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The secondary metabolites geosmin and 2-methylisoborneol (MIB) are known to taint fish with an undesirable, earthy-muddy taste and odor. In an earlier study on a zero-discharge recirculating aquaculture system (RAS), it was found that geosmin and MIB were removed by microbial communities residing in sludge from the digestion basin of the system. In the present study, 16S amplicon sequencing was used to identify changes in relative abundances of bacterial taxa in geosmin and MIB-enriched crude sludge. The removal of geosmin and MIB by the sludge was accompanied by increased abundances of 12 operational taxonomic units (OTUs). The most prominent increase in abundances was recorded for OTUs affiliated with bacterial genera known to harbor denitrifiers. Among these were the Betaproteobacteria genera Thauera, which utilizes terpenes to fuel denitrification, and Comamonas, which was previously isolated from the digestion basin of the same system and is capable of growth on geosmin and MIB as sole carbon and energy sources. Thus far, denitrification has been associated with bacteria capable of utilizing terpenes other than geosmin and MIB. The significant increase in the abundance of denitrifying bacterial genera in sludge in which geosmin and MIB comprised only 0.06% of the total carbon content might indicate that such bacteria play a major role in the removal of these compounds in anoxic environments.

Background Oysters in coastal environments are subject to fluctuating environmental conditions that may impact the ecosystem services they provide. Oyster-associated microbiomes are responsible for some of these services, particularly nutrient cycling in benthic habitats. The effects of climate change on host-associated microbiome composition are well-known, but functional changes and how they may impact host physiology and ecosystem functioning are poorly characterized. We investigated how environmental parameters affect oyster-associated microbial community structure and function along a trophic gradient in Narragansett Bay, Rhode Island, USA. Adult eastern oyster, Crassostrea virginica, gut and seawater samples were collected at 5 sites along this estuarine nutrient gradient in August 2017. Samples were analyzed by 16S rRNA gene sequencing to characterize bacterial community structures and metatranscriptomes were sequenced to determine oyster gut microbiome responses to local environments. Results There were significant differences in bacterial community structure between the eastern oyster gut and water samples, suggesting selection of certain taxa by the oyster host. Increasing salinity, pH, and dissolved oxygen, and decreasing nitrate, nitrite and phosphate concentrations were observed along the North to South gradient. Transcriptionally active bacterial taxa were similar for the different sites, but expression of oyster-associated microbial genes involved in nutrient (nitrogen and phosphorus) cycling varied throughout the Bay, reflecting the local nutrient regimes and prevailing environmental conditions. Conclusions The observed shifts in microbial community composition and function inform how estuarine conditions affect host-associated microbiomes and their ecosystem services. As the effects of estuarine acidification are expected to increase due to the combined effects of eutrophication, coastal pollution, and climate change, it is important to determine relationships between host health, microbial community structure, and environmental conditions in benthic communities.

Summary Marine Synechococcus thrive over a range of light regimes in the ocean. We examined the proteomic, genomic and physiological responses of seven Synechococcus isolates to moderate irradiances (5–80 μE m−2 s−1), and show that Synechococcus spans a continuum of light responses ranging from low light optimized (LLO) to high light optimized (HLO). These light responses are linked to phylogeny and pigmentation. Marine sub-cluster 5.1A isolates with higher phycouribilin: phycoerythrobilin ratios fell toward the LLO end of the continuum, while sub-cluster 5.1B, 5.2 and estuarine Synechococcus with less phycouribilin fell toward the HLO end of the continuum. Global proteomes were highly responsive to light, with > 50% of abundant proteins varying more than twofold between the lowest and highest irradiance. All strains downregulated phycobilisome proteins with increasing irradiance. Regulation of proteins involved in photosynthetic electron transport, carbon fixation, oxidative stress protection (superoxide dismutases) and iron and nitrogen metabolism varied among strains, as did the number of high light inducible protein (Hlip) and DNA photolyase genes in their genomes. All but one LLO strain possessed the photoprotective orange carotenoid protein (OCP). The unique combinations of light responses in each strain gives rise to distinct photophysiological phenotypes that may affect Synechococcus distributions in the ocean.

Larval oysters in hatcheries are susceptible to diseases caused by bacterial pathogens, including Vibrio spp. Previous studies have shown that daily addition of the probiotic Bacillus pumilus RI06-95 to water in rearing tanks increases larval survival when challenged with the pathogen Vibrio coralliilyticus. We propose that the presence of probiotics causes shifts in bacterial community structure in rearing tanks, leading to a net decrease in the relative abundance of potential pathogens. During three trials spanning the 2012-2015 hatchery seasons, larvae, tank biofilm, and rearing water samples were collected from control and probiotic-treated tanks in an oyster hatchery over a 12-day period after spawning. Samples were analyzed by 16S rRNA sequencing of the V4 or V6 regions followed by taxonomic classification, in order to determine bacterial community structures. There were significant differences in bacterial composition over time and between sample types, but no major effect of probiotics on the structure and diversity of bacterial communities (phylum level, Bray-Curtis k = 2, 95% confidence). Probiotic treatment, however, led to a higher relative percent abundance of Oceanospirillales and Bacillus spp. in water and oyster larvae. In the water, an increase in Vibrio spp. diversity in the absence of a net increase in relative read abundance suggests a likely decrease in the abundance of specific pathogenic Vibrio spp., and therefore lower chances of a disease outbreak. Co-occurrence network analysis also suggests that probiotic treatment had a systemic effect on targeted members of the bacterial community, leading to a net decrease in potentially pathogenic species.

Oligotrophic Lake Superior and mesotrophic Lake Erie are trophic end members of the hydrologically connected Laurentian Great Lakes system, and as such exhibit different profiles of dissolved nitrogen species. Nitrification in Lake Superior has led to increasing nitrate concentrations over the past century, as opposed to Erie, where nitrate inventories have declined due to denitrification. In this study, we examined the abundance and diversity of nitrifying microbes involved in the oxidation of ammonia to nitrite, and nitrite to nitrate. By in situ hybridization methods, we enumerated the major planktonic ammonia oxidizing bacteria (AOB) and archaea (AOA) during a July 2011 cruise from Lake Superior to Lake Erie. In Lake Superior, AOA dominated compared to AOB, typically exceeding 5 × 103 mL− 1, whereas in Erie, AOB were more abundant than AOA. These data parallel prior work on Lake Superior and Lake Erie sediments, in which AOA are far more abundant in Superior, but AOB dominate in Erie. The lakes were sampled during stratification, and AOA and AOB were largely restricted to the hypolimnion, consistent with the observation that ammonia oxidizers are photoinhibited in surface waters. In Lakes Superior and Erie, we also detected nitrite oxidizing bacteria (NOB) in a pattern paralleling AOA/AOB abundance. Phylogenetic analysis of archaeal 16S rRNA revealed that the planktonic archaea of Lake Superior are members of the ammonia oxidizing Group I.1a Thaumarchaeota most closely related to Nitrosoarchaeum limnia. These AOA are distinct from the Group I.1a AOA in Lake Superior sediments. The major AOB of Lake Erie form a subcluster within the genus Nitrosospira.

The vast majority of the phytoplankton communities in surface mixed layer of the oligotrophic ocean are numerically dominated by one of two ecotypes of Prochlorococcus, eMIT9312 or eMED4. In this study, we surveyed large latitudinal transects in the Atlantic and Pacific Ocean to determine if these ecotypes discretely partition the surface mixed layer niche, or if populations exist as a continuum along key environmental gradients, particularly temperature. Transitions of dominance occurred at approximately 19-21°C, with the eMED4 ecotype dominating the colder, and eMIT9312 ecotype dominating the warmer regions. Within these zones of regional dominance, however, the minority ecotype was not competed to extinction. Rather, a robust log-linear relationship between ecotype ratio and temperature characterized this stabilized coexistence: for every 2.5°C increase in temperature, the eMIT9312:eMED4 ratio increased by an order of magnitude. This relationship was observed in both quantitative polymerase chain reaction and in pyrosequencing assays. Water column stratification also contributed to the ecotype ratio along the basin-scale transects, but to a lesser extent. Finally, instances where the ratio of the eMED4 and eMIT9312 abundances did not correlate well with temperature were identified. Such occurrences are likely due to changes in water temperatures outpacing changes in community structure.

Southern Ocean primary productivity plays a key role in global ocean biogeochemistry and climate. At the Southern Ocean sea ice edge in coastal McMurdo Sound, we observed simultaneous cobalamin and iron limitation of surface water phytoplankton communities in late Austral summer. Cobalamin is produced only by bacteria and archaea, suggesting phytoplankton-bacterial interactions must play a role in this limitation. To characterize these interactions and investigate the molecular basis of multiple nutrient limitation, we examined transitions in global gene expression over short time scales, induced by shifts in micronutrient availability. Diatoms, the dominant primary producers, exhibited transcriptional patterns indicative of co-occurring iron and cobalamin deprivation. The major contributor to cobalamin biosynthesis gene expression was a gammaproteobacterial population, Oceanospirillaceae ASP10-02a. This group also contributed significantly to metagenomic cobalamin biosynthesis gene abundance throughout Southern Ocean surface waters. Oceanospirillaceae ASP10-02a displayed elevated expression of organic matter acquisition and cell surface attachment-related genes, consistent with a mutualistic relationship in which they are dependent on phytoplankton growth to fuel cobalamin production. Separate bacterial groups, including Methylophaga, appeared to rely on phytoplankton for carbon and energy sources, but displayed gene expression patterns consistent with iron and cobalamin deprivation. This suggests they also compete with phytoplankton and are important cobalamin consumers. Expression patterns of siderophore- related genes offer evidence for bacterial influences on iron availability as well. The nature and degree of this episodic colimitation appear to be mediated by a series of phytoplankton-bacterial interactions in both positive and negative feedback loops.

Cobalt-rich ferromanganese is an important seafloor mineral and is abundantly present in the seamount crusts. Such crusts form potential hotspots for biogeochemical activity and microbial diversity, yet our understanding of their microbial communities is lacking. In this study, a cultivation-independent approach was used to recover genomic information and derive ecological functions of the microbes in a sediment sample collected from the cobalt-rich ferromanganese crust of a seamount region in the central Pacific. A total of 78 distinct clones were obtained by fosmid library screening with a 16S rRNA based PCR method. Proteobacteria and MGI Thaumarchaeota dominated the bacterial and archaeal 16S rRNA gene sequence results in the microbial community. Nine fosmid clones were sequenced and annotated. Numerous genes encoding proteins involved in metabolic functions and heavy metal resistance were identified, suggesting alternative metabolic pathways and stress responses that are essential for microbial survival in the cobalt-rich ferromanganese crust. In addition, genes that participate in the synthesis of organic acids and exoploymers were discovered. Reconstruction of the metabolic pathways revealed that the nitrogen cycle is an important biogeochemical process in the cobalt-rich ferromanganese crust. In addition, horizontal gene transfer (HGT) events have been observed, and most of them came from bacteria, with some occurring in archaea and plants. Clone W4-93a, belonging to MGI Thaumarchaeota, contained a region of gene synteny. Comparative analyses suggested that a high frequency of HGT events as well as genomic divergence play important roles in the microbial adaption to the deep-sea environment.

Experiments were conducted in the Gulf of Aqaba to investigate top-down and bottom-up influences on microbial food webs. Experiments were run during winter mixing, spring bloom and summer stratification, periods with increasing nutrient limitation and with picophytoplankton (.98% of total year-round biomass) dominated by different species. Size fractionation (,64, ,6 and ,0.8 mm) was cross-classified with or without the addition of nutrients (0.3 mM NH4 þ and 0.19 mM PO4, equivalent to winter concentrations), using the same design for the six experiments. All experiments showed minimal top-down and strong bottom-up control, independent of season. Among phototrophs, small eukaryotic algae responded more strongly to nutrient addition within the 24-h experiment duration than did Synechococcus or Prochlorococcus. Ciliates responded most strongly to nutrient addition, becoming markedly more abundant in treatments with added nutrients relative to those without; a trend seen to a more limited degree in ciliate biomass, as smaller species responded faster. Treatments with added nutrients were also more dissimilar to one another than those without added nutrients: which species benefitted from added N and P appeared random. This could explain the high ciliate species diversity in the Gulf of Aqaba, if small nutrient pulses are rapidly transferred to different random collections of ciliate species in the vicinity of the pulse.

Protein markers of cyanobacterial stress Nutrients including iron, nitrogen, and phosphorus limit primary productivity in the oceans. Determining how abundant cyanobacteria such as Prochlorococcus adapt to nutrient stress across marine settings requires accurate molecular assays. Saito et al. developed a proteomic and metaproteomic approach capable of targeting specific metabolic biomarkers from mixed communities in seawater (see the Perspective by Moore). Prochlorococcus proteins are indicative of a major limiting nutrient across a wide transect in the Pacific Ocean; however, they also show that the limitation of multiple nutrients at overlapping biomes is an additional source of stress. Science, this issue p. 1173 ; see also p. 1120

Marine Synechococcus thrive over a range of light regimes in the ocean. We examined the proteomic, genomic and physiological responses of seven Synechococcus isolates to moderate irradiances (5-80 μE m

Heterotrophic bacteria play a key role in marine carbon cycling, and understanding their activities in polar systems is important for considering climate change impacts there. One goal of the ASPIRE project was to examine the relationship between the phytoplankton bloom and bacterial heterotrophy in the Amundsen Sea Polynya (ASP). Bacterial abundance, production (BP), respiration, growth efficiency, and extracellular enzyme activity (EEA) were compared to nutrient and organic matter inventories, chlorophyll a (Chl a), viral and microzooplankton abundance, and net primary production (NPP). Bacterial production and respiration clearly responded (0.04–4.0 and 10–53 µg C L−1 d−1, respectively) to the buildup of a massive Phaeocystis antarctica bloom (Chl a: 0.2–22 µg L−1), with highest rates observed in the central polynya where Chl a and particulate organic carbon (POC) were greatest. The highest BP rates exceeded those reported for the Ross Sea or any other Antarctic coastal system, yet the BP:NPP ratio (2.1–9.4%) was relatively low. Bacterial respiration was also high, and growth efficiency (2–27%; median = 10%) was similar to oligotrophic systems. Thus, the integrated bacterial carbon demand (0.8–2.8 g C m−2 d−1) was a high fraction (25–128%; median = 43%) of NPP during bloom development. During peak bloom, activity was particle-associated: BP and EEA correlated well with POC, and size fractionation experiments showed that the larger size fraction (> 3 µm) accounted for a majority (∼ 75%) of the BP. The community was psychrophilic, with a 5x reduction in BP when warmed to 20°C. In deeper waters, respiration remained relatively high, likely fueled by the significant downward particle flux in the region. A highly active, particle-associated, heterotrophic microbial community clearly responded to the extraordinary phytoplankton bloom in the ASP, likely limiting biological pump efficiency during the early season.

Yager, Patricia L. ... et al.-- Special feature: ASPIRE: The Amundsen Sea Polynya International Research Expedition.-- 36 pages, 11 figures, 9 tables, supplementary data http://www.bco-dmo.org/project/2132, Polynyas, or recurring areas of seasonally open water surrounded by sea ice, are foci for energy and material transfer between the atmosphere and the polar ocean. They are also climate sensitive, with both sea ice extent and glacial melt influencing their productivity. The Amundsen Sea Polynya (ASP) is the greenest polynya in the Southern Ocean, with summertime chlorophyll a concentrations exceeding 20 μg L−1. During the Amundsen Sea Polynya International Research Expedition (ASPIRE) in austral summer 2010–11, we aimed to determine the fate of this high algal productivity. We collected water column profiles for total dissolved inorganic carbon (DIC) and nutrients, particulate and dissolved organic matter, chlorophyll a, mesozooplankton, and microbial biomass to make a carbon budget for this ecosystem. We also measured primary and secondary production, community respiration rates, vertical particle flux and fecal pellet production and grazing. With observations arranged along a gradient of increasing integrated dissolved inorganic nitrogen drawdown (ΔDIN; 0.027–0.74 mol N m−2), changes in DIC in the upper water column (ranging from 0.2 to 4.7 mol C m−2) and gas exchange (0–1.7 mol C m−2) were combined to estimate early season net community production (sNCP; 0.2–5.9 mol C m−2) and then compared to organic matter inventories to estimate export. From a phytoplankton bloom dominated by Phaeocystis antarctica, a high fraction (up to ∼60%) of sNCP was exported to sub-euphotic depths. Microbial respiration remineralized much of this export in the mid waters. Comparisons to short-term (2–3 days) drifting traps and a year-long moored sediment trap capturing the downward flux confirmed that a relatively high fraction (3–6%) of the export from ∼100 m made it through the mid waters to depth. We discuss the climate-sensitive nature of these carbon fluxes, in light of the changing sea ice cover and melting ice sheets in the region, This project was funded by NSF Office of Polar Programs, Antarctic Organisms and Ecosystems (ANT-0839069 to PY, ANT-0838995 to RMS, ANT-0838975 to SS, ANT-0839012 to HWD, and ANT-0944727 to K Arrigo for support of ACA, GLvD and KEL). We also acknowledge support for participation of AFP by NSF (ANT-08-39012 to HWD and ANT 15-42220 to AFP) and for Swedish collaborators from the Swedish Research Council (#2008-6430 to S. Bertilsson and L. Riemann, and #2008-6429 to P.-O. Moksness and J. Havenhand, which supported participation of RS and SK)

Dispersal limitation in phyllosphere communities was measured on the leaf surfaces of salt-excreting Tamarix trees, which offer unique, discrete habitats for microbial assemblages. We employed 16S rRNA gene pyrosequencing to measure bacterial community dissimilarity on leaves of spatially dispersed Tamarix specimens in sites with uniform climatic conditions across the Sonoran Desert in the Southwestern United States. Our analyses revealed diverse bacterial communities with four dominant phyla that exhibited differential effects of environmental and geographic variables. Geographical distance was the most important parameter that affected community composition, particularly that of betaproteobacteria, which displayed a statistically significant, distance-decay relationship.

Atmospheric P solubility affects the amount of P available for phytoplankton in the surface ocean, yet our understanding of the timing and extent of atmospheric P solubility is based on short-term leaching experiments where conditions may differ substantially from the surface ocean. We conducted longer- term dissolution experiments of atmospheric aerosols in filtered seawater, and found up to 9-fold greater dissolution of P after 72 h compared to instantaneous leaching. Samples rich in anthropogenic materials released dissolved inorganic P (DIP) faster than mineral dust. To gauge the effect of biota on the fate of atmospheric P, we conducted field incubations with aerosol samples collected in the Sargasso Sea and Red Sea. In the Sargasso Sea phytoplankton were not P limited, and biological activity enhanced DIP release from aerosols, and aerosols induced biological mineralization of dissolved organic P in seawater, leading to DIP accumulation. However, in the Red Sea where phytoplankton were colimited by P and N, soluble P was rapidly consumed by phytoplankton following aerosol enrichment. Our results suggest that atmospheric P dissolution could continue over multiple days once reaching the surface ocean, and that previous estimates of atmospheric P deposition may underestimate the contribution from this source.

In the seasonally stratified Gulf of Aqaba Red Sea, both NO � 2 release by phytoplankton and NH þ 4 oxidation by nitrifying microbes contributed to the formation of a primary nitrite maximum (PNM) over different seasons and depths in the water column. In the winter and during the days immediately following spring stratification, NO � 2 formation was strongly correlated (R 2 = 0.99) with decreasing irradiance and chlorophyll, suggesting that incomplete NO � 3 reduction by light limited phytoplankton was a major source of

The above-ground surfaces of terrestrial plants, the phyllosphere, comprise the main interface between the terrestrial biosphere and solar radiation. It is estimated to host up to 10(26) microbial cells that may intercept part of the photon flux impinging on the leaves. Based on 454-pyrosequencing-generated metagenome data, we report on the existence of diverse microbial rhodopsins in five distinct phyllospheres from tamarisk (Tamarix nilotica), soybean (Glycine max), Arabidopsis (Arabidopsis thaliana), clover (Trifolium repens) and rice (Oryza sativa). Our findings, for the first time describing microbial rhodopsins from non-aquatic habitats, point towards the potential coexistence of microbial rhodopsin-based phototrophy and plant chlorophyll-based photosynthesis, with the different pigments absorbing non-overlapping fractions of the light spectrum.

The nitrogen (N) and oxygen (O) isotopic composition (δ 15 N and δ 18 O) of water soluble aerosol nitrate was measured in aerosol samples collected in Eilat, Israel, from August 2003 to November 2004. During this period δ 15 N values ranged from − 6.9‰ to + 1.9‰ and δ 18 O from + 65.1‰ to + 84.9‰ and exhibited strong seasonal variability with higher average δ 15 N values observed in the summer and higher δ 18 O values in the winter. Nitrate isotopic composition was compared with bulk chemical composition and extractable ion and trace metals on co-collected samples linking nitrate isotopic composition to various sources of aerosols to this region. Atmospheric processes impacting the isotopic signatures of nitrate were also considered. Based on back trajectory analyses, the majority of NO 3 − came from air masses originating over the Mediterranean Sea (34%), Western Europe (20%) and the local Negev desert (19%), which contain a larger anthropogenic imprint compared to southern and eastern air masses which are dominated by mineral dust. The potential role of reactive mineral dust aerosols as a regulator of NO 3 − isotopic composition is considered; however, based on factor analysis, neither δ 15 N nor δ 18 O were associated with mineral dust components (such as Fe or Al), but rather with anthropogenic indicators such as Cu, Cd, P and Pb. Seasonality in primary NO x cycling reactions driven by seasonal changes in solar radiation, relative humidity and temperature also influence the observed isotopic signatures. The isotope data, together with trace element analysis, suggests that seasonal variations in both δ 15 N NO3 and δ 18 O NO3 are related to both NO x source and transport processes as well as NO x chemical reactions in the atmosphere. The flux-weighted δ 15 N of aerosol NO 3 − in this area averaged − 2.6‰ making aerosol deposition a substantial contributor of low δ 15 N nitrogen to the oligotrophic waters of the Gulf of Aqaba. Thus, while the flux of atmospheric N to oligotrophic marine systems is smaller than the upward flux of NO 3 − from deep water, it nonetheless represents an important source of new N having a low δ 15 N. Further, if this low δ 15 N signature is not considered, it could interfere with N-fixation estimates based on isotopic composition of dissolved nitrate or particulate organic nitrogen. Thus, atmospheric deposition should be constrained for accurate estimates of marine N-fixation when based on δ 15 N in the ocean. Indeed, in the Gulf of Aqaba, low upper water δ 15 N NO3 values could be related to inputs of atmospheric NO 3 − as well as N-fixation.

Summary 1. It is an uncontested paradigm that an adequate supply of the macronutrients nitrogen (N) and phosphorus (P) is critical for maintaining phytoplankton primary production in aquatic ecosystems; it has also been suggested that there is an optimal total N : total P ratio for this globally significant process. 2. This ratio, normally assessed by chemical determination of the major dissolved N and P species, poses a dilemma: do chemical measurements actually reflect the bioavailable fraction of these nutrient pools? Accurate determination of the various N and P species and their fluxes into phytoplankton cells is notoriously difficult. 3. To provide a possible solution to this difficulty, we engineered strains of the cyanobacterium Synechococcus sp. strain PCC 7942 that ‘report’ on N and P bioavailability via a bioluminescent signal. These strains were used to quantify, for the first time, bioavailable concentrations of these essential macronutrients in a freshwater lake. 4. Only a small fraction (0.01–1%) of the chemically determined P may actually be bioavailable to this unicellular cyanobacterium and, by inference, to the phytoplankton community in general. In contrast, bioavailable N comprises most of the dissolved N pool. Consequently, bioavailable N : P ratios based on these assays are higher then those based on chemical determinations, indicating that P limitation in Lake Kinneret is more extensive then previously thought.

Ecological theory predicts that low productivity systems should have low biodiversity. However, despite the oligotrophic status of the Gulf of Aqaba (Northern Red Sea) ciliate species richness was unexpectedly high. In addition, phytoplankton, as main ciliate prey, was made up by only few genera, indicating a significant niche overlap among the grazers. Up to 97% of the ciliates were from the same taxonomic group and of the same size range, implying very similar food niches. Ciliate diversity was highest at times of lowest chlorophyll concentrations, during the period of stable abiotic conditions, but relatively high genetic diversity within the ciliate prey, notably among the cyanobacteria Synechococcus and Prochlorococcus. In the absence of disturbance and with little predation pressure, the alternate explanations for the observed ciliate diversity are either very fine niche partitioning by the ciliates, or their competitive equivalence resulting in a random assortment of species immigrating from a larger metacommunity, in accordance with Hubbell’s, (The unified neutral theory of biodiversity and biogeography. Princeton University Press, Princeton, 2001) neutral model. While the use of species abundance distributions (SAD’s) is far from definitive, the theoretical SAD’s that best fit the Gulf of Aqaba ciliate data was most often not that expected by neutral theory.

Using a polyphasic approach, we examined the presence of Archaea in the Gulf of Aqaba, a warm marine ecosystem, isolated from major ocean currents and subject to pronounced seasonal changes in hydrography. Catalyzed reported deposition FISH analyses showed that Archaea make up to >20% of the prokaryotic community in the Gulf. A spatial separation between the two major phyla of Archaea was observed during summer stratification. Euryarchaeota were found exclusively in the upper 200 m, whereas Crenarchaeota were present in greater numbers in layers below the summer thermocline. 16S rRNA gene-based denaturing gradient gel electrophoresis confirmed this depth partitioning and revealed further diversity of Crenarchaeota and Euryarchaeota populations along depth profiles. Phylogenetic analysis showed pelagic Crenarchaeota and Euryarchaeota to differ from coral-associated Archaea from the Gulf, forming distinct clusters within the Marine Archaea Groups I and II. Endsequencing of fosmid libraries of environmental DNA provided a tentative identification of some members of the archaeal community and their role in the microbial community of the Gulf. Incorporation studies of radiolabeled leucine and bicarbonate in the presence of different inhibitors suggest that the archaeal community participates in autotrophic CO(2) uptake and contributes little to the heterotrophic activity.

Atmospheric aerosol deposition is an important source of nutrients and trace metals to the open ocean that can enhance ocean productivity and carbon sequestration and thus influence atmospheric carbon dioxide concentrations and climate. Using aerosol samples from different back trajectories in incubation experiments with natural communities, we demonstrate that the response of phytoplankton growth to aerosol additions depends on specific components in aerosols and differs across phytoplankton species. Aerosol additions enhanced growth by releasing nitrogen and phosphorus, but not all aerosols stimulated growth. Toxic effects were observed with some aerosols, where the toxicity affected picoeukaryotes and Synechococcus but not Prochlorococcus . We suggest that the toxicity could be due to high copper concentrations in these aerosols and support this by laboratory copper toxicity tests preformed with Synechococcus cultures. However, it is possible that other elements present in the aerosols or unknown synergistic effects between these elements could have also contributed to the toxic effect. Anthropogenic emissions are increasing atmospheric copper deposition sharply, and based on coupled atmosphere–ocean calculations, we show that this deposition can potentially alter patterns of marine primary production and community structure in high aerosol, low chlorophyll areas, particularly in the Bay of Bengal and downwind of South and East Asia.

Cyanate, a by-product of urea decomposition, is a potential nitrogen (N) source in marine environments, but to date it has received scant attention. Cyanobacteria presumably acquire this compound via a substrate-specific ABC-type transporter (cynABD), and they convert it to ammonium and carbon dioxide by cyanase (cynS) activity. Participation of cyanate utilization genes in N-stress responses in cyanobacteria has been implied previously, but its ecological context has not been studied. We employed polymerase chain reaction protocols for cynA amplification to examine the potential for cyanate recruitment in the Gulf of Aqaba, northern Red Sea. We also monitored growth of cyanobacterial strains with different cynABDS complements on cyanate-containing media. We showed that cyanate is utilized as the sole N source by strains with the full gene complement, and residual growth, fed by natural decay of cyanate to ammonium, was observed in strains lacking any of these genes. Natural abundance of cynA products in the oligotrophic Gulf of Aqaba indicates that cyanate constitutes an essential N source for Prochlorococcus, but not for Synechococcus populations. Noncyanobacterial cynA sequences indicate cyanate utilization by a variety of other phototrophic microorganisms. We hypothesize that in stratified water bodies, cyanate utilization is confined to the N-deplete upper photic zone, where it plays a role in "regenerated" primary production.

Ciliate data were gathered along a 600 m deepwater column in central waters of the Gulf of Aqaba, Israel. Samples were taken during winter mixing, the onset of stratification in spring and during summer stratification (2003 to 2005). The phytoplankton community strongly differed in these 3 periods, from highest abundances of eukaryotic algae during mixing, followed by a Syne- chococcus bloom right after the onset of stratification and a Prochlorococcus-dominated community in summer. Ciliate abundance and biomass were high compared to chlorophyll standing stock, with maximal values of 3534 cells l -1 and 3554 ng C l -1 , respectively. Abundances were at times 5-fold higher than those found in comparable studies on nutrient-poor pelagic systems and approached those observed in coastal waters. Ciliate carbon:chlorophyll ratios of up to 26 were astonishingly high, as compared to values of 2 to 5 normally observed. This indicates a high efficiency of the cili- ates in utilizing the available food. The phytoplankton community in the Gulf is dominated by pico- cyanobacteria, and by repacking the algal prey into accessible particles, protozoans might be an important intermediate link in the pathway between phytoplankton and metazoan grazers. Olig- otrichs and nanociliates contributed up to 98 and 88% of ciliate total abundance, respectively. Small oligotrich species appeared to be the superior competitors in this oligotrophic marine system, being able to utilize the dominant picoautotrophs efficiently.

Natural populations of the marine cyanobacterium Prochlorococcus exist as two main ecotypes, inhabiting different layers of the ocean's photic zone. These so-called high light- (HL-) and low light (LL-) adapted ecotypes are both physiologically and genetically distinct. HL strains can be separated into two major clades (HLI and HLII), whereas LL strains are more diverse. Here, we used several molecular techniques to study the genetic diversity of natural Prochlorococcus populations during the Prosope cruise in the Mediterranean Sea in the summer of 1999. Using a dot blot hybridization technique, we found that HLI was the dominant HL group and was confined to the upper mixed layer. In contrast, LL ecotypes were only found below the thermocline. Secondly, a restriction fragment length polymorphism analysis of PCR-amplified pcb genes (encoding the major light-harvesting proteins of Prochlorococcus) suggested that there were at least four genetically different ecotypes, occupying distinct but overlapping light niches in the photic zone. At comparable depths, similar banding patterns were observed throughout the sampled area, suggesting a horizontal homogenization of ecotypes. Nevertheless, environmental pcb gene sequences retrieved from different depths at two stations proved all different at the nucleotide level, suggesting a large genetic microdiversity within those ecotypes.

Antarctica polynyas support intense phytoplankton blooms, impacting their environment by a substantial depletion of inorganic carbon and nutrients. These blooms are dominated by the colony-forming haptophyte Phaeocystis antarctica and they are accompanied by a distinct bacterial population. Yet, the ecological role these bacteria may play in P. antarctica blooms awaits elucidation of their functional gene pool and of the geochemical activities they support. Here, we report on a metagenome (῀160 million reads) analysis of the microbial community associated with a P. antarctica bloom event in the Amundsen Sea polynya (West Antarctica). Genomes of the most abundant Bacteroidetes and Proteobacteria populations have been reconstructed and a network analysis indicates a strong functional partitioning of these bacterial taxa. Three of them (SAR92, and members of the Oceanospirillaceae and Cryomorphaceae) are found in close association with P. antarctica colonies. Distinct features of their carbohydrate, nitrogen, sulfur and iron metabolisms may serve to support mutualistic relationships with P. antarctica. The SAR92 genome indicates a specialization in the degradation of fatty acids and dimethylsulfoniopropionate (compounds released by P. antarctica) into dimethyl sulfide, an aerosol precursor. The Oceanospirillaceae genome carries genes that may enhance algal physiology (cobalamin synthesis). Finally, the Cryomorphaceae genome is enriched in genes that function in cell or colony invasion. A novel pico-eukaryote, Micromonas related genome (19.6 Mb, ~94% completion) was also recovered. It contains the gene for an anti-freeze protein, which is lacking in Micromonas at lower latitudes. These draft genomes are representative for abundant microbial taxa across the Southern Ocean surface.

The vast majority of the phytoplankton communities in surface mixed layer of the oligotrophic ocean are numerically dominated by one of two ecotypes of Prochlorococcus, eMIT9312 or eMED4. In this study, we surveyed large latitudinal transects in the Atlantic and Pacific Ocean to determine if these ecotypes discretely partition the surface mixed layer niche, or if populations exist as a continuum along key environmental gradients, particularly temperature. Transitions of dominance occurred at approximately 19-21°C, with the eMED4 ecotype dominating the colder, and eMIT9312 ecotype dominating the warmer regions. Within these zones of regional dominance, however, the minority ecotype was not competed to extinction. Rather, a robust log-linear relationship between ecotype ratio and temperature characterized this stabilized coexistence: for every 2.5°C increase in temperature, the eMIT9312:eMED4 ratio increased by an order of magnitude. This relationship was observed in both quantitative polymerase chain reaction and in pyrosequencing assays. Water column stratification also contributed to the ecotype ratio along the basin-scale transects, but to a lesser extent. Finally, instances where the ratio of the eMED4 and eMIT9312 abundances did not correlate well with temperature were identified. Such occurrences are likely due to changes in water temperatures outpacing changes in community structure.

Marine Synechococcus are some of the most diverse and ubiquitous phytoplankton, and iron (Fe) is an essential micronutrient that limits productivity in many parts of the ocean. To investigate how coastal and oceanic Atlantic Synechococcus strains acclimate to Fe availability, we compared the growth, photophysiology, and quantitative proteomics of two Synechococcus strains from different Fe regimes. Synechococcus strain WH8102, from a region in the southern Sargasso Sea that receives substantial dust deposition, showed impaired growth and photophysiology as Fe declined, yet used few acclimation responses. Coastal WH8020, from the dynamic, seasonally variable New England shelf, displayed a multitiered, hierarchical cascade of acclimation responses with different Fe thresholds. The multitiered response included changes in Fe acquisition, storage, and photosynthetic proteins, substitution of flavodoxin for ferredoxin, and modified photophysiology, all while maintaining remarkably stable growth rates over a range of Fe concentrations. Modulation of two distinct ferric uptake regulator (Fur) proteins that coincided with the multitiered proteome response was found, implying the coastal strain has different regulatory threshold responses to low Fe availability. Low nitrogen (N) and phosphorus (P) availability in the open ocean may favor the loss of Fe response genes when Fe availability is consistent over time, whereas these genes are retained in dynamic environments where Fe availability fluctuates and N and P are more abundant.

We report results from a yearlong, moored sediment trap in the Amundsen Sea Polynya (ASP), the first such time series in this remote and productive ecosystem. Results are compared to a long-term (1992–2013) time series from the western Antarctic Peninsula (WAP). The ASP trap was deployed from December 2010 to December 2011 at 350 m depth. We observed two brief, but high flux events, peaking at 8 and 5 mmol C m−2 d−1 in January and December 2011, respectively, with a total annual capture of 315 mmol C m−2. Both peak fluxes and annual capture exceeded the comparable WAP observations. Like the overlying phytoplankton bloom observed during the cruise in the ASP (December 2010 to January 2011), particle flux was dominated by Phaeocystis antarctica, which produced phytodetrital aggregates. Particles at the start of the bloom were highly depleted in 13C, indicating their origin in the cold, CO2-rich winter waters exposed by retreating sea ice. As the bloom progressed, microscope visualization and stable isotopic composition provided evidence for an increasing contribution by zooplankton fecal material. Incubation experiments and zooplankton observations suggested that fecal pellet production likely contributed 10–40% of the total flux during the first flux event, and could be very high during episodic krill swarms. Independent estimates of export from the surface (100 m) were about 5–10 times that captured in the trap at 350 m. Estimated bacterial respiration was sufficient to account for much of the decline in the flux between 50 and 350 m, whereas zooplankton respiration was much lower. The ASP system appears to export only a small fraction of its production deeper than 350 m within the polynya region. The export efficiency was comparable to other polar regions where phytoplankton blooms were not dominated by diatoms.

© 2015 Mackey, Chien, Post, Saito and Paytan. Atmospheric deposition is a major source of trace metals in marine surface waters and supplies vital micronutrients to phytoplankton, yet measured aerosol trace metal solubility values are operationally defined, and there are relatively few multi-element studies on aerosol-metal solubility in seawater. Here we measure the solubility of aluminum (Al), cadmium (Cd), cobalt (Co), copper (Cu), iron (Fe), manganese (Mn), nickel (Ni), lead (Pb), and zinc (Zn) from natural aerosol samples in seawater over a 7 days period to (1) evaluate the role of extraction time in trace metal dissolution behavior and (2) explore how the individual dissolution patterns could influence biota. Dissolution behavior occurs over a continuum ranging from rapid dissolution, in which the majority of soluble metal dissolved immediately upon seawater exposure (Cd and Co in our samples), to gradual dissolution, where metals dissolved slowly over time (Zn, Mn, Cu, and Al in our samples). Additionally, dissolution affected by interactions with particles was observed in which a decline in soluble metal concentration over time occurred (Fe and Pb in our samples). Natural variability in aerosol chemistry between samples can cause metals to display different dissolution kinetics in different samples, and this was particularly evident for Ni, for which samples showed a broad range of dissolution rates. The elemental molar ratio of metals in the bulk aerosols was 23,189Fe: 22,651Al: 445Mn: 348Zn: 71Cu: 48Ni: 23Pb: 9Co: 1Cd, whereas the seawater soluble molar ratio after 7 days of leaching was 11Fe: 620Al: 205Mn: 240Zn: 20Cu: 14Ni: 9Pb: 2Co: 1Cd. The different kinetics and ratios of aerosol metal dissolution have implications for phytoplankton nutrition, and highlight the need for unified extraction protocols that simulate aerosol metal dissolution in the surface ocean.

Sampling ecosystems, even at a local scale, at the temporal and spatial resolution necessary to capture natural variability in microbial communities are prohibitively expensive. We extrapolated marine surface microbial community structure and metabolic potential from 72 16S rRNA amplicon and 8 metagenomic observations using remotely sensed environmental parameters to create a system-scale model of marine microbial metabolism for 5904 grid cells (49km 2 ) in the Western English Chanel, across 3 years of weekly averages. Thirteen environmental variables predicted the relative abundance of 24 bacterial Orders and 1715 unique enzyme-encoding genes that encode turnover of 2893 metabolites. The genes’ predicted relative abundance was highly correlated (Pearson Correlation 0.72, P-value o10 � 6 ) with their observed relative abundance in sequenced metagenomes. Predictions of the relative turnover (synthesis or consumption) of CO2 were significantly correlated with observed surface CO2 fugacity. The spatial and temporal variation in the predicted relative abundances of genes coding for cyanase, carbon monoxide and malate dehydrogenase were investigated along with the predicted inter-annual variation in relative consumption or production of B3000 metabolites forming six significant temporal clusters. These spatiotemporal distributions could possibly be explained by the co-occurrence of anaerobic and aerobic metabolisms associated with localized plankton blooms or sediment resuspension, which facilitate the presence of anaerobic micro-niches. This predictive model provides a general framework for focusing future sampling and experimental design to relate biogeochemical turnover to microbial ecology. The ISME Journal (2015) 9, 166–179; doi:10.1038/ismej.2014.107; published online 29 July 2014

Synechococcus-specific polymerase chain reaction (PCR) primers were developed and used to amplify fragments of the RNA polymerase core subunit gene rpoC1 of a set of 23 marine isolates from different oceanographic locations. Restriction fragment length polymorphism (RFLP) analysis of the PCR products differentiated between most strains and all phylogenetic clades, thus allowing the screening of large sample sets at high genetic resolution. The method was used to analyse changes in the genetic diversity of marine Synechococcus strains through an annual cycle at 100 m depth in the Gulf of Aqaba, Red Sea, and along the depth profile of a sample from June 1999. RFLP analysis of 36 clones of each of the 8 rpoC1 clone libraries screened showed that the Synechococcus population during summer and autumn was dominated by 1 to 3 genetically different clones, while maximum genetic richness was found during spring and winter. Analysis of the depth profile during the period of stratification revealed differences in relative abundance of particular RFLP types among depths (10, 30, 100 m), although the same genotypes dominated the assemblage throughout. Multivariate analysis of environmental variables and Synechococcus assemblage composition showed that Synechococcus genetic diversity was most closely, and significantly, correlated to a combination of temperature and NO3- (P = 0.645, p = 0.038). Phylogenetic analysis of the marine rpoC1 nucleotide sequences of the 40 different RFLP types identified in this study, and all Synechococcus rpoC1 sequences in the databases, demonstrates that the new PCR primers amplify rpoC1 gene fragments from all MC-A and some Cyanobium lineages of marine Synechococcus strains and are also useful for the identification of potentially novel lineages.

Proteomics has great potential for studies of marine microbial biogeochemistry, yet high microbial diversity in many locales presents us with unique challenges. We addressed this challenge with a targeted metaproteomics workflow for NtcA and P-II, two nitrogen regulatory proteins, and demonstrated its application for cyanobacterial taxa within microbial samples from the Central Pacific Ocean. Using METATRYP, an open-source Python toolkit, we examined the number of shared (redundant) tryptic peptides in representative marine microbes, with the number of tryptic peptides shared between different species typically being 1% or less. The related cyanobacteria Prochlorococcus and Synechococcus shared an average of 4.8 ± 1.9% of their tryptic peptides, while shared intraspecies peptides were higher, 13 ± 15% shared peptides between 12 Prochlorococcus genomes. An NtcA peptide was found to target multiple cyanobacteria species, whereas a P-II peptide showed specificity to the high-light Prochlorococcus ecotype. Distributions of NtcA and P-II in the Central Pacific Ocean were similar except at the Equator likely due to differential nitrogen stress responses between Prochlorococcus and Synechococcus. The number of unique tryptic peptides coded for within three combined oceanic microbial metagenomes was estimated to be ∼4 × 10(7) , 1000-fold larger than an individual microbial proteome and 27-fold larger than the human proteome, yet still 20 orders of magnitude lower than the peptide diversity possible in all protein space, implying that peptide mapping algorithms should be able to withstand the added level of complexity in metaproteomic samples.

Rising temperatures and changing winds drive the expansion of the highly productive polynyas (open water areas surrounded by sea ice) abutting the Antarctic continent. Phytoplankton blooms in polynyas are often dominated by the haptophyte Phaeocystis antarctica, and they generate the organic carbon that enters the resident microbial food web. Yet, little is known about how Phaeocystis blooms shape bacterial community structures and carbon fluxes in these systems. We identified the bacterial communities that accompanied a Phaeocystis bloom in the Amundsen Sea polynya during the austral summers of 2007–2008 and 2010–2011. These communities are distinct from those determined for the Antarctic Circumpolar Current (ACC) and off the Palmer Peninsula. Diversity patterns for most microbial taxa in the Amundsen Sea depended on location (e.g., waters abutting the pack ice near the shelf break and at the edge of the Dotson glacier) and depth, reflecting different niche adaptations within the confines of this isolated ecosystem. Inside the polynya, P. antarctica coexisted with the bacterial taxa Polaribacter sensu lato, a cryptic Oceanospirillum, SAR92 and Pelagibacter. These taxa were dominated by a single oligotype (genotypes partitioned by Shannon entropy analysis) and together contributed up to 73% of the bacterial community. Size fractionation of the bacterial community [3 μm (particle-associated bacteria)] identified several taxa (especially SAR92) that were preferentially associated with Phaeocystis colonies, indicative of a distinct role in Phaeocystis bloom ecology. In contrast, particle-associated bacteria at 250 m depth were enriched in Colwellia and members of the Cryomorphaceae suggesting that they play important roles in the decay of Phaeocystis blooms.

Determining the nitrogen (N) status of phytoplankton is important for understanding primary production and N cycling in marine ecosystems. We assayed transcript levels of the N regulatory gene ntcA to assess the physiological N status of Synechococcus populations exposed to different N regimes in the meso- to oligotrophic Gulf of Aqaba, Red Sea. Synechococcus populations were N sufficient even in low-N environments when the ratio of dissolved nitrogen to phosphorus indicated that overall phytoplankton biomass was constrained by N. Ammonium supported Synechococcus N requirements under most conditions, but during a massive spring bloom in April 2000 alternative N sources were utilized. Evidence from ntcA clone libraries indicates changes in the genotypic makeup of Synechococcus populations under different N regimes, suggesting that the Synechococcus genotypes present in N-poor waters were those adapted for life in these environments. Thus, the success of Synechococcus in the open oceans is likely to be at least partially due to the selection of genotypes suited to life under prevailing N conditions rather than to prolonged manifestation of the N stress response, mediated by ntcA, in less well-adapted genotypes. Low photosynthetic biomass, prevalent in vast oligotrophic expanses of the world’s oceans, is often attributed to 1

The marine unicellular cyanobacterium Prochlorococcus is the smallest-known oxygen-evolving autotroph1. It numerically dominates the phytoplankton in the tropical and subtropical oceans2,3, and is responsible for a significant fraction of global photosynthesis. Here we compare the genomes of two Prochlorococcus strains that span the largest evolutionary distance within the Prochlorococcus lineage4 and that have different minimum, maximum and optimal light intensities for growth5. The high-light-adapted ecotype has the smallest genome (1,657,990 base pairs, 1,716 genes) of any known oxygenic phototroph, whereas the genome of its low-light-adapted counterpart is significantly larger, at 2,410,873 base pairs (2,275 genes). The comparative architectures of these two strains reveal dynamic genomes that are constantly changing in response to myriad selection pressures. Although the two strains have 1,350 genes in common, a significant number are not shared, and these have been differentially retained from the common ancestor, or acquired through duplication or lateral transfer. Some of these genes have obvious roles in determining the relative fitness of the ecotypes in response to key environmental variables, and hence in regulating their distribution and abundance in the oceans.

Summary Marine cyanobacteria of the genus Prochlorococcus belong to one of two ecotypes that are specifically adapted to either low light (LL) or high light (HL) conditions. Previous analyses of the differences in pigmentation and gene complement revealed that LL-adapted ecotypes carry a gene cluster to produce a functional phycoerythrin, whereas in the fully sequenced genome of the HL-adapted strain MED4, only a single and free-standing cpeB gene occurs. This gene encodes a derived form of β-phycoerythrin, the function of which has remained enigmatic so far. Here, an analysis of HL-adapted Prochlorococcus strains from different ocean provinces revealed the presence of a cpeB gene highly similar to that of MED4. To investigate whether the presence of particular phycoerythrin genes is a common characteristic of the LL- and HL-adapted ecotypes, primer sets targeting specific motifs in LL-cpeB and HL-cpeB were designed for polymerase chain reaction (PCR) analysis of Red Sea phytoplankton. A major PCR product for Prochlorococcus HL-cpeB was obtained from samples taken at 5–70 m depth and for LL-cpeB from 70–125 m. The high sensitivity of this approach allowed the detection of HL-cpeB down to 100 m and LL-cpeB as deep as 175 m. DNA sequence and phylogenetic analysis of 70 individual clones for HL-cpeB and of 68 clones for LL-cpeB revealed a monophyletic origin for the HL and LL sequences respectively. This study shows that cpeB sequences are suitable as very sensitive molecular markers for the study of natural populations of Prochlorococcus. The low sequence divergence of HL-cpeB among Prochlorococcus strains, which have been isolated from the Mediterranean Sea, the Arabian Sea and the Southern Pacific Ocean as well as in populations from the Red Sea, suggests the HL-cpeB gene to be conserved and its product to be functional in Prochlorococcus.

Proteorhodopsins, ubiquitous retinylidene photoactive proton pumps, were recently discovered in the cosmopolitan uncultured SAR86 bacterial group in oceanic surface waters. Two related proteorhodopsin families were found that absorb light with different absorption maxima, 525 nm (green) and 490 nm (blue), and their distribution was shown to be stratified with depth. Using structural modeling comparisons and mutagenesis, we report here on a single amino acid residue at position 105 that functions as a spectral tuning switch and accounts for most of the spectral difference between the two pigment families. Furthermore, looking at natural environments, we found novel proteorhodopsin gene clusters spanning the range of 540-505 nm and containing changes in the same identified key switch residue leading to changes in their absorption maxima. The results suggest a simultaneous diversification of green proteorhodopsin and the new key switch variant pigments. Our observations demonstrate that this single-residue switch mechanism is the major determinant of proteorhodopsin wavelength regulation in natural marine environments.

In contrast to extensive studies of phosphorus, widely considered the main nutrient limiting phytoplankton biomass in freshwater ecosystems, there have been few studies on the role of nitrogen in controlling phytoplankton populations. This situation may be due partly to the complexity in estimating its utilization and bioavailability. In an attempt to provide a novel tool for this purpose, we fused the promoter of the glutamine synthetase-encoding gene, P glnA , from Synechococcus sp. strain PCC7942 to the luxAB luciferase-encoding genes of the bioluminescent bacterium Vibrio harveyi . The resulting construct was introduced into a neutral site on the Synechococcus chromosome to yield the reporter strain GSL. Light emission by this strain was dependent upon ambient nitrogen concentrations. The linear response range of the emitted luminescence was 1 mM to 1 μM for the inorganic nitrogen species tested (ammonium, nitrate, and nitrite) and 10- to 50-fold lower for glutamine and urea. When water samples collected from along a depth profile in Lake Kinneret (Israel) were exposed to the reporter strain, the bioluminescence of the reporter strain mirrored the total dissolved nitrogen concentrations determined for the same samples and was shown to be a sensitive indicator of the concentration of bioavailable nitrogen.

Photosynthetic microorganisms play a crucial role in the marine environment. In vast areas of the oceans, marine primary productivity is performed by cells smaller than 2-3 micro m (picoplankton). Here, we report on molecular analyses of the conserved photosynthetic psbA gene (coding for protein D1 of photosystem II reaction centre) as a diversity indicator of naturally occurring marine oxygenic picophytoplankton. The psbA genes proved to be good indicators of the presence of a wide variety of photosynthetic marine microbial groups, including new cyanobacterial groups and eukaryotic algae (prasinophytes). Furthermore, using environmental bacterial artificial chromosome (BAC) libraries, we were able to correlate psbA genes with small subunit rRNAs and, therefore, to confirm their phylogenetic affiliation.

The expression of ftsZ, encoding the initiating protein of the prokaryotic cell division was analysed in natural Prochlorococcus populations in the Gulf of Aqaba, northern Red Sea. During the seasonal Prochlorococcus bloom in September 2000, picoplankton was collected from the deep chlorophyll maximum (DCM) at 2-4 h intervals over 3 consecutive days. Flow cytometric measurements as well as DNA sequence analyses showed that Prochlorococcus was the dominant photosynthetic organism. Cell densities peaked as high as 1.4 x 10(5) cells ml(-1). This DCM population mainly consisted of brightly red fluorescing Prochlorococcus cells, corresponding to low light-adapted 'ecotypes' (sensu Moore et al., 1998, Nature 393: 464-467). Prochlorococcus populations grew in a highly synchronized fashion with DNA replication in the afternoon and cell division during the night. The ftsZ mRNA level reached maximum values within the replication phase between 14.00 and 16.00 hours, and minimum values between 02.00 and 06.00 hours. Thus, the transcriptional regulation of ftsZ could be a major factor triggering the synchronized cell division of Prochlorococcus populations. This is the first application of quantitative reverse transcriptase-coupled real-time polymerase chain reaction (PCR) to natural populations of an environmentally relevant marine organism.

The ntcA and amtl genes (encoding a transcriptional activator of genes involved in nitrogen metabolism and a high affinity ammonium transporter respectively) from the axenic Prochlorococcus strain PCC 9511 were amplified,cloned, and sequenced. Cultures of this strain were exposed to nitrogen deprivation, and the expression of ntcA and amtl was monitored along with cell density, photochemical quantum yield (as determined from F v /F m measurements), and cellular C:N ratios. Nitrogen deprivation led to arrested cell growth, reduced photochemical quantum yields, and increased cellular C:N ratios. Surprisingly, transcript accumulation patterns of ntcA and amtl were not correlated. High levels of amtl transcript were observed in both nitrogen-replete and -deplete conditions. Expression of amtl declined under severe deprivation of either phosphorus or nitrogen and showed a positive correlation with photochemical quantum yield. In contrast, ntcA expression was specifically enhanced in the absence of ammonium. High ntcA transcript levels were maintained over time and did not decline with low F v /F m values under severe N deprivation. Although ntcA transcript levels correlated somewhat with cellular C:N ratios, at the onset of nitrogen deprivation increases in ntcA mRNA levels occurred before increases in cellular C:N ratios. Constitutively high ammonium transporter expression together with low cellular ammonium requirements are likely to have important implications for the success of Prochlorococcus in the nitrogen-poor oligotrophic waters where they are most abundant. The function of NtcA in Prochlorococcus strain PCC 9511 remains unclear.

Prochlorococcus is the most abundant phytoplankter throughout the photic zone in stratified marine waters and experiences distinct gradients of light and nitrogen nutrition. Physiologically and genetically distinct Prochlorococcus ecotypes partition the water column: high-B/A (low-light adapted) ecotypes are generally restricted to the deep euphotic zone near or at the nitracline. Low-B/A (high-light adapted) ecotypes predominate in, but are not limited to, NO -depleted surface waters, where they outnumber coexisting Synechococcus populations. The niche parti2 3 tioning by different Prochlorococcus ecotypes begs the question of whether they also differ in their nitrogen (N) utilization physiology, especially with respect to NO utilization. To explore this possibility, we studied the capa2 3 bilities of different Prochlorococcus and Synechococcus strains to grow on a variety of N sources. We found that all the isolates grew well on NH and all were capable of urea utilization, occasionally at a lower growth rate. 1 4 None of the Prochlorococcus isolates were able to grow with NO . Four high-B/A Prochlorococcus isolates grew 2 3 on NO , but all others did not. Whole genome analysis of the low-B/A Prochlorococcus MED4 revealed that the 2 2 genes required for NO uptake and reduction were absent. The genome of the high-B/A ProchlorococcusMIT 9313 2 3 also lacked the NO utilization genes but has homologs of genes required for NO utilization consistent with its