Maria T. Maldonado

A mesoscale phytoplankton bloom in the polar Southern Ocean stimulated by iron fertilization

Changes in iron supply to oceanic plankton are thought to have a significant effect on concentrations of atmospheric carbon dioxide by altering rates of carbon sequestration, a theory known as the ‘iron hypothesis’. For this reason, it is important to understand the response of pelagic biota to increased iron supply. Here we report the results of a mesoscale iron fertilization experiment in the polar Southern Ocean, where the potential to sequester iron-elevated algal carbon is probably greatest. Increased iron supply led to elevated phytoplankton biomass and rates of photosynthesis in surface waters, causing a large drawdown of carbon dioxide and macronutrients, and elevated dimethyl sulphide levels after 13 days. This drawdown was mostly due to the proliferation of diatom stocks. But downward export of biogenic carbon was not increased. Moreover, satellite observations of this massive bloom 30 days later, suggest that a sufficient proportion of the added iron was retained in surface waters. Our findings demonstrate that iron supply controls phytoplankton growth and community composition during summer in these polar Southern Ocean waters, but the fate of algal carbon remains unknown and depends on the interplay between the processes controlling export, remineralisation and timescales of water mass subduction.

Importance of stirring in the development of an iron-fertilized phytoplankton bloom

The growth of populations is known to be influenced by dispersal, which has often been described as purely diffusive. In the open ocean, however, the tendrils and filaments of phytoplankton populations provide evidence for dispersal by stirring. Despite the apparent importance of horizontal stirring for plankton ecology, this process remains poorly characterized. Here we investigate the development of a discrete phytoplankton bloom, which was initiated by the iron fertilization of a patch of water (7 km in diameter) in the Southern Ocean. Satellite images show a striking, 150-km-long bloom near the experimental site, six weeks after the initial fertilization. We argue that the ribbon-like bloom was produced from the fertilized patch through stirring, growth and diffusion, and we derive an estimate of the stirring rate. In this case, stirring acts as an important control on bloom development, mixing phytoplankton and iron out of the patch, but also entraining silicate. This may have prevented the onset of silicate limitation, and so allowed the bloom to continue for as long as there was sufficient iron. Stirring in the ocean is likely to be variable, so blooms that are initially similar may develop very differently.

REDUCTION AND TRANSPORT OF ORGANICALLY BOUND IRON BY THALASSIOSIRA OCEANICA (BACILLARIOPHYCEAE)

Thalassiosira oceanica (Hustedt) Hasle et Heimdal (clone 1003) attained rapid rates of growth in low Fe seawater containing the siderophore ferrioxamine B (FeDFB) as the sole Fe source. Short‐term rates of Fe uptake were 109 times faster than those predicted from the equilibrium concentration of inorganic Fe, suggesting that FeDFB was the substrate for the Fe transport system. An extracellular reduction step, mediated by a cell surface reductase, preceded Fe transport from FeDFB and was induced under Fe limitation. The half‐saturation constant for the reduction was 0.68 μM. Iron reduction rates were two times faster than uptake rates, so that the activities of the reductase and the transporter were tightly coupled. The rates of Fe reduction of a number of Fe chelators, including synthetic organic ligands (nitrilotriacetate, diethylenetriaminepentaacetate, and EDTA) and fungal siderophores (desferrioxamine B and desferrioxamine E), were inversely proportional to the ratio of the stability constants of their Fe(III) and Fe(II) complexes and varied by a factor of two times, like the redox potentials of the Fe complexes. Platinum (II), a known inhibitor of Fe reductase activity, appeared to reduce the rates of Fe uptake from FeDFB but not from inorganic complexes. The results suggested that reoxidation of Fe(II) produced by reduction may be a necessary part of the Fe internalization reaction. Ferric reductase could be relevant to phytoplankton nutrition in the open sea where organic Fe complexes dominate the dissolved speciation and where the concentration of inorganic Fe is limiting.

Ferritin is used for iron storage in bloom-forming marine pennate diatoms

Primary productivity in 30–40% of the world’s oceans is limited by availability of the micronutrient iron. Regions with chronically low iron concentrations are sporadically pulsed with new iron inputs by way of dust or lateral advection from continental margins. Addition of iron to surface waters in these areas induces massive phytoplankton blooms dominated primarily by pennate diatoms. Here we provide evidence that the bloom-forming pennate diatoms Pseudo-nitzschia and Fragilariopsis use the iron-concentrating protein, ferritin, to safely store iron. Ferritin has not been reported previously in any member of the Stramenopiles, a diverse eukaryotic lineage that includes unicellular algae, macroalgae and plant parasites. Phylogenetic analyses suggest that ferritin may have arisen in this small subset of diatoms through a lateral gene transfer. The crystal structure and functional assays of recombinant ferritin derived from Pseudo-nitzschia multiseries reveal a maxi-ferritin that exhibits ferroxidase activity and binds iron. The protein is predicted to be targeted to the chloroplast to control the distribution and storage of iron for proper functioning of the photosynthetic machinery. Abundance of Pseudo-nitzschia ferritin transcripts is regulated by iron nutritional status, and is closely tied to the loss and recovery of photosynthetic competence. Enhanced iron storage with ferritin allows the oceanic diatom Pseudo-nitzschia granii to undergo several more cell divisions in the absence of iron than the comparably sized, oceanic centric diatom Thalassiosira oceanica. Ferritin in pennate diatoms probably contributes to their success in chronically low-iron regions that receive intermittent iron inputs, and provides an explanation for the importance of these organisms in regulating oceanic CO2 over geological timescales.

The fate of added iron during a mesoscale fertilisation experiment in the Southern Ocean

The first Southern Ocean Iron RElease Experiment (SOIREE) was performed during February 1999 in Antarctic waters south of Australia (61°S, 140°E), in order to verify whether iron supply controls the magnitude of phytoplankton production in this high nutrient low chlorophyll (HNLC) region. This paper describes iron distributions in the upper ocean during our 13-day site occupation, and presents a pelagic iron budget to account for the observed losses of dissolved and total iron from waters of the fertilised patch. Iron concentrations were measured underway during daily transects through the patch and in vertical profiles of the 65-m mixed layer. High internal consistency was noted between data obtained using contrasting sampling and analytical techniques. A pre-infusion survey confirmed the extremely low ambient dissolved (0.1 nM) and total (0.4 nM) iron concentrations. The initial enrichment elevated the dissolved iron concentration to 2.7 nM. Thereafter, dissolved iron was rapidly depleted inside the patch to 0.2-0.3 nM, necessitating three re-infusions. A distinct biological response was observed in iron-fertilised waters, relative to outside the patch, unequivocally confirming that iron limits phytoplankton growth rates and biomass at this site in summer. Our budget describing the fate of the added iron demonstrates that horizontal dispersion of fertilised waters (resulting in a quadrupling of the areal extent of the patch) and abiotic particle scavenging accounted for most of the decreases in iron concentrations inside the patch (31-58 and 12-49 of added iron, respectively). The magnitude of these loss processes altered towards the end of SOIREE, and on days 12-13 dissolved (1.1 nM) and total (2.3 nM) iron concentrations remained elevated compared to surrounding waters. At this time, the biogenic iron pool (0.1 nM) accounted for only 1-2 of the total added iron. Large pennate diatoms (> 20 μm) and autotrophic flagellates (2-20 μm) were the dominant algal groups in the patch, taking up the added iron and representing 13 and 39 of the biogenic iron pool, respectively. Iron regeneration by grazers was tightly coupled to uptake by phytoplankton and bacteria, indicating that biological Fe cycling within the bloom was self-sustaining. A concurrent increase in the concentration of iron-binding ligands on days 11-12 probably retained dissolved iron within the mixed layer. Ocean colour satellite images in late March suggest that the bloom was still actively growing 42 days after the onset of SOIREE, and hence by inference that sufficient iron was maintained in the patch for this period to meet algal requirements. This raises fundamental questions regarding the biogeochemical cycling of iron in the Southern Ocean and, in particular, how bioavailable iron was retained in surface waters and/or within the biota to sustain algal growth.

Retention of dissolved iron and Fe II in an iron induced Southern Ocean phytoplankton bloom

During the 13 day Southern Ocean Iron RE-lease Experiment (SOIREE), dissolved iron concentrations decreased rapidly following each of three iron-enrichments, but remained high (>1 nM, up to 80% as FeII) after the fourth and final enrichment on day 8. The former trend was mainly due to dilution (spreading of iron-fertilized waters) and particle scavenging. The latter may only be explained by a joint production-maintenance mechanism; photoreduction is the only candidate process able to produce sufficiently high FeII, but as such levels persisted overnight (8 hr dark period) —ten times the half—life for this species—a maintenance mechanism (complexation of FeII) is required, and is supported by evidence of increased ligand concentrations on day 12. The source of these ligands and their affinity for FeII is not known. This retention of iron probably permitted the longevity of this bloom raising fundamental questions about iron cycling in HNLC (High Nitrate Low Chlorophyll) Polar waters.

Co-limitation of phytoplankton growth by light and Fe during winter in the NE subarctic Pacific Ocean

Phytoplankton acclimate to low irradiance by increasing their cellular demand for Fe, to allow synthesis of additional light-harvesting pigments and Fe-containing redox proteins involved in photosynthesis. In the open NE subarctic Pacific, Fe concentrations limit primary productivity and irradiances may be suboptimal, particularly during winter. Phytoplankton thus may be unable to fulfill their extra Fe requirements for growth under these low-light conditions and become effectively co-limited. We tested this hypothesis by manipulating Fe and light in in vitro experiments at OSP (Ocean Station PAPA, 50°N 145°W) during winter 1997. The results show that metabolic rates, growth, and photosynthetic parameters of phytoplankton are enhanced in winter by increasing either irradiance or Fe. The greatest response occurs when Fe and light are amended concomitantly, confirming that the community is indeed co-limited by both resources. Analysis of environmental conditions (i.e. incident irradiance, mixed layer depth and Fe concentrations) in winter at OSP reveals that they are similar to those observed in the austral spring and fall at three sites in the Southern Ocean. Extrapolating our experimental field results to the Southern Ocean illustrates that co-limitation by light and Fe also may play an important role in regulating phytoplankton growth in this region.

Growth, nutrient uptake capacities and tissue constituents of the macroalgae Cladophora vagabunda and Gracilaria tikvahiae related to site-specific nitrogen loading rates

Cladophora vagabunda (L.) van den Hoek and Gracilaria tikvahiae (McLachlan) have become dominant components of the macroalgal assemblage in Waquoit Bay, a Massachusetts embayment, possibly due to nitrogen (N) enrichment from anthropogenic inputs transported via groundwater. During 1989–1993, we measured site-related growth, ammonium uptake rates and tissue constituents of these macroalgae from areas subject to high N loading rates (Childs River) and lower N loadings rates (Sage Lot Pond). We also conducted in situ and microcosm enrichment experiments to determine what limited algal growth throughout the year. Our results indicated that these species are strongly affected by and have a strong impact on the N environment of this embayment. For example, C. vagabunda and G. tikvahiae from Childs River had higher light-harvesting pigments and tissue-N concentrations than Sage Lot Pond populations. Additionally, both Childs River populations showed greater site-specific growth and N uptake rates, particularly during the summer period of peak growth. In fact, maximum uptake rates of 90 and over 140 μmol dry wt g-1 h-1 for Childs River C. vagabunda and G. tikvahiae, respectively, suggest that these species can remove substanital quantities of N from overlying waters, and may be responsible for low (often (<1 μM) water-column nutrient concentrations during summer. In situ and tank enrichment experiments indicated that growth rates were limited by available N during summer, while P may be limiting during a brief period toward the end of the annual growth cycle (autumn). Under experimental enrichment, growth rates of Sage Lot Pond algae were similar to values measured at the site receiving higher N inputs, and generally, G. tikvahiae showed growth enhancement (up to 0.2 doublings d-1) under light-saturating conditions (0.5 m) while C. vagabunda showed nutrient-enhanced growth at 2.5 m. The effects of available nutrients on algal growth were strongly influenced by irradiance and temperature, resulting in a complex seasonal interaction that emphasized the dynamic nature of species response to N loading. Dominance by these two macroalgae in Waquoit Bay, as in other areas undergoing eutrophication, is likely related to physiological strategies that enable these species to tolerate large environmental variations, to take advantage of greater N availability and to survive indirect effects of N loading (e.g. reduced irradiance, anoxia).

Diatom proteomics reveals unique acclimation strategies to mitigate Fe limitation.

Phytoplankton growth rates are limited by the supply of iron (Fe) in approximately one third of the open ocean, with major implications for carbon dioxide sequestration and carbon (C) biogeochemistry. To date, understanding how alteration of Fe supply changes phytoplankton physiology has focused on traditional metrics such as growth rate, elemental composition, and biophysical measurements such as photosynthetic competence (Fv/Fm). Researchers have subsequently employed transcriptomics to probe relationships between changes in Fe supply and phytoplankton physiology. Recently, studies have investigated longer-term (i.e. following acclimation) responses of phytoplankton to various Fe conditions. In the present study, the coastal diatom, Thalassiosira pseudonana, was acclimated (10 generations) to either low or high Fe conditions, i.e. Fe-limiting and Fe-replete. Quantitative proteomics and a newly developed proteomic profiling technique that identifies low abundance proteins were employed to examine the full complement of expressed proteins and consequently the metabolic pathways utilized by the diatom under the two Fe conditions. A total of 1850 proteins were confidently identified, nearly tripling previous identifications made from differential expression in diatoms. Given sufficient time to acclimate to Fe limitation, T. pseudonana up-regulates proteins involved in pathways associated with intracellular protein recycling, thereby decreasing dependence on extracellular nitrogen (N), C and Fe. The relative increase in the abundance of photorespiration and pentose phosphate pathway proteins reveal novel metabolic shifts, which create substrates that could support other well-established physiological responses, such as heavily silicified frustules observed for Fe-limited diatoms. Here, we discovered that proteins and hence pathways observed to be down-regulated in short-term Fe starvation studies are constitutively expressed when T. pseudonana is acclimated (i.e., nitrate and nitrite transporters, Photosystem II and Photosystem I complexes). Acclimation of the diatom to the desired Fe conditions and the comprehensive proteomic approach provides a more robust interpretation of this dynamic proteome than previous studies.

Role of dissolved and particulate cadmium in the accumulation of cadmium in cultured oysters (Crassostrea gigas)

Pacific oysters (Crassostrea gigas) collected on the coast of British Columbia, Canada have occasionally shown cadmium (Cd) concentrations at or above 2 microg g(-1) (wet weight), which has resulted in the loss of some international markets. This study investigated the source and transfer of Cd to oysters by focusing on the role of dissolved and particulate Cd in seawater. Parameters monitored for 1 year at two oyster farm sites on Vancouver Island included: oyster tissue mass and shell length, Cd in oysters, dissolved Cd, particulate Cd, temperature and salinity. Results show that dissolved Cd was the main source of Cd to the oysters and that Cd was mainly concentrated in the gut tissues. A seasonal trend was observed in Cd in oysters, in which levels were lowest during periods of higher temperatures. Results also indicate that the local oceanographic inputs and sediment diagenesis directly affect dissolved Cd and thereby influence the Cd levels in oysters. Particulate matter was not found to be a source of Cd in oysters, and was actually negatively correlated. This was likely due to the uptake of dissolved Cd by phytoplankton and the effect of phytoplankton on oyster tissue mass.