Marine fixation of atmospheric nitrogen is believed to be an important source of biologically useful nitrogen to ocean surface waters, stimulating productivity of phytoplankton and so influencing the global carbon cycle. The majority of nitrogen fixation in tropical waters is carried out by the marine cyanobacterium Trichodesmium, which supplies more than half of the new nitrogen used for primary production. Although the factors controlling marine nitrogen fixation remain poorly understood, it has been thought that nitrogen fixation is limited by iron availability in the ocean. This was inferred from the high iron requirement estimated for growth of nitrogen fixing organisms and the higher apparent densities of Trichodesmium where aeolian iron inputs are plentiful. Here we report that nitrogen fixation rates in the central Atlantic appear to be independent of both dissolved iron levels in sea water and iron content in Trichodesmium colonies. Nitrogen fixation was, instead, highly correlated to the phosphorus content of Trichodesmium and was enhanced at higher irradiance. Furthermore, our calculations suggest that the structural iron requirement for the growth of nitrogen-fixing organisms is much lower than previously calculated. Although iron deficiency could still potentially limit growth of nitrogen-fixing organisms in regions of low iron availability-for example, in the subtropical North Pacific Ocean-our observations suggest that marine nitrogen fixation is not solely regulated by iron supply.

The fresh water discharged by large rivers such as the Amazon is transported hundreds to thousands of kilometers away from the coast by surface plumes. The nutrients delivered by these river plumes contribute to enhanced primary production in the ocean, and the sinking flux of this new production results in carbon sequestration. Here, we report that the Amazon River plume supports N 2 fixation far from the mouth and provides important pathways for sequestration of atmospheric CO 2 in the western tropical North Atlantic (WTNA). We calculate that the sinking of carbon fixed by diazotrophs in the plume sequesters 1.7 Tmol of C annually, in addition to the sequestration of 0.6 Tmol of C yr ؊1 of the new production supported by NO 3 delivered by the river. These processes revise our current understanding that the tropical North Atlantic is a source of 2. diatom diazotroph associations ͉ nitrogen fixation ͉ new production ͉ river plumes ͉ Richelia D ownward vertical transport of organic carbon produced by phytoplankton, referred to as the biological pump, is a mechanism that transfers carbon from the surface to the deep ocean and regulates atmospheric CO 2 (1). The flux of nitrate (NO 3 ) from deep water to the photic zone can stimulate new phytoplankton production and export (2), but because the upwelling or diffusive flux of NO 3 is accompanied by a corresponding upward flux of CO 2 , its net contribution to removal of carbon from the atmosphere is much reduced. However, the sinking flux due to new production associated with nitrogenous inputs from rivers, atmospheric deposition, and N 2 fixation (diazotrophy), results in the net transport of atmospheric carbon to the deep ocean (3), or ''carbon sequestration'' (4).The Amazon River has the largest discharge of any river and accounts for 18% of all of the riverine input to the oceans. Between May and September, the Amazon plume covers up to 1.3 ϫ 10 6 km 2 with a freshwater lens of salinity Ͻ35 [supporting information (SI) Table S1], which accounts for 20% of the WTNA. Our understanding of the influence of the Amazon River on the carbon cycle in the WTNA has evolved significantly since Ryther et al. (5) first suggested that the Amazon River depressed the productivity of the region influenced by its plume. Several studies have focused on the nutrients delivered by the river to the inner shelf, the subsequent river-supported new production of 0. Fig. 1 and Table S2) complement earlier studies by examining the region of the plume starting 300 km north of the mouth of the river. We classified the stations into three categories based on sea surface salinity (SSS).¶ ¶ The ''low salinity'' group contained all of the stations with SSS Ͻ30. Stations that had SSS between 30 and 35 were classified as ''mesohaline,'' whereas those with SSS Ͼ35 were classified as ''oceanic.'' Surface NO 3 concentrations were below detection at most stations, with the highest value of 0.50 M recorded at the station with the lowest salinity of 24. DeMaster and Pope (7) found when plotting NO 3 vs...

The conventional model of iron uptake in marine eukaryotic phytoplankton-the FeЈ model-suggests a dependency of uptake rate on the concentration of unchelated iron species (FeЈ), and not the concentration of total iron or iron chelated with organic ligands. However, iron in seawater is bound by strong organic ligands that buffer such low FeЈ concentrations that they should not support phytoplankton growth. Studies that show uptake and extracellular reduction of siderophore-bound iron by diatoms and provide indications that the iron uptake system of phytoplankton may be similar to that of yeast in which extracellular reduction is a prerequisite for uptake, call for revisions of the FeЈ model. In this paper we propose a new model for iron uptake by diatoms in which extracellular reduction of all Fe species is a necessary step for iron acquisition. Experiments verifying the predictions of the model are presented. In particular we show data supporting the fact that Fe(II) is formed as an intermediate during Fe uptake from all experimental media, including those buffered by Fe(III)EDTA. This model reconciles the standing FeЈ model with new data and concepts on reduction of iron chelates and provides a convenient framework for designing and interpreting iron uptake experiments in a variety of natural and artificial media.A major role for iron in marine phytoplankton physiology, ecosystem structure, and the ocean carbon cycle is emerging from numerous oceanographic studies (e.g., Wells et al. 1994;Maldonado et al. 2001). Trace metal clean procedures and many innovative analytical techniques have provided accurate measurements of oceanic iron concentrations and are beginning to reveal the speciation and cycling of iron in the ocean (e.g., Johnson et al. 1997;Rue and Bruland 1997). In order to understand the import of these findings for phytoplankton physiology and ecology, we must understand the relationship between uptake mechanisms of phytoplankton and the concentration and speciation of iron in seawater. The conventional model of iron uptake by marine eukaryotic phytoplankton-the FeЈ model-suggests a dependency of uptake rate on the concentration of unchelated

Harmful algal blooms (HABs) cause significant economic and ecological damage worldwide. Despite considerable efforts, a comprehensive understanding of the factors that promote these blooms has been lacking, because the biochemical pathways that facilitate their dominance relative to other phytoplankton within specific environments have not been identified. Here, biogeochemical measurements showed that the harmful alga Aureococcus anophagefferens outcompeted co-occurring phytoplankton in estuaries with elevated levels of dissolved organic matter and turbidity and low levels of dissolved inorganic nitrogen. We subsequently sequenced the genome of A. anophagefferens and compared its gene complement with those of six competing phytoplankton species identified through metaproteomics. Using an ecogenomic approach, we specifically focused on gene sets that may facilitate dominance within the environmental conditions present during blooms. A. anophagefferens possesses a larger genome (56 Mbp) and has more genes involved in light harvesting, organic carbon and nitrogen use, and encoding selenium- and metal-requiring enzymes than competing phytoplankton. Genes for the synthesis of microbial deterrents likely permit the proliferation of this species, with reduced mortality losses during blooms. Collectively, these findings suggest that anthropogenic activities resulting in elevated levels of turbidity, organic matter, and metals have opened a niche within coastal ecosystems that ideally suits the unique genetic capacity of A. anophagefferens and thus, has facilitated the proliferation of this and potentially other HABs.

Environmental fluctuations affect distribution, growth and abundance of diatoms in nature, with iron (Fe) availability playing a central role. Studies on the response of diatoms to low Fe have either utilized continuous (24 hr) illumination or sampled a single time of day, missing any temporal dynamics. We profiled the physiology, metabolite composition, and global transcripts of the pennate diatom Phaeodactylum tricornutum during steady-state growth at low, intermediate, and high levels of dissolved Fe over light:dark cycles, to better understand fundamental aspects of genetic control of physiological acclimation to growth under Fe-limitation. We greatly expand the catalog of genes involved in the low Fe response, highlighting the importance of intracellular trafficking in Fe-limited diatoms. P. tricornutum exhibited transcriptomic hallmarks of slowed growth leading to prolonged periods of cell division/silica deposition, which could impact biogeochemical carbon sequestration in Fe-limited regions. Light harvesting and ribosome biogenesis transcripts were generally reduced under low Fe while transcript levels for genes putatively involved in the acquisition and recycling of Fe were increased. We also noted shifts in expression towards increased synthesis and catabolism of branched chain amino acids in P. tricornutum grown at low Fe whereas expression of genes involved in central core metabolism were relatively unaffected, indicating that essential cellular function is protected. Beyond the response of P. tricornutum to low Fe, we observed major coordinated shifts in transcript control of primary and intermediate metabolism over light:dark cycles which contribute to a new view of the significance of distinctive diatom pathways, such as mitochondrial glycolysis and the ornithine-urea cycle. This study provides new insight into transcriptional modulation of diatom physiology and metabolism across light:dark cycles in response to Fe availability, providing mechanistic understanding for the ability of diatoms to remain metabolically poised to respond quickly to Fe input and revealing strategies underlying their ecological success.

We quantified the relationships among the specific growth rate, intracellular iron content, and steady state iron uptake rate for cultures of the marine diazotrophic cyanobacterium Trichodesmium (IMS 101) grown under differing conditions of Fe and N availability. The Fe quotas necessary to support a moderately Fe-limited growth rate (70% max) of 0.1 d Ϫ1 under diazotrophy and ammonium were 38 and 8 mol mol Ϫ1 , indicating a fivefold increased Fe cost for diazotrophy. This increased demand reflects the influences of both the ninefold lower marginal use efficiency and the greater maintenance Fe : C requirement at zero growth rate under diazotrophy (13.5 versus 5.2 mol mol Ϫ1 ). For diazotrophic growth at ϭ 0.1 d Ϫ1 , we estimate that 19%-53% of the cellular Fe is bound in nitrogenase. Trichodesmium is capable of luxury uptake of at least 13-fold greater amounts of Fe than needed for moderately Fe-limited growth. At least half of the populations sampled from the continental shelf of northern Australia had Fe : C ratios within the range of iron limitation of laboratory cultures. Bottle incubations showed that, after 3-4 d with added iron, a low Fe : C population (21 mol mol Ϫ1 ) had higher N 2 fixation rates than controls, whereas a high Fe : C population (78 mol mol Ϫ1) showed no response. The cellular N : P quotas and N 2 fixation rates were compared to those previously reported for the Atlantic Ocean. Empirically, these data suggest a critical N : P ratio ϳ40-50 for the onset of P limitation of growth. Only 10% of the Trichodesmium populations from coastal Australia had N : P ratios within the putative P-limitation range (Ͼ40), whereas 40% the Atlantic populations fell within this range. These comparisons, along with those for N fixation versus colony Fe : C ratio, suggest that Fe limitation is more prevalent in the Australia Trichodesmium populations, while P limitation predominates in the Atlantic populations.Trichodesmium spp. is a genus consisting of five species of nitrogen-fixing cyanobacteria that inhabit (sub)tropical oligotrophic waters. These plankton provide new N for up to 50% of primary production in these waters (Karl et al. 1997) and, thus, can play a prominent role in the biogeochemical cycling of C and N (Carpenter and Romans 1991). N 2 fixation facilitates net C fixation and export from the euphotic zone (Karl et al. 1995) and may have pronounced global climate implications (Falkowski 1997). Iron has been proposed to be a primary factor limiting N 2 fixation and growth of Trichodesmium due to the high iron requirement for diazotrophic growth and the low availability of iron in oceanic 1 Present address: Department of Geosciences, Princeton University, Princeton, New Jersey 08544.2 Present address: Romberg Tiburon Center, San Francisco State University, 3152 Paradise Drive, Tiburon, California 94920. AcknowledgmentsThe crew of the R/V Maurice Ewing, as well as Jim King and Rachel Foster, were instrumental in implementing our field sampling campaign. We thank Tracy Villareal for providing ...

Diatoms are one of the most successful and ecologically important groups of eukaryotic phytoplankton in the modern ocean. Deciphering their genomes is a key step towards better understanding of their biological innovations, evolutionary origins, and ecological underpinnings. Here, we have used 90 RNA-Seq datasets from different growth conditions combined with published expressed sequence tags and protein sequences from multiple taxa to explore the genome of the model diatom Phaeodactylum tricornutum, and introduce 1,489 novel genes. The new annotation additionally permitted the discovery of extensive alternative splicing in diatoms, including intron retention and exon skipping, which increase the diversity of transcripts generated in changing environments. In addition, we have used up-to-date reference sequence libraries to dissect the taxonomic origins of diatom genes. We show that the P. tricornutum genome is enriched in lineage-specific genes, with up to 47% of the gene models present only possessing orthologues in other stramenopile groups. Finally, we have performed a comprehensive de novo annotation of repetitive elements showing novel classes of transposable elements such as SINE, MITE and TRIM/LARD. This work provides a solid foundation for future studies of diatom gene function, evolution and ecology.

The centric diatom Thalassiosira pseudonana Hasle et Heimdal and the pennate diatom Phaeodactylum tricornutum Bohlin possess genes with translated sequences homologous to high‐affinity ferric reductases present in model organisms. Thalassiosira pseudonana also possesses putative genes for membrane‐bound ferroxidase (TpFET3) and two highly similar iron (Fe) permeases (TpFTR1 and TpFTR2), as well as a divalent metal (M2+) transporter belonging to the NRAMP superfamily (TpNRAMP). In baker’s yeast, the ferroxidase–permease complex transports Fe(II) produced by reductases. We investigated transcript abundances of these genes as a function of Fe quota (QFe). Ferric reductase transcripts are abundant in both species (15%–60% of actin) under low QFe and are down‐regulated by 5‐ to 35‐fold at high QFe, suggesting Fe(III) reduction is a common, inducible strategy for Fe acquisition in marine diatoms. Permease transcript abundance was regulated by Fe status in T. pseudonana, but we did not detect significant differences in expression of the copper (Cu)‐containing ferroxidase. TpNRAMP showed the most dramatic regulation by QFe, suggesting a role in cellular Fe transport in either cell‐surface uptake or vacuolar mobilization. We could not identify ferroxidase or permease homologues in the P. tricornutum genome. The up‐regulation of genes in T. pseudonana that appear to be missing altogether from P. tricornutum as well as the finding that P. tricornutum seems to have an efficient system to acquire Fe′, suggest that diverse (and uncharacterized) Fe‐uptake systems may be at play within diatom assemblages. Different uptake systems among diatoms may provide a mechanistic basis for niche differentiation with respect to Fe availability in the ocean.