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.     

Diel changes in phytoplankton photosynthetic efficiency in Brackish waters

From 18 to 23 September 1974, investigations on the diel changes in phytoplankton were carried out in the Baltic Sea. Every 4 h, water samples were collected from 2 and 15 m, and PO₄, chlorophyll a, temperature, salinity, pH, phytoplankton composition and phytoplankton light photosynthesis relationship were determined. Continuous measurements of surface irradiance and some estimations of zooplankton were also made. P ^B (photosynthesis per unit chlorophyll a at low light levels of 2·10^-2 cal cm^-2 min^-1) revealed only random variation during the sampling period, i.e., 1.0 to 1.6 mg C (mg chlorophyll a)^-1 h^-1. P _m ^B (Light-saturated photosynthesis per unit of chlorophyll a) displayed pronounced diel fluctuations with the highest value of about 6 mg C (mg chlorophyll a)^-1 h^-1 around noon, and the lowest value of about 2.5 mg C (mg chlorophyll a)^-1 h^-1 during the night, during which latter period the value of P _m ^B was more or less constant. Reasons for the diel fluctuations are discussed, and an equation which describes these fluctuations is proposed. Using this equation, the daily phytoplankton production estimated in incubators by a previously described method can be corrected for the time of day at which samples are collected.     

Photoadaption in marine phytoplankton: Response of the photosynthetic unit

Some species of phytoplankton adapt to low light intensities by increasing the size of the photosynthetic unit (PSU), which is the ratio of light-harvesting pigments to P700 (reaction-center chlorophyll of Photosystem I). PSU size was determined for 7 species of marine phytoplankton grown at 2 light intensities: high (300 E m^-2 s^-1) and low (4 E m^-2 s^-1); PSU size was also determined for 3 species grown at only high light intensity. PSU size varied among species grown at high light from 380 for Dunaliella euchlora to 915 for Chaetoceros danicus. For most species grown at low light intensity, PSU size increased, while the percentage increase varied among species from 13 to 130%. No change in PSU size was observed for D. euchlora. Photosynthetic efficiency per chlorophyll a (determined from the initial slope of a curve relating photosynthetic rate to light intensity) varied inversely with PSU size. In contrast, photosynthetic efficiency per P700 was enhanced at larger PSU sizes. Therefore, phytoplankton species with intrinsically large PSU sizes probably respond more readily to the rapid fluctuations in light intensity that such organisms experience in the mixed layer.Contribution No. 1180 from the Department of Oceanography, University of Washington, Seattle, Washington, USA     

DCMU-enhanced fluorescence as an index of photosynthetic activity in phytoplankton

Tests have been carried out, both on phytoplankton cultures and in the field, on the relation between the ratio of 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU)-enhanced to normal fluorescence (F _D /F _N ) and the specific photosynthetic rate (P/B). In all cases observed, significant linear correlations were found between these two values. Differences in this relation were observed according to species and physiological conditions. Nutrient stress that occurs in batch cultures decreases both F _D /F _N and P/B, while lowered light intensity has a different effect on both, increasing F _D /F _N and decreasing P/B. This is interpreted as indicating that F _D /F _N is an index of photosynthetic efficiency at different light levels and an indicator of the specific photosynthetic rate (P/B) when physiological conditions vary at a given light intensity. The practical use of DCMU-enhanced fluorometry to estimate instantaneous P/B values in the field is discussed, stressing the frequent calibrations needed, as in all in situ fluorometry studies.     

Circadian variations in photosynthetic assimilation and estimation of daily phytoplankton production

In August 1984, hourly measurements of photosynthetic characteristics were carried out during 96 h, at 5 and 10 m, on a natural population of phytoplankton in the St Lawrence Estuary. Synchronous circadian variations of similar amplitude (max./min.: 2 to 3) were observed at the two depths in both the photosynthetic capacity (P _m ^B ) and the photosynthetic efficiency (^B). Maximum values occurred at around noontime and minima during the night. Estimates of daily specific productivity were computed with and without the observed circadian variability. Large differences (15 to 70%) were evidenced between estimates.Contribution to the programs of GIROQ (Groupe interuniversitaire de recherches océanographiques du Québec) and of the Maurice Lamontagne Institute (Department of Fisheries and Oceans)     

Light quality in relation to growth,photosynthetic rates and carbon metabolism in two species of marine plankton algae

The effect of light quality on growth, photosynthesis and carbon metabolism in two species of marine algae,Cyclotella nana (Hustedt) andDunaliella tertiolecta (Butcher), was examined. Relative growth constants forC. nana were 0.37, 0.29 and 0.25 in blue, white and green light, respectively. Corresponding constants were 0.41, 0.31 and 0.29 forD. tertiolecta. Photosynthetic rates in both species were higher in blue light and lower in green light compared with white light of the same intensity. More than 60% of¹⁴C assimilated byC. nana orD. tertiolecta grown in blue or green light was incorporated into the ethanol-insoluble fraction, compared with 10 to 30% in this fraction in white light. The relative importance of the various components within this fraction was independent of light quality. Although less¹⁴C was assimilated into the ethanol-soluble fraction in blue or green light, there was a relative increase in some amino acids and organic acids in this fraction and a decrease in sugars and sugar phosphates relative to white light of the same intensity. These differences were independent of light intensity, photosynthetic rate and cell density in the cultures.     

Light acclimation states of phytoplankton in the Southern Ocean, determined using photosynthetic pigment distribution

In high-latitude waters such as the Southern Ocean, the primary production of phytoplankton supports the ecosystem. To understand the photo-acclimation strategy of such phytoplankton within cold environments, the vertical distribution profile of photosynthetic pigments was analyzed in the Southern Ocean. Samples were taken along 110°E during the austral summer, and along 150°E and around the edge of the seasonal sea ice of the Antarctic Continent during the austral autumn. Pigment extraction methods were optimized for these samples. The standing crop of chlorophyll a was larger in the region along the edge of the seasonal sea ice than at sampling stations in open ocean areas. Chlorophyll concentration seemed to be dependent on the formation of thermo- and haloclines along the edge of the seasonal sea ice, but not in the open ocean where such clines are less pronounced. The marker pigments fucoxanthin and/or 19′-hexanoyloxyfucoxanthin were dominant at most sampling stations throughout the water column, while other marker pigments such as alloxanthin were quite low. This indicated that diatoms and/or haptophytes were the major phytoplankton in this area. Comparison of the relative ratio of fucoxanthin with that of 19′-hexanoyloxyfucoxanthin allowed some stations to be characterized as either diatom-dominant or haptophyte-dominant. The relative ratio of xanthophyll-cycle pigments (diadinoxanthin plus diatoxanthin) to chlorophyll a was high in surface waters and decreased gradually with depth. This suggests that near the ice edge during summer in the Southern Ocean, both diatoms and haptophytes acclimate to their light environments to protect their photosystems under high-light conditions.     

Buffer sensitivity of photosynthetic carbon utilisation in eight tropical seagrasses

Some of the mechanisms involved in inorganic carbon (Ci) acquisition by tropical seagrasses from the western Indian Ocean were described by Björk et al. (Mar Biol 129:363–366, 1997). However, since then, it has been found that an additional, buffer-sensitive, system of Ci utilisation may operate in some temperate seagrasses (Hellblom et al. in Aquat Bot 69:55–62, 2001, Hellblom and Axelsson in Photos Res 77:173–191, 2003); this buffer sensitivity indicates a mechanism in which electrogenic H⁺ extrusion may form acidic diffusion boundary layers, in which either HCO ₃ ^? –H⁺ is co-transported into the cells, or where HCO ₃ ^? is converted to CO₂ (as catalysed by carbonic anhydrase) prior to uptake of the latter Ci form. Because a buffer was used in the 1997 study, we found it important to reinvestigate those same eight species, taking into account the direct effect of buffers on this potential mode of Ci acquisition in these plants. In doing so, it was found that all seagrass species investigated except Cymodocea serrulata were sensitive to 50 mM TRIS buffer of the same pH as the natural seawater in which they grew (pH 8.0). Especially sensitive were Halophila ovalis, Halodule wrightii and Cymodocea rotundata, which grow high up in the intertidal zone (only ca. 50–65% of the net photosynthetic activity remained after the buffer additions), followed by the submerged Enhalus acoroides and Syringodium isoetifolium (ca. 75% activity remaining), while Thalassia hemprichii and Thalassodendron ciliatum, which grow in-between the two zones, were less sensitive to buffer additions (ca. 80–85% activity remaining). In addition to buffer sensitivity, all species were also sensitive to acetazolamide (AZ, an inhibitor of extracellular carbonic anhydrase activity) such that ca. 45–80% (but 90% for H. ovalis) of the net photosynthetic activity remained after adding this inhibitor. Raising the pH to 8.8 (in the presence of AZ) drastically reduced net photosynthetic rates (0–14% remaining in all species); it is assumed that this reduction in rates was due to the decreased CO₂ concentration at the higher pH. These results indicate that part of the 1997 results for the same species were due to a buffer effect on net photosynthesis. Based on the present results, it is concluded that (1) photosynthetic Ci acquisition in six of the eight investigated species is based on carbonic anhydrase catalysed HCO ₃ ^? to CO₂ conversions within an acidified diffusion boundary layer, (2) C. serrulata appears to support its photosynthesis by extracellular carbonic anhydrase catalysed CO₂ formation from HCO ₃ ^? without the need for acidic zones, (3) H. ovalis features a system in which H⁺ extrusion may be followed by HCO ₃ ^? –H⁺ co-transport into the cells, and (4) direct, non-H⁺-mediated, uptake of HCO ₃ ^? is improbable for any of the species.     

Differences in photosynthetic pigment signatures between phytoneuston and phytoplankton communities in a coastal lagoon of Baja California

In order to understand the relationships between the dynamics of phytoplankton populations in the surface microlayer (MIL) and in the water column below (SSW), this study used high-performance liquid chromatography-derived pigment markers in samples from a coastal lagoon of Baja California (Estero de Punta Banda, EPB) under summer (October 2003) and winter (December 2003) conditions. Photosynthetic pigment signatures of phytoplankton at the air–sea interface (phytoneuston) and subsurface measurements were related to bottom-up (temperature, salinity, nutrient concentrations) and top-down factors (zooplankton abundance). Slicks and scum layers were observed in the inner part of the lagoon and coincided with greater stratification of layers just below the sea surface and lower wind intensities. In general, spatial variability in pigment markers and ancillary data was very high and resulted in non-significant differences between MIL and subsurface samples when different regions of EPB or sampling dates were compared. However, different patterns were found between pigments and environmental factors of MIL and SSW samples when the relative numbers of stations with positive and negative differences (ΔX = X _MIL−X _SSW) were computed. For each survey, pigment markers of phytoneuston and phytoplankton samples were not necessarily correlated. Further analysis revealed that those markers (19′-butanoyloxyfucoxanthin, prasinoxanthin, divinil-chlorophyll a) corresponded to picophytoplankton groups (haptophyte, prasinophyte, and prochlorophyte). On both dates, the MIL was enriched in 19′-hexanoyloxyfucoxanthin (a marker for a type 4 haptophyte) and fucoxanthin (marker for bacillariophytes, haptophytes, and crysophytes) and depleted in peridinin (marker for dinophytes). Different zooplankton grazers accumulated in the MIL (loricate tintinnids) and in SSW (copepod nauplii).     

Measurement of photosynthetic production of a marine phytoplankton population using a stable 13C isotope

The use of stable isotope of carbon, ¹³C, for the determination of the photosynthetic rate of a marine phytoplankton population was examined. Particular concern was paid to the effects of non-phytoplanktonic organic carbon and the enrichment of inorganic carbon on the estimation of the photosynthetic rate. Photosynthetic rates determined by the ¹³C method showed a remarkable agreement with those determined by the ¹⁴C method. Insitu determinations of photosynthetic rate were made in three different water types: open ocean, coastal and neritic waters, which included oligo- and mesotrophic waters, by using the ¹³C method established.     

Release of photoassimilated carbon as dissolved organic matter by marine phytoplankton

The release of photoassimilated carbon as dissolved organic matter was studied in situ in oligotrophic and eutrophic marine waters and in axenic laboratory cultures. Percentage extracellular release (PER), integrated for the trophogenic zone, ranged from 6 to 12% in eutrophic waters and from 17 to 27% in oligotrophic seas. Most of the algal cultures released low amounts of dissolved organic matter (5%) during exponential growth. The highest levels of PER were observed in surface and deep samples, possibly as a result of elevated photorespiration and senescent cells. The generally lower values for extracellular release reported in this work as compared to other studies may be partly due to improved experimental techniques which minimized previously encountered artifacts.     

Observations on organic carbon utilization by photosynthetic marine microalgae

Organic carbon utilization was studied in 13 species of axenic unicellular marine algae. The compounds tested were 15 sugars, 19 acids, and 16 alcohols. Heterotrophy occurred in only three chlorophyte strains with acetate and ethanol; one strain with glucose. Mixotrophy was observed with the following compounds: glyceraldehyde, 1 species; acetate, 4 species; glycolic acid, 5 species; pyruvic acid, 9 species; caproic acid, 3 species; lactic acid, 1 species; ethanol, 3 species; glycerol, 9 species. Some ecological implications are discussed.     

Comparative study of the carbon cycle in Venus verrucosa fed on bacteria and phytoplankton

The aim of this, the third part of our study, was to compare the quantities of food consumed and assimilated by Venus verrucosa L. fed on two different living suspensions: the bacterium Lactobacillus sp. and the alga Pavlova lutheri. The experimental results have been presented in parts I and II of our study. The present paper describes a model constructed for each series of experiments with bacteria and with phytoplankton. An analog-computer model was succesfully applied and revealed that bacterial food was more easily consumed and assimilated than algal food, the dissolved form more easily assimilated than the living particulate form.     

Comparative study of the carbon cycle in Venus verrucosa fed on bacteria and phytoplankton

Following a previous study, this paper deals with the utilisation of a phytoplankton suspension (Pavlova lutheri) by the clam Venus verrucosa Linné, 1767 as a function of time. Experimental clams filtered and ingested 100% of the suspension after 2 h. The radioactivity recovered in the whole soft body of the clams was 83.8% after 2 h, 66.0% after 10 h and 46.0% after 41 h. The faeces contained 13.7% after 3 h and 34.3% after 41 h. Carbon-dioxide radioactivity (gas plus dissolved) increased slowly to 12.4% after 41 h. When the water was not changed after 17 h, the clams reabsorbed a great part of their dissolved products and faeces. Under these latter conditions the radioactivity recovered from the whole soft body of the clams was roughly the same as that recovered after the start of the experiment. The aim of this study was the comparison of the consumption of two kinds of food: bacteria and phytoplankton. The work was carried out over three years, from 1977 to 1980.     

A new method for estimating phytoplankton growth rates and carbon biomass

A new method is described for the determination of phytoplankton growth rates and carbon biomass. This procedure is easy to apply and utilizes the labeling of chlorophyll a (chl a) with ¹⁴C. Pure chl a is isolated using two-way thin-layer chromatography, and the specific activity of chl a carbon is determined. Data from laboratory cultures indicate that the specific activity of chl a carbon becomes nearly equal to that of total phytoplankton carbon in incubations lasting 6 to 12 h and can be used to calculate phytoplankton growth rates and carbon biomass. Application of the method to the phytoplankton community in an eutrophic estuary in Hawaii indicates that the cells are growing with a doubling time of about 2 d and that about 85% of the particulate carbon consists of phytoplankton carbon.     

Increased carbon dioxide availability alters phytoplankton stoichiometry and affects carbon cycling and growth of a marine planktonic herbivore

Rising levels of CO₂ in the atmosphere have led to increased CO₂ concentrations in the oceans. This enhanced carbon availability to the marine primary producers has the potential to change their nutrient stoichiometry, and higher carbon-to-nutrient ratios are expected. As a result, the quality of the primary producers as food for herbivores may change. Here, we present experimental work showing the effect of feeding Rhodomonas salina grown under different pCO₂ (200, 400 and 800 μatm) on the copepod Acartia tonsa. The rate of development of copepodites decreased with increasing CO₂ availability to the algae. The surplus carbon in the algae was excreted by the copepods, with younger stages (copepodites) excreting most of their surplus carbon through respiration and adult copepods excreting surplus carbon mostly as DOC. We consider the possible consequences of different excretory pathways for the ecosystem. A continued increase in the CO₂ availability for primary production, together with changes in the nutrient loading of coastal ecosystems, may cause changes in the trophic links between primary producers and herbivores.     

Nitrate uptake in marine phytoplankton: Energy sources and the interaction with carbon fixation

Field studies of whole natural phytoplankton communities from Knight Inlet, B. C., Canada and laboratory cultures of the diatom Skeletonema costatum indicate inorganic carbon fixation may be temporarily suppressed following 10 to 15% enrichment with NO ₃ ^- or NH ₄ ⁺ . (This effect is suggested to be due to competition between inorganic carbon and nitrogen for adenosine triphosphate (ATP), and is reduced when chlorophyll a is increased intracellularly after 6 to 8 h.) Results imply that the source of ATP for nitrate uptake is primarily from Photosystem I (cyclic photophosphorylation) in the presence of light. It would appear that a transient nutrient-adaptive response occurs upon addition of extracellular nitrogen.     

Comparative study of the carbon cycle inVenus verrucosa fed on bacteria and phytoplankton

The utilisation of a viable bacterial suspension (Lactobacillus sp.) by the clamVenus verrucosa Linné, 1767 has been investigated as a function of time. The clams filtered and ingested 95% of the suspension within 2 h. Radioactivity recovered from the soft-body of the clams measured 51% of the total radioactivity introduced at the start of the experiment after 2.5 h, 48.8% after 10 h, and 39.9% after 40 h. The dissolved organic matter remained nearly constant, with 27.5% after 2.5 h, 24.4% after 10 h, and 29.8% after 40 h. The faeces produced increased from 13.2% after 2.5 h to 21.9% after 10 h, and decreased until 40 h with 16.2%. The carbon-dioxide respired increased slowly to 5% after 40 h. These data on the functioning of the different compartments suggest an important recycling of the metabolic products of the bivalves by the bacteria of the suspension, demonstrating the role of bacteria as food for bivalves.     

Direct measurement of dissolved organic carbon release by phytoplankton and incorporation by microheterotrophs

It is now possible to divide particulate primary production into algal and heterotrophic components without physical separation. This depends on two innovations, the introduction of isotope in the form of labelled dissolved product(s) of primary production and the employment of a data analysis specifically designed for tracer kinetic incorporation experiments. The ¹⁴C technique described by Steemann Nielsen (1952) is inapplicable in the analyses of certain classes of systems and kinetic tracer incorporation experiments must be employed instead. We show that measurement of PDOC production rate requires such kinetic tracer analyses. Measurements made in the laboratory on water taken from 2 m depth in South West Arm of the Port Hacking estuary showed that: (1) the steady-state rate of PDOC production was 0.10 to 0.13 mg C.m^-3.h^-1; (2) the rate of PDOC incorporation into microheterotroph particulate organic carbon was 0.10 to 0.12 mg C.m^-3.h^-1; (3) the rate at which PDOC was respired to CO₂ was 0.001 to 0.003 mg C.m^-3.h^-1. (4) the PDOC makes up only about 0.1% of the total dissolved organic carbon. The size class of particles associated with PDOC production differed from the size class responsible for uptake of PDOC. More than 50% of the PDOC production was associated with particles having a nominal diameter range of 20 to 63 m, while this fraction was responsible for <10% of the incorporation.     

Respiration,photosynthesis and carbon metabolism in planktonic ciliates

Release of¹⁴C-labelled carbon dioxide from uniformly labelled cells was used to measure respiration by individual ciliates in 2-h incubations in 1989 and 1990. In a strictly heterotrophic ciliate,Strobilidium spiralis (Leegaard, 1915), release of labelled carbon dioxide was equivalent to ca. 2.8% of cell C h^–1 at 20°C, and there was no difference between rates in the dark and light. In the chloroplast-retaining ciliatesLaboea strobila Lohmann, 1908,Strombidium conicum (Lohmann, 1908) Wulff, 1919 andStrombidium capitatum (Leegaard, 1915) Kahl, 1932, release of labelled carbon dioxide was less in the light than in the dark in experiments done at 15°C. InL. strobila release of radiolabel as carbon dioxide was equivalent to ca. 2.4% of cell C h^–1 in the dark but ca. 1% at 50µE m^–2 s^–1, an irradiance limiting to photosynthesis. InS. conicum release of radiolabel as carbon dioxide was equivalent to ca. 4.4% of cell C h^–1 in the dark, but at an irradiance saturating to photosynthesis (250 to 300µE m^–2 s^–1) there was no detectable release of labelled carbon dioxide. InS. capitatum release of radiolabel as carbon dioxide was equivalent to ca. 4.3% of cell C h^–1 in the dark but at an irradiance saturating to photosynthesis was ca. 2.4% of cell C h^–1. These data, combined with data from photosynthetic uptake experiments, indicate that¹⁴C uptake underestimates the total benefit of photosynthesis by 50% or more in chloroplastretaining ciliates.Contribution no. 7510 from the Woods Hole Oceanographic Institution