An integrated response of Trichodesmium erythraeum IMS101 growth and photo-physiology to Iron, CO₂, and light intensity

We have assessed how varying CO 2 (180, 380, and 720 μatm) and growth light intensity (40 and 400 μmol photons m -2 s -1 ) affected Trichodesmium erythraeum IMS101 growth and photophysiology over free iron (Fe') concentrations between 20 and 9,600 pM. We found significant iron dependencies of growth rate and the initial slope and maximal relative PSII electron transport rates (rP m ). Under iron-limiting concentrations, high-light increased growth rates and rPm; possibly indicating a lower allocation of resources to iron-containing photosynthetic proteins. Higher CO 2 increased growth rates across all iron concentrations, enabled growth to occur at lower Fe' concentrations, increased rPm and lowered the iron half saturation constants for growth (K m ). We attribute these CO 2 responses to the operation of the CCM and the ATP spent/saved for CO 2 uptake and transport at low and high CO 2 , respectively. It seems reasonable to conclude that T. erythraeum IMS101 can exhibit a high degree of phenotypic plasticity in response to CO 2 , light intensity and iron-limitation. These results are important given predictions of increased dissolved CO 2 and water column stratification (i.e., higher light exposures) over the coming decades

Determination of Specific Iron Chelator by Using LC-ICP-MS and LC-ESI-MS

Identification and quantification of siderophore type chelate was undertaken using recently developed high-performance liquid chromatography-mass spectrometry methods. Five different siderophore type chelates were detected and the compounds comprised of two groups; ferrioxamines. In the dissolved phase, three types of hydroxamate siderophore have been identified; Ferioxamine B (FOB), Ferrioxamine G (FOG) and ferrioxamine E (FOE). Concentration of dissolved FOB, FOG and FOE are extremely low between 0–135 x 10-18 M, and their distribution is spatially and temporally variable in this region. The concentration and diversity of siderophore type chelates determined during this study, was lower than those reported previously for dissolved ferrioxamine siderophore concentrations at lower latitudes in the Atlantic Ocean. The initial data from this study suggested that dissolved siderophores in this region is not an important fraction (< 0.1%) of the natural organic Fe(III) binding ligand pool

Identification and quantification of marine siderophores using high-performance liquid chromatography with electrospray ionization mass spectrometry

Siderophores are low-molecular-weight organic compounds (500 - 1500 Da) produced by heterotrophic bacteria and highly selective for Fe(III). These complexation of iron by siderophores has the potential to affect both the solubility and bioavailability of iron (III) in seawater. We have determined the ferrioxamine siderophores in the seawater samples from high latitude North Atlantic Ocean The identification and quantification of this siderophore type chelate was done by using recently developed high-performance liquid chromatography-mass spectrometry methods. Five different siderophore type chelates were detected and the compounds comprised of two groups; ferrioxamines and amphibactins group. In the dissolved phase, three types of hydroxamate siderophore have been identified; Ferioxamine B (FOB), Ferrioxamine G (FOG) and ferrioxamine E (FOE). Concentration of dissolved FOB, FOG and FOE are extremely low between 0–135 x 10-18 M. Our present data indicated the presence of low concentrations of dissolved siderophores in the high latitude of the Atlantic Ocean and suggest that siderophore distributions are both spatially and temporally variable

Impact of impurities in bromocresol green indicator dye on spectrophotometric total alkalinity measurements

Due to its accurate and precise character, spectrophotometric pH detection is a common technique applied in measurement methods for carbonate system parameters. However, impurities in the used pH indicator dyes can influence the measurements quality. During our work described here, we focused on impacts of impurities in the pH indicator dye bromocresol green (BCG) on spectrophotometric seawater total alkalinity (AT) measurements. In order to evaluate the extent of such influences, purified BCG served as a reference. First, a high-performance liquid chromatography (HPLC) purification method for BCG was developed as such a method did not exist at the time of this study. An analysis of BCG dye from four different vendors with this method revealed different types and quantities of impurities. After successful purification, AT measurements with purified and unpurified BCG were carried out using the novel autonomous analyzer CONTROS HydroFIA® TA. Long-term measurements in the laboratory revealed a direct influence of impurity types and quantities on the drift behavior of the analyzer. The purer the BCG, the smaller was the AT increase per measurement. The observed drift is generally caused by deposits in the optical pathway mainly generated by the impurities. However, the analyzers drift behavior could not be fully overcome. Furthermore, we could show that a certain impurity type in some indicator dyes changed the drift pattern from linear to nonlinear, which can impair long-term deployments of the system. Consequently, such indicators are impractical for these applications. Laboratory performance characterization experiments revealed no improvement of the measurement quality (precision and bias) by using purified BCG as long as the impurities of the unpurified dye do not exceed a quantity of 2 % (relationship of peak areas in the chromatogram). However, BCG with impurity quantities higher than 6 % provided AT values which failed fundamental quality requirements. In conclusion, to gain optimal AT measurements especially during long-term deployments, an indicator purification is not necessarily required as long as the purchased dye has a purity level of at least 98 % and is free of the named impurity type. Consequently, high-quality AT measurements do not require pure but the purest BCG that is purchasable

Abundance of the iron containing biomolecule, heme b, during the progression of a spring phytoplankton bloom in a mesocosm experiment

Concentrations of heme b were determined in a mesocosm experiment situated in Gullmar Fjord off Sweden. The mesocosm experiment lasted for ca. one hundred days and was characterised by the growth of a primary nutrient replete and a secondary nutrient deplete phytoplankton bloom. Heme b varied between 40 ± 10 pmol L-1 in the prebloom period up to a maximum of 700 ± 400 pmol L-1 just prior to the time of the primary chlorophyll a maximum. Thereafter, heme b concentrations decreased again to an average of 120 ± 60 pmol L-1. When normalised to total particulate carbon, heme b was most abundant during the initiation of the nutrient replete spring bloom, when ratios reached 52 ± 24 μmol mol-1; ten times higher than values observed both pre and post the primary bloom. Concentrations of heme b correlated with those of chlorophyll a. Nevertheless, differences were observed in the relative concentrations of the two parameters, with heme b concentrations increasing relative to chlorophyll a during the growth of the primary bloom, decreasing over the period of the secondary bloom and increasing again through the latter period of the experiment. Heme b abundance was therefore influenced by nutrient concentrations and also likely by changing community composition. In half of the mesocosms, pCO2 was elevated and maintained at ca.1000 μatm, however we observed no significant differences between heme b in plus or ambient pCO2 mesocosms, either in absolute terms, or relative to total particulate carbon and chlorophyll a. The results obtained in this study contribute to our understanding of the distribution of this significant component of the biogenic iron pool, and provide an iron replete coastal water end member that aids the interpretation of the distributions of heme b in more iron deplete open ocean waters

Metallophores associated with Trichodesmium erythraeum colonies from the Gulf of Aqaba

Trichodesmium is a globally important marine nitrogen fixing cyanobacteria which forms colonies and utilizes atmospherically derived dust as a source for the limiting micro-nutrient iron. Here we report the identification of metallophores isolated from incubations of natural Trichodesmium colonies collected from the Gulf of Aqaba in the Red Sea. Three of our compounds were identified as the ferrioxamine siderophores B, E, and G. The remaining fifteen metallophores had mass to charge ratios that, to our knowledge, are not common to known siderophores. Putative sum formulas suggest most of these compounds were not structurally related to each other. We also found that the novel metallophores readily formed complexes with aluminium and were less specific for Fe than the ferrioxamines. In our incubations of Trichodesmium colonies, the abundance of ten of the novel metallophores positively correlated with Trichodesmium biomass, but not with bacterial biomass, whilst ferrioxamine siderophores were more strongly associated with bacterial biomass. We identified ferrioxamines and our novel metallophores in filtered surface seawater samples from the Gulf of Aqaba. However, our novel metallophores were only observed in the surface seawater sample collected at the time of highest Trichodesmium abundance, while ferrioxamines were observed even when Trichodesmium was not present. We hypothesize that the novel metallophores were specifically associated with Trichodesmium colonies. Together with the bacterially produced ferrioxamines they likely contribute to a distinctive “ligandosphere” surrounding the Trichodesmium colonies, with potential implications for metal homeostasis within the colony environment

Influence of ocean acidification on the organic complexation of iron and copper in Northwest European shelf seas; a combined observational and model study

The pH of aqueous solutions is known to impact the chemical speciation of trace metals. In this study we conducted titrations of coastal seawaters with iron and copper at pH 7.91, 7.37 and 6.99 (expressed on the total pH scale). Changes in the concentration of iron and copper that complexed with the added ligands 1-nitroso-2-napthol and salicylaldoxime respectively were determined by adsorptive cathodic stripping voltammetry - competitive ligand equilibrium (AdCSV-CLE). Interpretation of the results, assuming complexation by a low concentration of discrete ligands, showed that conditional stability constants for iron complexes increased relative to inorganic iron complexation as pH decreased by approximately 1 log unit per pH unit, whilst those for copper did not change. No trend was observed for concentrations of iron and copper complexing ligands over the pH range examined. We also interpreted our titration data by describing chemical binding and polyelectrolytic effects using non-ideal competitive adsorption in Donnan-like gels (NICA-Donnan model) in a proof of concept study. The NICA-Donnan approach allows for the development of a set of model parameters that are independent of ionic strength and pH, and thus calculation of metal speciation can be undertaken at ambient sample pH or the pH of a future, more acidic ocean. There is currently a lack of basic NICA-Donnan parameters applicable to marine dissolved organic matter (DOM) so we assumed that the measured marine dissolved organic carbon could be characterized as terrestrial fulvic acids. Generic NICA-Donnan parameters were applied within the framework of the software program visual MINTEQ and the metal –added ligand concentrations [MeAL] calculated for the AdCSV-CLE conditions. For copper, calculated [MeAL] using the NICA-Donnan model for DOM were consistent with measured [MeAL], but for iron an inert fraction with kinetically inhibited dissolution was required in addition to the NICA-Donnan model in order to approximate the trends observed in measured [MeAL]. We calculated iron and copper speciation in Northwest European shelf water samples at ambient alkalinity and projected increased pCO2 concentrations as a demonstration of the potential of the approach

Zinc oxide films grown by galvanic deposition from 99% metals basis zinc nitrate electrolyte

The use of relatively low purity zinc nitrate for electrochemical deposition of compact ZnO films is attractive for large scale production because of the cost saving potential. ZnO films were grown on SnO2:F and magnetron sputtered ZnO:Al templates using a three electrode potentiostatic system in galvanic mode. The electrolyte consisted of a 0.1 M zinc nitrate solution (either 99.998% or 99% purity) and 1 mM aluminium nitrate for extrinsic doping, when required. Moderate deposition rates of up to 0.9 nm s−1 were achieved on ZnO:Al templates with lower rates of up to 0.5 nm s−1 on SnO2:F templates. Observation of SEM images of the films revealed a wall-like morphology whose lateral thickness (parallel to the substrate) reduced as aluminium was added to the system either in the electrolyte or from the substrate. However, pre- deposition activation of the template by applying a negative voltage (approximately −2 V) allowed the growth of compact films even for the low purity electrolyte. The optical band gap energy of intrinsically doped films was lower than that of the Al doped films. The composite electrical conductivity of all the films studied, as inferred from sheet resistance and Hall effect measurements of the ZnO/template stacks was much less than that of the uncoated templates. A strong E2 (high) mode at around 437 cm−1 was visible in the Raman spectra for most films confirming the formation of ZnO. However, both the Raman modes and XRD reflections associated with wurtzite ZnO diminished for the Al doped films indicating a high level of mainly oxygen related defects. Based on these data, further studies are underway to improve the doping efficiency of aluminium, the crystalline structure and thus the conductivity of such films

Phosphorus Limitation Enhances Diazotroph Zinc Quotas

Trichodesmium spp. is a colonial diazotrophic cyanobacterium found in the oligotrophic (sub)tropical oceans, where dissolved inorganic phosphorus (DIP) can be depleted. To cope with low P concentrations, P can be scavenged from the dissolved organic P (DOP) pool. This requires the deployment of multiple enzymes activated by trace metals, potentially enhancing metal requirements under stronger P limitations. To test this, we grew Trichodesmium under trace-metal-controlled conditions, where P was supplied as either DIP or DOP (methylphosphonic acid). Mean steady-state biomass under the DOP treatment was only 40% of that grown under equivalent DIP supply, carbon normalized alkaline phosphorus activity was elevated 4-fold, and the zinc (Zn)–carbon ratio was elevated 3.5-fold. Our finding matches the known, dominant Zn requirement across a diversity of enzymes involved in P stress responses and supports an important interaction in the oceanic cycles of these two nutrients

Heme b distributions through the Atlantic Ocean: evidence for "anemic" phytoplankton populations

Heme b is an iron-containing cofactor in hemoproteins that participates in the fundamental processes of photosynthesis and respiration in phytoplankton. Heme b concentrations typically decline in waters with low iron concentrations but due to lack of field data, the distribution of heme b in particulate material in the ocean is poorly constrained. Here we report particulate heme b distributions across the Atlantic Ocean (59.9°N to 34.6°S). Heme b concentrations in surface waters ranged from 0.10 to 33.7 pmol L−1 (median = 1.47 pmol L−1, n = 974) and were highest in regions with a high biomass. The ratio of heme b to particulate organic carbon (POC) exhibited a mean value of 0.44 μmol heme b mol−1 POC. We identified the ratio of 0.10 µmol heme b mol−1 POC as the cut-off between heme b replete and heme b deficient (anemic) phytoplankton. By this definition, we observed anemic phytoplankton populations in the Subtropical South Atlantic and Irminger Basin. Comparison of observed and modelled heme b suggested that heme b could account for between 0.17–9.1% of biogenic iron. Our large scale observations of heme b relative to organic matter provide further evidence of the impact of changes in iron supply on phytoplankton iron status across the Atlantic Ocean