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Marine algae perform approximately half of global carbon fixation, but their growth is often limited by the availability of phosphate or other nutrients. As oceans warm, the area of phosphate-limited surface waters is predicted to increase, resulting in ocean desertification. Understanding the responses of key eukaryotic phytoplankton to nutrient limitation is therefore critical. We used advanced photo-bioreactors to investigate how the widespread marine green alga Micromonas commoda grows under transitions from replete nutrients to chronic phosphate limitation and subsequent relief, analysing photosystem changes and broad cellular responses using proteomics, transcriptomics and biophysical measurements. We find that physiological and protein expression responses previously attributed to stress are critical to supporting stable exponential growth when phosphate is limiting. Unexpectedly, the abundance of most proteins involved in light harvesting does not change, but an ancient light-harvesting-related protein, LHCSR, is induced and dissipates damaging excess absorbed light as heat throughout phosphate limitation. Concurrently, a suite of uncharacterized proteins with narrow phylogenetic distributions increase multifold. Notably, of the proteins that exhibit significant changes, 70% are not differentially expressed at the mRNA transcript level, highlighting the importance of post-transcriptional processes in microbial eukaryotes. Nevertheless, transcript–protein pairs with concordant changes were identified that will enable more robust interpretation of eukaryotic phytoplankton responses in the field from metatranscriptomic studies. Our results show that P-limited Micromonas responds quickly to a fresh pulse of phosphate by rapidly increasing replication, and that the protein network associated with this ability is composed of both conserved and phylogenetically recent proteome systems that promote dynamic phosphate homeostasis. That an ancient mechanism for mitigating light stress is central to sustaining growth during extended phosphate limitation highlights the possibility of interactive effects arising from combined stressors under ocean change, which could reduce the efficacy of algal strategies for optimizing marine photosynthesis.

The accuracy and precision of ion sensitive field effect transistor (ISFET) pH sensors have been well documented, but primarily by ocean chemistry specialists employing the technology at single locations. Here we examine their performance in a network context through comparison to discrete measurements of pH, using different configurations of the Honeywell DuraFET pH sensor deployed in six coastal settings by operators with a range of experience. Experience of the operator had the largest effect on performance. The average difference between discrete and ISFET pH was 0.005 pH units, but ranged from −0.030 to 0.083 among operators, with more experienced operators within ± 0.02 pH units of the discrete measurement. In addition, experienced operators achieved a narrower range of variance in difference between discrete bottle measurements and ISFET sensor readings compared to novice operators and novice operators had a higher proportion of data failing quality control screening. There were no statistically significant differences in data uncertainty associated with sensor manufacturer or deployment environment (pier-mounted, flowthrough system, and buoy-mounted). The variation we observed among operators highlights the necessity of best practices and training when instruments are to be used in a network where comparison across data streams is desired. However, while opportunities remain for improving the performance of the ISFET sensors when deployed by less experienced operators, the uncertainty associated with their deployment and validation was several-fold less than the observed natural temporal variability in pH, demonstrating the utility of these sensors in tracking local changes in acidification.

Marine algae perform approximately half of global carbon fixation, but their growth is often limited by the availability of phosphate or other nutrients

Increasing atmospheric carbon dioxide is driving a long-term decrease in ocean pH which is superimposed on daily to seasonal variability. These changes impact ecosystem processes, and they serve as a record of ecosystem metabolism. However, the temporal variability in pH is observed at only a few locations in the ocean because a ship is required to support pH observations of sufficient precision and accuracy. This paper describes a pressure tolerant Ion Sensitive Field Effect Transistor pH sensor that is based on the Honeywell Durafet ISFET die. When combined with a AgCl pseudoreference sensor that is immersed directly in seawater, the system is capable of operating for years at a time on platforms that cycle from depths of several km to the surface. The paper also describes the calibration scheme developed to allow calibrated pH measurements to be derived from the activity of HCl reported by the sensor system over the range of ocean pressure and temperature. Deployments on vertical profiling platforms enable self-calibration in deep waters where pH values are stable. Measurements with the sensor indicate that it is capable of reporting pH with an accuracy of 0.01 or better on the total proton scale and a precision over multiyear periods of 0.005. This system enables a global ocean observing system for ocean pH.

Eukaryotic algae within the picoplankton size class (≤2 μm in diameter) are important marine primary producers, but their spatial and ecological distributions are not well characterized. Here, we studied three picoeukaryotic prasinophyte genera and their cyanobacterial counterparts, Prochlorococcus and Synechococcus , during two cruises along a North Pacific transect characterized by different ecological regimes. Picoeukaryotes and Synechococcus reached maximum abundances of 1.44 × 10 5 and 3.37 × 10 5 cells · ml −1 , respectively, in mesotrophic waters, while Prochlorococcus reached 1.95 × 10 5 cells · ml −1 in the oligotrophic ocean. Of the picoeukaryotes, Bathycoccus was present at all stations in both cruises, reaching 21,368 ± 327 18S rRNA gene copies · ml −1 . Micromonas and Ostreococcus clade OI were detected only in mesotrophic and coastal waters and Ostreococcus clade OII only in the oligotrophic ocean. To resolve proposed Bathycoccus ecotypes, we established genetic distances for 1,104 marker genes using targeted metagenomes and the Bathycoccus prasinos genome. The analysis was anchored in comparative genome analysis of three Ostreococcus species for which physiological and environmental data are available to facilitate data interpretation. We established that two Bathycoccus ecotypes exist, named here BI (represented by coastal isolate Bathycoccus prasinos ) and BII. These share 82% ± 6% nucleotide identity across homologs, while the Ostreococcus spp. share 75% ± 8%. We developed and applied an analysis of ecomarkers to metatranscriptomes sequenced here and published -omics data from the same region. The results indicated that the Bathycoccus ecotypes cooccur more often than Ostreococcus clades OI and OII do. Exploratory analyses of relative transcript abundances suggest that Bathycoccus NRT2.1 and AMT2.2 are high-affinity NO 3 − and low-affinity NH 4 + transporters, respectively, with close homologs in multiple picoprasinophytes. Additionally, in the open ocean, where dissolved iron concentrations were low (0.08 nM), there appeared to be a shift to the use of nickel superoxide dismutases (SODs) from Mn/Fe/Cu SODs closer inshore. Our study documents the distribution of picophytoplankton along a North Pacific ecological gradient and offers new concepts and techniques for investigating their biogeography.

In July 2002, a combination of underway mapping and discrete profiles revealed significant along-shore variability in the concentrations of manganese and iron in the vicinity of Monterey Bay, California. Both metals had lower concentrations in surface waters south of Monterey Bay, where the shelf is about 2.5 km wide, than north of Monterey Bay, where the shelf is about 10 km wide. During non-upwelling conditions over the northern broad shelf, dissolvable iron concentrations measured underway in surface waters reached 3.5 nmol L−1 and dissolved manganese reached 25 nmol L−1. In contrast, during non-upwelling conditions over the southern narrow shelf, dissolvable iron concentrations in surface waters were less than 1 nmol L−1 and dissolved manganese concentrations were less than 5 nmol L−1. A pair of vertical profiles at 1000 m water depth collected during an upwelling event showed dissolved manganese concentrations of 10 decreasing to 2 nmol L−1, and dissolvable iron concentrations of 12–20 nmol L−1 in the upper 100 m in the north, compared to 3.5–2 nmol L−1 Mn and ∼0.6 nmol L−1 Fe in the upper 100 m in the south, suggesting the effect of shelf width influences the chemistry of waters beyond the shelf. These observations are consistent with current understanding of the mechanism of iron supply to coastal upwelling systems: Iron from shelf sediments, predominantly associated with particles greater than 20 μm, is brought to the surface during upwelling conditions. We hypothesize that manganese oxides are brought to the surface with upwelling and are then reduced to dissolved manganese, perhaps by photoreduction, following a lag after upwelling. Greater phytoplankton biomass, primary productivity, and nutrient drawdown were observed over the broad shelf, consistent with the greater supply of iron. Incubation experiments conducted 20 km offshore in both regions, during a period of wind relaxation, confirm the potential of these sites to become limited by iron. There was no additional growth response when copper, manganese or cobalt was added in addition to iron. The growth response of surface water incubated with bottom sediment (4 nmol L−1 dissolvable Fe) was slightly greater than in control incubations, but less than in the presence of 4 nmol L−1 dissolved iron. This may indicate that dissolvable iron is not as bioavailable as dissolved iron, although the influence of additional inhibitory elements in the sediment cannot be ruled out.

The availability of iron is known to exert a controlling influence on biological productivity in surface waters over large areas of the ocean and may have been an important factor in the variation of the concentration of atmospheric carbon dioxide over glacial cycles. The effect of iron in the Southern Ocean is particularly important because of its large area and abundant nitrate, yet iron-enhanced growth of phytoplankton may be differentially expressed between waters with high silicic acid in the south and low silicic acid in the north, where diatom growth may be limited by both silicic acid and iron. Two mesoscale experiments, designed to investigate the effects of iron enrichment in regions with high and low concentrations of silicic acid, were performed in the Southern Ocean. These experiments demonstrate iron's pivotal role in controlling carbon uptake and regulating atmospheric partial pressure of carbon dioxide.

We report measurements of dissolvable and particulate iron, particulate Al, nutrients and phytoplankton biomass in surface waters during the termination of one upwelling event and the initiation of a second event in August 2000. These events occurred in the area of the Ano Nuevo upwelling center off the coast of central California. The first event was observed after ∼8 days of continuous upwelling favorable winds, while the second event was observed through the onset of upwelling favorable winds to wind reversals ∼3 days later. Coincident with the upwelling signatures of low temperature and high salinity were significantly elevated concentrations of nitrate and silicate with average concentrations greater than 15 and 20 μM, respectively, during both upwelling events. Dissolvable Fe concentrations (TD-Fe) were significantly higher in the second event, 6.5 versus 1.2 nM Fe found in the first event. Nitrate was reduced by ∼5 μM day −1 within this second upwelled plume as compared to a drawdown of ∼2 μM day −1 within the first plume. Silicate was reduced in a ratio of 1.2 mol Si:mol NO 3 in the high Fe waters of the second plume as compared to a ratio of 2.2 in the lower Fe waters of the first plume. The observed differences in nutrient utilization are consistent with some degree of iron limitation. The area of increased dissolvable Fe in the second upwelling event was coincident with elevated particulate Fe concentrations, indicating the particulate pool as a possible source of the observed increase in TD-Fe. The elevated particulate Fe in surface waters was a result of resuspended sediments in the bottom boundary layer (BBL) of the shallow shelf being transported to the surface during upwelling. Particulate (and dissolvable) iron concentrations were significantly reduced as upwelling continued. This was most probably due to a decoupling of the BBL from upwelled source waters as the upwelling front moved offshore and/or reduced turbulence in the BBL as upwelling continued. The observed reduction in both particulate and dissolvable Fe, as upwelling continued to deliver macronutrients to surface waters, may result in varying levels of Fe limitation.

Iron has been measured for 16 months with ∼21 day resolution at three stations in the upwelling ecosystem of central California, providing the first detailed assessment of the annual iron cycle in the coastal zone. A large pulse of iron occurs during the first spring upwelling event of the year. Iron concentrations then decay up to 100-fold over several months, although upwelling continues. Excess surface nitrate and low iron are the result during the summer and fall at the two stations furthest offshore (20 and 45 km), while nitrate is depleted and iron high nearshore (5 km). Phytoplankton biomass, primary production and community structure appear to be controlled by iron concentrations in offshore waters during this period.

Manganese oxidation rates were determined at low (∼ 20 nM) concentrations in seawater by measuring dissolved manganese (Mn(II)) using flow injection analysis with chemiluminescence detection. Mn(II) was measured in samples that had been filtered (0.2 μm) and kept in the dark under controlled temperature and pH conditions for time periods up to 6 months. Eight 9 L carboys with mean pH values ranging from 8 to 8.7 were held at 25°C, another carboy (pH = 9.32) was kept at 5°C. Oxidation followed the Morgan (1967) homogeneous rate equation (d[Mn(II)]/dt = k1 [O2][OH−]2[Mn(II)]). The mean rate constant k1 = 1.7 ± 0.7 × 1012 M−3 d−1 (95% CI), determined using hydroxide ion activities determined with pH measurements on the NBS scale, was in agreement with work by Morgan (1967; k1 = 4 × 1012 M−3 d−1) and Davies and Morgan (1989; k1 = 1.1 × 1012 M−3 d−1) in dilute solutions. The rate constant at 5°C was 1.3 ± 0.3 × 1012 M−3 d−1. If free hydroxide concentrations (based on the free proton pH scale) are used, then the rate constant at 25°C was k 1 ∗ = 0.34 ± 0.14 × 10 12 M−3d−1. Autocatalytic increases in Mn(II) oxidation rates, as predicted by a heterogeneous reaction mechanism (Morgan, 1967) (d[Mn(II)]/dt= k2′[Mn(II)][MnO2]) were not observed, indicating that the homogeneous reaction dominates Mn(II) oxidation at low nM concentrations in seawater. Bacteria were enumerated by 4′,6-diamidino-2-phenylindole (DAPI) staining during the experiments. No significant correlation between bacterial concentrations and Mn(II) oxidation rates was found.

A sensitive technique for the shipboard determination of zinc in seawater has been developed. The technique couples flow injection analysis with fluorometric detection (FIA-FL). A cation exchange column was used to separate zinc from interering alkali and alkaline earth ions and to concentrate Zn from seawarer. The organic indicator ligan, p-tosyl-8-aminoquinoline, was used to form a complex with zinc, the fluorescence of which was determined with a flow-through fluorometer. The deterction limit (defined as three times the standard deviation of the blank, n=4) was 0.1 nM for a 4.4-mL sample

The idea that iron might limit phytoplankton growth in large regions of the ocean has been tested by enriching an area of 64 km2 in the open equatorial Pacific Ocean with iron. This resulted in a doubling of plant biomass, a threefold increase in chlorophyll and a fourfold increase in plant production. Similar increases were found in a chlorophyll-rich plume down-stream of the Galapagos Islands, which was naturally enriched in iron. These findings indicate that iron limitation can control rates of phytoplankton productivity and biomass in the ocean.

In situ measurements of dissolved manganese and total dissolved iron were conducted in hydrothermal plumes over the Juan de Fuca Ridge using a submersible chemical analyzer (Scanner). The Scanner was deployed as part of a CTD/transmissometer/rosette instrument package on both tow-yos and vertical casts during the VENTS Leg I cruise in 1989. Dissolved manganese and total dissolved iron concentrations, along with temperature and light attenuation anomalies, were determined over the ridge crest every 5 s. Discrete samples for laboratory analyses of dissolved iron II, total dissolved iron II+III and manganese were also collected. Metal to heat ratios (Me:Q) measured in situ were extremely variable in one steady state plume, while an event plume had constant Me:Q. Uniform values of Mn:Q in the event plume demonstrate that Mn behaves conservatively in the near-field plume. Variability in the Mn: Q ratios in a steady state plume indicated the presence of at least two hydrothermal sources with distinct Me:Q values. A simple mixing model shows that the contribution of Mn from high Me:Q sources, with a composition characteristic of black smoker vents, varies between 1% and 99% within the core of the steady state plume with an average value of 55%. On average, over 50% of the excess heat within the plume originates from low Me:Q ratio sources, with a composition characteristic of low-temperature, diffuse flow vent fluids. Less than 4% of the volume of hydrothermal fluids in the plume originates from black smokers. The Fe II concentrations were used to provide an estimate of plume age on a transect across the ridge axis. Plume ages were about 2.5 days on axis and >12 days off axis. These plume ages were modeled to provide estimates of plume transport and horizontal diffusion and show excellent agreement with ages determined using 222Rn.

The light is emitted by the reaction of brillant sulfoflavin with hydrogen peroxide and Fe(II) in a neutral medium. A cation-exchange column is used to preconcentrate the iron from seawater. The detection limit was 0.45 nmol/L when 4.4 mL of sample was passed through the column

[1] Seven years of observations of surface water iron concentrations in the Monterey Bay region of central California reveal a consistent annual cycle dominated by the injection of high concentrations of particulate iron each spring. A companion study of the water column near the upwelling center at the north end of the bay clearly indicates a sedimentary source for the iron. Local river discharge, during winter storm events, results in the deposition of a fine-grained sediment “fluff” layer along the shelf. The initial rapid shoaling of isotherms at the onset of upwelling in spring brings water from the fluff layer to the surface. Concentrations of iron in both surface and source waters are then quickly diminished although upwelling intensifies, bringing the highest concentration of nitrate to the surface ∼3 months later. The concentration of a number of constituents measured in bottom water samples collected during the companion study strongly suggests that the rapid decrease in surface water iron concentrations is due to the depletion of the fluff layer following initial isotherm shoaling. The decoupling of iron and nitrate supply sets up the potential for iron limitation. However, on the basis of average Fe/NO3 ratios, we conclude that the region is not often iron limited. Iron limitation was apparent only one summer at the offshore station. Summer chlorophyll concentrations are highly correlated to dissolvable (unfiltered) iron concentrations, evidence in support of the role of particulate iron in meeting the ecosystem's iron requirements.

[1] The flux of dissolved iron from sediments to the water column was measured with flux chambers along the California coast over a five-year period. High fluxes were observed from sediments on the continental shelf. The measured fluxes were an average of 75 times larger than flux values derived from pore-water iron gradients. The iron flux was significantly correlated with the oxidation of organic matter, which allows an extrapolation to the global shelf. The input from shelf sediments is at least as significant as the global input of dissolved iron from aerosols, which has been presumed to be the dominant external iron source. Evidence of this input is seen 100's of kilometers offshore where it can enable the high productivity of broad coastal regions seen in satellite images.

[1] Here we report measurements of iron and aluminum in surface and subsurface waters during late March and late May of 2001 on transects between central California and Hawaii. A large cloud of Asian dust was detected during April 2001, and there was a clear signal in surface water iron due to aerosol deposition on the May transect. Iron and aluminum concentrations increased synchronously by 0.5 and 2 nM along the southern portion of the transect, which includes the Hawaii Ocean Time series (HOT) station, from background values in March (0.1 to 0.2 nM Fe). These changes occurred in a ratio that is close to the crustal abundance ratio of the metals, which indicates a soil aerosol source. A vertical profile of dissolved iron was also measured at the HOT station in late April and this profile also shows a large increase near the surface. Direct observations of aerosol iron concentration at Mauna Loa Observatory on Hawaii indicate that aerosol concentrations were significantly lower than climatological values during this period. Soil aerosol concentrations along the transect were estimated using the real-time Navy Aerosol Analysis and Prediction System (NAAPS). The NAAPS results show a large meridional gradient with maximum concentrations in the boundary layer north of 30°N. However, the deposition of iron and aluminum to surface waters was highest south of 25°N, near Hawaii. There were only weak signals in the ecosystem response to the aerosol deposition.

The flux of manganese from continental margin sediments to the ocean was measured with a free-vehicle, benthic flux chamber in a transect across the continental shelf and upper slope of the California margin. The highest fluxes were observed on the shallow continental shelf. Manganese flux decreased linearly with bottom water oxygen concentration, and the lowest fluxes occurred in the oxygen minimum zone (at a depth of 600 to 1000 meters). Although the flux of manganese from continental shelf sediments can account for the elevated concentrations observed in shallow, coastal waters, the flux from sediments that intersect the oxygen minimum cannot produce the subsurface concentration maximum of dissolved manganese that is observed in the Pacific Ocean.