Christel S. Hassler

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.

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.

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.

Is arsenic biotransformation a detoxification mechanism for microorganisms

Arsenic (As) is extremely toxic to living organisms at high concentration. In aquatic systems, As exists in different chemical forms. The two major inorganic As (iAs) species are As(V), which is thermodynamically stable in oxic waters, and As(III), which is predominant in anoxic conditions. Photosynthetic microorganisms (e.g., phytoplankton and cyanobacteria) take up As(V), biotransform it to As(III), then biomethylate it to methylarsenic (MetAs) forms. Although As(III) is more toxic than As(V), As(III) is much more easily excreted from the cells than As(V). Therefore, majority of researchers consider the reduction of As(V) to As(III) as a detoxification process. The biomethylation process results in the conversion of toxic iAs to the less toxic pentavalent MetAs forms (monomethylarsonate; MMA(V), dimethylarsonate; DMA(V), and trimethylarsenic oxide; TMAO(V)) and trimethylarsine (TMAO(III)). However, biomethylation by microorganisms also produces monomethylarsenite (MMA(III)) and dimethylarsenite (DMA(III)), which are more toxic than iAs, as a result of biomethylation by the microorganisms, demonstrates the need to reconsider to what extent As biomethylation contributes to a detoxification process. In this review, we focused on the discussion of whether the biotransformation of As species in microorganisms is really a detoxification process with recent data.

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.

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.

Toxicity of arsenic species to three freshwater organisms and biotransformation of inorganic arsenic by freshwater phytoplankton (Chlorella sp. CE-35)

In the environment, arsenic (As) exists in a number of chemical species, and arsenite (As(III)) and arsenate (As(V)) dominate in freshwater systems. Toxicity of As species to aquatic organisms is complicated by their interaction with chemicals in water such as phosphate that can influence the bioavailability and uptake of As(V). In the present study, the toxicities of As(III), As(V) and dimethylarsinic acid (DMA) to three freshwater organisms representing three phylogenetic groups: a phytoplankton (Chlorella sp. strain CE-35), a floating macrophyte (Lemna disperma) and a cladoceran grazer (Ceriodaphnia cf. dubia), were determined using acute and growth inhibition bioassays (EC₅₀) at a range of total phosphate (TP) concentrations in OECD medium. The EC₅₀ values of As(III), As(V) and DMA were 27 ± 10, 1.15 ± 0.04 and 19 ± 3 mg L(-1) for Chlorella sp. CE-35; 0.57 ± 0.16, 2.3 ± 0.2 and 56 ± 15 mg L(-1) for L. disperma, and 1.58 ± 0.05, 1.72 ± 0.01 and 5.9 ± 0.1 mg L(-1) for C. cf. dubia, respectively. The results showed that As(III) was more toxic than As(V) to L. disperma; however, As(V) was more toxic than As(III) to Chlorella sp. CE-35. The toxicities of As(III) and As(V) to C. cf. dubia were statistically similar (p>0.05). DMA was less toxic than iAs species to L. disperma and C. cf. dubia, but more toxic than As(III) to Chlorella sp. CE-35. The toxicity of As(V) to Chlorella sp. CE-35 and L. disperma decreased with increasing TP concentrations in the growth medium. Phosphate concentrations did not influence the toxicity of As(III) to either organism. Chlorella sp. CE-35 showed the ability to reduce As(V) to As(III), indicating a substantial influence of phytoplankton on As biogeochemistry in freshwater aquatic systems.

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.

Optimization of iron-dependent cyanobacterial (Synechococcus, Cyanophyceae) bioreporters to measure iron bioavailability

Complex chemistry and biological uptake pathways render iron bioavailability particularly difficult to assess in natural waters. Bioreporters are genetically modified organisms that are useful tools to directly sense the bioavailable fractions of solutes. In this study, three cyanobacterial bioreporters derived from Synechococcus PCC 7942 were examined for the purpose of optimizing the response to bioavailable Fe. Each bioreporter uses a Fe‐regulated promoter (isiAB, irpA and mapA), modulated by distinct mechanisms under Fe deficiency, fused to a bacterial luciferase (luxAB). In order to provide a better understanding of the way natural conditions may affect the ability of the bioreporter to sense iron bioavailability, the effect of relevant environmental parameters on the response to iron was assessed. Optimal conditions (and limits of applicability) for the use of these bioreporters on the field were determined to be: a 12 h (12–24 h) exposure time, temperature of 15°C (15°C–22°C), photon flux density of 100 μmol photons·m−2·s−1 (37–200 lmol photons·m−2·s−1), initial biomass of 0.6–0.8 lg chlorophyll a (chl a)·L−1 (0.3–1.5 lg chl a·L−1) or approximately 105 bioreporter cells·mL−1, high phosphate (10 lM), and low micronutrients (absent). The measured luminescence was optimal with an exogenous addition of 60 lM aqueous decanal substrate allowing a 5 min reaction time in the dark before analysis. This study provides important considerations relating to the optimization in the use of bioreporters under field conditions that can be used for method development of other algal and cyanobacterial bioreporters in aquatic systems.

Primary productivity induced by iron and nitrogen in the Tasman Sea: An overview of the PINTS expedition

The Tasman Sea and the adjacent subantarctic zone (SAZ) are economically important regions, where the parameters controlling the phytoplankton community composition and carbon fixation are not yet fully resolved. Contrasting nutrient distributions, as well as phytoplankton biomass, biodiversity and productivity were observed between the North Tasman Sea and the SAZ. In situ photosynthetic efficiency (FV/FM), dissolved and particulate nutrients, iron biological uptake, and nitrogen and carbon fixation were used to determine the factor-limiting phytoplankton growth and productivity in the North Tasman Sea and the SAZ. Highly productive cyanobacteria dominated the North Tasman Sea. High atmospheric nitrogen fixation and low nitrate dissolved concentrations indicated that non-diazotroph phytoplankton are nitrogen limited. Deck-board incubations also suggested that, at depth, iron could limit eukaryotes, but not cyanobacteria in that region. In the SAZ, the phytoplankton community was dominated by a bloom of haptophytes. The low productivity in the SAZ was mainly explained by light limitation, but nitrogen, silicic acid as well as iron were all depleted to the extent that they could become co-limiting. This study illustrates the challenge associated with identification of the limiting nutrient, as it varied between phytoplankton groups, depths and sites.