Kinetics of nitrogen- or phosphorus-limited growth and effects of growth conditions on nutrient uptake inChattonella antiqua

Chattonella antiqua was grown in a nitrogen- or phosphorus-limited semicontinuous culture system. Using the cells in steady growth state, the relationship between growth rate and cell quota and effects of growth conditions on nitrate, ammonium and phosphate uptake were examined. Under nitrogen-limited conditions, growth rate as a function of nitrogen cell quota followed the empirical Droop equation and the uptake of nitrate and ammonium was not significantly affected by growth rate. Similarly, under phosphorus-limited conditions, the growth rate as a function of phosphorus cell quota also followed the Droop equation and phosphate uptake was not significantly affected by growth rate.Combining the results obtained in the present study with those from previous studies on nutrient uptake, half saturation constants for growth (K _g ) were calculated for nitrate, ammonium and phosphate. Comparisons ofK _g with nutrient concentrations in the Seto Inland Sea in summer, where red tides ofC. antiqua often occur, suggest that phosphate is one of the controlling factors for the population ofC. antiqua.     

The effects of light and nutrients on rates of ammonium transformation in a eutrophic river

Rates for nitrification, phytoplankton uptake of ammonium, and regeneration of ammonium were measured in the Delaware River as functions of irradiance and nutrient concentrations, using ¹⁵N labeling methods. Phytoplankton uptake increased and nitrification rates declined with increased light intensity. The irradiance level required for maximum uptake by phytoplankton was similar to that for maximal inhibition of nitrification (about 300μEm⁻² s⁻¹). Daily, water-column averaged rates, calculated by integration of the observed rate-intensity relationships, indicate that light plays a key role in regulating the balance between oxidation of NH₄⁺ by bacteria and assimilation by phytoplankton in the Delaware. The results show that uptake of ammonium by phytoplankton in the dark may exceed uptake in the light in optically thick systems.     

Chemical environment for red tides due toChattonella antiqua in the seto inland sea,Japan

Severe red tides due toChattonella antiqua occur sporadically during summer in the Seto Inland Sea, Japan, and cause significant damage to the fishing industry. In order to assess the chemical environment with respect to the outbreak ofC. antiqua, environmental factors that affect the growth ofC. antiqua were monitored around the Ie-shima Islands, the Seto Inland Sea, in the summer of 1986. In addition, a growth bioassay of the seawater usingC. antiqua was conducted under a semicontinuous culture system. Although temperature, salinity and light intensity were optimum for the growth ofC. antiqua, red tides by this species did not occur. Concentrations of NH ₄ ⁺ , NO ₃ ^? and PO ₄ ^3? were low (<0.4, <0.2, <0.06 µM, respectively) above the thermocline (8–12 m) and high below it (0.6–2, 4–8, 0.4–0.8 µM, respectively). Vitamin B₁₂ concentrations did not change significantly between the surface (0 m) and below the thermocline (25 m) in the level of 2–4 ng·l^?1. The growth bioassay revealed that in the surface waters, concentrations of N- as well as P- nutrients were too low to support a rapid growth ofC. antiqua. At the depth of 25 m, neither N, P nor B₁₂ limited the growth rate. In order to obtain more quantitative information on the growth rate as a function of the concentrations of N- and P- nutrients,C. antiqua was grown in a semicontinuous culture system by changing nutrient concentrations systematically. The observed growth rate (μ) can be approximated as follows: $$\mu = \mu _{\max } .\frac{{S_N }}{{K_g ^N + S_N }}.\frac{{S_{PO4} }}{{K_g ^P + S_{PO4} }},$$ whereS _N is the concentration of NO ₃ ^? plus NH ₄ ⁺ (0–6 µM),S _PO, the concentration of PO ₄ ^3? (0–0.6 µM), μ_max (0.97 d^?1) the maximal growth rate,K ₀ ^N (1.0 µM) andK ₀ ^P (0.11 µM) the half saturation constants for NO ₃ ^? and PO ₄ ^3? , respectively. Using the above equation with nutrient concentrations measured, the rate at which seawater supports the growth ofC. antiqua can be estimated and this can be used for the assessment of chemical environments with respect to the outbreak ofC. antiqua.     

Chemical environment for red tides due toChattonella antiqua

Environmental parameters that affect the growth ofChattonella antiqua were monitored throughout the outbreak period of this species around the Ie-shima Islands, the Seto Inland Sea, in the summer of 1987 (20 July–13 August). Averaged cell concentration ofC. antiqua over the water column (21 m) was below 10 cells· ml^–1 on 20 July, gradually increased to reach the maximum of 250 cells·ml^–1 on 7 August, and then rapidly decreased to the value of 30 cells·ml^–1 on 13 August.Thermal stratifications were prominent from 20 July to 3 August and were destroyed after 4 August. Temperature and salinity were optimum for the growth ofC. antiqua throughout the survey period.At the bloom initiation period (20–21 July), concentrations of N- and P-nutrients (S _N andS _P ) were high throughout the water column. From 22 July to 3 August, whenC. antiqua increased its populations,S _N andS _P at the depth of 0–5m were low but those at the depth of 10–20m kept a high value. After 4 August,S _n andS _P at the depth of 10–20m decreased rapidly due to wind mixing coupled with the nutrient uptake byC. antiqua. When the populations ofC. antiqua reached the maximum (7–9 August), N-nutrients were depleted throughout the water column but P-nutrients were not. Concentrations of vitamin B₁₂ were almost in the same range as those of the previous years and were optimum for the growth ofC. antiqua.GP- value (growth potential of the seawater with respect to nitrogen and phos-phorus) was higher than 0.6 even at the surface layer (0–5 m) at the bloom-initiation period. During the bloom development period (22 July–3 August), GP at the surface layer (0–5m) was low (<0.2), but GP at the depth of 10–20m kept a rather high value (>0.4).In situ growth rates ofC. antiqua at the depth of 0 and 5m estimated from bottle experiments coincided well with the values expected from GP. A high value of GP at the surface layer in the initiation period and a shallow GP-cline in the development period, combined with the ability of diurnal vertical migration seemed to be at least one reason that natural populations ofC. antiqua grew at a rather high rate and formed red tides in the summer of 1987.     

Ammonium uptake kinetics and interactions between nitrate and ammonium uptake inChattonella antiqua

Ammonium uptake kinetics and interactions between nitrate and ammonium uptake were examined inChattonella antiqua. After the addition of ammonium to the culture ofC. antiqua, the ammonium concentration decreased linearly with time. The ammonium uptake rate as a function of ammonium concentration followed the Michaelis-Menten equation; the maximal uptake rate was 2.0 pmol cell^–1hr^–1 and the half saturation constant, 2.2M. Although the ammonium uptake was not affected by nitrate, uptake of nitrate was rapidly (15min) suppressed by ammonium and a 50% reduction in nitrate uptake was observed at an ammonium concentration ofca. 2M.     

Nickel utilization in phytoplankton assemblages from contrasting oceanic regimes

In most oceanic environments, dissolved nickel (Ni) concentrations are drawn down in surface waters with increasing concentrations at depth, implying a role for biology in the geochemical distribution of Ni. Studies with phytoplankton isolates from the surface ocean have established the biochemical roles of Ni in the assimilation of urea and oxidative defense. To determine if these requirements are relevant in natural marine planktonic assemblages, bottle-based fertilization experiments were used to test the effects of low-level additions of Ni, urea, or both Ni and urea to surface waters at several locations offshore of Peru and California, as well as in the Gulf of California. Urea and Ni+urea additions consistently promoted phytoplankton growth relative to control and +Ni treatments, except in a coastal upwelling site and Peruvian water. No effect was observed in the upwelling site, but in Peruvian waters urea additions resulted in increased phytoplankton pigments and phosphate drawdown only when Ni was added concurrently, suggesting a biochemically dependent Ni–urea colimitation. In the Gulf of California, Ni additions without urea resulted in increased abundances of cyanobacteria, picoeukaryotes, and the corresponding pigments. As urea additions showed the overall phytoplankton community was also urea-limited, it appears that the cyanobacteria and potentially the picoeukaryotes were colimited by Ni and urea in a biochemically independent fashion. In parallel, radiotracer-based uptake experiments were used to study the kinetics and spatial variation of biological Ni assimilation. In these experiments, the added radiotracer rarely equilibrated with the natural Ni present, precluding estimates a determination of in situ Ni uptake rates and suggesting that much of the natural Ni was not bioavailable. The lack of equilibration likely did not preclude the measurement of community Ni uptake kinetics, nor the comparison of measured rates between locations. The highest V_maxK_ρ^?1 values, which reflect a competitive advantage in Ni acquisition at low concentrations, were observed in stratified nitrogen-deplete communities, potentially linking Ni and nitrogen biogeochemistry in a manner consistent with the biochemical utilization of Ni. Overall, uptake rates were higher in the euphotic rather than non-euphotic zone communities, directly reconciling the nutrient-like depth profile of Ni. The Ni uptake rates observed at the nitrate-replete Fe-deplete Peru stations were an order of magnitude lower than the other sites. This result agrees with calculations suggesting that saturation of the cell surface with Ni and iron (Fe) transporters may limit uptake rates in low Fe waters.     

Size-fractionated biological iron and carbon uptake along a coastal to offshore transect in the NE Pacific

Iron has been shown to limit phytoplankton growth in high-nutrient low-chlorophyll (HNLC) regions such as the NE subarctic Pacific. We report size-fractionated Fe-uptake rates by the entire plankton community in short (6–8 h) light and dark incubations along an E–W transect from P04 (a coastal ocean station) to OSP (an open-ocean HNLC station) during August–September 1997. Size-fractionated primary productivity and chl a were measured to monitor algal Fe : C uptake ratios and Fe-uptake relative to phytoplankton biomass. The >5.0 μm size-class, which consisted mostly of large diatoms, had the highest Fe-uptake rate at nearshore stations (P04 and P8), but Fe-uptake rates for this size class decreased despite increases in biomass and primary productivity when transecting westwards to HNLC waters. Fe-uptake rates of the small size class (0.2–1.0 μm, including heterotrophic bacteria and autotrophs) were inversely related to the >5.0 μm size-class uptake rates, in that stations with high dissolved Fe (DFe) concentrations had relatively low uptake rates compared to those in the low-Fe offshore region. The 1.0–5.0 μm size-class Fe-uptake rates were low, relatively invariant along the transect, and differed little between light and dark incubations. Dark Fe-uptake rates averaged 10–20% less than those in the light for the >5.0 μm size class. Dark uptake rates however, were higher than light uptake rates for the 0.2–1.0 μm size class at all stations. Fe : C uptake ratios were high for all size classes at P04, but decreased as DFe concentrations decreased offshore. The prokaryote-dominated 0.2–1.0 μm size class had the highest Fe : C uptake ratios at all stations. These data suggest that prokaryotic organisms make an important contribution to biological Fe uptake in this region. Our experiments support the results of previous culture work, suggesting higher Fe : C ratios in coastal phytoplankton compared to open-ocean species, and demonstrate that light can have a large effect on Fe partitioning between size classes in subarctic Pacific HNLC waters.     

Temporal and spatial variability in phytoplankton ammonium and nitrate uptake in the Delaware Estuary

Phytoplankton NH₄⁺ and NO₃⁻ uptake was examined along the longitudinal salinity gradient of the Delaware Estuary over several seasonal cycles using ¹⁵N-tracer techniques. Saturated nitrogen uptake rates increased directly with water temperature and reached a maximum of 380 nmol Nl⁻¹h⁻¹ during summer. This temperature dependence was related primarily to changes in the rate of maximum chlorophyll specific uptake, which varied exponentially between 2 and 70 nmol N [μg Chl h]⁻¹ over a temperature range of 2–28°C. Despite these high uptake rates, balanced growth (C:N7:1) could be maintained over the diel light cycle only by highly efficient nitrogen uptake at low light intensities and dark uptake below the photic zone and at night (dark UPTAKE=25% maximum uptake).Ammonium fulfilled 82% of the annual phytoplankton nitrogen demand in the estuary despite dominance of NO₃⁻ in the ambient dissolved inorganic nitrogen pool. The predominance of NH₄⁺ uptake occurred because of the general suppression of NO₃⁻ assimilation at NH₄⁺ concentrations in excess of 2 μ . This suppression, however, was not as universal as has been reported for other systems, and it is suggested that the extremely high NO₃⁻ concentrations found in the estuary contribute to this pattern. Nitrate was a significant source of nitrogen only during periods of high phytoplankton production in summer, and when NH₄⁺ concentrations were low towards the end of the spring bloom.     

Simulation of upper-ocean biogeochemistry with a flexible-composition phytoplankton model: C, N and Si cycling in the western Sargasso Sea

We report the first application of a biogeochemical model in which the major elemental composition of the phytoplankton is flexible, and responds to changing light and nutrient conditions. The model includes two phytoplankton groups: diatoms and non-siliceous picoplankton. Both fix C in accordance with photosynthesis-irradiance relationships used in other models and take up NO₃⁻ and NH₄⁺ (and Si(OH)₄ for diatoms) following Michaelis-Menten kinetics. The model allows for light dependence of photosynthesis and NO₃⁻ uptake, and for the observed near-total light independence of NH₄⁺ uptake and Si(OH)₄ uptake. It tracks the resulting C/N ratios of both phytoplankton groups and Si/N ratio of diatoms, and permits uptake of C, N and Si to proceed independently of one another when those ratios are close to those of nutrient-replete phytoplankton. When the C/N or Si/N ratio of either phytoplankton group indicates that its growth is limited by N, Si or light, uptake of non-limiting elements is controlled by the content of the limiting element in accordance with the cell-quota formulation of Droop (J. Mar. Biol. Ass. U.K 54 (1974) 825).We applied this model to the Bermuda Atlantic Time-series Study (BATS) site in the western Sargasso Sea. The model was tuned to produce vertical profiles and time courses of [NO₃⁻], [NH₄⁺] and [Si(OH)₄] that are consistent with the data, by adjusting the kinetic parameters for N and Si uptake and the rate of nitrification. The model then reproduces the observed time courses of chlorophyll-a, particulate organic carbon and nitrogen, biogenic silica, primary productivity, biogenic silica production and POC export with no further tuning. Simulated C/N and Si/N ratios of the phytoplankton indicate that N is the main growth-limiting nutrient throughout the thermally stratified period and that [Si(OH)₄], although always limiting to the rate of Si uptake by diatoms, seldom limits their growth rate. The model requires significant nitrification in the upper 200 m to yield realistic time courses and vertical profiles of [NH₄⁺] and [NO₃⁻], suggesting that NO₃⁻ is not supplied to the upper water column entirely by physical processes. A nitrification-corrected f-ratio (f_NC), calculated for the upper 200 m as: (NO₃⁻ uptake—nitrification)/(NO₃⁻ uptake+NH₄⁺ uptake) has annual values ranging from only 0.05–0.09, implying that 90–95% of the N taken up annually by phytoplankton is supplied by biological regeneration (including nitrification) in the upper 200 m. Reported discrepancies between estimates of organic C export based on seasonal chemical changes and POC export measured at the BATS site can be almost completely resolved if there is significant regeneration of NO₃⁻ via organic-matter decomposition in the upper 200 m.     

Growth characteristics ofChattonella antiqua (Raphidophyceae)

Chattonella antiqua (Raphidophyceae), which causes heavy red tides in the Seto Inland Sea, Japan, was placed in axenic clonal culture by micropipette washing. The effects of temperature, salinity, light intensity and pH on growth were monitored. Maximum growth occurred at 25°C, at salinities between 25 and 41‰, under light intensities above 0.04 ly min^?1. The pH effect was not significant in the pH range from 7.6 to 8.3. Comparisons of our results with those from field observations suggest that the development of theC. antiqua red tide is strongly temperature dependent.     

Chemical environment for red tides due toChattonella antiqua

In order to assess the roles of Fe and Cu in outbreaks ofChattonella antiqua red tide, concentrations of these metals in the surface seawater were monitored around the Ie-shima Islands in the Seto Inland Sea during the summers of 1986–1988. Bioassay of the surface seawater with respect to Fe and Cu was also conducted using a cultured strain ofC. antiqua.Concentrations of Fe and Cu in the filtered seawater (Fe^F and Cu^F) were in the range of 3.9–10.0 and 9.3–11.2 nM, respectively. The bioassay with respect to Fe revealed that Fe at the surface layer was usually insufficient to support the maximum growth rate ofC. antiqua, except whenC. antiqua was dominant in the field. However, correlations between Fe^F and the growth rate of the control cultures (Fe, EDTA=not enriched; N, P, B₁₂=enriched at optimum levels) were not apparent, probably because Fe^F did not reflect the concentration of available Fe.The bioassay with respect to Cu was coupled with the Cu^F values obtained. The results indicated that Cu at the surface layer was detoxified by complexation with natural organic ligand(s), and that pCu (=minus log of cupric ion activity) was 11.5–11.7, optimum for the growth ofC. antiqua, throughout the survey period. It is suggested that Fe, but not Cu, is a potentially important factor in regulating the natural populations ofC. antiqua in the Seto Inland Sea.     

Effects of simulated benthic fluxes on phytoplankton dynamic and photosynthetic parameters in a mesocosm experiment (Bay of Brest,France)

Benthic faunal activity and density play an important role in determining the rates of benthic nutrient fluxes, which enrich the water column and contribute to phytoplankton growth. The intensity of nutrient fluxes in the Bay of Brest depends on the density of the invasive gastropod, Crepidula fornicata. In order to study the impact of benthic fluxes on phytoplankton dynamics, realistic daily nutrient inputs simulating various densities of C. fornicata were added to six enclosures during three weeks. The increase in fertilization intensity influenced the phytoplankton biomass. A succession from Chaetoceros spp. to Pseudo-nitzschia spp. and Leptocylindrus danicus was observed in all enclosures, but the dynamics of successions were different. Pseudo-nitzschia spp. was favored in the three more fertilized enclosures, while Chaetoceros spp. persisted longer in less enriched enclosures. Despite an apparent nitrogen limitation, the quantum efficiency of PSII (F_v/F_m) was high (>0.5) and stable in all enclosures. The maximal photosynthetic capacity (P^Bmax) was also invariable and oscillated around an average value of 2.23 mg C (mg Chl a)⁻¹ h⁻¹. The stability of F_v/F_m and P^Bmax observed at different nutrient input intensities demonstrates that the daily inputs maintained the physiological balance of the microalgae. The maximal light utilization efficiency (α) and the light saturation parameter (E_k) were also quite stable after day 8, which reveals that photosynthetic parameters were driven by growth constraints due to nutrient availability and not by incident light or species successions. We suggest that our results correspond to an “E_k independent variation” regulation. We propose that such regulation of photosynthetic parameters appears when there are frequent nutrient additions which do not allow replete nutrient conditions to be reached but lead to physiological equilibrium.     

Seasonal and interannual variability in phytoplankton and nutrient dynamics along Line P in the NE subarctic Pacific

We analysed mixed-layer seasonal and interannual variability in phytoplankton biomass and macronutrient (NO₃ and Si(OH)₄) concentrations from three decades of observations, and nitrogen uptake rates from the 1990s along Line P in the NE subarctic Pacific. Chlorophyll a concentrations near 0.35 mg m⁻³ were observed year-round along Line P except at the nearshore station (P4) where chlorophyll a concentrations in spring were on average 2.4 times the winter values. In contrast, the temporal variability in carbon-to-chlorophyll ratios at the two main end members of Line P (P4 and OSP) was high. Large seasonal and interannual variability in NO₃ and Si(OH)₄ concentration were observed along Line P. Highest upper mixed-layer (top 15 m) nutrient concentrations occurred on the continental shelf in late summer and early fall due to seasonal coastal upwelling. Beyond the shelf, maximum nutrient concentrations increased gradually offshore, and were highest in late winter and early spring due to mixing by winter storms. Interannual variations in upper mixed-layer nutrient concentrations beyond the shelf (>128°W) were correlated with E-W winds and the PDO since 1988 but were not correlated with either climate index between 1973 and 1981. Despite differences in nutrient concentration, nutrient utilization (ΔNO₃ and ΔSi(OH)₄) during the growing season were about 7.5 μM at all offshore stations. Variations in ΔNO₃ were correlated with those of ΔSi(OH)₄. The annual cycle of absolute NO₃ uptake (ρNO₃) and NH₄ uptake (ρNH₄) rates by phytoplankton in the upper mixed-layer showed a weak increasing trend from winter to spring/summer for the period 1992-1997. Rates were more variable at the nearshore station (P4). Rates of ρNO₃ were low along the entire line despite abundant NO₃ and low iron (Fe), at the offshore portion of Line P and sufficient Fe at the nearshore station (P4). As a result, new production contributed on average to only 32 ± 15% of the total nitrogen (N) uptake along Line P. NO₃ utilization in the NE subarctic Pacific is probably controlled by a combination of environmental variables, including Fe, light and ambient NH₄ levels. Elevated ambient NH₄ concentrations seem to decrease the rates of new production (and f-ratios) in surface waters of the oceanic subarctic NE Pacific. Contrary to expectation, phytoplankton biomass, nutrient utilization (ΔNO₃ and ΔSi(OH)₄), and nitrogen uptake (ρNO₃ + ρNH₄) varied relatively little along Line P, despite significant differences in the factors controlling phytoplankton composition assemblages and production. Future studies would benefit from including other variables, especially light limitation, to improve our understanding of the seasonal and interannual variability in phytoplankton biomass and nutrients in this region.     

Phytoplankton spring bloom of the Gironde plume waters in the Bay of Biscay: early phosphorus limitation and food-web consequences

During the spring 1995 (2–25 May), a cruise was carried on the RV Poseidon (Germany) on the continental shelf of the south Bay of Biscay. The objective was a comprehensive study of the planktonic food web within the Gironde plume waters. In these waters phosphate was present at very low concentrations (undetectable to < 0.1 μmol.L⁻¹), whereas nitrate, silicate and ammonium concentrations were much higher (several μmol·L⁻¹ for nitrate and silicate and 0.5 to 1.0 μmol·L⁻¹ for ammonium). The size distribution of the phytoplankton biomass (estimated from chlorophyll a measurements by high performance liquid chromatography) and primary production (measured by ¹⁴C in situ method) showed a great proportion of small (40 to 70 % < 3 μm) and active autotrophic cells (growth rates estimated from 0.4 to 0.8 d⁻¹ for the entire euphotic layer). Considering the very high values of NO₃-N:PO₄-P ratios and the high C:P and N:P ratios for the particulate organic matter, it is suggested that an early phosphorus depletion limits the spring bloom phytoplankton and particularly the new production (nitrate uptake coming from the Gironde waters).From these results and other simultaneous observations on the heterotrophic processes (such as grazing of microzooplankton), we can conclude that the planktonic food web would be close to a maintenance system as defined by Platt et al. The possible generalisation of these results for each spring is discussed with respect to the scarcity of previous and reliable phosphate data.     

Early spring bloom phytoplankton-nutrient dynamics at the Celtic Sea Shelf Edge

Phytoplankton production was measured at the shelf edge region of the Celtic Sea in April/May 1994 at the beginning of the spring bloom. Size fractionated ¹⁴C uptake experiments showed that phytoplankton >2 μm dominated the bloom although, in the period immediately before the increase in phytoplankton biomass, picophytoplankton (<2 μm) was responsible for up to 42% of the production; in these late winter conditions, chlorophyll concentrations were generally <0.7 μg l^-1 and primary production was ca. 70 mmol C m^-2 d^-1. As the spring bloom developed, phytoplankton production rates of 120 mmol C m^-2 d^-1 were measured. Chlorophyll concentration increased to >2 μg l^-1 as a result of growth of larger phytoplankton, including diatoms, with large numbers of Nitzschia, Thalassionema and Chaetoceros dominating the assemblage. Picophytoplankton production declined as the spring bloom progressed. Nutrient concentrations were not depleted during the sampling period, and NO^-₃ concentrations were >6 μmol l^-1. Nutrient assimilation rates were measured at the same time as primary production was estimated. Before the development of any substantial phytoplankton biomass, the uptake rates for ammonium and nitrate were very similar, with f-ratios ranging from 0.5 to 0.6. Assimilation of ammonium remained relatively constant after the onset of stratification and bloom development, but nitrate uptake increased by a factor of 2 or more, resulting in f-ratios >0.8. There was significant phosphate uptake in the dark, which was generally ca. 50% of the rate in the light. The C : N : P assimilation ratios changed as the bloom developed; in the pre-bloom situation, when small phytoplankton cells dominated the assemblage, the C : N assimilation ratio was variable, with some stations having ratios less than (ca 2.5), and some higher than (ca. 9), the Redfield ratio. The most actively growing assemblages had N : P ratios close to the Redfield ratio, but the C : N ratios were consistently lower. New production was found to be closely correlated with the size of the species making up the phytoplankton assemblage, and high f ratios were measured when larger phytoplankton dominated the assemblage.     

Close coupling between ammonium uptake by phytoplankton and excretion by Antarctic krill, Euphausia superba

In this study we examined the hypothesis that, under conditions of replete macronutrients and iron in the Southern Ocean, phytoplankton abundance and specific N uptake rates are influenced strongly by the processes of grazing and NH₄ regeneration. NH₄ and NO₃ uptake rates by marine phytoplankton were measured to the northeast and northwest of the island of South Georgia during January-February 1998. Mean specific uptake rate for NO₃ (vNO₃) was 0.0026 h⁻¹ (range 0.0013-0.0065 h⁻¹) and for NH₄ (vNH₄) was 0.0097 h⁻¹ (0.0014-0.0376 h⁻¹). vNH₄ was related positively with NH₄ availability, which ranged from 0.1 to 1.5 mmol m⁻³ within the upper mixed layer. Ambient NH₄ concentrations and vNH₄ were both positively related to local krill biomass values, computed from mean values along acoustic transect segments within 2 km of the uptake measurement stations. These biomass values ranged from ∼1 g krill fresh mass m⁻² in the northwest to >4 kg krill wet mass m⁻² in the northeast. In contrast to the variability found with NH₄ concentrations and uptake rates, vNO₃ was more uniform across the sampling sites. Under these conditions, increasing NH₄ concentration appeared to represent an additional N resource. However, high vNH₄ tended to be found for stations with lower phytoplankton standing stocks, across a total range of 0.24-20 mg chlorophyll a m⁻³. These patterns suggest a coupling between phytoplankton biomass, vNH₄ and krill in this region of variable but high krill biomass. Locally high concentrations of krill in parts of the study area appeared to have two opposing effects. On the one hand they could graze down phytoplankton stocks, but on the other hand, their NH₄ excretion supported enhanced uptake rates by the remaining, ungrazed cells.     

Distribution and photo-physiological condition of phytoplankton in the tropical and subtropical North Pacific

To better understand the vertical distribution of phytoplankton in the tropical and subtropical North Pacific, we used fast repetition rate fluorometry to investigate the photo-physiological condition of the phytoplankton assemblage in this region between February and March 2007. Along 155°E, between the equator and 24°N, the peak of fluorescence (F _m), an indication of the deep chlorophyll maximum (DCM), was deeper than the top of the nitracline and occurred at the 2.4 ± 1.3 % (mean ± SD) light depth (relative to 0 m). The photochemical efficiency (F _v/F _m) and effective absorption cross-section of photosystem II (σ_PSII) were low at the surface but increased rapidly at depths between the top of the nitracline (40–138 m) and the DCM (70–158 m), an indication that the photo-physiological condition of the phytoplankton improved below the top of the nitracline. The depth of the maximal F _v/F _m [Z(F _v/F _{m max})] was 18–32 m deeper than the DCM and corresponded to the 0.8 ± 0.2 % light depth. The values of F _v/F _m at the Z(F _v/F _{m max}) were 20 % higher than those at the DCM and averaged 0.48 ± 0.01. These results suggest that the phytoplankton assemblage beneath the DCM had a high potential photosynthetic performance capacity and was growing by using the very low ambient light in this region.     

Growth characteristics ofChattonella antiqua

Nutrient requirements of a red tide flagellate,Chattonella antiqua, were investigated in a laboratory culture experiment. Growth ofC. antiqua was supported by nitrate and ammonium, and by urea to a limited extent, but not by glycine, alanine and glutamate. Orthophosphate served as a good phosphorus source but glycerophosphate did not. Fe³⁺ (1µM) fully promoted the flagellate's growth in the presence of 80µM of EDTA. The addition of Mn²⁺ (0–20µM), Zn²⁺ (0–10µM) and Co²⁺ (0–0.4µM) did not show any effect. Among three vitamins tested, only B₁₂ (6 ng 1^?1) served as a growth factor. Glucose, acetate and glycolate did not improve growth in the light nor did they support growth in darkness. The minimum cell quotas for nitrogen, phosphorus, iron and B₁₂ were estimated to be 11 pmoles ce^?1, 1.0, ～0.09 and 1.1 fg cell^?1, respectively.     

Carbon fluxes through major phytoplankton groups during the spring bloom and post-bloom in the Northwestern Mediterranean Sea

The carbon flux through major phytoplankton groups, defined by their pigment markers, was estimated in two contrasting conditions of the Northwestern Mediterranean open ocean ecosystem: the spring bloom and post-bloom situations (hereafter Bloom and Post-bloom, respectively). During Bloom, surface chlorophyll a (Chl a) concentration was higher and dominated by diatoms (53% of Chl a), while during Post-bloom Synechococcus (42%) and Prymnesiophyceae (29%) became dominant. The seawater dilution technique, coupled to high pressure liquid chromatography (HPLC) analysis of pigments and flow cytometry (FCM), was used to estimate growth and grazing rates of major phytoplankton groups in surface waters. Estimated growth rates were corrected for photoacclimation based on FCM-detected changes in red fluorescence per cell. Given the 30% average decrease in the pigment content per cell between the beginning and the end of the incubations, overlooking photoacclimation would have resulted in a 0.40 d^?1 underestimation of phytoplankton growth rates. Corrected average growth rates (μ_o) were 0.90±0.20 (SD) and 0.40±0.14 d^?1 for Bloom and Post-bloom phytoplankton, respectively. Diatoms, Cryptophyceae and Synechococcus were identified as fast-growing groups and Prymnesiophyceae and Prasinophyceae as slow-growing groups across Bloom and Post-bloom conditions. The higher growth rate during Bloom was due to dominance of phytoplankton groups with higher growth rates than those dominating in Post-bloom. Average grazing rates (m) were 0.58±0.20 d^?1 (SD) and 0.31±0.07 d^?1. The proportion of phytoplankton growth consumed by microzooplankton grazing (m/μ_o) tended to be lower in Bloom (0.69±0.34) than in Post-bloom (0.80±0.08). The intensity of nutrient limitation experienced by phytoplankton indicated by μ_o/μ_n (where μ_n is the nutrient-amended growth rate), was similar during Bloom (0.78) and Post-bloom (0.73). Primary production from surface water (PP) was estimated with ¹⁴C incubations. A combination of PP and Chl a synthesis rate yielded C/Chl a ratios of 34±21 and 168±75 (g:g) for Bloom and Post-bloom, respectively. Transformation of group-specific Chl a fluxes into carbon equivalents confirmed the dominant role of diatoms during Bloom and Synechococcus and Prymnesiophyceae during Post-bloom.     

Studies of nitrogen assimilation by marine phytoplankton and its implication in the nitrogen cycling in the sea

I present here a review of my work concerning nitrogen assimilation by marine phytoplankton. This opportunity was provided to me as the recipient of the Okada Prize for 1990 from the Oceanographical Society of Japan. Assimilation of nitrogenous nutrients by phytoplankton has received considerable research effort since it is an essential process in organic matter production in the sea surface. The use of¹⁵N technique is necessary for tracing nitrogen assimilation by natural marine phytoplankton, but nitrogen metabolism of heterogenous natural populations significantly complicates flow of isotope. Dilution of¹⁵N isotope by heterotrophic regeneration of ammonium causes underestimates of uptake rates. I made an evaluation of isotope dilution effects in available data sets of¹⁵N-ammonium uptake experiments in literature. Incorporated¹⁵N in particulates might revert back to dissolved organic or inorganic nitrogen. I conducted pulse-chase experiments which can quantify such loss of tracer. From these studies, a short term experiment with sufficient amount of tracer enrichment is found to overwhelm these problems. In such an experiment, however, the elevation of nutrient concentration by tracer addition may likely perturb the uptake process. An initial rapid uptake is expected if the population is nitrogen deficient, but I found that this phenomenon is not common to surface oligotrophic open oceans. Uptake rate from such an experiment, or capacity of nitrogen uptake, was obtained using surface waters from an extended area in the North Pacific, and its regional variability was discussed. In addition to overall¹⁵N uptake, time series analysis of intracellular¹⁵N partitioning between hot ethanol soluble and insoluble fractions was found to be useful. When¹⁵N-ammonium is added to nitrogen deficient cells of phytoplankton,¹⁵N is accumulated in the ethanol soluble fraction. Using cultured strains of marine phytoplankton, this accumulation was proved to be caused by the difference of rates of nitrogen uptake and nitrogenous macromolecule synthesis. Uptake rate per cell is relatively constant irrespective of nutritional status, but macromolecule synthesis decreases with nitrogen deficiency. This accumulation of¹⁵N in the ethanol soluble fraction was used as an index of nutritional status with respect to nitrogen of the natural populations of phytoplankton from the western North Pacific. The uptake capacity of nitrate was observed to be higher than that of ammonium in the regional upwelling around Izu Islands and during the spring bloom in Alaskan coastal water. The¹⁵N partitioning technique revealed that nitrate taken up was rapidly incorporated in the macromolecule fraction. This suggests that ammonium uptake is suppressed to be smaller than intracellular nitrogen assimilation, rather than that nitrate is taken up in excess and accumulates within the cell. Regulation of nitrate uptake by light intensity was also discussed in detail for the Alaskan data. Several other studies currently conducted are also mentioned.