Carbon-nitrogen relations at whole-cell and free-amino-acid levels during batch growth of Isochrysis galbana (Prymnesiophyceae) under conditions of alternating light and dark

Isochrysis galbana Parke, Strain CCAP 927/1, was grown in ammonium-limited batch culture under a 12 h light: 12 h dark illumination cycle. Samples were taken every 12 h over the 26 d period from lag phase through exponential into stationary phase (no net carbon fixation), with more frequent sampling at points of interest. Exponential cell-specific growth rate was 0.3 to 0.4d^-1. Cell division occurred during the dark phase, while cell volume increase, ammonium uptake, and pigment synthesis occurred during the light. Stationary phase cells were small, and the lag phase was long (5 d) even though the C:N ratio had returned from 18 to 6.5 within 2 d, followed by synthesis of chlorophyll a. Net chlorophyll synthesis ceased within 4 d of exhaustion of the nitrogen source. The chlorophyll c: chlorophyll a ratio remained constant during increasing nitrogen deprivation. Biovolume and carotenoids correlated with carbon biomass. Levels of chlorophyll a correlated poorly with carbon fixation and carbon biomass once the nitrogen source had been exhausted. Except after the addition of ammonium to nitrogen-deprived cells (refeeding), the content of intracellular glutamine and the glutamine: glutamate ratio were low during the dark phase, rising to a plateau within the first 1 h of illumination. Refeeding of cells which had only just exhausted the extracellular nitrogen source resulted in a much smaller increase in glutamine than refeeding of nitrogen-starved (stationary-phase) cells. Nitrogen biomass correlated with the presence of an unidentified intracellular amine.     

Kinetics of light-intensity adaptation in a marine planktonic diatom

The marine planktonic diatom Thalassiosira weisflogii was grown in turbidostat culture under both continuous and 12 hL: 12 hD illumination regimes in order to study the kinetics of adaptation to growth-irradiance levels. In both illumination regimes adaptation to a higher growth-irradiance level was accompanied by an increase in cell division rates and a decrease in chlorophyll a cell^-1. The rates of adaptation for both processes, derived from first order kinetic analysis, equaled each other in each experiment. The results suggest that during the transition from low-to-high growth-irradiance levels chlorophyll a is diluted by cell division and is not actively degraded. Introduction of a light/dark cycle lowered the rate of adaptation. In transitions from high-to-low growth-irradiance levels there was a sharp drop in growth rates and a slow increase in chlorophyll a cell^-1 under both continuous and intermittent illumination. In the 12 hL:12hD cycle there was a circadian rhythm in chlorophyll a cell^-1, where cellular chlorophyll contents increased during the light cycle and decreased during the dark cycle. This circadian rhythm was distinctly different from light intensity adaptation. For kinetic analysis of light intensity adaptation in a 12 hL: 12 hD cycle, the circadian periodicity was separated from the light intensity response by subjecting the data to a Kaiser window optimization digital filter. Kinetic parameters for light-intensity adaptation were resolved from the filtered data. The kinetics of lightintensity adaptation of marine phytoplankton are discussed in relation to their spatial variations and time scales of mixing.This research was performed at Brookhaven National Laboratory under the auspices of the United States Department of Energy under Contract No. DE-AC02-76 CH00016     

Effects of light quality on initiation and development of meroplanktonic diatom blooms in a eutrophic shallow sea

We investigated the effects of light quality on resting stage cell germination and vegetative cell growth of meroplanktonic diatoms in a small port in Hakata Bay, Japan and in the laboratory. During the investigation over the year of 2006, the meroplanktonic diatom bloom first occurred in the end of May and then repeated wane and wax until October in the small port. From late April to middle May, light penetrating the water column was often strong and attenuations of all spectral lights were low. During this period, Skeletonema costatum, Thalassiosira minima, and Chaetoceros sp. appeared frequently, followed by the blooms of S. costatum and Chaetoceros sp. in late May. Thereafter, S. costatum and Chaetoceros sp. bloomed in late June but not in middle June, when pigmented flagellates bloom appeared. The attenuation of short-wavelength light such as violet and blue lights was markedly high during these diatom and flagellate blooms; all blooms disappeared within several days. Vegetative cell strains of the three diatoms under light emitting diodes (LEDs) with six different spectra (violet, blue, green, orange, red, and near-infrared) grew at a higher rate under short-wavelength light, violet and blue. On the other hand, when suspensions of bottom sediments from Hakata Bay were cultured under the same LEDs and in the dark, vegetative cells of S. costatum appeared under all LEDs except for orange and near-infrared, vegetative cells of T. minima appeared under all LEDs but not in the dark, and vegetative cells of Chaetoceros sp. appeared under violet and blue LEDs. However, vegetative cell densities of the three diatoms increased much more under violet light than under other LEDs within a short period (6 days). Our study indicates that underwater penetration by short-wavelength light, such as violet and blue, may be an important factor in the initiation and development of meroplanktonic diatom blooms.     

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.     

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     

DNA synthesis and the cell cycle in the noxious red-tide dinoflagellateGymnodinium nagasakiense

Gymnodinium nagasakiense is a noxious red tide dinoflagellate often associated with damage to fisheries in Japan. DNA synthesis and the cell cycle in this organism were investigated from 1989 to 1990 by determining relative DNA contents of individual cells using an epifluorescence microscopy-based microfluorometry system. The nuclei were stained with the DNA-specific fluorochrome 4-6-diamidino-2-phenylindole (DAPI). Because photosynthetic pigment interferes with the fluorescence from the DAPI-DNA complex, the pigment was eliminated by methanol treatment as a first step in quantitative microfluorometry. Nuclear DNA contents, cell size distribution, cell density, and frequency of paired cells were determined every 2h for 24h using cells grown on a 12h light:dark cycle. DNA synthesis and cell division were tightly phased to a particular period of the light:dark cycle. DNA synthesis (S phase) occurred from 10:00 to 22:00 hrs and was followed by cytokinesis. The presence of such a distinct S phase strongly suggests thatG. nagasakiense has a typical eukaryotic cell cycle. This type of cell cycle makes it possible to estimate speciesspecific in situ growth rate based on the diel pattern of DNA synthesis.     

Carbon and nitrogen metabolism by Monochrysis lutheri: Measurement of growth-rate-dependent respiration rates

Dark respiration rates were measured and carbon-excretion rates calculated for a nitrate-limited population of the marine chrysophyte Monochrysis lutheri grown in continuous culture at 20°C on a 12 h light-12 h dark cycle of illumination and over a series of 4 growth rates. A significant (P<0.05) positive correlation was found between dark respiration rate and growth rate. From a simple linear fit to the data, the respiration rate at maximum growth rate was estimated to be roughly 10.5% of the maximum gross-carbon-production rate, and more than three times higher than the extrapolated respiration rate at zero net-growth rate. Carbon-excretion rates showed no significant correlation with growth rate, and averaged less than 5% of the maximum gross-carbon-production rate. Mean cell nitrogen to carbon ratios were correlated in a virtually linear manner (r=0.994) with growth rate, and at a given growth rate were consistently higher than nitrogen to carbon ratios for the same species grown on continuous light. A comparison of carbon and nitrogen quotas as a function of growth rate for M. lutheri and other species suggests that the increase of cellular nitrogen at high growth rates under nitrate-limited growth conditions may be associated with the storage of cellular protein or amino acids rather than the presence of an inorganic nitrogen reservoir. The maximum nitrate uptake rate per cell during the day changed very little over the range of growth rates studied, and was comparable to the maximum uptake rate found for cells grown on continuous light. However, the cell nitrogen quota increased steadily with growth rate, causing a reduction in the maximum specific-uptake rate of nitrate during the day at high growth rates. The dark nitrate-uptake capacity of the population was clearly exceeded by the supply rate at the two higher growth rates, leading to a buildup of nitrate during the night which amounted to as much as 21% of the particulate nitrogen in the growth chamber by morning.Hawaii Institute of Marine Biology Contribution No. 478.     

Growth and competition of the marine diatoms Phaeodactylum tricornutum and Thalassiosira pseudonana. II. Light limitation

The marine diatoms Phaeodactylum tricornutum (Bohlin) and Thalassiosira pseudonana (Hasle and Heimdal) were grown under both continous illumination and a 14 h light: 10 h dark cycle at light intensities ranging from 1.53×10^-4 to 2.95×10^-1 ly min^-1. Under both photoperiods, T. pseudonana exhibited higher division rates than P. tricornutum at high light intensities, but the reverse was true at all light intensities <3×10^-3 ly min^-1. Comparison of these results with available data on light-limited growth of other planktonic algae suggests that P. tricornutum may be unusually efficient at maintaining its cell division rate at low light intensity. This efficiency may contribute substantially to its success in turbid, nutrient-enriched mass algal culture systems, the only environments in which it is known to attain great numbers.Contribution No. 4086 from the Woods Hole Oceanographic Institution.     

Diel periodicity in cellular chlorophyll content in marine diatoms

Intracellular chlorophyll a content is one of the many measurable parameters which displays a diel rhythm in marine phytoplankton. In asynchronous laboratory cultures of the diatom Skeletonema costatum, cellular chlorophylls a and c exhibit periodicities which closely follow the light-dark cycle and are not the result of cell division. The laboratory cultures also exhibit diel rhythms in cellular flourescence properties and carbon: chlorophyll a ratios. The occurrence of similar patterns of cellular flourescence, carbon: chlorophyll a ratios, and in situ flourescence in diatom-dominated natural phytoplankton communities suggests the possibility of diel rhythms in cellular chlorophyl a content in diatoms in the sea. The data also suggest that the observed periodicity in cellular chlorophyll content is regulated by the diel light cycle and that the co-occurrence of synchronous or phased cell division would only modify the observed periodicity.This research was performed under the auspices of the United States Department of Energy under Contract No. EY-76-C-02-0016     

Differences between cell division and carbon fixation rates associated with light intensity and oxygen concentration: Implications in the cultivation of an estuarine diatom

The effect of light intensity and oxygen concentration on the growth of an estuarine diatom was investigated. Differences between rates of cell division and net carbon fixation were found to be dependent upon light intensity and oxygen concentration. Under conditions favoring large differences between cell division and net carbon fixation cultures of Thalassiosira pseudonana clone 3H depart from exponential and enter stationary phase at low cell concentrations. It is suggested that single cell algae may not be able to balance maintenance, growth, and division outside a fairly narrow range of environmental conditions.     

Cell cycle and toxin production in the benthic dinoflagellate Prorocentrum lima

Profiles of diarrhetic shellfish poisoning (DSP) toxins produced throughout the growth cycle and the cell cycle of the toxigenic marine dinoflagellate Prorocentrum lima were studied in triplicate unialgal batch cultures. Cells were pre-conditioned at 18 ± 1 °C, under a photon flux density (PFD) of 90 ± 5 μmol m⁻² s⁻¹ on a 14 h light:10 h dark photoperiod. In exponential growth phase, cultures were synchronized in darkness for 17 d. After dark synchronization, cultures were transferred back to the original photoperiod regime. Cells were harvested for DSP toxin analysis by LC-MS (liquid chromatography with mass spectrometry), and double-stranded (nuclear) DNA was quantified by flow cytometry. The cell populations became asynchronous within approximately 3 d after transition from darkness to the 14 h light:10 h dark photoperiod. This may be due to the prolonged division cycle (5 to 7 d) that is not tightly phased by the photoperiod. Unlike other planktonic Prorocentrum spp., cytokinesis in P. lima occurred early in the dark and ceased by “midnight”. Cellular levels of the four principal DSP toxins, okadaic acid (OA), OA C8-diol-ester (OA-D8), dinophysistoxin-1 (DTX1) and dinophysistoxin-4 (DTX4), ranged from 0.37 to 6.6, 0.02 to 1.5, 0.04 to 2.6, and 1.8 to 7.8 fmol cell⁻¹, respectively. No toxin production was evident during the extended period of dark synchronization nor during the initial period when NH₄ was consumed as the major nitrogen source. Soon after the cells were returned to the 14 h light:10 h dark cycle and they began to take up NO₃, cellular levels of all four toxins gradually increased. This increase in DSP toxins usually occurred in the light, marked by a rise in DTX4 levels that preceded an increase in the cellular concentration of OA and DTX1 (delayed by 3 to 6 h). Thus, DTX4 synthesis is initiated in the G1 phase of the cell cycle and persists into S phase (“morning” of the photoperiod), whereas OA and DTX1 production occurs later during S and G2 phases (“afternoon”). No toxin production was measured during cytokinesis, which happened early in the dark. The evidence indicates that toxin synthesis is restricted to the light period and is coupled to cell cycle events. Received: 3 September 1998 / Accepted: 30 March 1999     

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.     

Heterotrophic nutrition of the marine pennate diatom Nitzschia angularis var. affinis

The nutritional pattern for heterotrophic growth of Nitzschia angularis var. affinis (Grun.) Perag. is more complex than for other diatom species studied previously. This species grew slowly in the dark in the presence of single amino acids, either glutamate or alanine; other amino acids when supplied singly were not used as substrates. Carbon from glutamate was converted to cell carbon with an efficiency of 43%. Glutamine was inhibitory both in the light and in the dark, and aspartate inhibited heterotrophic growth on glutamate. Glucose and tryptone supplied singly did not support heterotrophic growth, but when combined, together they allowed for rapid growth of N. angularis (generation time of 16 h). Glucose in combination with glutamate, alanine, aspartate, or asparagine (but not with any other amino acids) also supported growth in the dark, at a rate considerably more rapid than with glutamate alone. In the presence of excess glucose and limiting concentrations of glutamate, approximately 50% of the cell carbon for heterotrophic growth came from glucose, while in combination with tryptone about 25% of the cell carbon came from glucose. Amino acids were taken up by cells grown either photoautrophically or in the dark in the presence or absence of organic substrates; uptake rates were some-what higher for dark-grown than for light-grown cells. Glucose was taken up only by dark-grown cells; induction of a glucose uptake system in the dark required the presence of glutamate but not of glucose. The rates of uptake of glutamate and glucose by cells incubated in the dark with glutamate were sufficiently high to account for the observed rates of growth on these substrates in the dark. The uptake systems of N. angularis have relatively high affinities for glucose (K _s =0.03 mM) and glutamate (K _s =0.02 mM).Contribution No. 890 from the Department of Oceanography, University of Washington, Seattle, Washington 98195, USA.     

The concept of light intensity adaptation in marine phytoplankton: Some experiments with Phaeodactylum tricornutum

The historical background on adaptation of algae to various light intensities is analysed. It is argued that there is little evidence to suggest that previous growth at low light intensities enhances the ability of an alga to utilize these low light levels. Rather, the published evidence suggests that the most general response to growth at sub-optimal light intensities is a reduced ability to utilize saturating levels. The present experiments with Phaeodactylum tricornutum Bohlin have tested this concept of light intensity adaptation. Changing photosynthetic abilities during batch growth depended on the light intensity used for growth and these changes affected interpretations of the data. When measurements were made intensities appeared to photosynthesize (at all intensities) better than did those grown at higher light levels. When the changes during batch growth were taken into account, or when the alga was grown in turbidostat cultures, a different picture was obtained. Growth at reduced light intensities was accompanied by (a) increased chlorophyll content, (b) decreased rate of light-saturated photosynthesis expressed on a chlorophyll, cell number or cell protein basis, and (c) decreased activity of RuDP carboxylase. One result suggested that growth at a suboptimal light intensity did enhance the ability to utilize lower light levels. The light-saturation curve of cells grown in batch culture at 0.7 klux showed higher slopes at the low light intensities than did those grown at 12 klux. This was most marked when photosynthesis was expressed per cell, but was also apparent when it was put on a per chlorophyll basis.     

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.     

Phosphorus metabolism of oceanic dinoflagellates: phosphate uptake,chemical composition and growth of Pyrocystis noctiluca

The phosphorus metabolism of Pyrocystis noctiluca Murray (Schuett) 1886 has characteristics which may enhance its potential for success in orthophosphate impoverished waters. The steady-state phosphate uptake rates were equal in the light and dark, and were directly proportional to both the phosphorus cell quota and the cell division rate. In contrast, nutrient-saturated uptake rates were multiphasic, faster in the light than the dark, 2 to 4 orders of magnitude greater than steady-state rates, and were inversely proportional to both the phosphorus cell quota and the cell division rate. These uptake characteristics suggest that P. noctiluca may take up phosphate coincidently at their typically low ambient concentrations as well as to exploit episodic nutrient events in nature. Cell division rates were a hyperbolic function of the ambient orthophosphate concentration. The shortest doubling time was 8.7 d, the phosphate concentration at half the maximum division rate was 0.15 M and the threshold, concentration for cell division was ca 0.05 M PO ₄ ^3- . Division rates of P. noctiluca in the ocean are much faster than predicted from the measured ambient orthophosphate concentrations. Since this dinoflagellate has high naturally occurring alkaline phosphatase activities, and can utilize organic-P compounds, we suggest that organic-P can be as important as orthophosphate in supporting the observed division rates of P. noctiluca in the sea.     

Allelopathic interactions between Skeletonema costatum and Alexandrium minutum

Potential allelopathic interactions between Skeletonema costatum and Alexandrium minutum were investigated using mixed cultures and culture filtrate in nutrient-replete medium. A. minutum growth was inhibited when grown in S. costatum filtrate, with the inhibitory effect directly proportional to the percentage of filtrate added. This demonstrates that the release of allelopathic compounds caused the growth inhibition. In contrast, the filtrate of A. minutum exerted no allelopathic activity on S. costatum. An autoinhibitory compound (15(S)-HEPE) extracted and purified from S. costatum culture was added to cultures of both S. costatum and A. mintum. This substance could depress S. costatum growth, but showed no significant inhibitory activity on A. minutum. This documented a second type of allelochemical interaction, termed auto-allelopathy, caused by a different compound from the one or ones that affected A. minutum in the co-cultures with added crude filtrate. Further studies are needed to explore the relative importance of these two types of allelopathy as factors influencing the competition between S. costatum and A. minutum in the field. Furthermore, given the observed decrease in diatom dominance relative to dinoflagellates with increasing eutrophication, one can predict that toxic species like A. minutum might become more prevalent in the future in the East China Sea if the trend of increasing pollution of coastal waters continues.     

Effects of light intensity on aging of the dinoflagellate Gonyaulax polyedra

Gonyaulax polyedra Stein grown in increasingly nutrientlimited batch culture undergoes the following changes (collectively termed aging): there is a decline in the intracellular concentrations of carbon, nitrogen and photosynthetic pigments; nitrate reductase activity decreases; rates of respiration and photosynthesis fall; and cell division virtually ceases (accompanied in bright light by a decrease in the volume of individual cells). The effect of light intensity on these aging events was tested by growing cells in either bright or dim light. The bright light (330 E m^-2 s^-1) was enough to saturate photosynthesis and the dim light (80 E m^-2 s^-1) was low enough to induce significant shade adaptation of photosynthesis without lowering growth rate. At both light intensities, a decline in carbon and nitrogen content preceded or accompanied all other monitored changes, and the sequence of aging events was similar. However the onset of the decline in intracellular nutrients and photosynthetic rate in low-light cells was delayed by a least one cell division time (i.e., to twice the cell density) in comparison to cells under bright light. At both light levels, pigment-protein complexes of the photosynthetic apparatus began to break down after intracellular carbon and nitrogen had been depleted to a critically low level. The beginning of the drop in pigmentation signalled the end of log-phase growth. It is suggested that the greater pigmentation of low-light cells may represent a larger nutrient supply than found in bright-light cells and could increase the survival time of nutrient-stressed populations.     

Effects of non-circadian light/dark cycles on the growth and moulting of Palaemon elegans reared in the laboratory

The growth and moulting of Palaemon elegans Rathke has been compared under a circadian (12 h light: 12 h dark) and two non-circadian (8 h light:8 h dark and random light:dark) light/dark cycles. In prawns reared individually from hatching to the late juvenile phase, growth, measured as increase in total length, was significantly retarded in the non-circadian regimes, during zoeal, post-larval and early juvenile development. This effect was primarily the result of reduced increments at moulting in the non-circadian regimes. Growth of prawns reared from hatching to the post-larval phase, measured as wet and dry weights, was similarly reduced in a non-circadian regime. These effects support the hypothesis that the integrity of an animal's physiology is partially dependent on maintaining, through the action of daily environmental cycles, correct timing relationships between its oscillating sub-systems.