Glen A. Tarran

High bacterivory by the smallest phytoplankton in the North Atlantic Ocean

Planktonic algae <5 μm in size are major fixers of inorganic carbon in the ocean. They dominate phytoplankton biomass in post-bloom, stratified oceanic temperate waters. Traditionally, large and small algae are viewed as having a critical growth dependence on inorganic nutrients, which the latter can better acquire at lower ambient concentrations owing to their higher surface area to volume ratios. Nonetheless, recent phosphate tracer experiments in the oligotrophic ocean have suggested that small algae obtain inorganic phosphate indirectly, possibly through feeding on bacterioplankton. There have been numerous microscopy-based studies of algae feeding mixotrophically in the laboratory and field, as well as mathematical modelling of the ecological importance of mixotrophy. However, because of methodological limitations there has not been a direct comparison of obligate heterotrophic and mixotrophic bacterivory. Here we present direct evidence that small algae carry out 40–95% of the bacterivory in the euphotic layer of the temperate North Atlantic Ocean in summer. A similar range of 37–70% was determined in the surface waters of the tropical North-East Atlantic Ocean, suggesting the global significance of mixotrophy. This finding reveals that even the smallest algae have less dependence on dissolved inorganic nutrients than previously thought, obtaining a quarter of their biomass from bacterivory. This has important implications for how we perceive nutrient acquisition and limitation of carbon-fixing protists as well as control of bacterioplankton in the ocean.

Picoplanktonic community structure on an Atlantic transect from 50°N to 50°S

Plankton samples were collected from 10 depths at 25 stations spaced at intervals of about 4° of latitude along a transect from the British Isles to the Falkland Islands. Four categories of picoplankton were discriminated: Synechococcus spp., Prochlorococcus spp., eukaryotic picophytoplankton and heterotrophic bacteria. The populations in each category in the samples were counted by flow cytometry and the mean size of bacterial cells was determined by fractionation through filters. Categories of phototrophic cells were discriminated by size and by the fluorescence of photosynthetic pigments; samples stained with the fluorochrome TOTO were used to enumerate heterotrophic bacteria (and Prochlorococcus in surface waters where their chlorophyll content was very small). The carbon biomass concentration of each category in each sample was calculated. Prochlorococcus was present at all stations between 47°N and 38°S, and reached peak population densities above 200,000 cells ml-1 in equatorial waters; the depth occupied by these cells increased in oligotrophic waters, where they dominated picophytoplankton biomass. Synechococcus reached high concentrations in the Mauritanian upwelling region and in the frontal region near the southern end of the transect, where they represented the largest single component of picophytoplankton biomass, but was almost absent in oligotrophic regions. Picoeukaryotes were present in low numbers at all latitudes, but they are larger cells and constituted a substantial part of the total picophytoplankton biomass at most latitudes. The depth-integrated (200 m) biomass of heterotrophic bacteria was nearly as great as that of the picophytoplankton at all latitudes, because substantial numbers of cells occurred at all depths. Numbers and biomass of these bacteria were maximal in the upwelling region and high at both ends of the transect. There was a clear contrast in the composition of the picoplankton community in both the North and South Atlantic between mesotrophic waters where Synechococcus and picoeukaryotes dominated the biomass, and oligotrophic waters where the smaller total biomass was dominated by Prochlorococcus.

Latitudinal distribution of prokaryotic picoplankton populations in the Atlantic Ocean

Members of the prokaryotic picoplankton are the main drivers of the biogeochemical cycles over large areas of the worlds oceans. In order to ascertain changes in picoplankton composition in the euphotic and twilight zones at an ocean basin scale we determined the distribution of 11 marine bacterial and archaeal phyla in three different water layers along a transect across the Atlantic Ocean from South Africa (32.9 degrees S) to the UK (46.4 degrees N) during boreal spring. Depth profiles down to 500 m at 65 stations were analysed by catalysed reporter deposition fluorescence in situ hybridization (CARD-FISH) and automated epifluorescence microscopy. There was no obvious overall difference in microbial community composition between the surface water layer and the deep chlorophyll maximum (DCM) layer. There were, however, significant differences between the two photic water layers and the mesopelagic zone. SAR11 (35 +/- 9%) and Prochlorococcus (12 +/- 8%) together dominated the surface waters, whereas SAR11 and Crenarchaeota of the marine group I formed equal proportions of the picoplankton community below the DCM (both approximately 15%). However, due to their small cell sizes Crenarchaeota contributed distinctly less to total microbial biomass than SAR11 in this mesopelagic water layer. Bacteria from the uncultured Chloroflexi-related clade SAR202 occurred preferentially below the DCM (4-6%). Distinct latitudinal distribution patterns were found both in the photic zone and in the mesopelagic waters: in the photic zone, SAR11 was more abundant in the Northern Atlantic Ocean (up to 45%) than in the Southern Atlantic gyre (approximately 25%), the biomass of Prochlorococcus peaked in the tropical Atlantic Ocean, and Bacteroidetes and Gammaproteobacteria bloomed in the nutrient-rich northern temperate waters and in the Benguela upwelling. In mesopelagic waters, higher proportions of SAR202 were present in both central gyre regions, whereas Crenarchaeota were clearly more abundant in the upwelling regions and in higher latitudes. Other phylogenetic groups such as the Planctomycetes, marine group II Euryarchaeota and the uncultured clades SAR406, SAR324 and SAR86 rarely exceeded more than 5% of relative abundance.

Isolation of viruses responsible for the demise of an Emiliania huxleyi bloom in the English Channel

This study used analytical flow cytometry (AFC) to monitor the abundance of phytoplankton, coccoliths, bacteria and viruses in a transect that crossed a high reflectance area in the western English Channel. The high reflectance area, observed by satellite, was caused by the demise of an Emiliania huxleyi bloom. Water samples were collected from depth profiles at four stations, one station outside and three stations inside the high reflectance area. Plots of transect data revealed very obvious differences between Station 1, outside, and Stations 2–4, inside the high reflectance area. Inside, concentrations of viruses were higher; E. huxleyi cells were lower; coccoliths were higher; bacteria were higher and virus:bacteria ratio was lower than at Station 1, outside the high reflectance area. This data can simply be interpreted as virus-induced lysis of E. huxleyi cells in the bloom causing large concentrations of coccoliths to detach, resulting in the high reflectance observed by satellite imagery. This interpretation was supported by the isolation of two viruses, Eh V84 and Eh V86, from the high reflectance area that lysed cultures of E. huxleyi host strain CCMP1516. Basic characterization revealed that they were lytic viruses approximately 170 nm–190 nm in diameter with an icosahedral symmetry. Taken together, transect and isolation data suggest that viruses were the major contributor to the demise of the E. huxleyi population in the high reflectance area. Close coupling between microalgae, bacteria and viruses contributed to a large organic carbon input. Consequent cycling influenced the succession of an E. huxleyi -dominated population to a more characteristic mixed summer phytoplankton community.

Mixotrophic basis of Atlantic oligotrophic ecosystems

Oligotrophic subtropical gyres are the largest oceanic ecosystems, covering >40% of the Earths surface. Unicellular cyanobacteria and the smallest algae (plastidic protists) dominate CO2 fixation in these ecosystems, competing for dissolved inorganic nutrients. Here we present direct evidence from the surface mixed layer of the subtropical gyres and adjacent equatorial and temperate regions of the Atlantic Ocean, collected on three Atlantic Meridional Transect cruises on consecutive years, that bacterioplankton are fed on by plastidic and aplastidic protists at comparable rates. Rates of bacterivory were similar in the light and dark. Furthermore, because of their higher abundance, it is the plastidic protists, rather than the aplastidic forms, that control bacterivory in these waters. These findings change our basic understanding of food web function in the open ocean, because plastidic protists should now be considered as the main bacterivores as well as the main CO2 fixers in the oligotrophic gyres.

Microbial community structure and standing stocks in the NE Atlantic in June and July of 1996

The standing stocks of nanophytoplankton and picoplankton in the northeast Atlantic Ocean in June and July 1996 were quantified using flow cytometry and microscopy. Diatoms and dinoflagellates were analysed by microscopy and coccolithophores, other nanophytoplankton, picoeukaryotic phytoplankton, cyanobacteria (Synechococcus spp.), prochlorophytes (Prochlorococcus spp.) and heterotrophic bacteria by flow cytometry. The research was divided into three components: a lagrangian study of a nutrient replete cold-core eddy centred around 59° 12′N 20° 12′W; a transect close to the 20°W meridian from 59° 18′N to 37°N, which passed through contrasting water masses; and a lagrangian study in oligotrophic waters, centred around 36° 42′N 19° 12′W. The eddy was characterised by a bloom of the coccolithophore Coccolithus pelagicus whose standing stocks averaged 4.26 g C m−2 over the upper 50 m. C. pelagicus and other nanophytoplankton (excluding diatoms and dinoflagellates) dominated the standing stocks of the microbial community, averaging approx. 70% of the total microbial standing stocks of the groups quantified. The majority of the remaining biomass was accounted for by the picoeukaryotic phytoplankton and heterotrophic bacteria. The microbial community immediately outside the eddy was significantly different in both composition and standing stocks. There were no C. pelagicus outside the eddy and fewer nanophytoplankton, resulting in microbial standing stocks of approx. one-third that found in the eddy. The transect was characterised by a frontal region at approx. 52° 30′N. There was a general decrease in the standing stocks of all components of the microbial community from the start of the transect to the front. Just to the south of the front, nanophytoplankton, Synechococcus spp. and heterotrophic bacteria showed marked increases in standing stocks, especially the nanophytoplankton, which increased from 3.43 to 7.90 g C m−2. The nanophytoplankton dominated the microbial standing stocks throughout the transect, even in the oligotrophic waters where the integrated carbon biomass was 4.58 g C m−2, representing 69% of the total microbial standing stocks. During the lagrangian study around 37°N the picoplanktonic community was dominated by heterotrophic bacteria. However, heterotrophic bacteria standing stocks decreased with time, along with Synechococcus spp. and picoeukaryotic phytoplankton. Peak biomass for these three groups shifted deeper down in the water column with time. Prochlorococcus spp. were only present towards the end of the transect and at the oligotrophic site. At the oligotrophic site their standing stocks increased, unlike other groups, so that they became the dominant picophytoplanktonic group.

The life-cycle of Emiliania huxleyi: A brief review and a study of relative ploidy levels analysed by flow cytometry

Emiliania huxleyi exists in several principal forms including the familiar coccolith-bearing C-cell, non-motile naked N-cells, and scale-bearing swarmers (S-cells), but the relationships between these cells are unclear. Flow cytometric analyses have been undertaken on whole cells using fluorochrome staining of the DNA in order to determine the relative DNA content and the relative GC content of the S- and C-cells of selected clones. Results showed that the DNA complement of the S-cells was half that of the C-cells and the two cell types are, therefore, haploid and diploid relative to each other. The S-cells may, therefore, represent a gametic stage, though processes such as sexual fusion and meiosis have not been observed.

Depth related amino acid uptake by Prochlorococcus cyanobacteria in the Southern Atlantic tropical gyre.

Ambient concentrations and turnover rates of two amino acids, leucine and methionine, by total bacterioplankton and Prochlorococcus cyanobacteria were determined along a latitudinal transect across the Southern Atlantic gyre using a combined isotopic dilution and flow cytometric sorting technique. The ambient concentrations of methionine (0.2-0.65 nM) were about 2 times higher than the concentrations of leucine, while the turnover rates of the two amino acids were remarkably similar (0.1-0.7 nM d(-1)). The concentrations of both amino acids did not vary significantly with depth between 3 and 150 m but their turnover rates were 1.5-2 times higher in the top 3-80 m. Prochlorococcus took up amino acids in situ at high rates. Using a representative (35)S-methionine precursor, about 25% of total bacterioplankton consumption of amino acids could be assigned to Prochlorococcus with low red fluorescence (Pro LRF) inhabiting the surface mixed layer down to 80 m and about 50% assigned to Prochlorococcus with high red fluorescence (Pro HRF) living below 100 m. In the same deep waters the cellular amino acid uptake of Pro LRF was less than 6% of that of the Pro HRF, indicating declining metabolic activity of the former. The mean cellular uptake rate of Pro HRF at depths below 120 m was 2.5 amol cell(-1) d(-1), 4 times higher than the rates of Pro LRF in the top 80 m. The difference could be partially explained by Pro HRF cellular biomass being twice that of Pro LRF. The biomass specific rates of Prochlorococcus were comparable or higher (particular of the Pro HRF) than that of other bacterioplankton. The reported findings could explain ecological success of mixotrophic Prochlorococcus cyanobacteria over both strictly autotrophic algae and heterotrophic bacteria in oligotrophic regions sustained by nutrient remineralisation.

Comparable light stimulation of organic nutrient uptake by SAR11 and Prochlorococcus in the North Atlantic subtropical gyre

Subtropical oceanic gyres are the most extensive biomes on Earth where SAR11 and Prochlorococcus bacterioplankton numerically dominate the surface waters depleted in inorganic macronutrients as well as in dissolved organic matter. In such nutrient poor conditions bacterioplankton could become photoheterotrophic, that is, potentially enhance uptake of scarce organic molecules using the available solar radiation to energise appropriate transport systems. Here, we assessed the photoheterotrophy of the key microbial taxa in the North Atlantic oligotrophic gyre and adjacent regions using 33P-ATP, 3H-ATP and 35S-methionine tracers. Light-stimulated uptake of these substrates was assessed in two dominant bacterioplankton groups discriminated by flow cytometric sorting of tracer-labelled cells and identified using catalysed reporter deposition fluorescence in situ hybridisation. One group of cells, encompassing 48% of all bacterioplankton, were identified as members of the SAR11 clade, whereas the other group (24% of all bacterioplankton) was Prochlorococcus. When exposed to light, SAR11 cells took 31% more ATP and 32% more methionine, whereas the Prochlorococcus cells took 33% more ATP and 34% more methionine. Other bacterioplankton did not demonstrate light stimulation. Thus, the SAR11 and Prochlorococcus groups, with distinctly different light-harvesting mechanisms, used light equally to enhance, by approximately one-third, the uptake of different types of organic molecules. Our findings indicate the significance of light-driven uptake of essential organic nutrients by the dominant bacterioplankton groups in the surface waters of one of the less productive, vast regions of the world’s oceans—the oligotrophic North Atlantic subtropical gyre.

Diel rhythmicity in amino acid uptake by Prochlorococcus.

The marine cyanobacterium Prochlorococcus, the most abundant phototrophic organism on Earth, numerically dominates the phytoplankton in nitrogen (N)-depleted oceanic gyres. Alongside inorganic N sources such as nitrite and ammonium, natural populations of this genus also acquire organic N, specifically amino acids. Here, we investigated using isotopic tracer and flow cytometric cell sorting techniques whether amino acid uptake by Prochlorococcus is subject to a diel rhythmicity, and if so, whether this was linked to a specific cell cycle stage. We observed, in contrast to diurnally similar methionine uptake rates by Synechococcus cells, obvious diurnal rhythms in methionine uptake by Prochlorococcus cells in the tropical Atlantic. These rhythms were confirmed using reproducible cyclostat experiments with a light-synchronized axenic Prochlorococcus (PCC9511 strain) culture and (35)S-methionine and (3)H-leucine tracers. Cells acquired the tracers at lower rates around dawn and higher rates around dusk despite >10(4) times higher concentration of ammonium in the medium, presumably because amino acids can be directly incorporated into protein. Leucine uptake rates by cells in the S+G(2) cell cycle stage were consistently 2.2 times higher than those of cells at the G(1) stage. Furthermore, S+G(2) cells upregulated amino acid uptake 3.5 times from dawn to dusk to boost protein synthesis prior to cell division. Because Prochlorococcus populations can account from 13% at midday to 42% at dusk of total microbial uptake of methionine and probably of other amino acids in N-depleted oceanic waters, this genus exerts diurnally variable, strong competitive pressure on other bacterioplankton populations.