Utilization of glutamate and glucose for heterotrophic growth by the marine pennate diatom Nitzschia laevis

Nitzschia laevis Hustedt grew in the dark in the presence of either glutamate or glucose as substrate. Complex mixtures of yeast extract or tryptone plus lactate also supported good heterotrophic growth, while tryptone alone only supported very slow growth in the dark. The observed growth rates of N. laevis in the dark at different concentrations of glutamate or glucose could be accounted for by the measured uptake rates of these compounds. The affinity of the uptake systems for glutamate and glucose (K _s =0.03 mM for each) was quite high, and similar for dark- and light-grown cells. The lack of a lag-phase when cells were transferred from photoautotrophic to heterotrophic growth conditions can be explained by the presence of uptake systems for glutamate and glucose in ligh--grown cells, as well as in dark-grown cells. However, the uptake capacity was generally higher in the latter than the former. N. laevis also took up alanine and lactate according to Michaelis-Menten kinetics, with a K _s for alanine of 0.02 mM and for lactate of 0.4 mM. Malate and glycerol were not taken up to a significant extent by the cells. Cells grown in continous light had a doubling time of 18 h. The shortest doubling time observed in the dark on glutamate was 48 h and on glucose 24 h. Glutamate was used for heterotrophic growth with an efficiency of 43% and glucose with an efficiency of 48%.Contribution No., 945 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA.     

Photosynthetic characteristics of phycoerythrin-containing marine Synechococcus spp.

Marine Synechococcus spp. are sufficiently abundant to make a significant contribution to primary productivity in the ocean. They are characterized by containing high cellular levels of phycoerythrin which is highly fluorescent in vivo. We sought (Jan.–Apr., 1984) to determine the adaptive photosynthetic features of two clonal types of Synechococcus spp., and to provide a reliable physiological basis for interpreting remote sensing data in terms of the biomass and productivity of this group in natural assemblages. It was found that the two major clonal types optimize growth and photosynthesis at low photon flux densities by increasing the numbers of photosynthetic units per cell and by decreasing photosynthetic unit size. The cells of clone WH 7803 exhibited dramatic photoinhibition of photosynthesis and reduction in growth rate at high photon flux densities, accompanied by a large and significant increase in phycoerythrin fluorescence. Maximal photosynthesis of cells grown under 10–50 E m^-2 s^-1 was reduced by 20 to 30% when the cells were exposed to photon flux densities greater than 150 E m^-2 s^-1. However, steady-state levels of photosynthesis maintained for brief periods under these conditions were higher than those of cells grown continuously at high photon flux densities. No photoinhibition occurred in clone WH 8018 and rates of photosynthesis were greater than in WH 7803. Yields of in-vivo phycoerythrin fluorescence under all growth photon flux densities were lower in clone WH 8018 compared to clone WH 7803. Since significant inverse correlations were obtained between phycoerythrin fluorescence and P_max and for both clones grown in laboratory culture, it may be possible to provide a reliable means of assessing the physiological state, photosynthetic capacity and growth rate of Synechococcus spp. in natural assemblages by remote sensing of phycoerythrin fluorescence. Poor correlations between phycoerythrin fluorescene and pigment content indicate that phycoerythrin fluorescence may not accurately estimate Synechococcus spp. biomass based on pigment content alone.     

Development of the ability to take up L-lysine by the diatom Phaeodactylum tricornutum

Freshly harvested cells of Phaeodactylum tricornutum Bohlin grown with nitrate, ammonium or lysine as a sole nitrogen source had a low ability to take up lysine or arginine, but this ability increased when cells were deprived, over 48 h, of either nitrogen or carbon. The effects of nitrogen and carbon deprivation were additive, and the uptake ability was greatest in cells incubated in darkness in nitrogen-free medium. Uptake ability increased in cells illuminated in the presence of 10^-5 M 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) an inhibitor of photosynthetic electron transport. An inverse relationship between rate of development of the uptake system and rate of photosynthesis was also established. Development of the uptake system was prevented by cycloheximide or by anaerobiosis. Following transfer to a normal nitrate medium, illuminated cells lost the lysineuptake system by dilution as the cells grew. There was a linear and positive correlation between the initial rate of uptake of lysine and the maximum concentration which was maintained in the cells when equilibrium was reached, indicating that transinhibition of lysine uptake may occur and that the extent of this inhibition is related to the size of the internal amino acid pool. The relevance of the findings to the growth of phytoplankton in natural waters is discussed.     

Photosynthetic characteristics of marine Synechococcus spp. with special reference to light environments near the bottom of the euphotic zone of the open ocean

The present study aimed to resolve the question why marine Synechococcus spp. abundantly occur even at the bottom of the euphotic zone in the Kuroshio are. Photosynthesis under such conditions was examined using simulated blue-green model light (BGL). Results indicated that photosynthesis of marine Synechococcus spp. under BGL is as active enough to support growth of these organisms. Examination of light-harvesting under BGL indicated that active photosynthesis is permitted by an unusually high abundance of phycoerythrin (PE), which is the main light-harvesting pigment for photosystem II (PSII), due to a phycobilisome (PBS) structure which is different from ordinary hemidiscoidals. Although the absorption maximum of PE is located at longer wavelengths than the energy maximum of BGL, PE was found to absorb BGL significantly. Thus, BGL cannot be a typical photosystem I (PSI) light. PSII is also significantly excited by BGL. Carotenoids, which largely absorb BGL, were found to be effective in light-harvesting for PSI. Based on the results obtained, possible reasons why marine Synechococcus spp. commonly occur in warm waters were discussed. Two strains of Synechococcus spp. isolated from the Gulf Stream in 1981 and from Kuroshio, Japan in 1983 were used in the present study.     

Different responses of photosynthesis and flow cytometric signals to iron limitation and nitrogen source in coastal and oceanic Synechococcus strains (Cyanophyceae)

Iron plays an important role in marine primary productivity, and Synechococcus species as major contributors to the total photosynthetic biomass in the world’s oceans might be limited by iron supply in some regions. The present study aimed to compare the photosynthesis and flow cytometric signals of four Synechococcus strains grown under different iron concentrations with either nitrate or ammonium as the sole nitrogen source. Two oceanic strains were much more sensitive to iron limitation than two coastal strains. The inhibition of iron limitation on the growth, maximal PSII photochemical yield, maximal rate of relative electron transport and photochemical quenching of the two oceanic strains was higher than for their coastal counterparts. Under iron limitation condition, the connectivity factor between individual photosynthetic units (ρ) increased for the two coastal strains, while decreased for the two oceanic strains. Furthermore, iron limitation accelerated the Q _A re-oxidation of the two oceanic strains and the PQ pool re-oxidation of the two coastal strains. Under iron limitation condition, the cell size of the two coastal strains and intracellular pigment concentrations of the two oceanic strains decreased, while the side light scatter/front light scatter (SS/FS) ratio of the two coastal strains increased. In contrast to iron limitation, nitrogen source only marginally affected the photosynthesis of the four Synechococcus strains. Ammonium enhanced the growth of the two coastal strains under iron-replete condition. For the two oceanic strains, ammonium increased their cell size and decreased their SS/FS ratio and intracellular pigment concentrations under iron-deplete and iron-replete conditions.     

Cycling of selenite and selenate in marine phytoplankton

The marine phytoplankton Dunaliella tertiolecta, Cachonina niei, Thalassiosira nordenskioldii, Phaeodactylum tricornutum, and Chaetoceros sp. were incubated with a range of molar concentrations of sodium-selenite (Na₂-SeIVO₃) and sodium-selenate (Na₂-SeVIO₄) to examine further their role in metabolic cycling of selenium in ocean waters. At low selenium concentrations, approaching those found naturally in seawater (10^-10 to 10^-9 M), all species distinguished between selenite and selenate, and actively concentrated selenite from the incubating medium while only marginally accumulating selenate. At much higher concentrations (10^-8 to 10^-6 M), selenate was also taken up. At the highest concentration tested, i.e., 10^-5 M with C. niei, after an immediate rapid uptake in the first 24 h, the intracellular selenite and selenate levels dropped to about 35 to 50% of the initial peak values. These observations suggest an uptake mechanism in these algae which, at normal ambient concentrations of selenium (10^-9 M), preferentially selects selenite and excludes selenate. At much higher concentrations (10^-8 M), the mechanism becomes overloaded and both selenium species enter the cells. Intracellularly, selenite became associated primarily with protein and amino acid fractions, in approximately equal proportions, while only ca. 4% of total intracellular selenium was found in the lipid fraction. Trace amounts of selenate that entered the cells, mainly during the first minutes of exposure, also entered the protein and amino acid components, but over time were increasingly associated with the protein fraction only. At the end of a 10-d incubation of algal cells in selenite-spiked medium, less than 25% of total Se in the medium could in fact be identified analytically as selenite. This suggests the presence of a non-selenite metabolite, possibly released back into the medium from the algae.     

Cultures of the pelagic cyanophytes Trichodesmium erythraeum and T. thiebautii in synthetic medium

Trials for determination of culture conditions for the marine cyanophytes of Trichodesmium erythraeum and T. thiebautii were made with use of a synthetic medium. The Aquil medium, either with or without combined nitrogen, brought about stable growth of the two strains, T. erythraeum and T. thiebautii. However, they failed to grow in an ASP₇ medium. The failure was found to be due to the toxic effect of Tris-aminomethane, the pH-buffer in this medium. Two important chemical conditions for the stable growth of Trichodesmium spp. were revealed. (1) Stable growth was supported by Ca²⁺ at high concentrations; in a concentration lower than 0.9 mM, cell-lysis promptly occurred, while the cells could grow without cell-lysis at Ca²⁺ concentrations higher than 7.5 mM even at a salinity as low as 19 S. Ca²⁺ is probably essential for the osmotic regulation in this organism. (2) Phosphate-toxicity at high concentrations was at least partly due to heavy metal(s) contaminating the reagent of inorganic phosphate. After treatment with a Chelex-100 column, phosphate concentration could be increased up to four times the previous concentrations without toxicity.     

Aerobic nitrogenase activity measured as acetylene reduction in the marine non-heterocystous cyanobacterium Trichodesmium spp. grown under artificial conditions

Aerobic nitrogenase activity in the marine non-heterocystous cyanobacterium Trichodesmium spp. NIBB 1067, isolated off the Izu Peninsula, Japan in 1983 and grown under artificial conditions, was assayed by the acetylene reduction method. This strain exhibited acetylene reduction activity under aerobic conditions when cells had been grown in the medium free of combined nitrogen. Activity was markedly enhanced by light, and dependent on the growth phase being higher during the exponential growth phase and lower during the late linear and stationary growth phases. Since typical colony formation occurred during the last growth phase, the present results contradict the idea that N₂-fixation depends on colony formation. The photosynthesis inhibitor DCMU at 10^-6 M inhibited light-dependent acetylene reduction completely. Acetylene reduction by Trichodesmium spp. was tolerant of O₂ as strongly as that in the heterocystous cyanobacteria. Even at a partial pressure of oxygen (p_{O 2}) of 3 atm, the activity still remained as high as half of the maximum. It was almost under anaerobic conditions. Maximum activity was obtained at pO₂ of ca. 0.1 atm.     

Vertical distributions and photosynthetic action spectre of two oceanic picophytopianlcters,Prochlorococcus marinus andSynechococcus sp.

Two picophytoplankters,Prochlorococcus marinus andSynechococcus sp., were isolated from the bottom of the euphotic zone (150 m depth) in the western Pacifie Ocean. The concentration ofP. marinus at this depth was more than 10⁴ cells ml^–1 while that ofSynechococcus sp. was less than 10² cells ml^–1. TheP. marinus isolate has a high divinyl-chlorophylla:b ratio similar to that of the Mediterranean strain, while theSynechococcus sp. isolate is of the phycourobilinrich type. The growth rate ofP. marinus was higher thanSynechococcus sp. when both were cultured under weak blue-green to blue-violet light (ca. 2 E m^–2 s^–1). While the chlorophyll-specific absorption spectra showed higher values inSynechococcus sp., the photosynthetic action spectre revealed thatP. marinus was able to use blue-violet light, whereasSynechococcus sp. was able to use blue-green light, more efficiently for photosynthesis. The photosynthetic quantum yield ofP. marinus was higher than that ofSynechococcus sp. at any wavelength between 400 and 700 nm. The calculated in situ photosynthesis rates per Gell volume forP. marinus were estimated to be higher than forSynechococcus sp. at 50 and 150 m depth. These results indicate thatP. marinus photosynthetically surpassesSynechococcus sp. in the blue-light-rieh environment of the oceanic euphotic zone. This may be why the former predominates at depths in temperate to tropical open ocean waters.     

Phosphate uptake by intertidal algae in relation to zonation and season

The removal of phosphate from ambient seawater by whole plants of five species of fucoid algae, collected from the east coast of N. Ireland in 1988 and 1989, was followed over 6-h periods. A transient uptake pattern was observed forPelvetia canaliculata (L.) Dcne. et Thuret,Fucus spiralis L.,F. vesiculosus L. andF. serratus L., consisting of an initial period of high uptake, followed by a phase of zero uptake and then a period at an intermediate rate.Ascophyllum nodosum (L.) Le Jolis had a constant slow rate of uptake over 6 h. The initial uptake rate ofF. spiralis was significantly greater than that of any other species. Phosphate uptake over a 2-h period was measured at concentrations ranging from that of ambient seawater to 25µg-at. l^–1 for whole plants ofF. spiralis andF. serratus, using a large scale batch method. A small scale batch method was used for whole plants ofP. canaliculata and sections of the other four species investigated. Uptake abilities of the algae at low concentrations of phosphate were compared using the parameterV ₁ (the uptake rate at 1µg-at. l^–1) and at high concentrations usingV _max, the maximum uptake rate. These kinetic parameters of uptake were calculated using a method that avoids bias and permits statistical evaluation of the results. The fucoid algae studied could be divided into two distinct groups on the basis of their abilities to take up phosphate from seawater.P. canaliculata andA. nodosum had low values ofV ₁ in winter, which were also correlated with their positions on the shore and did not vary between winter and summer. TheFucus species had higher values ofV ₁ in winter, which were also correlated with their positions on the shore. In summer, however,V ₁-values for these species decreased and no longer correlated with their shore heights. TheV _max-value forF. spiralis was higher in winter than in summer but was signifcantly greater than that of any other species at all times of year. The ecological significance ofV _max is discussed in relation to nutrient limitation and the possible occurrence of patches of high nutrient concentration in the intertidal environment.     

Dark CO2 uptake by the diatom Chaetoceros simplex in response to nitrogen pulsing

In a continuing investigation of dark CO₂ uptake by nitrogen-limited cultures of the marine diatom Chaetoceros simplex (Bbsm), we expanded on several of our earlier conclusions regarding the potential application of this physiological response for measuring the degree and type of nitrogen limitation in phytoplankton populations. First, the duration over which the maximal enhancement of dark ¹⁴CO₂ uptake was sustained after NH ₄ ⁺ enrichment was a function both of the concentration of added NH ₄ ⁺ and the standing crop of phytoplankton nitrogen — in effect, the total N demand. Second, pulsing with NH ₄ ⁺ for a given degree of N-limitation always produced the same level of enhanced dark CO₂ uptake regardless of whether the cultures were preconditioned with oxidized or reduced nitrogen. In contrast, urea pulsing led to reduced dark CO₂ uptake, but the effect was most pronounced in cells grown on NO ₃ ^– . And third, the assay could be used to distinguish readily between no, moderate, and severe N limitation. The degree of severe N limitation was quantitatively correlated with the degree of enhanced dark CO₂ uptake, but this relationship was not so clear in the region of moderate N limitation. The main advantage of the assay is that it is a relatively simple and effective alternative to more complicated techniques for gauging the degree and form of N limitation in phytoplankton. Further evaluation will be required, both in the laboratory and field, before the assay can be calibrated for quantitative use.Contribution No. 5982 from the Woods Hole Oceanographic Institution     

Effect of UV-B radiation on the marine diatoms Lauderia annulata and Thalassiorsira rotula grown in different salinities

The marine diatoms Lauderia annulata Cleve and Thalassiosira rotula Meunier were grown at different salinities (20, 35 and 45) and exposed to different levels of midultraviolet, UV-B) 439, 717 and 1230 J m^-2 d^-1, weighted) for 2 d. A low UV-B dose (439 J m^-2 d^-1) usually caused a slight increase in biomass production (dry weight) compared to non-UV-B irradiated cells. Enhanced UV-B radiation (717 J m^-2 d^-1) depressed protein and pigment content (chlorophyll a, chlorophyll c ₁+c₂ and carotenoids), especially in algae grown at 20 or 35 salt concentration of the nutrient solution. The effect of UV-B radiation (717 J m^-2 d^-1) on the pattern and concentration of amino acids was species-dependent. Aspartic acid was reduced in all tested diatoms. A drastic increase in glutamine and a reduction in glutamic acid pools could be observed in L. annulata samples, but no significant variation of the impact of UV-B was found in dependence on the salt concentration of the nutrient medium. T. rotula cells grown at 35 S showed an increase of glutamic acid and a decrease of glutamine levels after UV-B radiation. The results are discussed in relation to the impact of UV-B upon carbon and nitrogen metabolism.     

Growth,photosynthesis and carbon metabolism in the temperate marine diatom Skeletonema costatum adapted to low temperature and low photon-flux density

The temperate diatom Skeletonema costatum (Grev.) Cleve was grown in low temperature and/or low light conditions. The cultures were acclimatized for at least three months before experiments were begun. Our data indicate that the initial slope of the photosynthesis vs irradiance curve () is controlled predominantly by light history and the light-saturated photosynthesis (P _max) by temperature. The number of divisions per day decreased with decreasing light intensity, but was identical for cultures grown at 3° or 18°C. The metabolic pathways of inorganic carbon fixation were not fundamentally affected by low temperature or low light intensity, but both these factors increased labelling of C₃ compounds, synthesized by the Calvin-Benson cycle, and decreased that of phosphoenolpyruvate (PEP) and other metabolites. This indicates an enhancement of ribulose-1,5-bisphosphate (RuBP) carboxylase activity, which is the first step in the C₃ pathway (3-phosphoglycerate and sugar phosphate synthesis); this may optimize cell functions. At low temperatures, a seven-fold increase in RuBP carboxylase activity per cell was observed. S. costatum is able to adapt to low irradiance by increasing and decreasing I _k (the ratio of P _max:, light intensity at onset of light saturation), and to low temperature by increasing its cellular chlorophyll a and RuBP carboxylase content. However, in the latter case, adaptation is not optimal. This study revealed two main features: (1) there is evidence that RuBP carboxylase has a key function in adjustment to high rates of photosynthesis at suboptimal temperatures or irradiances; (2) adaptive mechanisms are dynamic processes and the role of the time scale in physiological adaptation should be considered.     

Resting spore formation and phosphorus composition of the marine diatom Chaetoceros pseudocurvisetus under various nutrient conditions

The marine diatom Chaetoceros pseudocurvisetus was used for a study of resting spore formation and cellular phosphorus composition. Resting spores were found in any culture medium with ample silica, including nitrogen limited, phosphorus limited and nutrient replete conditions. Resting spores protected themselves with thick silica walls, so that vegetative cells required about 3 pmol cell^-1 of additional silica to form resting spores. Phosphorus compounds in the cells were divided into eight fractions: nucleotide-P, orthophosphate, acid soluble polyphosphate, sugar phosphate, nucleic acid-P, acid insoluble polyphosphate, lipid-P and residual-P. The sum of orthophosphate, sugar phosphate and nucleic acid-P comprised over 65% of the total phosphorus in cells under any culture conditions. Sugar phosphate was the most variable component, being most abundant in vegetative cells and least abundant in resting spores.     

Different patterns of growth and nitrogen uptake in two clones of marineSynechococcus spp.

We grew marineSynechococcus Clones WH7803 and WH8018 at 150µE m^–2 s^–1 in dilute batch cultures with NH ₄ ⁺ as the limiting nutrient. The maximal uptake capacities for NH ₄ ⁺ and NO ₃ ^- were measured in frequent experiments during log and stationary phases of growth. Clone WH7803, originally isolated from oceanic waters, had a specific uptake rate of NH ₄ ⁺ that approximated the maximum (log phase) specific growth rate (ca ~ 0.025 h^–1). NO ₃ ^- uptake was observed only after nitrogen in the media was depleted; the NO ₃ ^– uptake capacity was ca 12% the capacity for NH ₄ ⁺ uptake throughout the nitrogen depleted period. Growth was arrested upon nitrogen depletion, but resumed soon after reinoculation into fresh media, even after 5 d of starvation. Clone WH8018, originally isolated from coastal waters, revealed a five-fold enhancement in the NH ₄ ⁺ uptake rate relative to growth rate at the time of nitrogen depletion. As nitrogen starvation proceeded, this enhancement was reduced. This clone, too, was able to take up NO ₃ ^- once nitrogen in the media was depleted, but only after ca 20 h. Growth continued for a limited period during nitrogen depletion, but nitrogen-starved cells were slow to recover upon reinoculation into fresh media. We speculate that clonal differences may reflect differences in the molecular regulation of nitrogen assimilation.     

Genetic differentiation in two sibling species of the brackish-water polychaete Hediste japonica complex (Nereididae)

Genetic divergence among ten populations of small- and large-egg forms of the brackish-water polychaete Hediste japonica complex was investigated on 14 isozyme loci by electrophoretic analysis. The two forms were distinguishable by complete allele substitutions at five loci, resulting in high genetic differentiation (Nei's D: 0.533 to 0.662). No genetic evidence of hybridization between the two forms was detected in sympatric populations in three rivers. These results indicate that the two forms are reproductively isolated, clearly showing that the two forms are distinct species. The genetic differentiation among populations was higher in the large-egg form (D: 0.005 to 0.111, G _ST: 0.435) than that in the small-egg form (D: 0.000 to 0.001, G _ST: 0.020). This genetic difference between the two forms seems to be attributable to a difference in their life histories. The average expected heterozygosity was low in populations of both the large-egg form (0.005 to 0.068) and the small-egg form (0.014 to 0.038) in comparison with other marine invertebrates. Received: 11 April 1997 / Accepted: 8 September 1997     

Phosphate acquisition in the giant clam-zooxanthellae symbiosis

The effect of phosphate on the giant clam Tridacna gigas and on its symbiotic dinoflagellate Symbiodinium sp. was compared with that on cultured Symbiodinium sp. originally isolated from the same clarn species. Incubation of whole clams in elevated phosphate (10 M) reduced their capacity for phosphate uptake, but the uptake capacity of the clam's zooxanthellae population was not influenced. In addition, there was no change in the zooxanthellae density and the N:P ratio, of these algae., On the other hand, cultured zooxanthellae were influenced by the phosphate regimen of their culture medium. Compared with controls (0 M P), addition of 10 M phosphate to the culture medium caused an increase of 100% in cell density and decreases of 50% in the N:P ratio, and 80% in the phosphate-uptake capacity of the zooxanthellae. Zooxanthellae freshly isolated from the clams exhibited properties similar to those of zooxanthellae cultured in the absence of phosphate. These results demonstrate that the zooxanthellae population of T. gigas have limited access to the inorganic phosphate in sea water and the phosphate reserves within the animal host.     

Sodium-dependent amino acid transport in the chlorophyte Platymonas subcordiformis

Sodium-dependent transport of alanine and serine by Platymonas subcordiformis was demonstrated by evaluating the kinetics of entry of these substrates over a range of concentrations of ambient sodium. The reciprocal of the concentration at which entry rate was half maximal (K _t ) was linearly related to ambient sodium concentration; maximum entry (V _max) was not affected. Entry of labeled amino acids as measured by determining radioactivity in the medium was close to but slightly less than rates of net entry as determined by high-performance liquid chromatography. This difference presumably reflects extrusion of labeled carbon by the cells in a form not detectable by the chromatographic techniques employed. The coupling coefficient (sodium ions per amino acid molecule) for entry of alanine and serine was evaluated and determined to be 2. Methylaminoisobutyric acid was not taken up by P. subcordiformis and had no inhibitory effect on uptake of alanine or serine. B-2-aminobicycloheptane-2-carboxylic acid competitively inhibited uptake of both substrates. The internal sodium concentration of P. subcordiformis was measured, and the maximum gradient energetically favorable for amino acid transport was calculated from data in the present work and drawn from the literature. Assuming that uptake of amino acids is strictly sodium-dependent, an amino acid concentration gradient of the order of 10⁶ (cell:medium) can be achieved. This concentration differential permits net uptake of amino acids by P. subcordiformis from ambient amino acid levels in the nanomolar range.     

Marine diatoms grown in chemostats under silicate or ammonium limitation. III. Cellular chemical composition and morphology of Chaetoceros debilis,Skeletonema costatum,and Thalassiosira gravida

Three marine diatoms, Skeletonema costatum, Chaetoceros debilis, and Thalassiosira gravida were grown under no limitation and ammonium or silicate limitation or starvation. Changes in cell morphology were documented with photomicrographs of ammonium and silicate-limited and non-limited cells, and correlated with observed changes in chemical composition. Cultures grown under silicate starvation or limitation showed an increase in particulate carbon, nitrogen and phosporus and chlorophyll a per unit cell volume compared to non-limited cells; particulate silica per cell volume decreased. Si-starved cells were different from Si-limited cells in that the former contained more particulate carbon and silica per cell volume. The most sensitive indicator of silicate limitation or starvation was the ratio C:Si, being 3 to 5 times higher than the values for non-limited cells. The ratios Si:chlorophyll a and S:P were lower and N:Si was higher than non-limited cells by a factor of 2 to 3. The other ratios, C:N, C:P, C:chlorophyll a, N:chlorophyll a, P:chlorophyll a and N:P were considered not to be sensitive indicators of silicate limitation or starvation. Chlorophyll a, and particulate nitrogen per unit cell volume decreased under ammonium limitation and starvation. NH₄-starved cells contained more chlorophyll a, carbon, nitrogen, silica, and phosphorus per cell volume than NH₄-limited cells. N:Si was the most sensitive ratio to ammonium limitation or starvation, being 2 to 3 times lower than non-limited cells. Si:chlorophyll a, P:chlorophyll a and N:P were less sensitive, while the ratios C:N, C:chlorophyll a, N:chlorophyll a, C:Si, C:P and Si:P were the least sensitive. Limited cells had less of the limiting nutrient per unit cell volume than starved cells and more of the non-limiting nutrients (i.e., silica and phosphorus for NH₄-limited cells). This suggests that nutrient-limited cells rather than nutrient-starved cells should be used along with non-limited cells to measure the full range of potential change in cellular chemical composition for one species under nutrient limitation.Contribution No. 943 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA.     

Uptake and release of phosphorus by phytoplankton in the Chesapeake Bay estuary,USA

The phytoplankton uptake and release rates for inorganic phosphate, dissolved organic phosphate and polyphosphate were estimated during 5 cruises on the Chesapeake Bay over a 9-month period. Phosphorus in all pools turned over in several minutes to 100 h, and each soluble pool appeared to contain fractions which were metabolically useful to the phytoplankton. Maximal uptake rates (V _m ) for orthophosphate ranged from 0.02 to 2.95 μg-at P (1.h)^-1 with half saturation constants (K _s ) between 0.09 and 1.72 μg-at P l^-1. At low soluble reactive phosphorus concentrations, the uptake rate of trace ³²P orthophosphate was initially rapid, but declined after 15 to 60 min incubation. The data suggest that the initial uptake phase was dominated by exchange of ³²PO₄ ^≡ for ³¹PO₄ ^≡ in the membrane transport systems whereas the subsequent phase represented the net incorporation of orthophosphate into phytoplankton cells.