Products of photosynthesis by marine phytoplankton: the effect of environmental factors on the relative rates of protein synthesis

A method is presented by which the gross pattern of photosynthetic carbon-dioxide fixation in marine phytoplankton can be determined. It depends on differential solvent extraction yielding an ethanol-soluble, a hot TCA-soluble (polysaccharide) and a residue (protein) fraction. Using this fractionation technique, the effects of various environmental factors on the pattern of photosynthesis by the marine diatom Phaeodactylum tricornutum (Bohlin) have been investigated. Low light intensities and increasing degrees of nitrogen limitation in a chemostat increase markedly the relative rates of protein synthesis. Growth of the alga at lower temperatures also increases the proportion of carbon incorporated into the protein fraction. This increased protein syntheses is generally at the expense of the polysaccharide fraction. Preliminary experiments have established the suitability of this fractionation method for natural populations of phytoplankton and have shown similar effects of light intensity on the relative rates of protein synthesis.     

Decomposition of urea associated with photosynthesis of phytoplankton in coastal waters

Decomposition of urea in seawater was studied in Mikawa Bay, a shallow eutrophic bay on the southern coast of central Japan. The urea concentration in seawater ranged from 1.3 to 5.9 μg-at. N/1 and comprised 12 to 40% of the dissolved organic nitrogen. Using ¹⁴C labelled urea, the rate of CO₂ liberation from urea and the incorporation rate of urea carbon into the particulate organic matter were determined. For the surface samples, high rates of CO₂ liberation from urea as well as the incorporation of urea carbon into the particulate organic matter were observed in the light, while much lower rates were obtained in the dark. Incubation experiments with exposure to different light intensities revealed that the rate of CO₂ liberation from urea and the incorporation of urea carbon into particulate organic matter changed with light intensity, showing a pattern similar to that of photosynthesis. The highest liberation and incorporation rates were observed at 12,000 lux. Incubation in light and in dark produced marked decreases and increases, respectively, in urea and ammonia, while no appreciable changes were observed for nitrate and nitrite. It is suggested that urea decomposition associated with photosynthetic activity of phytoplankton is one of the major processes of urea decomposition, and that it plays a significant role in the nitrogen supply for phytoplankton in coastal waters.     

Significance of active and passive depuration in the clearance of naphthalene from the tissues of Hemigrapsus nudus (Crustacea: Decapoda)

Photosynthate incorporation into lipids, low molecular weight compounds, polysaccharides and proteins by individual phytoplankton species isolated from natural populations is described. This sensitive method uses serial solvent extraction and liquid scintillation counting and gives results indentical to those obtained from filtering large numbers of cells from suspension. The time course of incorporation of ¹⁴C into these polymers for algae isolated from estuarine and coastal populations shows (i) considerable differences in the carbon metabolism among species, (ii) the pattern of incorporation by any individual species may not reflect that of the phytoplankton community and (iii) significant reallocation of cell carbon among carbon pools and net protein synthesis at night. This technique permits the in-situ carbon metabolism of individual species to be examined.     

Interspecific differences in the photosynthetic carbon metabolism of marine phytoplankton

Six species of marine phytoplankton of different sizes and taxonomic categories were grown in microcosms under identical experimental conditions; the species cultured were: Pavlova lutheri (Prymnesiophyceae), Dunaliella tertiolecta (Chlorophyceae), Phaeodactylum tricornutum (Baciollariophyceae), Eutreptiella sp. (Euglenophyceae), Alexandrium tamarense (Dinophyceae), and Phaeocystis pouchetii (Prymnesiophyceae). The photosynthetic carbon metabolism of these phytoplankton was studied throughout the exponential and lag phases of growth after nutrient depletion. The relative incorporation of carbon into protein was positively correlated with phytoplankton growth, while carbon assimilation into low molecular weight metabolites (LMWM) and storage products, i.e., lipid and polysaccharides, generally increased under nutrient-limiting conditions. Clear taxonspecific differences were observed in the proportions of carbon incorporated into cell constituents. A significant linear relationship was consistently found between the relative synthesis of protein to LMWM, and both the production normalised to chlorophyll (P:B) and the phytoplankton growth rate. However, ANCOVA revealed significant, interspecific differences in these relationships.     

Photosynthesis and chlorophyll a fluorescence rhythms of marine phytoplankton

Circadian rhythms in photosynthesis were defined in field populations of phytoplankton. Measurements of carbon-dioxide fixation rates demonstrated that a diurnal periodicity of photosynthesis in samples incubated under natural light-dark (LD) cycles also were observed to continue in similar samples which had been photoadapted to constant dim light (LL) for 48 h. These changes in photosynthetic rates preceded sunset and sunrise, had daily amplitudes that ranged from 1.5 to 2.0, appeared to be independent of light-intensity, and displayed maxima about midday, while rates of dark fixation of carbon dioxide and the photosynthetic pigment content per cell were constant over the circadian cycle. Similar rhythmicity also was detected in room-temperature (22°C) chlorophyll a fluorescence yield, in both the obsence and presence of the photosynthesis inhibitor DCMU [3-(3,4-dichlorophenyl)-1, 1-dimethylurea]. However, the magnitude and timing of the fluorescence rhythm maxima seem to depend on wavelengths monitored and, in part, on the measuring technique used. Also, the circadian changes in the fluorescence intensity were abolished at low temperature (-60°C), and the shape of the emission spectra of chlorophyll fluorescence of cells in LD and LL did not change over time. The significance of the fluorescence rhythms with regard to chlorophyll a determinations and photosynthetic rates is discussed. It was concluded that there was sufficient similarity between circadian rhythms of photosynthesis in natural phytoplankton populations and in laboratory cultures of dinoflagellates to suggest that the mechanism of regulation may be the same for both of them.     

Size fractionation of natural aquatic populations associated with autotrophic and heterotrophic carbon uptake

Natural phytoplankton samples from the Gulf of California were incubated with radioactive bicarbonate or glucose, with or without the addition of gentamycin, a broad spectrum antibiotic which inhibits bacterial activity but has little effect on many algae. The relative distribution of chlorophyll and radioactivity in particles retained by filters of 3 and 0.45 μ mean pore size was measured. From 13 to 83% of the ¹⁴C-bicarbonate assimilated by natural populations appeared to be associated with organisms having a smallest dimension less than 3 μ. These were probably algae, since the distribution of chlorophyll on the filters was similar to that of the radioactivity. Furthermore, gentamycin treatment did not change the distribution of radioactivity from ¹⁴C-bicarbonate. In these waters, considerable photosynthesis may be carried out by ultra-nannoplankton (3 to 1 μ). Glucose was assimilated mainly by bacteria, of which more than 90% passed through 3 μ filters. Gentamycin significantly reduced glucose uptake in most cases. Although this method cannot be adopted uncritically for the estimation of carbon flux from photoautotrophs to heterotrophs, differential filtration can be used to elucidate the size distribution of various metabolic groups in the plankton.     

Utilization of L-lysine and L-arginine by the diatomPhaeodactylum tricornutum

The rate of growth ofPhaeodactylum tricornutum Bohlin with L-lysine as sole nitrogen source is about half that with ammonium or nitrate, and only one of the two nitrogen atoms in the lysine molecule appears to be used for growth. The organism cannot grow heterotrophically with lysine as carbon source. The rate of lysine metabolism is slow and most of that utilized is incorporated into protein, this process being faster in light than in darkness. The dark incorporation of lysine-carbon into protein is stimulated by addition of ammonium, whereas incorporation in light is unaffected. Arginine-carbon is also mainly incorporated into protein. Light has little effect on this incorporation, and addition of ammonium decreases it both in light and darkness. There is no appreciable conversion of the carbon of either amino acid to carbon dioxide. Although, under anaerobiosis, lysine is taken up and accumulated, negligible metabolism of lysine occurs.     

Observations of heterotrophic activity on photoassimilated organic matter

The natural heterotrophic utilization of released and particulate photoassimilated compounds by natural phytoplankton populations was followed during the summer of 1975 in the Kiel Bight (Western Baltic Sea). Phytoplankton exudates (2 to 21% of primary production) seem to represent an important substrate for heterotrophic bacteria: uptake rates between 8 and 17,5%/h were observed. Samples incubated with antibiotics indicated that the heterotrophic uptake of phytoplankton exudates during photosynthesis could lead to underestimation of the release rate. Production of 14_{CO 2}was used as an estimate of the mineralization of particulate photoassimilated matter. The mineralization rates measured were in the range of 6 to 20% after 20 h. In order to confirm the quantitative measurements, micro-autoradiographic examinations of the samples were performed.     

Modeling the seasonal autochthonous sources of dissolved organic carbon and nitrogen in the upper Chesapeake Bay

In this paper we investigate the seasonal autochthonous sources of dissolved organic carbon (DOC) and nitrogen (DON) in the euphotic zone at a station in the upper Chesapeake Bay using a new mass-based ecosystem model. Important features of the model are: (1) carbon and nitrogen are incorporated by means of a set of fixed and varying C:N ratios; (2) dissolved organic matter (DOM) is separated into labile, semi-labile, and refractory pools for both C and N; (3) the production and consumption of DOM is treated in detail; and (4) seasonal observations of light, temperature, nutrients, and surface layer circulation are used to physically force the model. The model reasonably reproduces the mean observed seasonal concentrations of nutrients, DOM, plankton biomass, and chlorophyll a. The results suggest that estuarine DOM production is intricately tied to the biomass concentration, ratio, and productivity of phytoplankton, zooplankton, viruses, and bacteria. During peak spring productivity phytoplankton exudation and zooplankton sloppy feeding are the most important autochthonous sources of DOM. In the summer when productivity peaks again, autochthonous sources of DOM are more diverse and, in addition to phytoplankton exudation, important ones include viral lysis and the decay of detritus. The potential importance of viral decay as a source of bioavailable DOM from within the bulk DOM pool is also discussed. The results also highlight the importance of some poorly constrained processes and parameters. Some potential improvements and remedies are suggested. Sensitivity studies on selected parameters are also reported and discussed.     

Influence of inter- and intra-daily light-field variability on photosynthesis by coastal phytoplankton

An investigation was carried out to determine the relationships between the principal photosynthetic parameters of natural marine phytoplankton and the properties of the irradiance of the preceding five days and the intra-daily ambient irradiance. Samples used in the study were collected from Strangford Lough, Northern Ireland during both winter and summer between 1979 and 1981. Photosynthetic parameters were determined for both constant and ambient incubation light fields. The fluctuation properties of the irradiance field exerted little influence on the photosynthetic parameters. In summer populations, the parameters obtained from morning samples were strongly influenced by the total irradiance recorded on the fourth day previous to experimentation, irrespective of the incubation irradiance. In contrast, the corresponding parameters from afternoon samples were most influenced by the total irradiance received during incubation. The influence of these two separate factors over a day may contribute to variability in the magnitude of diel changes in marine phytoplankton photosynthesis.     

Time-course of photoadaptation in the photosynthesis-irradiance relationship of a dinoflagellate exhibiting photosynthetic periodicity

Cultures of the marine dinoflagellate Glenodinium sp. were light-shifted and rates of photoadaptation determined by monitoring changes in cell volume, growth rate, pigmentation, parameters of the photosynthesisirradiance (P-I) curves and respiration. To approximate physiological conditions of field populations, cells were cultured on an alternating light-dark cycle of 12hL:12hD, which introduced a daily periodicity of photosynthesis. One result of the present study was to demonstrate how specific parameters of the P-I relationship influenced by periodicity of the light: dark cycle are distinguished from photosynthetic parameters influenced by changes in light level. Under steady-state conditions, rates of both light-saturated (P_max) and light-limited photosynthesis changed in unison over the day; these changes were not related to pigmentation, and displayed their maxima midday. This close relationship between P_max and the slope (a) of the cellular P-I curves in steadystate conditions was quickly adjusted when growth illumination was altered. Rates of light-limited photosynthesis were increased under low light conditions and the periodicity of cellular photosynthesis was maintained. The short-term responses of the P-I relationship to changing light level was different, depending on (1) whether the light shift was from high to low light or vice versa, and (2) whether the high light levels were sufficient to promote maximal photosynthesis rates. Major increases in the photosynthetic carotenoid peridinin, associated with a single type of light-harvesting chromo protein in the chloroplast, was observed immediately upon shifting high light cultures to low light conditions. Following pigment synthesis, significant increases in rates of light-limited photosynthesis were observed in about one-tenth the generation time, while cellular photosynthetic potential was unaffected. it is suggested that general results were consistent with suggested that general results were consistent with earlier reports that the major photoadaptive strategy of Glenodinium sp. is to alter photosynthetic unit (PSU) size. Photoadaptive response times to high light were light-dependent, but appeared to be shower than photoadaptive responses to low light. If light intensities were bright enough to maximize growth rates, photosynthetic response times were on the order of a generation period and pigmentation fell quickly as cells divided at a faster rate. If light-intensities were not sufficient to maximize growth rates, then pigment content did not decline, while rates of light-limited photosynthesis declined quickly. In all cases, photoadaptation was followed best by monitoring fast changes in half saturation constants for photosynthesis, rather than fluctuating changes in pigmentation. Results compared well with time-course phenomena reported for other groups of phytoplankton. Overall, results suggest phytoplankton can bring about photo-induced changes in photosynthesis very quickly and thus accommodate widely fluctuating light regimes over short periods of time.     

Light quality effects on carbon metabolism and allocation in Gracilaria verrucosa

Under conditions of new nitrogen availability and low light intensities, light quality (blue, green, or red light) was not found to affect carbon fixation patterns into major metabolic fractions (total ethanol soluble, carbohydrate, and protein) in preconditioned nitrogen enriched or limited apical tips of Gracilaria verrucosa Papenfuss. Within the ethanol soluble fractions of both nitrogen enriched and limited tips, blue light treatment led to a greater percentage of ¹⁴ carbon in amino acids as compared to non-ninhydrin reactive metabolites. A lesser effect was observed with red light, and green light did not appear to stimulate amino acid synthesis appreciably. The low intensity blue light effect in G. verrucosa appears to be an enhancement of nonphotosynthetic carbon incorporation into amino acids, possibly through some form of the urea-ornithine cycle.Harbor Branch Foundation Contribution No. 237     

Production of picoplankton and small nanoplankton in the Celtic Sea

A significant proportion of the total primary production in the Celtic Sea (50°30′N; 07°00′W) has been found to be due to picoplankton and small nanoplankton. In July, August and October, 1982, 20 to 25% of the ¹⁴C fixed in primary production was in organisms >5 μm, 35 to 40% was in organisms <5–1 μm and 20 to 30% was in organisms<1 μm. Bacterial production was estimated by the incorporation of ³H and would account for less than 10% of the production in the <1–>0.2 μm fraction; therefore, production in the <1–>0.2 μm fraction was the result of photosynthesis per se by picoplankton and could not have been due to heterotrophic bacteria utilizing exudates from larger phytoplankton. Time-course experiments demonstrated some transfer of label from the <1–>0.2μm fraction to the >5 μm fraction, presumably by grazing, but again most of the production in this fraction was the result of photosynthesis by organisms larger than 5 μm and was not due to grazing by heterotrophic microflagellates on smaller phytoplankton.     

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.     

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.     

Photosynthetic studies of Chondrus crispus

The net photosynthesis of intertidal, subtidal, carposporic, tetrasporic, and winter versus summer acclimatized plants of Chondrus crispus Stackhouse were evaluated under different temperatures and quantities of light. The optimum temperature and light conditions for net photosynthesis of C. crispus are seasonally and spatially variable, and there is an adaptive shift in the photosynthetic capacity at different seasons and positions on the shore. Plants collected during the fall and winter had lower light optima (465 to 747 ft-c) for net photosynthesis than spring and summer specimens (about 1000 ft-c). Intertidal populations exhibited a higher rate of net photosynthesis between 250 and 2819 ft-c than subtidal plants. Summer materials have a greater tolerance to high temperatures and a higher temperature optimum than winter materials. Shallow subtidal populations (-6m) exhibited a higher temperature optimum than deep subtidal plants (-12m). Tetrasporic plants (diploid) showed a higher rate of net photosynthesis than carposporic plants (haploid). It is suggested that the diploid plants of C. crispus may extend deeper in the subtidal zone, because they have a higher rate of net photosynthesis than carposporic plants. The results of the present studies are compared with previous physiological studies of C. crispus.Published with the approval of the Director of the New Hampshire Agriculture Experiment Station as Scientific Contribution Number 742.     

Phytoplankton production modelling in three marine ecosystems—static versus dynamic approach

Phytoplankton productivity is usually determined from water samples incubated at a number of irradiance levels during several hours. The resultant productivity-irradiance (P–E) curves are then used to estimate local and/or global phytoplankton production. However, there is growing evidence that these curves, referred as static, underestimate phytoplankton photosynthesis to a great deal, by assuming a stable response to light over the incubation period. One of the drawbacks of static P–E curves is the overestimation of photoinhibition.In this work, three one-dimensional vertically resolved models were developed as simply as possible, to investigate differences between static and dynamic phytoplankton productivity in three marine ecosystems: a turbid estuary, a coastal area and an open ocean ecosystem. The results show that, when photoinhibition development time is considered (dynamic model), the primary production estimates are always higher than when calculated with the static model. The quantitative importance of these differences varies with the type of ecosystem and it appears to be more important in coastal areas and estuaries (from 21 to 72%) than in oceanic waters (10%). Thus, these results suggest that primary production estimates, obtained under the assumption of a static behaviour response to light, may underestimate the real values of global phytoplankton primary production. Calculations suggest that the quantitative importance of this underestimation may be larger than the global missing carbon sink.     

Photosynthesis and sink activity of wasp-induced galls in Acacia pycnantha

Although insect galls are widely known to influence source-sink relationships in plants, the relationship between photosynthesis and gall activity has not been extensively studied. In this study we used 14CO2, photosynthesis, and respiration measurements to examine the capacity of bud galls induced by the wasp Trichilogaster signiventris (Pteromalidae) as carbon sinks in Acacia pycnantha. Galls of this species develop either in vegetative or reproductive buds, depending on the availability of tissues at different times of the year, and effectively eliminate seed production by the plant. Photosynthetic rates in phyllodes subtending clusters of galls were greater than rates in control phyllodes, a result we attributed to photosynthesis compensating for increased carbon demand by the galls. Contrary to previous studies, we found that photosynthesis within galls contributed substantially to the carbon budgets of the galls, particularly in large, mature galls, which exhibited lower specific respiration rates allowing for a net carbon gain in the light. To determine the sink capacity and competitive potential of galls, we measured the proportion of specific radioactivity in galls originating from either vegetative or reproductive buds and found no difference between them. The proportion of the total amount of phyllode-derived 14C accumulated in both clustered and solitary galls was less than that in fruits. Galls and fruits were predominantly reliant on subtending rather than on distant phyllodes for photosynthate. Solitary galls that developed in vegetative buds constituted considerably stronger sinks than galls in clusters on inflorescences where there was competition between galls or fruits for resources from the subtending phyllode. Wasps developing in solitary vegetative galls were correspondingly significantly larger than those from clustered galls. We conclude that, in the absence of inflorescence buds during summer and fall, the ability of the wasps to cause gall formation in vegetative tissues tempers intraspecific competition and substantially increases the availability of plant resources for the development of wasps in such galls.     

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.     

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.