Chromium Cycling in Redox-Stratified Basins Challenges δ53Cr Paleoredox Proxy Applications.

Chromium stable isotope composition (δ53Cr) is a promising tracer for redox conditions throughout Earth's history; however, the geochemical controls of δ53Cr have not been assessed in modern redox-stratified basins. We present new chromium (Cr) concentration and δ53Cr data in dissolved, sinking particulate, and sediment samples from the redox-stratified Lake Cadagno (Switzerland), a modern Proterozoic ocean analog. These data demonstrate isotope fractionation during incomplete (non-quantitative) reduction and removal of Cr above the chemocline, driving isotopically light Cr accumulation in euxinic deep waters. Sediment authigenic Cr is isotopically distinct from overlying waters but comparable to average continental crust. New and published data from other redox-stratified basins show analogous patterns. This challenges assumptions from δ53Cr paleoredox applications that quantitative Cr reduction and removal limits isotope fractionation. Instead, fractionation from non-quantitative Cr removal leads to sedimentary records offset from overlying waters and not reflecting high δ53Cr from oxidative continental weathering.

Exopolymeric Substances Control Microbial Community Structure and Function by Contributing to both C and Fe Nutrition in Fe-Limited Southern Ocean Provinces.

Organic ligands such as exopolymeric substances (EPS) are known to form complexes with iron (Fe) and modulate phytoplankton growth. However, the effect of organic ligands on bacterial and viral communities remains largely unknown. Here, we assessed how Fe associated with organic ligands influences phytoplankton, microbial, and viral abundances and their diversity in the Southern Ocean. While the particulate organic carbon (POC) was modulated by Fe chemistry and bioavailability in the Drake Passage, the abundance and diversity of microbes and viruses were not governed by Fe bioavailability. Only following amendments with bacterial EPS did bacterial abundances increase, while phenotypic alpha diversity of bacterial and viral communities decreased. The latter was accompanied by significantly enhanced POC, pointing toward the relief of C limitation or other drivers of the microbial loop. Based on the literature and our findings, we propose a conceptual framework by which EPS may affect phytoplankton, bacteria, and viruses. Given the importance of the Southern Ocean for Earth's climate as well as the prevalence of viruses and their increasingly recognized impact on marine biogeochemistry and C cycling; the role of microbe-virus interactions on primary productivity in the Southern Ocean needs urgent attention.

Elemental Stoichiometry and Photophysiology Regulation of Synechococcus sp. PCC7002 Under Increasing Severity of Chronic Iron Limitation.

Iron (Fe) is an essential cofactor for many metabolic enzymes of photoautotrophs. Although Fe limits phytoplankton productivity in broad areas of the ocean, phytoplankton have adapted their metabolism and growth to survive in these conditions. Using the euryhaline cyanobacterium Synechococcus sp. PCC7002, we investigated the physiological responses to long-term acclimation to four levels of Fe availability representative of the contemporary ocean (36.7, 3.83, 0.47 and 0.047 pM Fe'). With increasing severity of Fe limitation, Synechococcus sp. cells gradually decreased their volume and growth while increasing their energy allocation into organic carbon and nitrogen cellular pools. Furthermore, the total cellular content of pigments decreased. Additionally, with increasing severity of Fe limitation, intertwined responses of PSII functional cross-section (σPSII), re-oxidation time of the plastoquinone primary acceptor QA (τ) and non-photochemical quenching revealed a shift in the photophysiological response between mild to strong Fe limitation compared with severe limitation. Under mild and strong Fe limitation, there was a decrease in linear electron transport accompanied by progressive loss of state transitions. Under severe Fe limitation, state transitions seemed to be largely supplanted by alternative electron pathways. In addition, mechanisms to dissipate energy excess and minimize oxidative stress associated with high irradiances increased with increasing severity of Fe limitation. Overall, our results establish the sequence of physiological strategies adopted by the cells under increasing severity of chronic Fe limitation, within a range of Fe concentrations relevant to modern ocean biogeochemistry.

Nutrient uplift in a cyclonic eddy increases diversity, primary productivity and iron demand of microbial communities relative to a western boundary current.

The intensification of western boundary currents in the global ocean will potentially influence meso-scale eddy generation, and redistribute microbes and their associated ecological and biogeochemical functions. To understand eddy-induced changes in microbial community composition as well as how they control growth, we targeted the East Australian Current (EAC) region to sample microbes in a cyclonic (cold-core) eddy (CCE) and the adjacent EAC. Phototrophic and diazotrophic microbes were more diverse (2-10 times greater Shannon index) in the CCE relative to the EAC, and the cell size distribution in the CCE was dominated (67%) by larger micro-plankton [Formula: see text], as opposed to pico- and nano-sized cells in the EAC. Nutrient addition experiments determined that nitrogen was the principal nutrient limiting growth in the EAC, while iron was a secondary limiting nutrient in the CCE. Among the diazotrophic community, heterotrophic NifH gene sequences dominated in the EAC and were attributable to members of the gamma-, beta-, and delta-proteobacteria, while the CCE contained both phototrophic and heterotrophic diazotrophs, including Trichodesmium, UCYN-A and gamma-proteobacteria. Daily sampling of incubation bottles following nutrient amendment captured a cascade of effects at the cellular, population and community level, indicating taxon-specific differences in the speed of response of microbes to nutrient supply. Nitrogen addition to the CCE community increased picoeukaryote chlorophyll a quotas within 24 h, suggesting that nutrient uplift by eddies causes a 'greening' effect as well as an increase in phytoplankton biomass. After three days in both the EAC and CCE, diatoms increased in abundance with macronutrient (N, P, Si) and iron amendment, whereas haptophytes and phototrophic dinoflagellates declined. Our results indicate that cyclonic eddies increase delivery of nitrogen to the upper ocean to potentially mitigate the negative consequences of increased stratification due to ocean warming, but also increase the biological demand for iron that is necessary to sustain the growth of large-celled phototrophs and potentially support the diversity of diazotrophs over longer time-scales.

Southern Ocean phytoplankton physiology in a changing climate.

The Southern Ocean (SO) is a major sink for anthropogenic atmospheric carbon dioxide (CO2), potentially harbouring even greater potential for additional sequestration of CO2 through enhanced phytoplankton productivity. In the SO, primary productivity is primarily driven by bottom up processes (physical and chemical conditions) which are spatially and temporally heterogeneous. Due to a paucity of trace metals (such as iron) and high variability in light, much of the SO is characterised by an ecological paradox of high macronutrient concentrations yet uncharacteristically low chlorophyll concentrations. It is expected that with increased anthropogenic CO2 emissions and the coincident warming, the major physical and chemical process that govern the SO will alter, influencing the biological capacity and functioning of the ecosystem. This review focuses on the SO primary producers and the bottom up processes that underpin their health and productivity. It looks at the major physico-chemical drivers of change in the SO, and based on current physiological knowledge, explores how these changes will likely manifest in phytoplankton, specifically, what are the physiological changes and floristic shifts that are likely to ensue and how this may translate into changes in the carbon sink capacity, net primary productivity and functionality of the SO.

The impact of iron limitation on the physiology of the Antarctic diatom Chaetoceros simplex.

Iron availability strongly governs the growth of Southern Ocean phytoplankton. To investigate how iron limitation affects photosynthesis as well as the uptake of carbon and iron in the Antarctic diatom Chaetocerossimplex, a combination of chlorophyll a fluorescence measurements and radiotracer incubations in the presence and absence of chemical inhibitors was conducted. Iron limitation in C. simplex led to a decline in growth rates, photochemical efficiency and structural changes in photosystem II (PSII), including a reorganisation of photosynthetic units in PSII and an increase in size of the functional absorption cross section of PSII. Iron-limited cells further exhibited a reduced plastoquinone pool and decreased photosynthetic electron transport rate, while non-photochemical quenching and relative xanthophyll pigment content were strongly increased, suggesting a photoprotective response. Additionally, iron limitation resulted in a strong decline in carbon fixation and thus the particulate organic carbon quotas. Inhibitor studies demonstrated that, independent of the iron supply, carbon fixation was dependent on internal, but not on extracellular carbonic anhydrase activity. Orthovanadate more strongly inhibited iron uptake in iron-limited cells, indicating that P-type ATPase transporters are involved in iron uptake. The stronger reduction in iron uptake by ascorbate in iron-limited cells suggests that the re-oxidation of iron is required before it can be taken up and further supports the presence of a high-affinity iron transport pathway. The measured changes to photosystem architecture and shifts in carbon and iron uptake strategies in C. simplex as a result of iron limitation provide evidence for a complex interaction of these processes to balance the iron requirements for photosynthesis and carbon demand for sustained growth in iron-limited waters.

Iron Biogeochemistry in Aquatic Systems: From Source to Bioavailability.

Iron (Fe) is an essential trace element for several key metabolic processes in phytoplankton; however Fe is present in low concentration in many aquatic systems including vast oceanic regions and large lakes. In these systems, Fe can limit the growth of phytoplankton and atmospheric carbon dioxide biological fixation. Indeed Fe limitation exerts a global impact on the carbon cycle and the imprint of aquatic systems on our climate. In order to understand how aquatic systems function and increase our ability to predict their response to changing conditions, it is therefore paramount to understand when and how Fe controls operate. This review presents the complex relationship between Fe chemistry and the biology of surface waters to highlight the parameters defining the forms of Fe that are accessible for phytoplankton growth (or bioavailable). Particular attention is given to the identification of Fe sources and Fe organic complexation as these, in conjunction with biological recycling and remineralisation, mostly control Fe residence time, chemistry and bioavailability.

Iron limitation modulates ocean acidification effects on southern ocean phytoplankton communities.

The potential interactive effects of iron (Fe) limitation and Ocean Acidification in the Southern Ocean (SO) are largely unknown. Here we present results of a long-term incubation experiment investigating the combined effects of CO2 and Fe availability on natural phytoplankton assemblages from the Weddell Sea, Antarctica. Active Chl a fluorescence measurements revealed that we successfully cultured phytoplankton under both Fe-depleted and Fe-enriched conditions. Fe treatments had significant effects on photosynthetic efficiency (Fv/Fm; 0.3 for Fe-depleted and 0.5 for Fe-enriched conditions), non-photochemical quenching (NPQ), and relative electron transport rates (rETR). pCO2 treatments significantly affected NPQ and rETR, but had no effect on Fv/Fm. Under Fe limitation, increased pCO2 had no influence on C fixation whereas under Fe enrichment, primary production increased with increasing pCO2 levels. These CO2-dependent changes in productivity under Fe-enriched conditions were accompanied by a pronounced taxonomic shift from weakly to heavily silicified diatoms (i.e. from Pseudo-nitzschia sp. to Fragilariopsis sp.). Under Fe-depleted conditions, this functional shift was absent and thinly silicified species dominated all pCO2 treatments (Pseudo-nitzschia sp. and Synedropsis sp. for low and high pCO2, respectively). Our results suggest that Ocean Acidification could increase primary productivity and the abundance of heavily silicified, fast sinking diatoms in Fe-enriched areas, both potentially leading to a stimulation of the biological pump. Over much of the SO, however, Fe limitation could restrict this possible CO2 fertilization effect.

Exploring the Link between Micronutrients and Phytoplankton in the Southern Ocean during the 2007 Austral Summer.

Bottle assays and large-scale fertilization experiments have demonstrated that, in the Southern Ocean, iron often controls the biomass and the biodiversity of primary producers. To grow, phytoplankton need numerous other trace metals (micronutrients) required for the activity of key enzymes and other intracellular functions. However, little is known of the potential these other trace elements have to limit the growth of phytoplankton in the Southern Ocean. This study, investigates whether micronutrients other than iron (Zn, Co, Cu, Cd, Ni) need to be considered as parameters for controlling the phytoplankton growth from the Australian Subantarctic to the Polar Frontal Zones during the austral summer 2007. Analysis of nutrient disappearance ratios, suggested differential zones in phytoplankton growth control in the study region with a most intense phytoplankton growth limitation between 49 and 50°S. Comparison of micronutrient disappearance ratios, metal distribution, and biomarker pigments used to identify dominating phytoplankton groups, demonstrated that a complex interaction between Fe, Zn, and Co might exist in the study region. Although iron remains the pivotal micronutrient for phytoplankton growth and community structure, Zn and Co are also important for the nutrition and the growth of most of the dominating phytoplankton groups in the Subantarctic Zone region. Understanding of the parameters controlling phytoplankton is paramount, as it affects the functioning of the Southern Ocean, its marine resources and ultimately the global carbon cycle.

PHYTOPLANKTON SELENIUM REQUIREMENTS: THE CASE FOR SPECIES ISOLATED FROM TEMPERATE AND POLAR REGIONS OF THE SOUTHERN HEMISPHERE(1).

A series of laboratory culture experiments was used to investigate the effect of selenium (Se, 0-10 nM) on the growth, cellular volume, photophysiology, and pigments of two temperate and four polar oceanic phytoplankton species [coccolithophore Emiliania huxleyi (Lohmann) W. W. Hay et H. P. Mohler, cyanobacterium Synechococcus sp., prymnesiophyte Phaeocystis sp., and three diatoms-Fragilariopsis cylindrus (Grunow) Kriegar, Chaetoceros sp., and Thalassiosira antarctica G. Karst.]. Only Synechoccocus sp. and Phaeocystis sp. did not show any requirement for Se. Under Se-deficient conditions, the growth rate of E. huxleyi was decreased by 1.6-fold, whereas cellular volume was increased by 1.9-fold. Se limitation also decreased chl a (2.5-fold), maximum relative electron transport rate (1.9-fold), and saturating light intensity (2.8-fold), suggesting that Se plays a role in photosynthesis or high-light acclimation. Pigment analysis for Antarctic taxa provided an interesting counterpoint to the physiology of E. huxleyi. For all Se-dependent Antarctic diatoms, Se limitation decreased growth rate and chl a content, whereas cellular volume was not affected. Pigment analysis revealed that other pigments were affected under Se deficiency. Photoprotective pigments increased by 1.4-fold, while diadinoxanthin:diatoxanthin ratios decreased by 1.5- to 4.9-fold under Se limitation, supporting a role for Se in photoprotection. Our results demonstrate an Se growth requirement for polar diatoms and indicate that Se could play a role in the biogeochemical cycles of other nutrients, such as silicic acid in the Southern Ocean. Se measurements made during the austral summer in the Southern Ocean and Se biological requirement were used to discuss possible Se limitation in phytoplankton from contrasting oceanographic regions.

Saccharides enhance iron bioavailability to Southern Ocean phytoplankton.

Iron limits primary productivity in vast regions of the ocean. Given that marine phytoplankton contribute up to 40% of global biological carbon fixation, it is important to understand what parameters control the availability of iron (iron bioavailability) to these organisms. Most studies on iron bioavailability have focused on the role of siderophores; however, eukaryotic phytoplankton do not produce or release siderophores. Here, we report on the pivotal role of saccharides--which may act like an organic ligand--in enhancing iron bioavailability to a Southern Ocean cultured diatom, a prymnesiophyte, as well as to natural populations of eukaryotic phytoplankton. Addition of a monosaccharide (>2 nM of glucuronic acid, GLU) to natural planktonic assemblages from both the polar front and subantarctic zones resulted in an increase in iron bioavailability for eukaryotic phytoplankton, relative to bacterioplankton. The enhanced iron bioavailability observed for several groups of eukaryotic phytoplankton (i.e., cultured and natural populations) using three saccharides, suggests it is a common phenomenon. Increased iron bioavailability resulted from the combination of saccharides forming highly bioavailable organic associations with iron and increasing iron solubility, mainly as colloidal iron. As saccharides are ubiquitous, present at nanomolar to micromolar concentrations, and produced by biota in surface waters, they also satisfy the prerequisites to be important constituents of the poorly defined "ligand soup," known to weakly bind iron. Our findings point to an additional type of organic ligand, controlling iron bioavailability to eukaryotic phytoplankton--a key unknown in iron biogeochemistry.

A novel in situ tool for the exposure and analysis of microorganisms in natural aquatic systems.

To evaluate the effects of contaminants or nutrient limitation in natural waters, it is often desirable to perform controlled exposures of organisms. While in situ exposures are routine for caged organisms or macrophytes, they are extremely difficult to perform for microorganisms, mainly due to difficulties in designing an exposure device that isolates the cells while allowing rapid equilibration with the external media. In this paper, a stirred underwater biouptake system (SUBS) based on the diffusion of chemicals across a semipermeable membrane housing a controlled population of microorganisms is reported. Cd diffusion through the semipermeable membrane was evaluated by voltammetry using a microelectrode. Comparison of stirred and unstirred solutions demonstrated a significantly increased diffusive flux in the presence of stirring. Lab tests using Chlamydomonas reinhardtii showed that diffusion across the semipermeable membrane was not limiting with respect to the biouptake of Cd. The SUBS device was field tested and the results of viability studies and trace metal biouptake by C. reinhardtii are reported. No diffusion limitation due to the SUBS was observed for Cd under the tested field conditions. The SUBS device was also shown to be useful for field exposures and subsequent measurements of trace metal uptake and viability. The results support the future use of the SUBS for the in situ measurement of phytochelatin/metallothionein production, photosynthetic efficiency, or reporter gene induction of controlled organisms.

Application of the biotic ligand model to explain potassium interaction with thallium uptake and toxicity to plankton.

Competitive interaction between TI(I) and K was successfully predicted by the biotic ligand model (BLM) for the microalga Chlorella sp. (Chlorophyta; University of Toronto Culture Collection strain 522) during 96-h toxicity tests. Because of a greater affinity of T1(I) (log K = 7.3-7.4) as compared to K (log K = 5.3-6.3) for biologically sensitive sites, an excess of 40- to 160-fold of K is required to suppress T1(I) toxic effects on Chlorella sp., regardless of [T1(I)] in solution. Similar excess of K is required to suppress T1(I) toxicity to Synechococcus leopoliensis (Cyanobacteria; University of Texas Culture Collection strain 625) and Brachionus calyciflorus (Rotifera; strain AB-RIF). The mechanism for the mitigating effect of K on T1(I) toxicity was investigated by measuring 204T1(I) cellular uptake flux and efflux in Chlorella sp. Potassium shows a competitive effect on T1(I) uptake fluxes that could be modeled using the BLM-derived stability constants and a Michaelis-Menten relationship. A strong T1 efflux dependent only on the cellular T1 concentration was measured. Although T1 efflux does not explain the effect of K on T1(I) toxicity and uptake, it is responsible for a high turnover of the cellular T1 pool (intracellular half-life = 12-13.5 min). No effect of Na+, Mg2+, or Ca2+ was observed on T1+ uptake, whereas the absence of trace metals (Cu, Co, Mo, Mn, Fe, and Zn) significantly increased T1 uptake and decreased the mitigating effect of K+. The importance of K+ in determining the aquatic toxicity of T1+ underscores the use of ambient K+ concentration in the establishment of T1 water-quality guidelines and the need to consider K in predicting biogeochemical fates of T1 in the aquatic environment.

Bioavailability of iron sensed by a phytoplanktonic Fe-bioreporter.

This study describes a short-term (12 h) evaluation of iron (Fe) bioavailability to an Fe-dependent cyanobacterial bioreporter derived from Synechococcus PCC 7942. Several synthetic ligands with variable conditional stability constants for Fe(lll) (K* of 10(19.8) to 10(30.9)), in addition to several defined natural Fe-binding ligands and a fulvic acid of aquatic origin (Suwannee River), were used to elucidate the forms of Fe that are discerned by this phytoplanktonic microbe: Fe-HEBD (log conditional stability constant, K*, = 28.1, HEBD = N,N'-di(2-hydroxybenzyl)ethylenediamine-N,N'-diacetic acid monohydrochloride hydrate), Fe-HDFB (K* = 30.9, DFB = desferroxamine B), Fe-ferrichrome (K* = 23.2), Fe-DTPA (K* = 21.1, DTPA = diethylenetrinitrilopentaacetic acid), Fe-(8HQS)2 (K* = 20.4, 8HQS = 8-hydroxyquinoline-5-sulfonic acid), Fe-CDTA (K* = 19.8, CDTA = trans-1,2-cyclohexylenedinitrilotetraacetic acid), and Fe-EDTA (K* = 19.2). Iron bioavailability sensed by the bioreporter was related to diffusion limitation and activity of high-affinity transporters rather than by siderophore secretion. Iron complexed with a K* < 23.2 contributes to the bioavailable pool; bioavailability could be explained by disjunctive ligand exchange considerations and fully, partially, and nonbioavailable complexes could be distinguished according to their conditional stability constant. The use of Fe-bioreporters provides a relevant measurement of bioavailability to an important group of primary producers in freshwaters (cyanobacteria) and is thus a promising technique for understanding Fe cycling in aquatic systems.

Impact of zinc acclimation on bioaccumulation and homeostasis in Chlorella kesslerii.

Growth curves, cellular Zn contents and cellular protein expression were examined for the green alga, Chlorella kesslerii, as a function of different Zn growth regimes (growth in 16 pM, 1.7 nM or 1.6 microM calculated Zn2+). Zn homeostasis was responsible for observed differences in the capacity of the organism to accumulate Zn. The rapid acclimation that occurred in response to a Zn deficiency was likely due to the production of Zn transport sites. No differences were observed among cellular phytochelatin contents or efflux rate constants, although efflux did play an important role in regulating Zn cellular content. A long-term adaptation to Zn was not thought to occur since bioaccumulation and biological responses were similar for four successive cultures (30-40 days, 16-19 cell cycles) at different [Zn2+]. Among proteins that were influenced by the Zn growth regime, the Rubisco and histone H3 proteins were identified as being induced in the presence of 1.6 microM Zn2+ as compared with 1.7 nM Zn2+. The impact of the Zn preconditioning demonstrated that the concentrations of essential metals in the algal growth media would have an important, if not predominant effect on toxicity or bioaccumulation assessments. Furthermore, the high regulation of Zn transport and intracellular events by the microorganisms will likely preclude the use of simple metal uptake models including the free ion activity model and the biotic ligand model to predict either bioaccumulation or toxicological effects of Zn and perhaps other essential metals.

Some fundamental (and often overlooked) considerations underlying the free ion activity and biotic ligand models.

Trace metal bioavailability is often evaluated on the basis of steady-state models such as the free ion activity model (FIAM) and the biotic ligand model (BLM). Some of the assumptions underlying these models were verified by examining Pb and Zn uptake by the green microalga Chlorella kesslerii. Transporter bound metal ([M-Rcell]) and free ion concentrations ([M(Z+)]) were related to experimentally determined uptake fluxes (Jint). Although the BLM and FIAM correctly predicted Pb uptake in the absence of competing ions, they failed to predict competitive interactions with Ca2+, likely because of modifications of the algal surface charge and the active nature of Ca2+ transport. Zinc transport is also active; in this case, both the internalization rate constant (kint) and the equilibrium constant for the binding of Zn to the transport sites (K(Zn-Rcell)) varied as a function of [Zn2+] in the bulk solution. For this reason, Zn uptake could not be modeled by the steady-state models either in the presence or absence of competitors (Cd and Ca). Furthermore, the role of Cu on Pb and Zn adsorption and uptake could not be predicted by either model because of secondary effects on the algal physiology and membrane permeability. Finally, a 17 degree C reduction in temperature resulted in a two- to fivefold decrease in membrane permeability of the metals, an observation that also is unaccounted for by either the FIAM or BLM. This paper emphasizes the limitations of the models in well-controlled laboratory systems with the goal of extrapolating the results to complex environmental systems.

Failure of the biotic ligand and free-ion activity models to explain zinc bioaccumulation by Chlorella kesslerii.

Zinc accumulation by Chlorella kesslerii (Chlorophycee) was studied for [Zn2+] ranging from 4 pM to 1 mM. A first-order uptake flux as predicted by the free ion activity model (FIAM) and the biotic ligand model (BLM) was not observed. Furthermore, when algae were preconditioned in slightly limiting (10 pM) versus optimal (1 nM) free zinc concentrations, the internalization flux increased and was nearly constant over the range of [Zn2+] examined. This observation was attributed to the synthesis of membrane-bound zinc transporters. Neither solution Zn chemistry nor surface-bound Zn was a good predictor of Zn uptake fluxes. Several hypotheses were examined to explain the failure of the steady-state uptake models. Although zinc excretion and a Zn diffusion limitation were observed, neither result could explain the majority of observed deviation from the models. Bioaccumulation experiments performed with inhibitors (vanadate and carbonyl cyanide m-chlorophenylhydrazone [CCCP]) demonstrated clearly that zinc transport is an energy-dependent process in Chlorella kesslerii. The presence of an active transport signifies that Zn uptake may function independently of the electrochemical Zn gradient and that, in some cases, both uptake fluxes and receptor-bound Zn may be independent of solution chemistry.