μmol O₂ · g dwt^?1 · h^?1 and for P. torta was 90–240 μmol O₂ · g dwt^?1 · h^?1. Maximal photosynthesis for P. “pseudolinearis” occurred at 7°C and 250 μmol photons · m^?2 · s^?1 at 30 psu, but P_max did not change much with temperature. Maximal photosynthesis for P. torta occurred at 15°C, 200 μmol photons · m^?2 · s^?1, and 30 psu. Photosynthesis rates for all species declined at salinities <25 or >35 psu. Estimated compensation irradiances (I_c) were relatively low (3–5 μmol · photons · m^?2 · s^?1) for intertidal macrophytes. Porphyra conchocelis had lower respiration rates at 7°C than at 11°C or 15°C. All three species exhibited minimal respiration rates at salinities between 25 and 35 psu.     

Uptake kinetics and storage capacity of dissolved inorganic phosphorus and corresponding N:P dynamics in Ulva lactuca (Chlorophyta)

Dissolved inorganic phosphorus (DIP ) is an essential macronutrient for maintaining metabolism and growth in autotrophs. Little is known about DIP uptake kinetics and internal P‐storage capacity in seaweeds, such as Ulva lactuca (Chlorophyta). Ulva lactuca is a promising candidate for biofiltration purposes and mass commercial cultivation. We exposed U. lactuca to a wide range of DIP concentrations (1–50 μmol · L^?1) and a nonlimiting concentration of dissolved inorganic nitrogen (DIN ; 5,000 μmol · L^?1) under fully controlled laboratory conditions in a “pulse‐and‐chase” assay over 10 d. Uptake kinetics were standardized per surface area of U. lactuca fronds. Two phases of responses to DIP ‐pulses were measured: (i) a surge uptake (V_S ) of 0.67 ± 0.10 μmol · cm^?2 · d^?1 and (ii) a steady state uptake (V_M ) of 0.07 ± 0.03 μmol · cm^?2 · d^?1. Mean internal storage capacity (ISC_P ) of 0.73 ± 0.13 μmol · cm^?2 was calculated for DIP . DIP uptake did not affect DIN uptake. Parameters of DIN uptake were also calculated: V_S  = 12.54 ± 1.90 μmol · cm^?2 · d^?1, V_M  = 2.26 ± 0.86 μmol · cm^?2 · d^?1, and ISC_N  = 22.90 ± 6.99 μmol · cm^?2. Combining ISC and V_M values of P and N, nutrient storage capacity of U. lactuca was estimated to be sufficient for ~10 d. Both P and N storage capacities were filled within 2 d when exposed to saturating nutrient concentrations, and uptake rates declined thereafter at 90% for DIP and at 80% for DIN . Our results contribute to understanding the ecological aspects of nutrient uptake kinetics in U. lactuca and quantitatively evaluating its potential for bioremediation and/or biomass production for food, feed, and energy.     

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

SUMMARY.

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

     

LOW THERMAL LIMIT OF GROWTH RATE OF SYMBIODINIUM CALIFORNIUM (DINOPHYTA) IN CULTURE MAY RESTRICT THE SYMBIONT TO SOUTHERN POPULATIONS OF ITS HOST ANEMONES (ANTHOPLEURA SPP.; ANTHOZOA,CNIDARIA)1

Symbiodinium californium (#383, Banaszak et al. 1993 ) is one of two known dinoflagellate symbionts of the intertidal sea anemones Anthopleura elegantissima, A. xanthogrammica, and A. sola and occurs only in hosts at southern latitudes of the North Pacific. To investigate if temperature restricts the latitudinal distribution of S. californium, growth and photosynthesis at a range of temperatures (5°C–30°C) were determined for cultured symbionts. Mean specific growth rates were the highest between 15°C and 28°C (μ 0.21–0.26 · d^?1) and extremely low at 5, 10, and 30°C (0.02–0.03 · d^?1). Average doubling times ranged from 2.7 d (20°C) to 33 d (5, 10, and 30°C). Cells cultured at 10°C had the greatest cell volume (821 μm³) and the highest percentage of motile cells (64.5%). Growth and photosynthesis were uncoupled; light‐saturated maximum photosynthesis (P_max) increased from 2.9 pg C · cell^?1 · h^?1 at 20°C to 13.2 pg C · cell^?1 · h^?1 at 30°C, a 4.5‐fold increase. Less than 11% of daily photosynthetically fixed carbon was utilized for growth at 5, 10, and 30°C, indicating the potential for high carbon translocation at these temperatures. Low temperature effects on growth rate, and not on photosynthesis and cell morphology, may restrict the distribution of S. californium to southern populations of its host anemones.     

Factors underlying genotypic differences in the induction of photosynthesis in soybean [Glycine max (L.) Merr.]

Crop leaves are subject to continually changing light levels in the field. Photosynthetic efficiency of a crop canopy and productivity will depend significantly on how quickly a leaf can acclimate to a change. One measure of speed of response is the rate of photosynthesis increase toward its steady state on transition from low to high light. This rate was measured for seven genotypes of soybean [Glycine max (L.) Merr.]. After 10 min of illumination, cultivar ‘UA4805’ (UA) had achieved a leaf photosynthetic rate (P_n) of 23.2 μmol · m^?2 · s^?1, close to its steady‐state rate, while the slowest cultivar ‘Tachinagaha’ (Tc) had only reached 13.0 μmol · m^?2 · s^?1 and was still many minutes from obtaining steady state. This difference was further investigated by examining induction at a range of carbon dioxide concentrations. Applying a biochemical model of limitations to photosynthesis to the responses of P_n to intercellular CO₂ concentration (C_i), it was found that the speed of apparent in vivo activation of ribulose‐1:5‐bisphosphate carboxylase/oxygenase (Rubisco) was responsible for this difference. Sequence analysis of the Rubisco activase gene revealed single nucleotide polymorphisms that could relate to this difference. The results show a potential route for selection of cultivars with increased photosynthetic efficiency in fluctuating light.     

Carbonate ions appear to neither inhibit nor stimulate use of bicarbonate ions in photosynthesis by Ulva lactuca

Rates of photosynthesis by the marine macroalga Ulva lactuca were measured in a factorial experiment at five concentrations of HCO₃^? and CO₃^2- between 0·20 and 1·26 mol m^?3, but very low concentrations of CO₂. The results demonstrated that HCO₃^? was available for use, but an analysis of variance showed that CO₃^2- had neither an inhibiting nor a stimulating effect on rates of photosynthesis over this concentration range. Over the experiment, pH varied from 8·46 to 10·06 and this also had no significant effect on rates of photosynthesis. The lack of a stimulatory effect of high concentrations of CO₃^2- on the rate of photosynthesis at low concentrations of HCO₃^? was taken as circumstantial evidence for direct uptake of HCO₃^? rather than proton extrusion and external production of CO₂. In the rockpools in which U. lactuca grows, pH values up to 10·35 have been recorded, and for much of the time, CO₃^2- was the major form of inorganic carbon available. The apparent lack of an ability to use CO₃^2- under these conditions suggests that direct use of CO₃^2- as a source of inorganic carbon for photosynthesis is unlikely to be widespread.     

RELATIONSHIP BETWEEN PHOTOSYNTHESIS AND CELL SIZE OF MARINE DIATOMS12

Three photosynthetic parameters of 7 species of marine diatoms were studied using Na₂¹⁴CO₃ at 5–8 C using log phase axenic cultures. The cell volumes of the different species varied from 70 μm³ to 40 × 10⁵μm³. The present experiment is consistent with the interpretation that the initial slope α (mg C · [mg chl a]^?1· h^?1· w^?1· m²) of photosynthesis vs. light curves is controlled by self-shading of chlorophyll a in the cell. Pm, the rate of photosynthesis at light saturation (mg C · [mg cell, C]^?1· h^?1) and R, the intercept at zero light intensity (mg C · [mg cell C]^?1· H^?1) are both dependent on the ratio of surface area to volume of cell.     

The CO2‐concentrating mechanism in the bloom‐forming cyanobacterium Microcystis aeruginosa (Cyanophyceae) and effects of UVB radiation on its operation1

The bloom‐forming cyanobacterium Microcystis aeruginosa (Kütz.) Kütz. 854 was cultured with 1.05 W · m^?2 ultraviolet‐B radiation (UVBR) for 3 h every day, and the CO₂‐concentrating mechanism (CCM) within this species as well as effects of UVBR on its operation were investigated. Microcystis aeruginosa 854 possessed at least three inorganic carbon transport systems and could utilize external HCO₃^? and CO₂ for its photosynthesis. The maximum photosynthetic rate was approximately the same, but the apparent affinity for dissolved inorganic carbon was significantly decreased from 74.7 μmol · L^?1 in the control to 34.7 μmol · L^?1 in UVBR‐treated cells. At 150 μmol · L^?1 KHCO₃ and pH 8.0, Na⁺‐dependent HCO₃^? transport contributed 43.4%–40.2% to the photosynthesis in the control and 34.5%–31.9% in UVBR‐treated cells. However, the contribution of Na⁺‐independent HCO₃^? transport increased from 8.7% in the control to 18.3% in UVBR‐treated cells. The contribution of CO₂‐uptake systems showed little difference: 47.9%–51.0% in the control and 49.8%–47.2% in UVBR‐treated cells. Thus, the rate of total inorganic carbon uptake was only marginally affected, although UVBR had a differential effect on various inorganic carbon transporters. However, the number of carboxysomes in UVBR‐treated cells was significantly decreased compared to that in the control.     

A COMPARISON OF PHOTORESPONSE AMONG TEN DIFFERENT KARENIA BREVIS (DINOPHYCEAE) ISOLATES1

Many laboratories have solely used the Wilson isolate to physiologically characterize the harmful algal bloom (HAB) dinoflagellate Karenia brevis (C. C. Davis) G. Hansen et Moestrup. However, analysis of one isolate may lead to misinterpretations when extrapolating measurements to field populations. In this study, pulse‐amplitude‐modulated chlorophyll fluorometer (PAM‐FL) relative electron transport rate (ETR), F_v/F_m, and chl were compared with traditional techniques, such as ¹⁴C photosynthesis versus irradiance (P–E) curves, DCMU [3‐(3′,4′‐dichlorophenyl)‐1,1‐dimethyl urea] F_v/F_m, and extracted chl. The DCMU and PAM‐FL values of F_v/F_m (r² = 0.51) and chl (r² = 0.58) were in good agreement. There was no correlation between ¹⁴C and PAM‐FL α, P_max, and β parameters because PAM‐FL ETR was only a relative measurement. The PAM‐FL techniques were then used to investigate P–E curves, quantum yield of PSII (F_v/F_m), and chl from 10 K. brevis isolates to determine whether one or all isolates would better represent the species. Comparisons were made with a radial photosynthetron, which allowed for controlled conditions of light and temperature. Isolate α, P_max, and β varied between 0.097 and 0.204 μmol e^? · m^?2 · s^?1 · (μmol quanta · m^?2 · s^?1)^?1, 80.41 and 241 μmol e^? · m^?2 · s^?1, and 0.005 and 0.160 μmol e^? · m^?2 · s^?1 · (μmol quanta · m^?2 · s^?1)^?1, respectively. Either carbon limitation and/or bacterial negative feedback were implicated as the cause of the P–E parameter variability. Furthermore, these results directly contradicted some literature suggestions that K. brevis is a low‐light‐adapted dinoflagellate. Results showed that K. brevis was more than capable of utilizing and surviving in light conditions that may be present on cloudless days off Florida.     

PHOTOSYNTHETIC PROPERTIES OF PROTOPLASTS,AS COMPARED WITH THALLI,OF ULVA FASCIATA (CHLOROPHYTA)1

Protoplasts were prepared from Ulva fasciata Delile, and their photosynthetic performance was measured and compared with that of thalli discs. These protoplasts maintained maximal rates of photosynthesis as high as those of thalli (up to 300 μmol O₂·mg chlorophyll^?1·h^?1) for several hours after preparation and were therefore considered suitable for kinetic studies of inorganic carbon utilization. The photosynthetic K_1/2(inorganic carbon) at pH 6.1 was 3.8 μM and increased to 67, 158, and 1410 μM at the pH values 7.0, 7.9, and 8.9, respectively. Compared with these protoplasts, thalli had a much lower affinity for CO₂ but approximately the same affinity for HCO₃^?. Comparisons between rates of photosynthesis and the spontaneous dehydration of HCO₃^? (at 50 μM inorganic carbon) revealed that photosynthesis of both protoplasts (which lacked apparent activity of extracellular/surface-bound carbonic anhydrase) and thalli (which were only 25% inhibited by the external carbonic anhydrase inhibitor acetazolamide) could not be supported by CO₂ formation in the medium at the higher pH values, indicating HCO₃^? uptake. Since both protoplasts and thalli were sensitive to 4,4′-diisothiocyanostilbene-2,2′-disulfonate, we suggest that HCO₃^? transport was facilitated by the membrane-located anion exchange protein recently reported to function in certain Ulva thalli. These findings suggest that the presence of a cell wall may constitute a diffusion barrier for CO₂, but not for HCO₃^?, utilization under natural seawater conditions.     

GROWTH AND CARBON CONTENT OF THREE DIFFERENT‐SIZED DIAZOTROPHIC CYANOBACTERIA OBSERVED IN THE SUBTROPICAL NORTH PACIFIC1

To develop tools for modeling diazotrophic growth in the open ocean, we determined the maximum growth rate and carbon content for three diazotrophic cyanobacteria commonly observed at Station ALOHA (A Long‐term Oligotrophic Habitat Assessment) in the subtropical North Pacific: filamentous nonheterocyst‐forming Trichodesmium and unicellular Groups A and B. Growth‐irradiance responses of Trichodesmium erythraeum Ehrenb. strain IMS101 and Crocosphaera watsonii J. Waterbury strain WH8501 were measured in the laboratory. No significant differences were detected between their fitted parameters (±CI) for maximum growth rate (0.51 ± 0.09 vs. 0.49 ± 0.17 d^?1), half‐light saturation (73 ± 29 vs. 66 ± 37 μmol quanta · m^?2 · s^?1), and photoinhibition (0 and 0.00043 ± 0.00087 [μmol quanta · m^?2 · s^?1]^?1). Maximum growth rates and carbon contents of Trichodesmium and Crocosphaera cultures conformed to published allometric relationships, demonstrating that these relationships apply to oceanic diazotrophic microorganisms. This agreement promoted the use of allometric models to approximate unknown parameters of maximum growth rate (0.77 d^?1) and carbon content (480 fg C · μm^?3) for the uncultivated, unicellular Group A cyanobacteria. The size of Group A was characterized from samples from the North Pacific Ocean using fluorescence‐activated cell sorting and real‐time quantitative PCR techniques. Knowledge of growth and carbon content properties of these organisms facilitates the incorporation of different types of cyanobacteria in modeling efforts aimed at assessing the relative importance of filamentous and unicellular diazotrophs to carbon and nitrogen cycling in the open ocean.     

Maximum sustainable speed, energetics and swimming kinematics of a tropical carangid fish, the green jack Caranx caballus

Maximum sustained swimming speeds, swimming energetics and swimming kinematics were measured in the green jack Caranx caballus (Teleostei: Carangidae) using a 41 l temperature‐controlled, Brett‐type swimming‐tunnel respirometer. In individual C. caballus [mean ±s.d. of 22·1 ± 2·2 cm fork length (L_F), 190 ± 61 g, n = 11] at 27·2 ± 0·7° C, mean critical speed (U_crit) was 102·5 ± 13·7 cm s^?1 or 4·6 ± 0·9 L_F s^?1. The maximum speed that was maintained for a 30 min period while swimming steadily using the slow, oxidative locomotor muscle (U_max,c) was 99·4 ± 14·4 cm s^?1 or 4·5 ± 0·9 L_F s^?1. Oxygen consumption rate (M in mg O₂ min^?1) increased with swimming speed and with fish mass, but mass‐specific M (mg O₂ kg^?1 h^?1) as a function of relative speed (L_F s^?1) did not vary significantly with fish size. Mean standard metabolic rate (R_S) was 170 ± 38 mg O₂ kg^?1 h^?1, and the mean ratio of M at U_max,c to R_S, an estimate of factorial aerobic scope, was 3·6 ± 1·0. The optimal speed (U_opt), at which the gross cost of transport was a minimum of 2·14 J kg^?1 m^?1, was 3·8 L_F s^?1. In a subset of the fish studied (19·7–22·7 cm L_F, 106–164 g, n = 5), the swimming kinematic variables of tailbeat frequency, yaw and stride length all increased significantly with swimming speed but not fish size, whereas tailbeat amplitude varied significantly with speed, fish mass and L_F. The mean propulsive wavelength was 86·7 ± 5·6 %L_F or 73·7 ± 5·2 %L_T. Mean ±s.d . yaw and tailbeat amplitude values, calculated from lateral displacement of each intervertebral joint during a complete tailbeat cycle in three C. caballus (19·7, 21·6 and 22·7 cm L_F; 23·4, 25·3 and 26·4 cm L_T), were 4·6 ± 0·1 and 17·1 ± 2·2 %L_T, respectively. Overall, the sustained swimming performance, energetics, kinematics, lateral displacement and intervertebral bending angles measured in C. caballus were similar to those of other active ectothermic fishes that have been studied, and C. caballus was more similar to the chub mackerel Scomber japonicus than to the kawakawa tuna Euthynnus affinis.     

COMPARATIVE PHOTOSYNTHESIS OF TWO SPECIES OF INTERTIDAL EPIPHYTIC MACROALGAE ON MANGROVE ROOTS DURING SUBMERSION AND EMERSION

Photosynthesis and dark respiration rates were measured in water and in air, and the capacity to recover photosynthetic activity from emersion stress was examined for two species of intertidal, epiphytic macroalgae—Bostrychia calliptera (Montagne) Montagne and Caloglossa leprieurii (Montagne) J. Agardh—collected on prop roots of the red mangrove Rhizophora mangle L. in Buenaventura Bay, Pacific coast of Colombia. In both species, net photosynthetic rates were significantly higher under submersed conditions. Maximum photosynthetic rates (P_max) in water and in air were highest in B. calliptera, 126 ± 4 versus 52 ± 9 μmol O₂·mg chl a⁻¹·h⁻¹, respectively. In C. leprieurii, P_max of submerged plants in water and in air were 98 ± 9 versus 30 ± 11 μmol O₂·mg chla⁻¹·h⁻¹. The photoinhibition model of Platt et al. (1980) was used to fit the experimental data in both water and air for both species. Photoinhibition occurred at irradiance as low as 200 μmol·m⁻²·s⁻¹. The photosynthesis–light response curves demonstrated an adaptation to shaded habitats for both species, as light compensation points in water and air for both species were below 17 ± 5 μmol·m⁻²·s⁻¹. The rate of dehydration was significantly lower in thalli of B. calliptera compared to C. leprieurii. An increase of photosynthetic activity in B. calliptera was evident between 5% and 15% water loss, but rates decreased thereafter with declining water content. In C. leprieurii, desiccation negatively influenced photosynthetic rates that significantly decreased linearly with declining water content. In B. calliptera, net photosynthesis reached zero only at a water content between 29% and 35%, whereas in C. leprieurii no net photosynthesis occurred in plants containing less than about 50% of their relative water content. Resubmerged plants ofB. calliptera exhibited 100% photosynthetic recovery after 45 min, whereas C. leprieurii recovered 100% at about 120 min. On the basis of the comparison of rates of light-saturated net photosynthesis for B. calliptera in air versus in water, aerial photosynthetic activity ranged from 35% to 42% of that in water, whereas the emersed photosynthetic capacity of C. leprieurii ranged from 24% to 29% of that in water. Using tidal predictions and the emersed photosynthetic rates, a carbon balance model was constructed for both species over a single daylight period. The calculations indicated that emersed photosynthesis increased average daily carbon production of B. calliptera by 17% and C. leprieuri by 12%. The physiological responses to desiccation stress and the photosynthetic recovery capacities between species correlated with, and may determine, their vertical distribution in the mangrove habitats of Buenaventura Bay.     

Cholecystokinin and tryptic activity in the gut and body of developing Atlantic halibut larvae: evidence for participation in the regulation of protein digestion

At 7 days after first feeding (DAFF), the peptide hormone cholecystokinin (CCK) content (fmol individual^?1) and the tryptic activity [μmol arginine‐methyl‐coumarinyl‐7‐amide (MCA) min^?1 individual^?1] per individual gut of Atlantic halibut Hippoglossus hippoglossus larvae were low: 0·2 ± 0·1 and 0·14 ± 0·10, respectively. Thereafter, both parameters increased with the increase in gut mass and reached 19·67 ± 5·58 and 2·71 ± 0·64 at 26 DAFF, respectively. Due to the small sample size, the dry mass (M_G, mg) of the individual gut could not be determined accurately at 7 DAFF. At 13 DAFF M_G represented 5·5% of whole body dry mass (M_w, mg) while at 26 DAFF it had increased to 23%. The mass specific tryptic activity [μmol MCA min^?1 per mg dry mass (M)] in the gut increased from 2·74 ± 1 ± 98 at 13 DAFF to 5·00 ± 0·78 at 26 DAFF. There was more individual variation in the mass specific CCK content (fmol M^?1) but no significant differences were found, although the data indicated an increase (from 23·38 ± 11·26 at 13 DAFF to 36·27 ± 8·96 fmol M^?1 at 26 DAFF). At 7 DAFF the CCK content of the gut represented c. 2% of the whole body CCK content while it increased to c. 62% of the whole body CCK content at 26 DAFF. This demonstrates that it is necessary to separate neural and gastrointestinal sources of CCK in order to determine its alimentary role in fish larvae. Trypsin activity was only found in the gut compartment. In larvae aged 45 DAFF dietary proteins delivery into the gut by tube‐feeding appeared to stimulate post‐prandial secretion of CCK from the gut as well as stimulate pancreatic trypsin secretion, suggesting that both factors contribute to protein digestion.