Diel variation of the cellular carbon to nitrogen ratio of Chlorella autotrophica (Chlorophyta) growing in phosphorus‐ and nitrogen‐limited continuous cultures

We investigated the relationship between daily growth rates and diel variation of carbon (C) metabolism and C to nitrogen (N) ratio under P‐ and N‐limitation in the green algae Chlorella autotrophica. To do this, continuous cultures of C. autotrophica were maintained in a cyclostat culture system under 14:10 light:dark cycle over a series of P‐ and N‐limited growth rates. Cell abundance, together with cell size, as reflected by side scatter signal from flow cytometric analysis demonstrated a synchronized diel pattern with cell division occurring at night. Under either type of nutrient limitation, the cellular C:N ratio increased through the light period and decreased through the dark period over all growth rates, indicating a higher diel variation of C metabolism than that of N. Daily average cellular C:N ratios were higher at lower dilution rates under both types of nutrient limitation but cell enlargement was only observed at lower dilution rates under P‐limitation. Carbon specific growth rates during the dark period positively correlated with cellular daily growth rates (dilution rates), with net loss of C during night at the lowest growth rates under N‐limitation. Under P‐limitation, dark C specific growth rates were close to zero at low dilution rates but also exhibited an increasing trend at high dilution rates. In general, diel variations of cellular C:N were low when dark C specific growth rates were high. This result indicated that the fast growing cells performed dark C assimilation at high rates, hence diminished the uncoupling of C and N metabolism at night.     

PHOSPHATE‐LIMITED GROWTH OF PAVLOVA LUTHERI (PRYMNESIOPHYCEAE) IN CONTINUOUS CULTURE: DETERMINATION OF GROWTH‐RATE‐LIMITING SUBSTRATE CONCENTRATIONS WITH A SENSITIVE BIOASSAY PROCEDURE1

The relationship between steady‐state growth rate and phosphate concentration was studied for the marine prymnesiophyte Pavlova lutheri (Droop) J. C. Green grown in a chemostat at 22°C under continuous irradiance. A bioassay procedure involving short‐term uptake of 10 picomolar spikes of ³³P‐labeled phosphate was used to estimate the concentration of phosphate in the growth chamber. The relationship between growth rate and phosphate was well described by a simple rectangular hyperbola with a half‐saturation constant of 2.6 nM. The cells were able to take up micromolar spikes of phosphate at rates two to three orders of magnitude higher than steady‐state uptake rates. The kinetics of short‐term uptake displayed Holling type III behavior, suggesting that P. lutheri may have multiple uptake systems with different half‐saturation constants. Chl a:C ratios were linearly related to growth rate and similar to values previously reported for P. lutheri under nitrate‐limited conditions. C:N ratios, also linearly related to growth rate, were consistently lower than values reported for P. lutheri under nitrate‐limited conditions, a result presumably reflecting luxury assimilation of nitrogen under phosphate‐limited conditions. C:P ratios were linearly related to growth rate in a manner consistent with the Droop equation for growth rate versus cellular P:C ratio.     

CHANGES IN NUTRIENT CONTENT OF PALMARIA PALMATA IN RESPONSE TO VARIABLE LIGHT AND UPWELLING IN NORTHERN SPAIN1

Light has been identified as one of the main factors affecting seaweed ecophysiology. We investigated the dependence of nutrient metabolism on sun and shade light conditions and whether episodes of upwelling of nutrient‐rich subsuperficial water could reduce the summer nutrient limitation driving physiological changes in Palmaria palmata (L.) Kuntze. We measured the major nutrient pools, photosynthetic pigments, and light curves, under sun and shade conditions during a summer period when one upwelling was recorded. The redundancy analysis (RDA) produced two clear groups: sun‐ and shade‐acclimated algae. Light was the major predictive factor. Sun‐acclimated algae exhibited higher carbon (C) and lower nitrogen (N) and phosphorus (P) content in association with the storage of floridoside (main C reserve) to benefit from higher irradiance (under nutrient limitation). Among N pools, N reserves (phycoerythrin, nitrate) were a lower proportion of the total N in sun‐acclimated algae, suggesting their degradation to fulfill the N demands of the cell. The orthophosphate content was also lower in sun‐acclimated algae, indicating its utilization as a nutrient reserve. In contrast, N within cell walls and membranes and chl a contributed to a similar proportion of the total N in sun‐ and shade‐acclimated algae, suggesting a response to sustain cell integrity. Transient high nutrient concentration due to the upwelling was unrelated to the nutrient content of the thallus. The storage of C as floridoside from high light exposure was shown to be the driving force for the metabolic adjustment of P. palmata at the end of summer before the onset of dormancy.     

FERRIC CHELATE REDUCTASE ACTIVITY AS AFFECTED BY THE IRON‐LIMITED GROWTH RATE IN FOUR SPECIES OF UNICELLULAR GREEN ALGAE (CHLOROPHYTA)1

Four species of green algae (Chlorella kessleri Fott et Nováková, Chlorococcum macrostigmatum Starr, Haematococcus lacustris[Girod‐Chantrans] Rostaf., Stichococcus bacillaris Näg.) were grown in iron‐limited chemostats and under phosphate limitation and iron (nutrient) sufficiency. For all four species, steady‐state culture density declined with decreasing degree of iron limitation (increasing iron‐limited growth rate), whereas chl per cell or biovolume increased. Plasma membrane ferric chelate reductase activity was enhanced by iron limitation in all species and suppressed by phosphate limitation and iron sufficiency. These results confirm previous work that C. kessleri uses a reductive mechanism of iron acquisition and also suggest that the other three species use the same mechanism. Although imposition of iron limitation led to enhanced activities of ferric chelate reductase in all species, the relationship between ferric chelate reductase activity and degree of iron limitation varied. Ferric chelate reductase activity in C. macrostigmatum and S. bacillaris was an inverse function of the degree of iron limitation, with the most rapidly growing iron‐limited cells exhibiting the highest ferric chelate reductase activity. In contrast, ferric chelate reductase activity was only weakly affected by the degree of iron limitation in C. kessleri and H. lacustris. Calculation of ferric reductase activity per unit chl allowed a clear differentiation between iron‐limited and iron‐sufficient cells. The possible extension of the ferric chelate reductase assay to investigate the absence or presence of iron limitation in natural waters may be feasible, but it is unlikely that the assay could be used to estimate the degree of iron limitation.     

Mixotrophy in gyrodinium galatheanum (DINOPHYCEAE): grazing responses to light intensity and inorganic nutrients*

This paper presents results of field and laboratory studies on mixotrophy in the estuarine dinoflagellate Gyrodinium galatheanum (Braarud) Taylor. We tested the hypotheses that this primarily photosynthetic organism becomes phagotrophic when faced with suboptimal light and/or nutrient environments. In Chesapeake Bay, incidence of feeding of this species on cryptophytes is positively correlated with prey density and concentrations of nitrate and nitrite, but negatively correlated with depth, salinity, and phosphate concentration. Feeding in natural assemblages and cultures increased hyperbolically with light intensity. The stoichiometric proportions of dissolved inorganic P and N (DIP:DIN) at the stations where G. galatheanum was present were far below the optimal growth P:N (1:10). Incidence of feeding was negatively related to the ratio of DIP to DIN, suggesting that P limitation may have induced feeding. Addition of nitrate, or addition of both nitrate and phosphate, inhibited feeding in a natural population, indicating that N limitation may also induce feeding. Ingestion of the cryptophyte, Storeatula major, by cultured G. galatheanum was higher in media low in nitrate or phosphate or both, but moderate rates of feeding occurred in nutrient‐replete cultures. When cells were grown in media with varying concentrations of nitrate and phosphate, N deficiency resulted in greater cellular N and Chl a losses than did P deficiency, but P deficiency stimulated feeding more than N deficiency. Both N and P deficiency, or P:N ratios that deviated greatly from 1:10, result in an increase of cellular carbon content and an increase in propensity to feed. Our results suggest that feeding in G. galatheanum is partly a strategy for supplementing major nutrients (N and P) that are needed for photosynthetic carbon assimilation. Feeding in G. galatheanum may also be a strategy for supplementing C metabolism or acquiring trace organic growth factors, since feeding occurs, although at a reduced rate, in nutrient‐replete cultures.     

EFFECT OF NUTRIENT AVAILABILITY ON THE BIOCHEMICAL AND ELEMENTAL STOICHIOMETRY IN THE FRESHWATER DIATOM STEPHANODISCUS MINUTULUS (BACILLARIOPHYCEAE)*

The objective of this study was to examine the differences in the biochemical and elemental stoichiometry of a freshwater centric diatom, Stephanodiscus minutulus (Grun.), under various nutrient regimes. Stephanodiscus minutulus was grown at μ_max or 22% of μ_max under limitation by silicon, nitrogen, or phosphorus. Cell sizes for nutrient‐limited cultures were significantly smaller than the non‐limited cell sizes, with N‐limited cells being significantly smaller than all other treatments. Compared with the nutrient‐replete treatment, both carbohydrates and lipids increased in Si‐ and P‐limited cells, whereas carbohydrates increased but proteins decreased in N‐limited cells. All of the growth‐limited cells showed an increase of carbohydrate and triglyceride, and a decrease of cell size and polar lipids as a percentage of total lipids. The non‐limited cells also had a significantly higher chl a concentration and galactolipids as a percentage of total lipids than any of the limited treatments, and the low‐Si and low‐P cells had significantly higher values than the low‐N cells. The particulate C concentrations showed significant differences between treatments, with the Si‐ and P‐limited treatments being significantly higher than the N‐ and non‐limited treatments. Particulate Si did not show a strong relationship with any of the parameters measured, and it was the only parameter with no differences between treatments. The low‐Si cells had a significantly higher P content (about two times more) than any other treatment, presumably owing to the luxury consumption of P, and a correspondingly high phospholipid concentration. The elemental data showed that S. minutulus had a high P demand with low optimum N:P (4) and Si:P (10) ratios and a C:N:P ratio of 109:16:2.3. The particulate C showed a positive relationship with POM (r = 0.93), dry weight (r = 0.88), lipid (r = 0.87) and protein (r = 0.84, all P < 0.0001). Particulate N showed a positive relationship with galactolipids (r = 0.95), protein (r = 0.90), dry weight (r = 0.78), lipid (r = 0.75), and cell volume (r = 0.64, all P < 0.0001). It is evident that nutrient limitation in the freshwater diatom S. minutulus has pronounced effects on its biochemical and elemental stoichiometry.     

ELEVATED ATMOSPHERIC CARBON DIOXIDE INCREASES ORGANIC CARBON FIXATION BY EMILIANIA HUXLEYI (HAPTOPHYTA), UNDER NUTRIENT‐LIMITED HIGH‐LIGHT CONDITIONS1

Phytoplankton play a key role in determining the partitioning of CO₂ between the atmosphere and the ocean on seasonal, interannual, and millennial time scales. The magnitude of biological draw‐down of atmospheric CO₂ and C storage in the oceans is affected by concurrent changes in other environmental factors, like nutrient supply. Furthermore, variations in carbon‐to‐nitrogen (C:N) and carbon‐to‐phosphorus (C:P) assimilation ratios modify the oceanic CO₂ storage capacity. Here we show that increased atmospheric CO₂ concentration enhances CO₂ fixation into organic matter by a noncalcifying strain of Emiliania huxleyi (Lohmann) Hay & Mohler only under certain conditions, namely high light and nutrient limitation. Enhanced organic matter production was accompanied by marked deviations of the C:N:P ratio from the canonical stoichiometry of marine particulate matter of 106:16:1 (C:N:P) known as the Redfield ratio. Increased cell organic carbon content, C:N, and C:P were observed at high light when growth was either nitrogen or phosphorus limited. Elevated CO₂ led to further increases in the particulate C:N and C:P ratios. Enhanced CO₂ uptake by phytoplankton such as E. huxleyi, in response to elevated atmospheric CO₂, could increase carbon storage in the nitrogen‐limited regions of the oceans and thus act as a negative feedback on rising atmospheric CO₂ levels.     

UNCOUPLING OF SILICON COMPARED WITH CARBON AND NITROGEN METABOLISMS AND THE ROLE OF THE CELL CYCLE IN CONTINUOUS CULTURES OF THALASSIOSIRA PSEUDONANA (BACILLARIOPHYCEAE) UNDER LIGHT,NITROGEN, AND PHOSPHORUS CONTROL1

The elemental composition and the cell cycle stages of the marine diatom Thalassiosira pseudonana Hasle and Heimdal were studied in continuous cultures over a range of different light‐ (E), nitrogen‐ (N), and phosphorus‐ (P) limited growth rates. In all growth conditions investigated, the decrease in the growth rate was linked with a higher relative contribution of the G2+M phase. The other phases of the cell cycle, G1 and S, showed different patterns, depending on the type of limitation. All experiments showed a highly significant increase in the amount of biogenic silica per cell and per cell surface with decreasing growth rates. At low growth rates, the G2+M elongation allowed an increase of the silicification of the cells. This pattern could be explained by the major uptake of silicon during the G2+M phase and by the independence of this process on the requirements of the other elements. This was illustrated by the elemental ratios Si/C and Si/N that increased from 2‐ to 6‐fold, depending of the type of limitation, whereas the C/N ratio decreased by 10% (E limitation) or increased by 50% (P limitation). The variations of the ratios clearly demonstrate the uncoupling of the Si metabolism compared with the C and N metabolisms. This uncoupling enabled us to explain that in any of the growth condition investigated, the silicification of the cells increased at low growth rates, whereas carbon and nitrogen cellular content are differently regulated, depending of the growth conditions.     

NUTRIENT LIMITATION AND HIGH IRRADIANCE ACCLIMATION REDUCE PAR AND UV‐INDUCED VIABILITY LOSS IN THE ANTARCTIC DIATOM CHAETOCEROS BREVIS (BACILLARIOPHYCEAE)1

The effects of high PAR (400–700 nm), UVA (315–400 nm), and UVB (280–315 nm) radiation on viability and photosynthesis were investigated for Chaetoceros brevis Schütt. This Antarctic marine diatom was cultivated under low, medium, and high irradiance and nitrate, phosphate, silicate, and iron limitation before exposure to a simulated surface irradiance (SSI) treatment, with and without UVB radiation. Light‐harvesting and protective pigment composition and PSII parameters were determined before SSI exposure, whereas viability was measured by flow cytometry in combination with a viability stain after the treatment. Recovery of PSII efficiency was measured after 20 h in dim light in a separate experiment. In addition, low and high irradiance acclimated cells were exposed outdoors for 4 h to assess the effects of natural PAR, UVA, and UVB on viability. Low irradiance acclimated cells were particularly sensitive to photo induced viability loss, whereas no viability loss was found after acclimation to high irradiance. Furthermore, nutrient limitation reduced sensitivity to photo induced viability loss, relative to nutrient replete conditions. No additional viability loss was found after UVB exposure. Sunlight exposed cells showed no additional UVB effect on viability, whereas UVA and PAR significantly reduced the viability of low irradiance acclimated cells. Recovery of PSII function was nearly complete in cultures that survived the light treatments. Increased resistance to high irradiance coincided with an increased ratio between protective‐ and light‐harvesting pigments before the SSI treatment, demonstrating the importance of nonphotochemical quenching by diatoxanthin for survival of near‐surface irradiance. We conclude that a sudden transfer to high irradiance can be fatal for low irradiance acclimated C. brevis.     

Phosphate‐limited growth of the marine diatom Thalassiosira weissflogii (Bacillariophyceae): evidence of non‐monod growth kinetics1

The marine diatom Thalassiosira weissflogii (Grunow) G. A. Fryxell & Hasle was grown in a chemostat over a series of phosphate‐limited growth rates. Ambient substrate concentrations were determined from bioassays involving picomolar spikes of ³³P‐labeled phosphate, and maximum uptake rates were determined from analogous bioassays that included the addition of micromolar concentrations of unlabeled phosphate and tracer concentrations of ³³P. The relationship between cell phosphorus quotas and growth rates was well described by the Droop equation. Maximum uptake rates of phosphate spikes were several orders of magnitude higher than steady state uptake rates. Despite the large size of the T. weissflogii cells, diffusion of phosphate through the boundary layer around the cells had little effect on growth kinetics, in part because the cellular N:P ratios exceeded the Redfield ratio at all growth rates. Fitting the Monod equation to the experimental data produced an estimate of the nutrient‐saturated growth rate that was ~50% greater than the maximum growth rate observed in batch culture. A modified hyperbolic equation with a curvature that is a maximum in magnitude at positive growth rates gave a better fit to the data and an estimate of the maximum growth rate that was consistent with observations. The failure of the Monod equation to describe the data may reflect a transition from substrate to co‐substrate limitation and/or the presence of an inducible uptake system.     

NITRATE AND PHOSPHATE UPTAKE INTERACTIONS IN A MARINE PRYMNESIOPHYTE1

The short- and long-term uptake of nitrate and phosphate ions, and their interactions, were studied as functions of the preconditioning of Pavlova lutheri (Droop) Green. Populations were preconditioned in continuous culture at a variety of growth rates and N:P supply ratios. The maximum uptake rates cell^?1 for nitrate and phosphate were of similar magnitudes, in spite of the forty-fold smaller requirement for phosphorus. Short-term phosphate uptake was independent of the nitrate concentration, but the short-term nitrate uptake rate was reduced in the presence of phosphate. The severity of inhibition of nitrate uptake by phosphate was positively correlated with the preconditioning N:P supply ratio and the preconditioning growth rate. In response to large additions of nutrients, P. lutheri was able to increase its phosphorus content sixty-fold, but was only able to take up enough nitrate to double its nitrogen content. The high rate of phosphate uptake relative to its requirement, the inhibition of nitrate uptake by phosphate, and the large capacity for phosphorus storage relative to its requirement, all of which were observed even under N limitation, may imply that even where nitrogen is limiting there can be interspecific competition for available phosphate.     

NEUTRAL LIPID AND CARBOHYDRATE PRODUCTIVITIES AS A RESPONSE TO NITROGEN STATUS IN ISOCHRYSIS SP. (T‐ISO; HAPTOPHYCEAE): STARVATION VERSUS LIMITATION1

Partitioning of the carbon (C) fixed during photosynthesis between neutral lipids (NL) and carbohydrates was investigated in Isochrysis sp. (Haptophyceae) in relation to its nitrogen (N) status. Using batch and nitrate‐limited continuous cultures, we studied the response of these energy reserve pools to both conditions of N starvation and limitation. During N starvation, NL and carbohydrate quotas increased but their specific growth rates (specific rates of variation, μ_CAR and μ_NL) decreased. When cells were successively deprived and then resupplied with NO₃, both carbohydrates and neutral lipids were inversely related to the N quota (N:C). These negative relationships were not identical during N impoverishment and replenishment, indicating a hysteresis phenomenon between N and C reserve mobilizations. Cells acclimated to increasing degrees of N limitation in steady‐state chemostat cultures showed decreasing NL quota and increasing carbohydrate quota. N starvation led to a visible but only transient increase of NL productivity. In continuous cultures, the highest NL productivity was obtained for the highest experimented dilution rate (D = 1.0 d^?1; i.e., for non N‐limited growth conditions), whereas the highest carbohydrate productivity was obtained at D = 0.67 d^?1. We used these results to discuss the nitrogen conditions that optimize NL productivities in the context of biofuel production.     

Nutrient limitation in boreal rich‐fen vegetation: A fertilization experiment

Abstract. Rich‐fen vegetation influenced by hay‐making in the Sølendet Nature Reserve, Central Norway, was fertilized with N, P and K in a full‐factorial fertilization experiment to investigate the nutrient limitation of plant growth at both community and species levels. Above‐ground biomass, shoot density and nutrient concentration were measured in several species and groups of species at three sites after two years of fertilization. At the community level, the results indicate multiple limitation by N and P in the two least productive rich‐fen communities: one characterized by small sedges and herbs, and the other by high abundance of Menyanthes trifoliata and tall sedges. Increased nutrient availability had no effect on a more highly productive, tall‐growing, spring‐influenced community, indicating no nutrient limitation. The results at the species level correspond well with those at the community level, indicating multiple limitation by N and P in most of the dominant and sub‐dominant species. However, P seems to limit growth more than N in Succisa pratensis, and N seems to limit growth more than P in Carex panicea. Furthermore, Eriophorum angustifolium seems to be limited by K. The results did not show which nutrient limits the growth of Carex dioica, C. lasiocarpa and Trichophorum cespitosum. Indications that growth in low‐productive, boreal rich‐fen communities is generally limited by P was not confirmed.     

N‐ASSIMILATION IN THE NOXIOUS FLAGELLATE HETEROSIGMA CARTERAE (RAPHIDOPHYCEAE): DEPENDENCE ON LIGHT,N‐SOURCE,AND PHYSIOLOGICAL STATE1

The capabilities of the diel vertically migrating flagellate Heterosigma carterae Hulburt for assimilating ammonium and nitrate into cell‐N in light and in darkness were studied using cells of different N‐status. Ammonium utilization in darkness, except by N‐replete cells, attained>50% of use in the light. However, the capacity to use nitrate was poor in darkness, and less than 20% of nitrate‐N that was taken up in darkness was then actually incorporated into cell‐N. The ability to assimilate N in darkness improved as N‐status (N:C) declined, concurrent with an increasing content of water‐soluble carbohydrate. This carbohydrate was used to support dark N‐assimilation. Cells held in darkness for over a day and that had halted nitrate‐uptake were still capable of taking up ammonium. Furthermore, the act of taking up ammonium appeared to make available a source of C to support nitrate uptake that was previously unavailable. The implications of these results for the ecophysiology of this organism and for the construction of mathematical models of algal growth are considered.     

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.     

Differential recovery of above‐ and below‐ground rich fen vegetation following fertilization

Abstract. For seven years we studied the recovery of vegetation in a Belgian P limited rich fen (Caricion davallianae), which had been fertilized with nitrogen (200 g.m^?2) and phosphorus (50 g.m^?2) in 1992. The vegetation in this fen has low above‐ground biomass production (< 100 g m^?2) due to the strong P limitation. Above‐ground biomass was harvested from 1992 to 1998 and P and N concentrations measured. In 1998, below‐ground biomass was also harvested. The response to fertilization differed markedly between below‐ and above‐ground compartments. Above‐ground, P was the single most important factor that continued to stimulate growth 7 yr after fertilization. Below‐ground, N tended to decrease live root biomass and increase dead root biomass and seemed to have a toxic effect on the roots. In the combined NP treatment the stimulating effect of P (an increase of live root biomass) was countered by N. The 1998 soil analysis showed no difference in soil P in the plots. Thus, Fe hydroxides are not capable of retaining P in competition with fen vegetation uptake. However, higher capture of P in root Fe coatings from N plots may partially explain this negative N effect. The results suggest that N root toxicity will be amplified in strongly P limited habitats but that its persistence will be less influenced by P availability. This mechanism may be a competitive advantage for N₂ fixing species that grow in strongly P limited wetlands.     

INTERACTING EFFECTS OF LIGHT AND NUTRIENT LIMITATION ON THE GROWTH RATE OF SYNECHOCOCCUS LINEARIS (CYANOPHYCEAE)1

The blue-green alga Synechococcus linearis (Naeg.) Kom. was grown in P- and N-limited chemostats over a range of potentially limiting irradiances in order to determine the combined effects of light and nutrient limitation on some aspects of the composition and metabolism of this alga. Over a narrow range of low irradiances, simultaneous limitation of growth rate by light and either N or P was shown. This simultaneous limitation of growth rate by a nutrient and a physical factor can be explained by the ability of an increased supply of one to compensate in part for a decreased supply of the other. At all irradiances, the internal concentration of the limiting nutrient increased with increasing dilution rate, and the results could be fitted to the Droop relationship. With decreasing irradiance, the internal concentration of the limiting nutrient increased. There appeared to be little or no effect of light on the minimum internal concentration of P but that of N increased with decreasing light. Both chlorophyll a and biliprotein per unit particulate C increased with increasing dilution rate and decreasing irradiance. The critical N/P ratio increased with decreasing light as the N requirement of N-limited cells increased faster than did the P requirement of P-limited cells. The composition of exponentially growing cells in complete medium varied much less with light. Neither dilution rate nor irradiance during growth had a great effect on saturated rates of P or N uptake or alkaline phosphatase activity. Calculated assimilation ratios increased with light and dilution rate. The role of the flexibility of nutrient composition in adaptation to adverse conditions and the implications of the results for the use of physiological indicators of nutrient status are discussed.     

GROWTH AND DOMOIC ACID PRODUCTION BY PSEUDO‐NITZSCHIA SERIATA (BACILLARIOPHYCEAE) UNDER PHOSPHATE AND SILICATE LIMITATION1

Pseudo‐nitzschia seriata (Cleve) H. Peragallo isolated from Scottish west coast waters was studied in batch culture with phosphate (P) or silicate (Si) as the yield‐limiting nutrient at 15°C. This species produced the neurotoxin domoic acid (DA) when either nutrient was limiting but produced more when stressed by Si limitation during the stationary phase. Under P‐limiting conditions, exponential growth stopped after P was reduced to a low threshold concentration. Under Si‐limiting conditions, fast exponential growth was followed by a period of slower exponential growth, until Si became exhausted. A stationary phase was observed in the P‐limited but not the Si‐limited cultures, the latter showing a rapid decrease in cell density after the second exponential growth phase. Si‐limited cultures exhibited a further period of active metabolism (as indicated by increases in chl and carbon per cell) late in the experiment, presumably fueled by regenerated Si. DA production was low in exponential phase under both conditions. In P‐limited cultures, most DA was produced during the immediate postexponential phase, with little or no new DA produced during later cell senescence. In contrast, although a substantial amount of DA was produced during the slower exponential phase of the Si‐limited cultures, DA production was even greater near the end of the experiment, coincident with the period of chl synthesis and increase in carbon biomass. Comparison of the magnitude of toxin production in the two nutrient regimes indicated a greater threat of P. seriata‐generated amnesic shellfish poisoning events under Si rather than P nutrient limitation.