Photosynthetic Picoeukaryotes Research Articles

Structure and seasonal dynamics of the eukaryotic picophytoplankton community in a wind‐driven coastal upwelling ecosystem

We studied the structure and seasonal dynamics of the photosynthetic picoeukaryote community at a coastal site off central Chile that is strongly affected by seasonal wind‐driven upwelling. We determined the picoeukaryote phylogenetic diversity by cloning and sequencing the 18S ribosomal ribonucleic acid (rRNA) gene amplified with Chloroplastida‐biased primers. Chloroplastida made up 100% (autumn), 61% (spring), and 20% (summer) of the clone libraries. Most sequences belonged to Mamiellophyceae and clustered with Micromonas (clades A.BC.1, B.E.3, C.D.5), Ostreococcus (clade A), and Crustomastix (clade A). Fluorescent in situ hybridization coupled with tyramide signal amplification (FISH–TSA) was used to determine the time variability in abundance of the total picoeukaryotic community, the Chloroplastida, and the genera Micromonas, Ostreococcus, and Bathycoccus over a period of 2 yr with nearly monthly sampling. The abundance of photosynthetic picoeukaryotes was low during the upwelling season (spring and summer) and high during the nonupwelling season (late fall and winter). Chloroplastida made up, on average, 74% of the total picoeukaryotic community, and the sum of the three genera probed accounted for 100% of the Chloroplastida abundances. Ostreococcus dominated the picophytoplanktonic community numerically throughout the year and, thus, appears to be a key component of the upwelling picoplanktonic community in the eastern South Pacific.

Invariable biomass‐specific primary production of taxonomically discrete picoeukaryote groups across the Atlantic Ocean

Oceanic photosynthetic picoeukaryotes (< 3 µm) are responsible for > 40% of total primary production at low latitudes such as the North-Eastern tropical Atlantic. In the world ocean, warmed by climate changes, the expected gradual shift towards smaller primary producers could render the role of photosynthetic picoeukaryotes even more important than they are today. Little is still known, however, about how the taxonomic composition of this highly diverse group affects primary production at the basin scale. Here, we combined flow cytometric cell sorting, NaH¹⁴CO₃ radiotracer incubations and class-specific fluorescence in situ hybridization (FISH) probes to determine cell- and biomass-specific inorganic carbon fixation rates and taxonomic composition of two major photosynthetic picoeukaryote groups on a ∼7500-km-long latitudinal transect across the Atlantic Ocean (Atlantic Meridional Transect, AMT19). We show that even though larger cells have, on average, cell-specific CO₂ uptake rates ∼5 times higher than the smaller ones, the average biomass-specific uptake is statistically similar for both groups. On the other hand, even at a high taxonomic level, i.e. class, the contributions to both groups by Prymnesiophyceae, Chrysophyceae and Pelagophyceae are significantly different (P < 0.001 in all cases). We therefore conclude that these group's carbon fixation rates are independent of the taxonomic composition of photosynthetic picoeukaryotes across the Atlantic Ocean. Because the above applies across different oceanic regions the diversity changes seem to be a secondary factor determining primary production.

Analysis of photosynthetic picoeukaryote community structure along an extended Ellett Line transect in the northern North Atlantic reveals a dominance of novel prymnesiophyte and prasinophyte phylotypes

Photosynthetic picoeukaryotes (PPEs) of a size <3 μm can contribute significantly to primary production. Here, PPE community structure was analysed along an extended Ellett Line transect, an area in the North Atlantic well studied by physical oceanographers but largely neglected in the field of microalgal ecology. Distribution patterns of specific PPE classes were determined using dot-blot hybridization analysis, while the taxonomic composition of specific PPE classes was revealed by phylogenetic analysis of plastid 16S rRNA gene sequences. In addition, we performed fluorescent in situ hybridization (FISH) analysis of seawater samples collected along the transect to provide a PCR-independent survey of class level PPE distribution patterns. We found the PPE community was dominated by members of the Prymnesiophyceae, Prasinophyceae and Mamiellophyceae. Interestingly, phylogenetic analysis revealed several novel Prymnesiophyceae and Prasinophyceae phylotypes (with only 85–96% identity to neighbouring sequences) within lineages for which cultured counterparts are unknown.

Plastid 16S rRNA Gene Diversity among Eukaryotic Picophytoplankton Sorted by Flow Cytometry from the South Pacific Ocean

The genetic diversity of photosynthetic picoeukaryotes was investigated in the South East Pacific Ocean. Genetic libraries of the plastid 16S rRNA gene were constructed on picoeukaryote populations sorted by flow cytometry, using two different primer sets, OXY107F/OXY1313R commonly used to amplify oxygenic organisms, and PLA491F/OXY1313R, biased towards plastids of marine algae. Surprisingly, the two sets revealed quite different photosynthetic picoeukaryote diversity patterns, which were moreover different from what we previously reported using the 18S rRNA nuclear gene as a marker. The first 16S primer set revealed many sequences related to Pelagophyceae and Dictyochophyceae, the second 16S primer set was heavily biased toward Prymnesiophyceae, while 18S sequences were dominated by Prasinophyceae, Chrysophyceae and Haptophyta. Primer mismatches with major algal lineages is probably one reason behind this discrepancy. However, other reasons, such as DNA accessibility or gene copy numbers, may be also critical. Based on plastid 16S rRNA gene sequences, the structure of photosynthetic picoeukaryotes varied along the BIOSOPE transect vertically and horizontally. In oligotrophic regions, Pelagophyceae, Chrysophyceae, and Prymnesiophyceae dominated. Pelagophyceae were prevalent at the DCM depth and Chrysophyceae at the surface. In mesotrophic regions Pelagophyceae were still important but Chlorophyta contribution increased. Phylogenetic analysis revealed a new clade of Prasinophyceae (clade 16S-IX), which seems to be restricted to hyper-oligotrophic stations. Our data suggest that a single gene marker, even as widely used as 18S rRNA, provides a biased view of eukaryotic communities and that the use of several markers is necessary to obtain a complete image.

Water‐column stratification governs the community structure of subtropical marine picophytoplankton

The increase in the areal extent of the subtropical gyres over the past decade has been attributed to a global tendency towards increased water-column stratification. Here, we examine how vertical stratification governs the community structure of the picophytoplankton that dominate these vast marine ecosystems. We analysed phytoplankton community composition in the three Southern Subtropical basins of the Pacific, Indian and Atlantic Oceans using a variety of methods and show that the distributions of picocyanobacteria and photosynthetic picoeukaryotes (PPEs) are strongly correlated with depth and strength of vertical mixing: the changes in community structure occur at various taxonomic levels. In well-mixed waters, PPEs, in particular haptophytes, dominate, whereas in strongly stratified waters, picocyanobacteria of the genus Prochlorococcus are prevalent, regardless of whether the relative contributions to total biomass are assessed in terms of pigment or of carbon. This ecological diochotomy within the picophytoplankton supports the hypothesis that genomic streamlining provides a selective advantage for Prochlorococcus in highly stable, oligotrophic systems, but may restrict their ability to dominate in regions subject to dynamic mixing.

Global distribution patterns of distinct clades of the photosynthetic picoeukaryote Ostreococcus

Ostreococcus is a marine picophytoeukaryote for which culture studies indicate there are 'high-light' and 'low-light' adapted ecotypes. Representatives of these ecotypes fall within two to three 18S ribosomal DNA (rDNA) clades for the former and one for the latter. However, clade distributions and relationships to this form of niche partitioning are unknown in nature. We developed two quantitative PCR primer-probe sets and enumerated the proposed ecotypes in the Pacific Ocean as well as the subtropical and tropical North Atlantic. Statistical differences in factors such as salinity, temperature and NO(3) indicated the ecophysiological parameters behind clade distributions are more complex than irradiance alone. Clade OII, containing the putatively low-light adapted strains, was detected at warm oligotrophic sites. In contrast, Clade OI, containing high-light adapted strains, was present in cooler mesotrophic and coastal waters. Maximal OI abundance (19 555±37 18S rDNA copies per ml) was detected in mesotrophic waters at 40 m depth, approaching the nutricline. OII was often more abundant at the deep chlorophyll maximum, when nutrient concentrations were significantly higher than at the surface (stratified euphotic zone waters). However, in mixed euphotic-zone water columns, relatively high numbers (for example, 891±107 18S rDNA copies per ml, Sargasso Sea, springtime) were detected at the surface. Both Clades OI and OII were found at multiple euphotic zone depths, but co-occurrence at the same geographical location appeared rare and was detected only in continental slope waters. In situ growth rate estimates using these primer-probes and better comprehension of physiology will enhance ecological understanding of Ostreococcus Clades OII and OI which appear to be oceanic and coastal clades, respectively.

Basin‐scale distribution patterns of photosynthetic picoeukaryotes along an Atlantic Meridional Transect

Photosynthetic picoeukaryotes (PPEs) of a size < 3 µm play a crucial role in oceanic primary production. However, little is known of the structure of the PPE community over large spatial scales. Here, we investigated the distribution of various PPE classes along an Atlantic Meridional Transect sampled in boreal autumn 2004 that encompasses a range of ocean provinces (gyres, upwelling, temperate regions), using dot blot hybridization technology targeting plastid 16S rRNA gene amplicons. Two algal classes, Prymnesiophyceae and Chrysophyceae, dominated the PPE community throughout the Atlantic Ocean, over a range of water masses presenting different trophic profiles. However, these classes showed strongly complementary distributions with Chrysophyceae dominating northern temperate waters, the southern gyre and equatorial regions, while prymnesiophytes dominated the northern gyre. Phylogenetic analyses using both plastid and nuclear rRNA genes revealed a high diversity among members of both classes, including sequences contained in lineages with no close cultured counterpart. Other PPE classes were less prevalent along the transect, with members of the Cryptophyceae, Pelagophyceae and Eustigmatophyceae essentially restricted to specific regions. Multivariate statistical analyses revealed strong relationships between the distribution patterns of some of these latter PPE classes and temperature, light intensity and nutrient concentrations. Cryptophyceae, for example, were mostly found in the upwelling region and associated with higher nutrient concentrations. However, the key classes of Prymnesiophyceae and Chrysophyceae were not strongly influenced by the variables measured. Although there appeared to be a positive relationship between Chrysophyceae distribution and light intensity, the complementary distributions of these classes could not be explained by the variables recorded and this requires further explanation.

Plankton metabolism in surface waters of the tropical and subtropical Pacific Ocean

AME Aquatic Microbial Ecology Contact the journal Facebook Twitter RSS Mailing List Subscribe to our mailing list via Mailchimp HomeLatest VolumeAbout the JournalEditorsSpecials AME 62:1-12 (2011) - DOI: https://doi.org/10.3354/ame01451 FEATURE ARTICLE Plankton metabolism in surface waters of the tropical and subtropical Pacific Ocean Donn A. Viviani, Karin M. Björkman, David M. Karl, Matthew J. Church* Department of Oceanography, University of Hawaii, Honolulu, Hawaii 96822, USA *Corresponding author. Email: mjchurch@hawaii.edu ABSTRACT: We measured plankton metabolism and evaluated changes in plankton community structure within the central Pacific Ocean during a trans-equatorial cruise from the South Pacific Subtropical Gyre (SPSG) to the North Pacific Subtropical Gyre (NPSG). Rates of net community production (NCP) and respiration (R) were determined in near-surface ocean waters, based on light–dark bottle measurements of oxygen production and consumption. Gross primary production (GPP) was computed, where GPP = NCP + R. Rates of GPP and R in the oligotrophic SPSG and NPSG were tightly coupled, resulting in NCP that was either zero or slightly negative. These subtropical waters were characterized by very low concentrations of inorganic nutrients (e.g. nitrate+nitrite range: 0.004 to 0.033 µmol l–1). Within the relatively nutrient-enriched equatorial waters (e.g. nitrate+nitrite concentrations range: 0.67 to 5.7 µmol l–1), GPP exceeded R by up to 5-fold, resulting in positive NCP. GPP varied ~16-fold (0.25 to 4.05 µmol O2 produced l–1 d–1) while R varied ~5-fold (0.31 to 1.56 µmol O2 consumed l–1 d–1). Picoplankton cell abundances and diagnostic pigment biomarkers were used to determine regional differences in phytoplankton community structure. In the near-equatorial waters, abundances of Synechococcus and photosynthetic picoeukaryotes increased up to 100- and 10-fold, respectively, and diagnostic pigment biomarkers for eukaryotic phytoplankton (including diatoms, pelagophytes, and haptophytes) also increased. These results indicate that variability in nutrient availability constitutes an important regulator of net productivity and plankton community structure in large regions of the open ocean. KEY WORDS: Gross primary production · Respiration · Net community production · Equatorial Pacific · Subtropical gyres Full text in pdf format Information about this Feature Article NextCite this article as: Viviani DA, Björkman KM, Karl DM, Church MJ (2011) Plankton metabolism in surface waters of the tropical and subtropical Pacific Ocean. Aquat Microb Ecol 62:1-12. https://doi.org/10.3354/ame01451 Export citation RSS - Facebook - Tweet - linkedIn Cited by Published in AME Vol. 62, No. 1. Online publication date: January 04, 2011 Print ISSN: 0948-3055; Online ISSN: 1616-1564 Copyright © 2011 Inter-Research.

Grazing suppression of dimethylsulphoniopropionate (DMSP) accumulation in iron-fertilised, sub-Antarctic waters

The impact of in situ iron fertilisation on the production of particulate dimethylsulphoniopropionate (DMSPp) and its breakdown product dimethyl sulphide (DMS) was monitored during the SOLAS air–sea gas exchange experiment (SAGE). The experiment was conducted in the high nitrate, low chlorophyll (HNLC) waters of the sub-Antarctic Southern Ocean (46.7°S 172.5°E) to the south-east of New Zealand, during March–April, 2004. In addition to monitoring net changes in the standing stocks of DMSPp and DMS, a series of dilution experiments were used to determine the DMSPp production and consumption rates in relation to increased iron availability. In contrast to previous experiments in the Southern Ocean, DMS concentrations decreased over the course of the 15-d iron-fertilisation experiment, from an integrated volume-specific concentration in the mixed layer on day 0 of 0.78 nM (measured values 0.65–0.91 nM) to 0.46 nM (measured values 0.42–0.47 nM) by day 15, in parallel with the surrounding waters. DMSPp, chlorophyll a and the abundance of photosynthetic picoeukaryotes exhibited indiscernible or only moderate increases in response to the raised iron availability, despite an obvious physiological response by the phytoplankton. High specific growth rates of DMSPp, equivalent to 0.8–1.2 doublings d −1, occurred at the simulated 60% light level of the dilution experiments. Despite the high production rates, DMSPp accumulation was suppressed in part by microzooplankton grazers who consumed between 61% d −1 and 126% d −1 of the DMSPp production. Temporal trends in the rates of production and consumption illustrated a close coupling between the DMSP-producing phytoplankton and their microzooplankton grazers. Similar grazing and production rates were observed for the eukaryotic picophytoplankton that dominated the phytoplankton biomass, partial evidence that picoeukaryotes contributed a substantial proportion of the DMSP synthesis. These rates for DMSPp and picoeukaryotes were considerably higher than for chlorophyll a, indicating higher cycling rates of the DMSP-producing taxa than for the bulk phytoplankton community. When compared to the total phytoplankton community, there was no evidence of selection against the DMSP-containing phytoplankton by the microzooplankton grazers; the opposite appeared to be the case. SAGE demonstrated that increased iron availability in the HNLC waters of the Southern Ocean does not invariably lead to enhanced DMS sea–air flux. The potential suppression of DMSPp accumulation by grazers needs to be taken into account in future attempts to elevate DMS emission through in situ iron fertilisation and in understanding the hypothesised link between levels of Aeolian iron deposition in the Southern Ocean, DMS emission and global albedo.

Diversity of active marine picoeukaryotes in the Eastern Mediterranean Sea unveiled using photosystem-II psbA transcripts

In vast areas of the oceans, most of the primary production is performed by cells smaller than 2-3 mum in diameter (picophytoplankton). In recent years, several in situ molecular studies showed a broad genetic diversity of small eukaryotes by sequencing 18S rRNA genes. Compared with photosynthetic cyanobacteria that are dominated by two genera, Prochlorococcus and Synechococcus, marine photosynthetic picoeukaryotes (PPEs) are much more diverse, with virtually every algal class being represented. However, the genetic diversity and ecology of PPEs are still poorly described. Here, we show using in situ molecular analyses of psbA transcripts that PPEs in the Eastern Mediterranean Sea are highly diverse, probably very active, and dominated by groups belonging to the red algal lineages, Haptophyta, Heterokontophyta (also called Stramenopiles), and Cryptophyta.

Photosynthetic picoeukaryote community structure in the South East Pacific Ocean encompassing the most oligotrophic waters on Earth

Photosynthetic picoeukaryotes (PPEs), comprising organisms < 3 mum in size, are important primary producers in marine food webs and include representatives from all known algal lineages. Little is known, however, regarding the composition and distribution of PPE communities, particularly at large spatial scales, or in relation to the underlying biotic and abiotic factors that influence this structure. Here, we analysed PPE community structure along a transect in the South East Pacific Ocean (BIOSOPE cruise) that encompassed a large trophic gradient, including hyper-oligotrophic waters in the South Pacific Gyre (SPG), considered to be some of the 'clearest' natural waters on Earth. Using dot blot hybridizations with 16S rRNA oligonucleotide probes, we established that the PPE community was dominated by members of the classes Prymnesiophyceae and Chrysophyceae throughout the transect. Moreover, clone library construction followed by phylogenetic analysis of sequenced clones revealed several novel 16S rRNA gene lineages, including new clades of prymnesiophytes (designated Prym 16S-III) and prasinophytes (Pras 16S-VIII). Pras 16S-VIII was found at all five stations at which clone libraries were constructed, representing a range of trophic conditions, including the South Pacific Gyre, suggesting members of this clade have a broad distribution in this part of the South East Pacific at least. In contrast, Prym 16S-III sequences were largely restricted to oligotrophic stations of the SPG. Subsequent multivariate statistical analyses showed that, within the measured factors, chemical and biological factors seem to influence PPE community structure more than physical parameters. However, more than 50% of the variation in distribution of PPE classes remained unexplained.

Groups without Cultured Representatives Dominate Eukaryotic Picophytoplankton in the Oligotrophic South East Pacific Ocean

BackgroundPhotosynthetic picoeukaryotes (PPE) with a cell size less than 3 µm play a critical role in oceanic primary production. In recent years, the composition of marine picoeukaryote communities has been intensively investigated by molecular approaches, but their photosynthetic fraction remains poorly characterized. This is largely because the classical approach that relies on constructing 18S rRNA gene clone libraries from filtered seawater samples using universal eukaryotic primers is heavily biased toward heterotrophs, especially alveolates and stramenopiles, despite the fact that autotrophic cells in general outnumber heterotrophic ones in the euphotic zone.Methodology/Principal FindingsIn order to better assess the composition of the eukaryotic picophytoplankton in the South East Pacific Ocean, encompassing the most oligotrophic oceanic regions on earth, we used a novel approach based on flow cytometry sorting followed by construction of 18S rRNA gene clone libraries. This strategy dramatically increased the recovery of sequences from putative autotrophic groups. The composition of the PPE community appeared highly variable both vertically down the water column and horizontally across the South East Pacific Ocean. In the central gyre, uncultivated lineages dominated: a recently discovered clade of Prasinophyceae (IX), clades of marine Chrysophyceae and Haptophyta, the latter division containing a potentially new class besides Prymnesiophyceae and Pavlophyceae. In contrast, on the edge of the gyre and in the coastal Chilean upwelling, groups with cultivated representatives (Prasinophyceae clade VII and Mamiellales) dominated.Conclusions/SignificanceOur data demonstrate that a very large fraction of the eukaryotic picophytoplankton still escapes cultivation. The use of flow cytometry sorting should prove very useful to better characterize specific plankton populations by molecular approaches such as gene cloning or metagenomics, and also to obtain into culture strains representative of these novel groups.

Photosynthetic picoplankton in Lake Tanganyika: biomass distribution patterns with depth, season and basin

Photosynthetic picoplankton (PPP), particularly picocyanobacteria (PCya), are known to be a major component of phytoplankton in oligotrophic waters. We investigated the dynamics of PPP and heterotrophic bacteria (HBact) in Lake Tanganyika during the dry and rainy seasons of 2004 to 2007, in the two basins of this large lake. Flow cytometry analyses showed that PPP was mainly composed by PCya of the phycoerythrin-rich Synechococcus type, with maximal abundances (2.3 × 10 4 -8.2 × 10 5 cells mL -1 ) found in the mixolimnion (10-20 m depth). PPP biomass integrated over the water column depth ranged between 0. 41 and 3. 09 g C m - 2 , with maximal values in the south basin during the dry season (2.28 ± 0. 62 g C m -2 on average). The contribution of PPP to total phytoplankton biomass ranged from 41 to 99%, with highest values in the south basin in the dry season. Cellular measurements by image analysis of epifluorescence microscopy images showed a significant increase of the cell volume of the PCya during this period. Flow cytometry also allowed enumeration of photosynthetic picoeukaryotes. Assuming a conversion factor of 530 fg C cell - 1 , they contributed on average to 6% to PPP biomass, except during the dry season in the south basin, where their contribution increased to up to 20% of PPP biomass. Integrated over a 100-m water column depth, PCya biomass was on average 1.4 times higher than HBact biomass. This study establishes reference values for the biomass contribution of this part of the microbial food web, covering for the first time, spatial (different sites), vertical (profiles at different depths) and seasonal variations on a multi-year basis. The results strengthen the view of a major role of PPP in the pelagic food web of large tropical Lake Tanganyika.

Late summer phytoplankton distribution along a 3500 km transect in Canadian Arctic waters: strong numerical dominance by picoeukaryotes

A number of recent studies showed that photosynthetic picoeukaryotes are an active and often dominant component of Arctic algal assemblages. In order to place these observations in a large-scale context, samples were collected in the euphotic zone along a 3500 km transect across northern Baffin Bay, the Northwest Passage and the Beaufort Sea during late summer 2005. Picophy- toplankton ( 2 µm were identified and counted by light microscopy. Pigment composition of the total community was assessed by reverse-phase HPLC to determine the relative contribution of different algal groups. The spatial distribution of phytoplankton was heterogeneous along the transect. The highest abundance of picophytoplankton was observed in the Beaufort Sea/Northwest Passage region, whereas nanophytoplankton increased numerically toward the east- ern Canadian Arctic. Picophytoplankton abundance reached a maximum of 18 400 cells ml -1 and accounted for >70% of total cell counts in two-thirds of the samples. The <2 µm size fraction held a similar share of total chl a, which reached a maximum of 6 µg l -1 . Overall, the picophytoplankton community was strongly dominated by eukaryotes (presumably the Prasinophyceae Micromonas). Maximum abundances of picocyanobacteria (120 cells ml -1 ) were observed in brackish waters of the Beaufort Sea. These results confirm that picophytoplankton can dominate not only in warm oligo- trophic waters, but also in a perennially cold ocean during late summer.

Growth rate peaks at intermediate cell size in marine photosynthetic picoeukaryotes

We have performed an in situ test of Raven's prediction that there is a reversal of the relationship between cell size and maximum achievable growth rate in unicellular algae at the low end of size classes. In a natural population of marine phytoplankton, including the smallest picoeukaryote known to date, and under both nutrient sufficiency and deficiency, we find a maximum in growth rate (4.8 and 3.3 divisions d−1, respectively) in the 2–3–µm size class represented by coccoid Chlorella‐like cells, with lower growth rates in both higher and lower size classes. This 2–3–µm size class is also the most robust under nutrient deficiency, reducing its growth rate by 14% only relative to nutrient‐sufficient conditions, versus 50‐60% for the lowermost and uppermost size classes, respectively.

Molecular analysis of photosynthetic picoeukaryote community structure along an Arabian Sea transect

We developed oligonucleotide probes, based on plastid 16S ribosomal DNA (rDNA) sequences, to target 10 different algal classes (Chlorarachniophyceae, Chrysophyceae, Cryptophyceae, Eustigmatophyceae, Pavlovophyceae, Pelagophyceae, Pinguiophyceae, Prasinophyceae [clade VI], Prymnesiophyceae, and Trebouxiophyceae), for use with dot blot hybridization technology. These class‐specific probes were then used to investigate the community structure of photosynthetic picoeukaryotes (≪3 µm) along a transect in the Arabian Sea during September 2001, using hybridization to plastid 16S rDNA polymerase chain reaction amplicons. The transect ranged from oligotrophic open‐ocean waters, just south of the Equator, through mesotrophic and eutrophic waters toward the north coast of Oman, crossing a region of monsoonal upwelling off the Omani northeast coast. Photosynthetic picoeukaryotes in the surface‐mixed layer (SML) and deep chlorophyll maximum ranged from 1.0 x 103 cells mL‐1 in southern oligotrophic waters to almost 30 x 103 cells mL‐1 at the region of upwelling. Chrysophytes were abundant along most of the transect throughout much of the euphotic zone. Prymnesiophytes were abundant in surface waters along much of the transect. By contrast, trebouxiophytes were confined to deeper waters, below the SML. Pelagophytes were found in surface waters that were more mesotrophic, whereas cryptophytes were only detected in the more nutrient‐rich waters at the northern end of the transect, between depths of 20‐30 m. Pinguiophytes were also detected, but only in the warm surface waters off the north coast of Oman. Chlorarachniophytes, eustigmatophytes, pavlovophytes, and clade VI prasinophytes were essentially below detection limits for the entire transect.

Analysis of photosynthetic picoeukaryote diversity at open ocean sites in the Arabian Sea using a PCR biased towards marine algal plastids

Marine photosynthetic picoeukaryotes (PPEs), representing organisms <3 µm in size, are major contributors to global carbon cycling. However, the key members of the PPE community and hence the major routes of carbon fixation, particularly in the open ocean environment, are poorly described. Here, we have accessed PPE community structure using the plastid encoded 16S rRNA gene. Plastid 16S rRNA genes were sequenced from 65 algal cultures, about half being PPEs, representing 14 algal classes. These included sequences from 5 classes where previously no such sequences from cultured representatives had been available (Bolidophyceae, Dictyochophyceae, Eustigmatophyceae, Pelagophyceae and Pinguiophyceae). Sequences were also obtained for 6 of the 7 (according to 18S rRNA gene sequence) prasinophyte clades. Phylogenetic analysis revealed plastids from the same class as clustering together. Using all the obtained sequences, as well as plastid sequences currently in public databases, a non-degenerate marine algal plastid-biased PCR primer, PLA491F, was developed to minimize amplification of picocyanobacteria, which often dominate numerically environmental samples. Clone libraries subsequently constructed from the pico-sized fraction from 2 open ocean sites in the Arabian Sea, revealed an abundance of 16S rRNA gene clones phylogenetically related to chrysophytes, whilst prymnesiophyte, clade II prasinophyte (Ostreococcus-like) and pelagophyte clones were also well represented. The finding of a wealth of novel clones related to the Chrysophyceae highlights the utility of a PCR biased towards marine algal plastids as a valuable complement to 18S rDNA based studies of PPE diversity.

Microbial community structure in the sea surface microlayer at two contrasting coastal sites in the northwestern Mediterranean Sea

In an attempt to compare the microbial community structure between the sea surface microlayer (SML) and subsurface waters (SSW), we determined the enrichment factors (EF: the ratio of abundance or activity in the SML to abundance or activity in SSW) of 13 biological parameters. Samples were taken at 2 contrasting coastal sites in the Mediterranean Sea, corresponding to a high (Barcelona, Spain) and low (Banyuls-sur-Mer, France) urbanized area. Principal component analysis showed that temporal variability was much higher at Barcelona than at Banyuls, and that the characteristics of the SML and SSW samples were more closely related at Barcelona. At both sites, the SML was weakly enriched in heterotrophic bacteria (on average 1.1-fold), Synechococcus spp. (on average 1.1-fold), and photosynthetic picoeukaryotes (on average 1.5-fold) relative to SSW. In contrast, the SML was considerably enriched in chlorophyll a (chl a) (on average 1.9-fold), phaeophytin a (on average 7.4-fold), autotrophic (on average 6.1-fold) and heterotrophic nanoflagellates (on average 5.1-fold) relative to SSW. Enrichments in bacterial production and culturable bacteria were highly variable. EF were significantly different between the 2 sites only for concentrations of chl a, b, and c and the abundance of autotrophic nanoflagellates, with higher EF at the Barcelona site. Except for autotrophic flagellates, the abundance or activity of the parameters determined in the SML was highly correlated with that determined in SSW, suggesting that enrichment of the SML results mainly from upward transport of microorganisms attached to buoyant particles or bubble scavenging. In contrast, the high contribution of autotrophic nanoflagellates to overall phytoneuston biomass (mean = 26%) is likely due to their rapid colonization of the SML. The high abundances of auto- and heterotrophic nanoflagellates in the SML indicated that these organisms play a key role in the functioning of the microbial food webs at the air–sea interface.

Late summer community composition and abundance of photosynthetic picoeukaryotes in Norwegian and Barents Seas

We investigated marine picoeukaryotic diversity (cells <3 µm) in samples collected in late summer 2002 at the boundary between the Norwegian, Greenland, and Barents Seas. The two main Arctic and Atlantic water masses in this region are separated by the polar front. We combined total counts of picoeukaryotes assemblages by flow cytometry and epifluorescence microscopy with taxa detection by tyramide signal amplification-fluorescent in situ hybridization (TSAFISH) and high performance liquid chromatography (HPLC) pigment analyses. The picoeukaryotic community was primarily composed of photoautotrophs (75% of the cells on average). Members of the division Chlorophyta, in particular the species Micromonas pusilla (Butcher) Manton and Parke, were the major components in truly Arctic waters (32% of the picoeukaryotes, maximum 3,200 cells ml21). M. pusilla was also well represented in coastal waters and at the polar front (25% of the picoeukaryotes, maximum 9,100 cells ml21). Haptophyta were prominent in more typical Atlantic waters (up to 35% of the picoeukaryotes, maximum 4,500 cells ml21). Quantification of haptophyte biomass by HPLC pigment analyses and CHEMTAX, and haptophyte abundances by TSA-FISH were in good agreement. This confirms previous studies, which suggested that M. pusilla is a dominant contributor of picoeukaryotic communities in both coastal and nutrient rich environments, whereas haptophytes seem to be more important in open seawaters.

DNA LIBRARIES FOR SEQUENCING THE GENOME OF OSTREOCOCCUS TAURI (CHLOROPHYTA, PRASINOPHYCEAE): THE SMALLEST FREE‐LIVING EUKARYOTIC CELL1

Ostreococcus tauri is a marine photosynthetic picoeukaryote presenting a minimal cellular organization with one nucleus, one chloroplast, and one mitochondrion. It has the smallest genome described among free‐living eukaryotic cells, and we showed by pulsed‐field gel electrophoresis (PFGE) that it is divided between 15 bands ranging from 1.2 to 0.15 Mb, giving a total size of 9.7 Mb. A Bacterial Artificial Chromosome (BAC) library was prepared from genomic DNA extracted from a culture of O. tauri. A total of 2457 clones was obtained with an average insert size of around 70 kb, representing an 18‐fold coverage of the genome. The library was spotted on high density filters, and several probes of coding sequences were hybridized to both the high density BAC library filters and directly to the dried PFGE gels of the O. tauri genomic DNA. These hybridizations allowed a preliminary organization of the library and the localization of several markers on the chromosomes. Randomly selected fragments were also sequenced, representing 12% of the O. tauri genome. Many sequences showed significant similarities in data banks, mainly with plant and algae sequences. About 1000 coding sequences could be identified. These data confirmed the position of O. tauri in the green lineage and the hypothesis of a very compact organization of its genome.