Carbohydrate metabolism in the marine diatom Skeletonema costatum

This is a study on the metabolism of the algal biochemical constituents of a marine diatom, Skeletonema costatum. The carbohydrates, protein, and lipid, of the diatom grown under light and dark conditions were analyzed. The composition of the organic compounds did not vary in the alga grown under continuous light conditions throughout the experiment, although a little accumulation of carbohydrate and lipid was observed in the stationary phase of the algal growth. When the diatom, at maximum growth stage, was transferred to darkness, 44.4% of the carbohydrate, 57.8% of the protein, and 27.0% of the lipid were used by respiration within a few days. Detailed analysis of the algal carbohydrate demonstrated that low molecular weight carbohydrates, consisting of glucose, and -1,3-glucan, were readily metabolized by algal respiration, leaving cell-wall carbohydrates consisting of mannan and pentosan which are immune to algal respiration. The respiratory utilization of carbohydrates is discussed in relation to the rapid decay of glucan from marine particulate matter during the course of its sinking to further depths in the ocean, as observed previously by the present author.     

Ecotypic differentiation in the marine diatom Skeletonema costatum: influence of light intensity on the photosynthetic apparatus

Three genetically distinct clones of Skeletonema costatum (Grev.) Cleve were grown at 20°C under high (274 E m^-2 s^-1) and low (27 E m^-2 s^-1) light conditions and their photoadaptive photosynthetic responses compared. When all three clones were grown under low light, pigment analyses and fluorescence excitation spectra demonstrated that the accessory pigments, chlorophyll c and fucoxanthin, became more important in light-harvesting compared to chlorophyll a. Photosynthetic unit sizes increased for Photosystems I and II in low light, but photosynthesis vs irradiance characteristics were not reliable predictors of photosynthetic unit features. Fluorescence excitation spectra and photosynthesis vs irradiance (P-I) relationships indicated that changes in energy transfer occurred independent of changes in pigment content. Large increases in accessory pigment content were not accompanied by large increases in excitation from these pigments. Changes in energy transfer properties were as important as changes in PSU size in governing the photoadaptive responses of S. costatum. When the three clones were grown under identical conditions, each had a separate and distinct pattern of photoadaptation. Significant differences among clones were found for pigment ratios, photosynthetic unit sizes for Photosystems I and II and efficiency of energy transfer between pigments. These strikingly different photoadaptive strategies among clones may partially account for the great ecological success of the diatom species. This is the first quantitative investigation of the importance of both chlorophyll c and fucoxanthin to the adaptive responses of diatoms to light intensity, and represents the most complete characterization of the photoadaptive responses of a single species of marine phytoplankter to differences in light environment.     

Diel variation in chlorophyll a,carbohydrate and protein content of the marine diatom Skeletonema costatum

Skeletonema costatum (Greville) Cleve isolated from Narragansett Bay, USA, was incubated at 3 light intensities (ca. 0.008, 0.040 and 0.075 ly min^-1) under a 12 h light: 12 h dark (12L:12D) photoperiod at 2°, 10° and 20°C. Cellular chlorophyll a increased at intensities less than ca. 0.040 ly min^-1; increases occured within one photoperiod at temperatures above 10°C. Cellular carbohydrate increased with light intensity at all temperatures; increases during the photophase were due to net production of the dilute acid-soluble fraction. Cellular protein increased during the photoperiod at 10° and 20°C; there was little difference in cellular protein among all cultures after one photoperiod. The rate at which cellular chlorophyll a increased in response to a decrease in light suggests that diel variation in cellular chlorophyll a is temperature-dependent in S. costatum. Protein: carbohydrate ratios ranged from ca. 0.5 to 2.0 over a diel cycle; ratios increased at lower intensities and higher temperatures. The diel range in protein:carbohydrate ratios equals that in cultures developing nitrogen deficiency; thus, use of this ratio as an index to phytoplankton physiological state must account for diel light effects.     

The rapid response of the marine diatom Skeletonema costatum to changes in external and internal nutrient concentration

A nitrogen-deficient batch culture of the marine diatom Skeletonema costatum, when resupplied with a mixture of nitrate and ammonium, showed an initial enhanced nitrate uptake rate leading to a large internal concentration (pool) of nitrate. Following this initial nitrate uptake event, nitrate uptake ceased, and nitrate assimilation was inhibited until the ammonium present was used. At this point, nitrate uptake resumed and nitrate assimilation began. No internal ammonium pool was observed during nitrate utilization, but a large nitrate pool remained throughout the utilization of external nitrate. The internal nitrate pool decreased rapidly after exhaustion of nitrate from the culture medium, but growth of cellular particulate nitrogen continued for about 24 h. A mathematical simulation model was developed from these data. The model cell consisted of a nitrate pool, ammonium pool, dissolved organic nitrogen pool, and particulate nitrogen. It was found that simple Michaelis-Menten functions for uptake and assimilation gave inadequate fit to the data. Michaelis-Menten functions were modified by inclusion of inhibitory and stimulatory feedback from the internal pools to more accurately represent the observed nutrient utilization.     

Implications of salinity fluctuation for growth and nitrogen metabolism of the marine diatom Ditylum brightwellii in comparison with Skeletonema costatum

In 1987 effects of salinity fluctuations on growth of Ditylum brightwellii (West) Grunow, isolated from the Eastern Scheldt estuary (SW Netherlands) in 1981, were studied. D. brightwellii was grown in a 12 h light: dark cycle at constant salinity in brackish media. Ammonium-limited cultures were subjected to a salinity fluctuation. By decreasing the salinity to 4.8 photosynthesis and cell division were inhibited; cells were deformed. Protein and carbohydrate contents increased slightly, dark respiration was stimulated and cellular levels of glucose decreased at low salinity; this indicated a possible role of sugars in osmoregulation. Ammonium was accumulated in cultures, amino acids may have been stored; the role of the vacuole as a storage compartment was discussed. Both the ammonium uptake capacity and the affinity for ammonium decreased. Nitrogen limitation was relieved in the transient state. [With the activity of the nitrogen assimilation enzymes glutamine synthetase (GS) and glutamate synthase (GOGAT) being uninhibited by lower salinity.] Recovery from hypo-osmotic stress during a salinity increase was initiated by stimulated photosynthesis; chlorophyll a increased, but persistant contractions of cytoplasm (with chloroplasts) may have delayed cell growth. The glutamate dehydrogenase (GDH) activity decreased further whereas the cellular level of alanine increased in the presence of large ammonium pools; this may indicate a temporary activity of ADH (alanine dehydrogenase). Skeletonema costatum (Greville) Cleve, recovered faster from hypoosmotic stress than did D. brightwellii. Due to an osmotic shock from 13.6 to 7.1 S both species excreted amino acids and glucose; S. costatum accumulated more glucose, D. brightwellii accumulated more amino acids. S. costatum may with the competition for nitrogen in waters with an unstable salinity; it will replace D. brightwellii.Contribution no. 427 Delta Institute for Hydrobiological Research, Yerseke, The Netherlands     

Impact of a temporal salinity decrease on growth and nitrogen metabolism of the marine diatom Skeletonema costatum in continuous cultures

In 1987 effects of salinity fluctuations on growth of the centric diatom Skeletonema costatum (Greville) Cleve, isolated from the brackish Krammer estuary (SW Netherlands) in 1981, were investigated. Continuous cultures (12 h light: dark cycle) of S. costatum were adapted to constant salinity in natural (16.1) and synthetic (13.5) media. For several days the ammonium-limited cultures were exposed to a salinity fluctuation (minimum 4.8). Decreasing salinity caused an inhibition of photosynthesis, dark respiration and cell growth. Cellular pools of glucose decreased. While the carbohydrate content remained constant, the protein content increased slightly. Net carbon fixation was more inhibited than nitrogen assimilation. Ammonium accumulated during a salinity decrease; a total decline of the overcapacity of ammonium uptake was noticed and nitrogen limitation was relieved. Amino acid pools decreased, probably as a result of excretion (osmoregulation). The enzymes invoilved in ammonium assimilation showed an increased activity. Cellular activities were resumed during a salinity increase. Chlorophyll a increased; photosynthesis, ammonium uptake and growth were stimulated. The ammonium uptake capacity recovered completely; glutamic acid accumulation and increased glutamate-dehydrogenase (GDH) activity indicated supplementary ammonium assimilation via GDH. The activities of glutamine synthetase/glutamate synthase (GS/GOGAT) and GDH stabilized, and the cells returned to steady state under ammonium limitation.Communication no. 426 Delta Institute for Hydrobiological Research, Yerseke, The Netherlands     

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.     

Effect of heat stress on polyamine content and protein pattern in Skeletonema costatum

In the present paper we investigated the polyamine content and protein expression pattern in laboratory-reared Skeletonema costatum (Grev.) Cleve under temperature stress conditions. Putrescine, spermidine and to a lesser extent spermine were present in this alga. Free putrescine and spermidine increased after heat shock while a concomitant decrease in trichloroacetic acid (TCA)-soluble bound putrescine and spermidine occurred. After 40 min of 30 °C stress both were no longer detectable, whereas TCA-soluble bound spermine was present. The appearance of 83, 42, 40.7 and 35 kDa polypeptides was observed when S. costatum cells were subjected to heat treatment. Western blotting experiments revealed the presence in this diatom of ubiquitin and ubiquitin conjugates. Heat stress strongly increased the number of ubiquitin conjugate polypeptides of high molecular weight. No variation in the carbohydrate content was observed in response to stress. A possible role for these different metabolic events in the adaptive response of S. costatum to rapid temperature shifts has been hypothesized.     

Diurnal and circadian rhythms in the turbidity of growing Skeletonema costatum cultures

Skeletonema costatum (Grev.) Cleve grown in batch culture at low light intensity under a 14 h light: 10 h dark photocycle showed exponential cell proliferation (1.1 doublings d^-1) without significant phasing of the cell division by the light: dark cycle. The growth in carbon concentration was, however, restricted to the light period. The turbidity of the culture closely followed the carbon oattern, and was not affected by the increase in the cell number during the dark period. It was found that a trustule suspension had only approximately 1% of the turbidity of the corresponding intact algae. Culture turbidity was therefore regarded as a biomass parameter similar to the carbon concentration, without direct correlation to the timing of the cell division. The short-time variations in the turbidity of growing algal cultures were further studied in a cage culture turbidostat. The growth rate (based on turbidity) increased rapidly during the first half of the light period, decreased slightly towards the evening and was zero throughout the dark period. When transformed to continuous light, the growth of the culture continued to show damped oscillations for up to 1 wk, but with a period of 26.7 h instead of 24 h. The same circadian rhythm was observed in chlorophyll content, and is thus possibly a reflection of a freely oscillating internal biological clock. The cage culture turbidostat was found to be a suitable device for studies of the photocycle related regulation of biosynthesis in S. costatum.     

钻井液及其组分对中肋骨条藻的毒性效应（Toxic Effects of Water-Based Drilling Fluid and Its Components on Skeletonema costatum）

为探讨钻井液及其组分的毒性效应,采用荧光技术研究了一种水基钻井液WBF及其7种主要组分(HXJ、PAC141、KPAM、PolyAL、XC、PolyA、Mud)对海洋微藻中肋骨条藻(Skeletonema costatum)的毒性作用,以未加入WBF或其组分的中肋骨条藻的叶绿素荧光强度为参照,计算得到钻井液及其组分对中肋骨条藻不同时间的EC50值.结果表明:WBF的7种单组分中,PolyA对中肋骨条藻的96h EC50=2×103mg·L-1,为有毒组分,PolyAL的96h EC50=3×104mg·L-1,为低毒组分,其余5种组分无毒;WBF的有毒和低毒组分复配时其毒性与作用时间有关,PolyA与PolyAL在24h之内表现为协同作用,之后主要表现为拮抗作用;WBF的毒性主要取决于其有毒组分,而低毒和无毒组分对钻井液的毒性有协同增强作用.     

Allelopathic inhibition on red tide microalgae Skeletonema costatum by five macroalgal extracts

This study aims to identify effective antialgal allelochemicals from marine macroalgae that inhibit the growth of red tide microalgae. Practically, new algicidal agents were developed to control red tide. The growth inhibitory effects of 5 marine macroalgae Porphyra tenera, Laminaria japonica, Ulva pertusa, Enteromorpha clathrata, and Undaria pinnatifida on Skeletonema costatum were evaluated by adding crude seawater extracts of macroalgal dry tissue into the culture medium containing S. costatum. The half-effective concentrations at 120 h (EC_{50, 120 h}) of the seawater extracts were 0.6, 0.9, 1.0, 1.0, and 4.7 g/L for the five macroalgae above, respectively. E. clathrata, L. japonica and U. pertusa showed strong allelopathic effect on the growth of S. costatum. There have been no previous reports with regard to the allelopathic effects of the former two macroalgae so far. The possible allelochemicals of 21 compounds of the E. clathrata were detected using Gas chromatography-mass spectrometry (GC-MS) analysis. Unsaturated fatty acids, acrylic acid (C₃H₄O₂), and linolenic acid (C₁₈H₃₀O₂) were the most likely allelochemicals in E. clathrata.     

营养盐浓度对中肋骨条藻优势种群富集多溴联苯醚的影响

以中肋骨条藻（Skeletonema costatum）和东海原甲藻（Prorocentrum donghaiense）组成中肋骨条藻优势种群的海洋微藻为研究对象,以我国近海水体中存在的3种丰度较高的多溴联苯醚同系物为代表,包括2,4,4＇-三溴联苯醚（BDE-28）、2,2’,4,4’-四溴联苯醚（BDE-47）和2,2＇,4,4＇,5-五溴联苯醚（BDE-99）,研究了硝酸盐浓度（0、128、512μmol·L-1）和磷酸盐浓度（0、8、32μmol·L-1）对中肋骨条藻优势种群生化成分和富集3种多溴联苯醚的影响,分析了中肋骨条藻优势种群对多溴联苯醚的富集量与生化成分的关系。结果表明,随着营养盐浓度的升高,单位藻细胞的富集量呈现降低趋势,硝酸盐浓度为0μmol·L-1时,3种多溴联苯醚同系物每106cells的富集量为3.93-5.01 ng,而硝酸盐浓度为512μmol·L-1时,每106cells富集量为3.53-3.94 ng,降低了10%-23%;不同硝酸盐浓度下单位体积藻液富集量差异较小,随硝酸盐浓度的升高,BDE-99和47富集量分别从0.180和0.179 ng·mL^-1降低到0.167和0.150 ng·mL^-1,BDE-28没有出现明显差异。与硝酸盐相似,多溴联苯醚单位藻细胞富集量也随着磷酸盐浓度的升高而呈现出降低的趋势,每106cells的富集量从磷为0μmol·L-1的3.04-3.28 ng降低到32μmol·L-1时的2.27-2.73 ng;不同磷酸盐浓度下单位体积藻液的富集量差异不显著,其中BDE-99富集量范围在0.188-0.190 ng·mL^-1,随磷酸盐浓度的升高,BDE-47和28富集量略有降低,从0.20 ng·mL^-1降低到约0.16 ng·mL^-1。中肋骨条藻优势种群单位细胞对多溴联苯醚富集随着营养盐浓度的增加而降低,其原因是营养盐浓度影响了其细胞生化成分,其中脂类含量与富集量存在正相关关系。对3种同系物富集量的比较发现,随多溴联苯醚同系物的Kow降低,富集量总体上呈现降低的趋势。     

Influence of temperature on the growth rate of Skeletonema costatum in response to variations in daily light intensity

Daily light intensities (I _o) can vary 10-fold during the winter-spring and late-summer diatom blooms in New England, USA, coastal waters. Laboratory cultures and natural populations incubated in dialysis sacs were examined to determine the time course of growth rate in Skeletonema costatum (Greville) Cleve in response to variations in daily light intensity during two bloom periods in Narragansett Bay, Rhode Island, USA. Log-phase cultures of S. costatum require 2 d to attain maximum growth rates at 2°C following transfer to saturating intensities. At 20°C, only 1 d is required. As temperature increases, Detonula confervacea (Cleve) Gran, Thalassiosira nordenskiöldii Cleve and Ditylum brightwellii (West) Grunow also exhibit rapid increases in mean daily division rates (K) following transfer to saturating light intensities. Thalassiosira pseudonana Hustedt, however, did not alter the time required to achieve maximum K as temperature varied. Natural populations of S. costatum did not show a well-defined relationship between K and light. Throughout a winterspring bloom, K was limited by low temperatures and exhibited no clear response to variations in I _o. A change in K in response to variation in I _o may occur on a daily basis during the summer, when temperatures are near 20°C; this has yet to be verified for in situ populations.     

Action de métaux lourds à des doses sublétales sur les caractéristiques de la croissance chez la diatomée Skeletonema costatum

Sublethal effects of mercury, cadmium and copper on the diatom Skeletonema costatum (Grev.) Cleve, grown in batch and bacteria-free culture are studied. Division rate, maximum yield growth, mean cell volume, particulate carbon and nitrogen, and ¹⁴C-bicarbonate uptake are used as toxic impairment criteria. Division rate is the first-affected and most sensitive parameter, but algal responses vary according to the metal. Hg produces an acute decrease in division rate, followed by a temporary recovery of growth capacity within the first 48 h after metal addition. Cd, on the other hand, increases division rate, followed by an obvious decrease. Cu reduces division rate slowly or quickly, depending on the metal concentration. Cell synthesis capacity (culture biovolume, particulate carbon and nitrogen, carbon assimilation) is less affected than division rate, especially with Hg. The C:N cell ratio is unchanged at sublethal concentrations, even when production is reduced. The mean cell volume is slightly affected: the variations are not greater than those of the control during its growth phases. Markedly teratological forms are never observed. In the authors' opinion, these results confirm that many parameters and growth kinetic aspects must be considered to fully appreciate the effects of sublethal concentrations of heavy metals. It would also be of advantage to develop a better methodology for such research, applying, for example, techniques already employed in enzymology.     

Effects of ethyl 2-methyl acetoacetate (EMA) on the growth of Phaeodactylum tricornutum and Skeletonema costatum

We investigated the effects of ethyl 2-methyl acetoacetate (EMA) on growth of the marine diatom algae Phaeodactylum tricornutum (P. tricornutum) and Skeletonema costatum (S. costatum). Growth of P. tricornutum was significantly inhibited by the minimum concentration (3.5 mmol·L ^?1) of EMA at lower initial algal densities (IADs) (3.6×10⁴ and 3.3×10⁵ cells·mL ^?1). However, at the highest IAD, significant growth inhibition was found at above 7 mmol·L ^?1 of EMA exposure. In S. costatum, EMA concentrations of 10.5 mmol·L ^?1 or more significantly inhibited growth at lower IAD (3×10⁴ and 1.8×10⁵ cells·mL ^?1); at the highest IAD, only EMA concentrations above 14 mmol·L ^?1 obviously inhibited the growth of S. costatum. Changes in specific growth rates and pigment were consistent with algal growth, but only at higher EMA concentrations or lower IAD values was the ratio of chlorophyll a (Chla) to carotenoid significantly lower than the control. Medium effective concentration (EC ₅₀) values were in the order 4.07, 8.03 and 12.27 mmol·L ^?1 for P. tricornutum and 7.48, 11.92 and 17.22 mmol·L ^?1 for S. costatum. All these results show that the effect of EMA on the growth of algae was species specific and mainly depended on IAD, which might be an important factor to influence algal growth.     

Membrane voltage of marine phytoplankton,measured in the diatom Coscinodiscus radiatus

In order to investigate the role of the ionic relations in buoyancy of marine phytoplankton, voltage recordings have been made on the planktonic diatom Coscinodiscus radiatus using conventional glass microelectrode techniques. The most negative Nernst equilibrium voltage in C. radiatus is E _K, the potassium equilibrium voltage of around-85 mV. Accordingly, stable voltages of-40 to-80 mV were recorded from C. radiatus which conforms to the general theory of electro-diffusion of ions through membranes (voltage range V _d). In addition, membrane voltages much more negative, e.g. up to-140 mV (voltage range V _p), have been recorded in C. radiatus; these voltages demonstrate the operation of an electrogenic pump. Within the voltage range V _d, light-on and-off (microscope illumination) caused weak hyper- and depolarizations by about 2 mV with a time constant of about 10 s. Also within V _d, spontaneous oscillations could be observed with a frequency of about 0.03 Hz and irregular amplitudes up to 30 mV. These phenomena are simulated by a model for electrocoupling of the major ion transporters in plants, as worked out for guard cells with their subtle osmoregulatory system. Equivalent mechanisms are suggested to operate in planktonic diatoms for adjustment of buoyancy by appropriate uptake and release of ions.     

Grazing of Acartia hudsonica (A. clausi) on Skeletonema costatum in Narragansett Bay (USA): Influence of food concentration and temperature

Grazing experiments were performed with temperatureacclimated Acartia hudsonica fed the diatom Skeletonema costatum in concentrations ranging from 50 to 3×10⁴ cell ml^-1 at 5°, 10° and 15°C. The ingestion data were best fit by an Ivlev equation. Feeding threshold values of 39 and 59 cells ml^-1 were not significantly different from zero; however, filtration rates were depressed at low food concentrations. Maximum filtration rates increased exponentially with temperature, reaching a maximum with copepods collected at 14°–15°C, and then declining. Both the increase in ingestion rate with increasing food concentration and the maximum ingestion rate were significantly greater as experimental temperature was increased. Maximum ingestion rates were reached at concentrations greater than 6×10³ cells ml^-1. Percent of body carbon ingested per day at 5 g C L^-1 increased from 1.5% at 5°C to 6.7% at 15°C. At 500 g C L^-1, the ingestion increased from 84% (5°C) to 660% (15°C). Percent of body nitrogen at 0.5 g N L^-1 increased from 0.6% per day at 5°C to 2.5% per day at 15°C. At 50 g N L^-1, the ingestion was 42% body nitrogen at 5°C and 250% at 15°C. The influence of grazing by A. hudsonica on phytoplankton in Narragansett Bay, USA was estimated for 1972–1977. The percent of standing stock removed by grazing rarely exceeded 5% per day except during the late spring when S. costatum growth becomes nutrient limited and higher temperatures favor the rapid population growth of A. hudsonica.     

Marine diatoms grown in chemostats under silicate or ammonium limitation. II. Transient response of Skeletonema costatum to a single addition of the limiting nutrient

Skeletonema costatum was grown at different steady-state growth rates in ammonium or silicate-limited chemostats. The culture was perturbed from its steady-state condition by a single addition of the limiting nutrients ammonium or silicate. The transient response was followed by measuring nutrient disappearance of the liliting perturbation experiment indicate that three distinct modes of uptake of the limiting nutrient can be distinguished; surge uptake (V _s ), internally controlled uptake (V _i ), and externally controlled uptake (V _e ). An interpretation of these three modes of uptake is given and their relation to control of uptake of the limiting nutrient is discussed. The uptake rates of the non-limiting nutrients were shown to be depressed during the surge of the uptake of the limiting nutrient. Kinetic uptake parameters, K _s and V _max, were obtained from data acquired during the externally controlled uptake segment, V _e . The same V _max value of 0. 12 h^-1, was obtained under either silicate or ammonium limitation. Estimates of K _s were 0.4 g-at NH₄-N l^-1 and 0.7 g-at Si l^-1. Short-term ¹⁵N uptake-rate measurements conducted on nitrogen-limited cultures appear to be a combination of V _s or V _i , or at lower substrate concentrations V _s and V _e . It is difficult to separate these different uptake modes in batch or tracer experiments, and ensuing problems in interpretation are discussed.Contribution No. 882 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA. This work represents portion of three dissertations submitted to the Department of Oceanography, University of Washington, Seattle, in partial fulfillment of the requirements for the Ph.D. degree.     

Marine diatoms grown in chemostats under silicate or ammonium limitation. I. Cellular chemical composition and steady-state growth kinetics of Skeletonema costatum

Skeletonema costatum was grown in chemostats under ammonium or silicate limitation to examine its growth kinetics and changes in cellular chemical composition at different steady-state growth rates. When the relationship between the effluent limiting substrate concentration and steady-state growth rates was examined, deviations from the simple hyperbolic form of the Monod growth equation were noted at low and high dilution rates. The data from the plot of growth rate and substrate concentration were divided into 4 regions and the relationship of these region to cell quota is discussed. Two physiological states were identified. All populations grown at D<0.05 h^-1, regardless of the size of the cells or the magnitude of Q, exhibited a maximal growth rate of approximately 0.05 h^-1, while populations grown at higher dilution rates (D>0.06 h^-1 to 0.14 h^-1). The maximal value of growth rate is obtained only in cultures grown at very high dilution rates where nutrient shift-up appears to occur, the cell quota approaches a maximum and the heterogeneous cell population becomes more homogeneous.Contribution No. 881 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA. This paper represents a portion of two dissertations submitted by P.J.H. and H.L.C. to the Department of Oceanography, University of Washington, Seattle, in partial fulfillment of the requirements for the Ph.D. degree.     

Life history of the temperate subtidal gastropod Umbonium costatum

The life history and reproductive biology of the trochid sand snail Umbonium costatum (Kiener) were investigated on a subtidal sandy shore in Hakodate Bay, Japan between 1988 and 1991. Female U. costatum mature at 1 yr of age (shell diameter-11 mm), reproduce twice (in June-July and September-October) in successive years, grow to a maximum size (shell diameter=22 mm) at age 8 yr, increase annual fecundity with age from 2000 (age 1 yr) to 37000 (age 8 yr), and show a maximum monthly gonad somatic index of 8% which is constant among ages. In comparison to a previously studied life history of a tropical Umbonium vestiatium, temperate U. costatum shows more sustained growth and a longer life span after maturation. This could be explained by: (1) the optimal size model concerned with resource investment, in gametes (Sebens 1987); and by (2) bet hedging to compensate large variability in larval success at high latitudes. These two hypotheses are not mutually exclusive, but both are based on season-related extremes of environment at high latitudes where the period suitable for reproduction is short.