Uptake kinetics of nitrate,ammonia and phosphate by the dinoflagellate Heterocapsa circularisquama isolated from Hiroshima Bay,Japan

Heterocapsa circularisquama is a harmful dinoflagellate whose first bloom in Hiroshima Bay, Japan, appeared in 1992. As suggested by the authors’ group, in the Seto Inland Sea including Hiroshima Bay, oligotrophication particularly the reduction of phosphate starting 1980 is severe. The bloom caused serious damage to the bay's extensive oyster culture. In the present study, the uptake kinetics of nitrate, ammonia, and phosphate by this species were experimentally investigated. The maximum uptake rate (ρ_max) and the half‐saturation constant (Ks) were 0.41 pmol cell^?1 h^?1 and 4.45 μM, respectively, for nitrate, 2.02 pmol cell^?1 h^?1 and 11.1 μM for ammonium, and 0.079 pmol cell^?1 h^?1 and 1.79 μM for phosphate. The maximum specific uptake rates (V_max) for nitrate, ammonia, and phosphate were estimated to be 8.95, 44.1, and 21.3 day^?1, respectively. A comparison of V_max/Ks, which is also an index of affinity to nutrients, between this species and others suggested that H. circularisquama can utilize nitrate and ammonia efficiently, but not phosphate. Considering both reports describing that H. circularisquama has the ability to utilize dissolved organic phosphorus (DOP) and the DOP concentration is higher than phosphate in Hiroshima Bay, it was concluded that H. circularisquama became dominant due to the phosphate reduction measure.     

Bottom-up controls on a mixed-species HAB assemblage: A comparison of sympatric Chattonella subsalsa and Heterosigma akashiwo (Raphidophyceae) isolates from the Delaware Inland Bays, USA

Recent novel mixed blooms of several species of toxic raphidophytes have caused fish kills and raised health concerns in the highly eutrophic Inland Bays of Delaware, USA. The factors that control their growth and dominance are not clear, including how these multi-species HAB events can persist without competitive exclusion occurring. We compared and contrasted the relative environmental niches of sympatric Chattonella subsalsa and Heterosigma akashiwo isolates from the bays using classic Monod-type experiments. C. subsalsa grew over a temperature range from 10 to 30 °C and a salinity range of 5–30 psu, with optimal growth occurring from 20 to 30 °C and 15 to 25 psu. H. akashiwo had similar upper temperature and salinity tolerances but also lower limits, with growth occurring from 4 to 30 °C and 5 to 30 psu and optimal growth between 16 and 30 °C and 10 and 30 psu. These culture results were confirmed by field observations of bloom occurrences in the Inland Bays. Maximum nutrient-saturated growth rates (μ_max) for C. subsalsa were 0.6 d⁻¹ and half-saturation concentrations for growth (K_s) were 9 μM for nitrate, 1.5 μM for ammonium, and 0.8 μM for phosphate. μ_max of H. akashiwo (0.7 d⁻¹) was slightly higher than C. subsalsa, but K_s values were nearly an order of magnitude lower at 0.3 μM for nitrate, 0.3 μM for ammonium, and 0.2 μM for phosphate. H. akashiwo is able to grow on urea but C. subsalsa cannot, while both can use glutamic acid. Cell yield experiments at environmentally relevant levels suggested an apparent preference by C. subsalsa for ammonium as a nitrogen source, while H. akashiwo produced more biomass on nitrate. Light intensity affected both species similarly, with the same growth responses for each over a range from 100 to 600 μmol photons m⁻² s⁻¹. Factors not examined here may allow C. subsalsa to persist during multi-species blooms in the bays, despite being competitively inferior to H. akashiwo under most conditions of nutrient availability, temperature, and salinity.     

pH affects ammonium,nitrate and proton fluxes in the apical region of conifer and soybean roots

The effect of pH on nitrate and ammonium uptake in the high‐affinity transport system and low‐affinity transport system ranges was compared in two conifers and one crop species. Many conifers grow on acidic soils, thus their preference for ammonium vs nitrate uptake can differ from that of crop plants, and the effect of pH on nitrogen (N) uptake may differ. Proton, ammonium and nitrate net fluxes were measured at seedling root tips and 5, 10, 20 and 30 mm from the tips using a non‐invasive microelectrode ion flux measurement system in solutions of 50 or 1500 µM NH₄NO₃ at pH 4 and 7. In Glycine max and Pinus contorta, efflux of protons was observed at pH 7 while pH 4 resulted in net proton uptake in some root regions. Pseudotsuga menziesii roots consistently showed proton efflux behind the root tip, and thus appear better adapted to maintain proton efflux in acid soils. P. menziesii's ability to maintain ammonium uptake at low pH may relate to its ability to maintain proton efflux. In all three species, net nitrate uptake was greatest at neutral pH. Net ammonium uptake in G. max and net nitrate uptake in P. menziesii were greatly reduced at pH 4, particularly at high N concentration, thus N concentration should be considered when determining optimum pH for N uptake. In P. menziesii and G. max, net N uptake was greater in 1500 than 50 µM NH₄NO₃ solution, but flux profiles of all ions varied among species.     

NITROGENOUS NUTRITION OF ALEXANDRIUM CATENELLA (DINOPHYCEAE) IN CULTURES AND IN THAU LAGOON,SOUTHERN FRANCE1

Alexandrium catenella (Whedon et Kofoid) Balech was isolated from Thau lagoon (northern Mediterranean) and its growth and uptake characteristics measured for nitrate, ammonium, and urea. Although affinity constants did not indicate a preference for ammonium over nitrate, there was a strong inhibition of nitrate uptake by ammonium when both nitrogen (N) sources were present. Nitrogen budgets during growth in cultures revealed major imbalances between decreases in dissolved N and increases in particulate N, indicating excretion of dissolved organic N during the early part of the growth phase and uptake during the later part. A quasi‐unialgal bloom in November 2001 (4×10⁶ cells·L^?1) allowed measurements of uptake of nitrate, nitrite, ammonium, and urea; net and gross growth rate of A. catenella; and grazing rates on this organism. The affinity constants indicate that it is not a strong competitor for the N nutrients tested when these are in low concentrations (<10 μgat N·L^?1), compared with other members of the phytoplankton community. Indirect evidence from cultures indicate that dissolved organic N compounds could be important in triggering those blooms. Finally, the strongly unbalanced growth observed in the field indicates that A. catenella exhibits a storage rather than a growth response to a nutrient pulse and is adapted to low frequency events such as the passage of frontal disturbances. The disappearance of A. catenella was due to grazing that balanced growth at the peak of the bloom.     

Growth, uptake, and assimilation of ammonium, nitrate, and urea, by three strains of Karenia brevis grown under low light

Observations of near-bottom populations of Karenia brevis suggest that these cells may derive nutrients from the sediment–water interface. Cells undergoing a metabolic-mediated migration may be in close proximity to enhanced concentrations of nutrients associated with the sediment during at least a fraction of their diel cycle. In this study, the growth, uptake and assimilation rates of ammonium, nitrate, and urea by K. brevis were examined on a diel basis to better understand the potential role of these nutrients in the near-bottom ecology of this species. Three strains of K. brevis, C6, C3, and CCMP 2229, were grown under 12:12 light dark cycle under 30 μmol photons m⁻² s⁻¹ delivered to the surface plain of batch cultures. Nitrogen uptake was evaluated using ¹⁵N tracer techniques and trichloroacetic acid extraction was used to evaluate the quantity of nitrogen (N) assimilated into cell protein. Growth rates ranged from a low of 0.12 divisions day⁻¹ for C6 and C3 grown on nitrate to a high of 0.18 divisions day⁻¹ for C3 grown on urea. Diurnal maximum uptake rates, ρ_max, varied from 0.41 pmol-N cell⁻¹ h⁻¹ for CCMP 2229 grown on nitrate, to 1.29 pmol-N cell⁻¹ h⁻¹ for CCMP 2229 grown on urea. Average nocturnal uptake rates were 29% of diurnal rates for nitrate, 103% of diurnal uptake rates for ammonium and 56% of diurnal uptake rates for urea. Uptake kinetic parameters varied between substrates, between strains and between day and night measurements. Highest maximum uptake rates were found for urea for strains CCMP2229 and C3 and for ammonium for strain C6. Rates of asmilation into protein also varied day and night, but overall were highest for urea. The comparison of maximal uptake rates as well as assimilation efficiencies indicate that ammonium and urea are utilized (taken up and assimilated) more than twice was fast as nitrate on a diel basis.     

Phytoplankton nitrogen demand and the significance of internal and external nitrogen sources in a large shallow lake (Lake Balaton,Hungary)

Since the middle of 1990s the trend of Lake Balaton towards an increasingly trophic status has been reversed, but N₂-fixing cyanobacteria are occasionally dominant, endangering water quality in summer. The sources of nitrogen and its uptake by growing phytoplankton were therefore studied. Experiments were carried out on samples collected from the middle of the Eastern (Siófok) and Western (Keszthely) basins between February and October 2001. Ammonium, urea and nitrate uptake and ammonium regeneration were measured in the upper 5-cm layer of sediment using the ¹⁵N-technique. Ammonium was determined by an improved microdiffusion assay. N₂ fixation rates were measured by the acetylene-reduction method. Ammonium regeneration rates in the sediment were similar in the two basins. They were relatively low in winter (0.13 and 0.16 μg N cm^?3 day^?1 in the Eastern and Western basin, respectively), increased slowly in the spring (0.38 and 0.45 μg N cm^?3 day^?1) and peaked in late summer (0.82 and 1.29 μg N cm^?3 day^?1, respectively). Ammonium uptake was predominant in spring in the Eastern basin and in summer in the Western basin, coincident with the cyanobacterial bloom. The amount of N₂ fixed was less than one third of the internal load during summer when external N loading was insignificant. Potentially, the phytoplankton N demand could be supported entirely by the internal N load via ammonium regeneration in the water column and sediment. However, the quantity of N from ammonium regeneration in the upper layer of sediment combined with that from the water column would limit the standing phytoplankton crop in spring in both basins and in late summer in the Western basin, especially when the algal biomass increases suddenly.     

Nitrate assimilatory properties of barley grown under long-term N limitation: Effects of local nitrate supply in split-root cultures

The effect of nitrate availability on characteristics of the nitrate assimilatory system was investigated in N-limited barley (Hordeum valgare L. cv. Golf), grown with the seminal root system split into initially equal-sized halves. The cultures were continuously supplied with nitrate-N at a relative addition rate (RA) of 0.09 day^?1, which resulted in relative growth rates (RG) that were ca 85% of those observed under surplus nitrate nutrition. The total N addition was divided between the subroots in ratios of 100:0, 80:20, 70:30, 60:40, and 50:50. For comparison, standard cultures were grown at RAs ranging from 0.03 to 0.18 day^?1. Initially, biomass and N partitioning to the subroots responded strongly and proportionally to the nitrate distribution ratio. After 12-14 days no further effect was observed. The V_max for net nitrate uptake and in vitro nitrate reductase (NR) activity were measured in acclimated plants, i.e., after > 14 days under a certain nitrate regime. In subroots fed from 20 to 100% of the total N addition, V_max for net nitrate uptake increased slightly, whereas NR activity was unaffected. Uptake and NR activities were insignificant in the 0%-subroot. Uneven nitrate supply to individual subroots had negligible effect on the whole-plant ability for nitrate uptake, and the relative V_max (unit N taken up per unit N in whole plant tissue and time) remained about 7-fold in excess of the demand set by growth. Balancing nitrate concentrations (the resulting external nitrate concentrations at a certain RA) generally ranged between 2 and 10 μM at growth-limiting RA, both when predicted from uptake kinetics and when actually measured. When comparing split root and standard cultures when acclimated, it appears that uptake and NR activities in roots respond more strongly to over-all nitrate availability than to nitrate availability to individual subroots.     

NITROGEN UPTAKE AND ASSIMILATION KINETICS IN ALEXANDRIUM MINUTUM (DYNOPHYCEAE): EFFECT OF N‐LIMITED GROWTH RATE ON NITRATE AND AMMONIUM INTERACTIONS1

Uptake and assimilation kinetics of nitrate and ammonium were investigated along with inhibition of nitrate uptake by ammonium in the harmful dinoflagellate Alexandrium minutum Halim at different nitrogen (N)–limited growth rates. Alexandrium minutum had a strong affinity for nitrate and ammonium (K_s=0.26±0.03 and 0.31±0.04 μmol·L^?1, respectively) whatever the degree of N deficiency of the cells. Ammonium was always the preferred form of nitrogen taken up (=0.42–0.50). In the presence of both forms, nitrate uptake was inhibited by ammonium, and inhibition was particularly marked in N‐sufficient cells (I_max～0.9 and K_i=0.31–0.56 μmol·L^?1). In the case of N assimilation, ammonium was also the preferred form in N‐deficient cells (=0.54–0.72), whereas in N‐sufficient cells, both N sources were equally preferred (=0.90–1.00). The comparison of uptake and assimilation rates highlighted the ability of A. minutum to significantly store in 1 h nitrate and ammonium in amounts sufficient to supply twice the daily N requirements of the slowest‐growing N‐deficient cells. Nitrogen uptake kinetic parameters of A. minutum and their ecological implications are discussed.     

Do cyanobacteria dominate in eutrophic lakes because they fix atmospheric nitrogen?

1. The sources of nitrogen for phytoplankton were determined for a bloom‐prone lake as a means of assessing the hypothesis that cyanobacteria dominate in eutrophic lakes because of their ability to fix nitrogen when the nitrogen : phosphorous (N : P) supply ratio is low and nitrogen a limiting resource. 2. Nitrogen fixation rates, estimated through acetylene reduction with ¹⁵N calibration, were compared with ¹⁵N‐tracer estimates of ammonium and nitrate uptake monthly during the ice‐free season of 1999. In addition, the natural N stable isotope composition of phytoplankton, nitrate and ammonium were measured biweekly and the contribution of N₂ to the phytoplankton signature estimated with a mixing model. 3. Although cyanobacteria made up 81–98% of phytoplankton biomass during summer and autumn, both assays suggested minimal N acquisition through fixation (<9% for the in‐situ incubations; <2% for stable isotope analysis). Phytoplankton acquired N primarily as ammonium (82–98%), and secondarily as nitrate (15–18% in spring and autumn, but <5% in summer). Heterocyst densities of <3 per 100 fixer cells confirmed low reliance on fixation. 4. The lake showed symptoms of both light and nitrogen limitation. Cyanobacteria may have dominated by monopolizing benthic sources of ammonium, or by forming surface scums that shaded other algae.     

ON PIGMENTS,GROWTH, AND PHOTOSYNTHESIS OF PHAEODACTYLUM TRICORNUTUM12

A maximum growth rate with doubling time of 18 hr at 18 C could be maintained. Continuous cultures at about half maximum growth rate provided cells for study of pigments and photosynthesis. The light intensity curve of photosynthesis had no unusual features and showed light-saturated rates of 30-35 μl O₂/mrn³-hr at 18 C. Pigment analysis showed chlorophylls a and c (a/c ratio = 4), fucoxanthin, β-carotene, and diadinoxanthin. Growth under red light (±660 mμ) altered pigments only by decrease in chlorophyll c to about one-half the content obtained under clear tungsten lamps. The large and anomalous spectral shift in fucoxanthin following organic solvent extraction runs confirmed, but efforts to isolate a native fucoxanthin were unsuccessful. Spectral analysis of acetone extracts and sonicated cell preparations allowed estimate of fractional absorption by each component pigment. The analysis shows that chlorophyll a and fucoxanthin are the principal light absorbing pigments and that absorption by other carotenoids is very small.     

Nitrogen isotope effects in the assimilation of inorganic nitrogenous compounds by marine diatoms

Nitrogen isotope fractionation in the assimilation of inorganic nitrogenous compounds was studied using marine diatoms (Phaeodactylum tricornutum and Chaetoceros sp.). The isotopic composition (δ¹⁵N) of the diatoms ranged from 7 to ‐18‰ relative to that of the nitrogen source, i.e., ammonium, nitrite, or nitrate. When the growth was light‐limited, the isotope fractionation in nitrate assimilation was inversely correlated with the growth rate. The highest fractionation factor of 1.016 was obtained when the growth rate was as low as 0.025 day^‐1. Fractionation was negligible when the growth, rate was higher than 1 day^‐1. A steady‐state kinetic model was applied to explain the isotope fractionation in nitrate assimilation. The nitrogen isotope fractionation primarily takes place at the step of N‐O bond breaking in nitrate reduction to nitrite. The extent of the isotope fractionation associated with the nitrate uptake is very small, and barely exceeds the limit of detection.     

VARIABILITY OF NITRATE UPTAKE CAPACITY IN MACROCYSTIS PYRIFERA (LAMINARIALES,PHAEOPHYTA) WITH NITRATE AND LIGHT AVAILABILITY1

The nitrate uptake capacity of mature blade tissue of the giant kelp, Macrocystis pyrifera (L.) C. Ag., was examined as a function of the availability of light and nitrate. Time course measurements indicated that nitrate uptake rate, as measured by the incorporation of ¹⁵N, was significantly increased by N starvation. The response was linear over the first hour of exposure regardless of the N status of the tissue indicating that surge uptake was not responsible for the increase. The Michaelis-Menten parameters V_max and K_s, however, were not significantly changed by either growth nitrate concentration or growth irradiance as a result of high variability among blades. Similarly, the initial slope (α) of the nitrate uptake kinetics curves was unaffected. Concentration of photosynthetic pigments increased in response to increased nitrate availability but not to increased growth irradiance. Time course and pigment data demonstrated that mature blade tissue responds to increased N availability by decreasing its capacity to take up nitrate and by increasing its investment in photosynthetic pigments, perhaps for N storage or enhanced light-harvesting capabilities and the increase in reducing power available for N assimilation. This study provides evidence for a dynamic regulatory system that responds to changes in nitrate availability in an integrated manner.     

Effect of NH4 +:NO3 - ratios on growth and nitrogen uptake by onions

The modelling of ion uptake by plants requires the measurement of kinetic and growth parameters under specific conditions. The objective of this study was to evaluate the effect of nine NH _inf4 ^sup+ :NO _inf3 ^sup− ratios on onions (Allium cepa L.). Twenty-eight to 84 day-old onion plants were treated with NH _inf4 ^sup+ :NO_f3/sup− ratios ranging from 0 to 100% of each ionic species in one mM solutions in a growth chamber. Maximum N influx (I_max) was assessed using the N depletion method. Except at an early stage, ionic species did not influence significantly I_max, the Michaelis constant (K_m) and the minimum concentration for net uptake (C_min). I_max for ammonium decreased from 101 to 59 pmole cm^-2 s^-1 while I_max for nitrate increased from 26 to 54 pmole cm^-2 s^-1 as the plant matured. On average, K_m and C_min values were 14.29 μM, and 5.06 μM for ammonium, and 11.90 μM and 4.54 μM for nitrate, respectively. In general, the effect of NH₄ ⁺:NO₃ ^- ratios on root weight, shoot weight and total weight depended on plant age. At an early stage, maximum plant growth and N uptake were obtained with ammonium as the sole source of N. At later stages, maximum plant growth and N uptake were obtained as the proportion of nitrate increased in the nutrient solution. The was no apparent nutrient deficiency whatever NH₄ ⁺:NO₃ ^- ratio was applied, although ammonium reduced the uptake of cations and increased the uptake of phosphorus. The research was supported by the Natural Sciences and Engineering Research Council of Canada.     

Nonlinear response of N2O flux to incremental fertilizer addition in a continuous maize (Zea mays L.) cropping system

The relationship between nitrous oxide (N₂O) flux and N availability in agricultural ecosystems is usually assumed to be linear, with the same proportion of nitrogen lost as N₂O regardless of input level. We conducted a 3‐year, high‐resolution N fertilizer response study in southwest Michigan USA to test the hypothesis that N₂O fluxes increase mainly in response to N additions that exceed crop N needs. We added urea ammonium nitrate or granular urea at nine levels (0–292 kg N ha^?1) to four replicate plots of continuous maize. We measured N₂O fluxes and available soil N biweekly following fertilization and grain yields at the end of the growing season. From 2001 to 2003 N₂O fluxes were moderately low (ca. 20 g N₂O‐N ha^?1 day^?1) at levels of N addition to 101 kg N ha^?1, where grain yields were maximized, after which fluxes more than doubled (to >50 g N₂O‐N ha^?1 day^?1). This threshold N₂O response to N fertilization suggests that agricultural N₂O fluxes could be reduced with no or little yield penalty by reducing N fertilizer inputs to levels that just satisfy crop needs.     

Nutrition and growth kinetics in nitrogen- and phosphorus-limited cultures of the novel red tide flagellate Chattonella ovata (Raphidophyceae)

In the summer of 2004, a harmful bloom caused by Chattonella ovata (Raphidophyceae) occurred over almost the entire area of the Seto Inland Sea and caused fishery damages. This incident was the first record of a bloom and damage to the fisheries caused by this species in Japanese waters. In order to elucidate the mechanism of the bloom outbreak, we examined the nutrition and the growth kinetics in nitrogen (N)- and phosphorus (P)-limited semi-continuous cultures of this species. Inorganic N compounds, such as nitrate, nitrite, and ammonium, were found to be good nitrogen sources for the growth of C. ovata, while organic nitrogen (urea and uric acid) was not utilized. This species was capable of using ATP, ADP and inorganic phosphorus compounds, but did not utilize phosphate monoesters as a sole P source. Under both N-limited and P-limited steady state conditions, the growth rate as a function of cell nitrogen and phosphorus quota, respectively, followed the Droop equation. Kinetic parameters μ_m (maximum growth rate) and k_q (minimum cell quota) obtained for N- and P-limited cultures were 0.79 day⁻¹ and 5.5 pmol N cell⁻¹ and 0.86 day⁻¹ and 0.48 pmol P cell⁻¹, respectively. The minimum cell quotas were 23–30% lower than those of C. antiqua. The nutrient availability and kinetic parameters of C. ovata are compared with other harmful algae and the ecological implications of these characteristics discussed.     

Nitrate regulation of nitrate uptake and nitrate reductase expression in barley grown at different nitrate:ammonium ratios at constant relative nitrogen addition rate

Barley (Hordeum vulgare L. cv. Golf) was cultured using the relative addition rate technique, where nitrogen is added in a fixed relation to the nitrogen already bound in biomass. The relative rate of total nitrogen addition was 0.09 day^?1 (growth limiting by 35%), while the nitrate addition was varied by means of different nitrate: ammonium ratios. In 3- to 4-week-old plants, these ratios of nitrate to ammonium supported nitrate fluxes ranging from 0 to 22 μmol g^?1 root dry weight h^?1, whereas the total N flux was 21.8 ± 0.25 μmol g^?1 root dry weight h^?1 for all treatments. The external nitrate concentrations varied between 0.18 and 1.5 μM. The relative growth rate, root to total biomass dry weight ratios, as well as Kjeldahl nitrogen in roots and shoots were unaffected by the nitrate:ammonium ratio. Tissue nitrate concentration in roots were comparable in all treatments. Shoot nitrate concentration increased with increasing nitrate supply, indicating increased translocation of nitrate to the shoot. The apparent V_max for net nitrate uptake increased with increased nitrate fluxes. Uptake activity was recorded also after growth at zero nitrate addition. This activity may have been induced by the small, but detectable, nitrate concentration in the medium under these conditions. In contrast, nitrate reductase (NR) activity in roots was unaffected by different nitrate fluxes, whereas NR activity in the shoot increased with increased nitrate supply. NR-mRNA was detected in roots from all cultures and showed no significant response to the nitrate flux, corroborating the data for NR activity. The data show that an extremely low amount of nitrate is required to elicit expression of NR and uptake activity. However, the uptake system and root NR respond differentially to increased nitrate flux at constant total N nutrition. It appears that root NR expression under these conditions is additionally controlled by factors related to the total N flux or the internal N status of the root and/or plant. The method used in this study may facilitate separation of nitrate-specific responses from the nutritional effect of nitrate.     

Soil fertility and fine root dynamics in response to 4 years of nutrient (N,P, K) fertilization in a lowland tropical moist forest,Panama

The question of how tropical trees cope with infertile soils has been challenging to address, in part, because fine root dynamics must be studied in situ. We used annual fertilization with nitrogen (N as urea, 12.5 g N m^?2 year^?1), phosphorus (P as superphosphate, 5 g P m^?2 year^?1) and potassium (K as KCl, 5 g K m^?2 year^?1) within 38 ha of old‐growth lowland tropical moist forest in Panama and examined fine root dynamics with minirhizotron images. We expected that added P, above all, would (i) decrease fine root biomass but, (ii) have no impact on fine root turnover. Soil in the study area was moderately acidic (pH = 5.28), had moderate concentrations of exchangeable base cations (13.4 cmol kg^?1), low concentrations of Bray‐extractable phosphate (PO₄ = 2.2 mg kg^?1), and modest concentrations of KCl‐extractable nitrate (NO₃ = 5.0 mg kg^?1) and KCl‐extractable ammonium (NH₄ = 15.5 mg kg^?1). Added N increased concentrations of KCl‐extractable NO₃ and acidified the soil by one pH unit. Added P increased concentrations of Bray‐extractable PO₄ and P in the labile fraction. Concentrations of exchangeable K were elevated in K addition plots but reduced by N additions. Fine root dynamics responded to added K rather than added P. After 2 years, added K decreased fine root biomass from 330 to 275 g m^?2. The turnover coefficient of fine roots <1 mm diameter ranged from 2.6 to 4.4 per year, and the largest values occurred in plots with added K. This study supported the view that biomass and dynamics of fine roots respond to soil nutrient availability in species‐rich, lowland tropical moist forest. However, K rather than P elicited root responses. Fine roots smaller than 1 mm have a short lifetime (<140 days), and control of fine root production by nutrient availability in tropical forests deserves more study.     

KINETICS OF CELL DEATH IN THE CYANOBACTERIUM ANABAENA FLOS-AQUAE AND THE PRODUCTION OF DISSOLVED ORGANIC CARBON1

Kinetics of cell death and the production of dissolved organic carbon (DOC) were investigated in Anabaena flos-aquae (Lyngb.) Bréb grown on three different N sources (N₂nitrate, and ammonium) in a phosphorus (P)-limited chemostat. The fraction of live cells in the total population increased as growth rate increased with decreasing P limitation. Cell death was less in nitrate and ammonium media than in N₂. The specific death rate (γ), when calculated as the slope ofv^?1_x vs. D^?1, where v_xand D are live cell fraction (or cell viability) and dilution rate, respectively, was 0. 0082 day^?1 in N₂and 0.0042 day^?1 in nitrate. The slope of the plot in ammonium culture was not significant; however, the value of the live cell fraction was within the range for the NO^?₃culture. The fraction of live vegetative cells in N₂ culture was constant at all growth rates and the increase in the overall live cell fraction with growth rate was due entirely to an increase in live heterocysts. Live heterocysts comprised 3.5% of the total cells at a growth rate of 0.25 day^?1 and increased to 6.3% at 0.75 day^?1 with the ratio of live heterocysts to live vegetative cells linearly increasing with growth rate. The fraction of live vegetative cells was invariant in nitrate cultures us in N₂cultures. The live heterocysts fraction also increased with growth rate in nitrate cultures, along with the live heterocysts : live vegetative cells ratio, but the level was lower than in N₂cultures. DOC released from dead cells increased inversely with growth rate in N₂from 36.4% of the total DOC at a growth rate of 0.75 day^?1 to 54.15% at 0.25 day^?1. The contribution of cell death to the total DOC production in nitrate and ammonium media was significantly less than that under N₂DOC from dead cells consisted mainly of high-molecular-weight compounds, whereas DOC excreted from live cells was largely of low molecular weight.     

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.