The acidification caused by the dissolution of anthropogenic carbon dioxide (CO2) in the ocean changes the chemistry and hence the bioavailability of iron (Fe), a limiting nutrient in large oceanic regions. Here, we show that the bioavailability of dissolved Fe may decline because of ocean acidification. Acidification of media containing various Fe compounds decreases the Fe uptake rate of diatoms and coccolithophores to an extent predicted by the changes in Fe chemistry. A slower Fe uptake by a model diatom with decreasing pH is also seen in experiments with Atlantic surface water. The Fe requirement of model phytoplankton remains unchanged with increasing CO2. The ongoing acidification of seawater is likely to increase the Fe stress of phytoplankton populations in some areas of the ocean.

Abstract. The control of key chemical parameters in phytoplankton cultures, such as pCO 2 , pH and (the saturation state of calcium carbonate), is made difficult by the interdependence of these parameters and by the changes resulting from the growth of the organisms, such as CO 2 fixation, nutrient uptake and, for coccolithophores, calcite precipitation. Even in cultures where pCO 2 or pH is maintained constant, other chemical parameters change substantially at high cell densities. Experimentally we observed that various methods of adjustment of pCO 2 /pH -acid or base addition, use of buffers or pH-stats, or bubbling of CO 2 -enriched air -can be used, the choice of one or the other depending on the goals of the experiments. At seawater pH, we measured the same growth rates in cultures of the diatom Thalassiosira weissflogii where the pCO 2 /pH was controlled by these different methods. The pH/pCO 2 control method also did not affect the rates of growth or calcification of the coccolithophore Emiliania huxleyi at seawater pH. At lower pH/higher pCO 2 , in the E. huxleyi strain PLY M219, we observed increases in rates of carbon fixation and calcification per cell, along with a slight increase in growth rate, except in bubbled cultures. In our hands, the bubbling of cultures seemed to induce more variable results than other methods of pCO 2 /pH control. While highly convenient, the addition of pH buffers to the medium apparently induces changes in trace metal availability and cannot be used under trace metal-limiting conditions.

Dissolution of anthropogenic CO 2 increases the partial pressure of CO 2 (pCO 2 ) and decreases the pH of seawater. The rate of Fe uptake by the dominant N 2 -fixing cyanobacterium Trichodesmium declines as pH decreases in metal-buffered medium. The slower Fe-uptake rate at low pH results from changes in Fe chemistry and not from a physiological response of the organism. Contrary to previous observations in nutrient-replete media, increasing pCO 2 /decreasing pH causes a decrease in the rates of N 2 fixation and growth in Trichodesmium under low-Fe conditions. This result was obtained even though the bioavailability of Fe was maintained at a constant level by increasing the total Fe concentration at low pH. Short-term experiments in which pCO 2 and pH were varied independently showed that the decrease in N 2 fixation is caused by decreasing pH rather than by increasing pCO 2 and corresponds to a lower efficiency of the nitrogenase enzyme. To compensate partially for the loss of N 2 fixation efficiency at low pH, Trichodesmium synthesizes additional nitrogenase. This increase comes partly at the cost of down-regulation of Fe-containing photosynthetic proteins. Our results show that although increasing pCO 2 often is beneficial to photosynthetic marine organisms, the concurrent decreasing pH can affect primary producers negatively. Such negative effects can occur both through chemical mechanisms, such as the bioavailability of key nutrients like Fe, and through biological mechanisms, as shown by the decrease in N 2 fixation in Fe-limited Trichodesmium.climate change | cyanobacteria | iron limitation A bout one-third of the anthropogenic CO 2 released into the atmosphere dissolves into the surface ocean, increasing the partial pressure of CO 2 , pCO 2 , and lowering the pH. This ocean acidification has been shown to have various consequences for marine phytoplankton (1-5). Organisms that invest a large amount of energy in the operation of a carbon-concentrating mechanism (CCM) are expected to be particularly sensitive to changes in pCO 2 . This is the case for marine cyanobacteria, which must elevate the CO 2 concentration at the site of carbon fixation as a result of the poor affinity for CO 2 of their carboxylating enzyme, ribulose bisphosphate carboxylase oxygenase (RubisCO) (6). Of particular interest is the effect of ocean acidification on the N 2 -fixing filamentous cyanobacterium Trichodesmium, which is responsible for a major fraction of all marine N 2 fixation and thus plays a prominent role in the biogeochemical cycling of C and N (7). This bloom-forming diazotroph thrives throughout the oligotrophic tropical and subtropical oceans where P and/or Fe often limit its growth and N 2 fixation (8-10).In the past few years, the effects of ocean acidification on Trichodesmium have been studied extensively in combination with those of other environmental variables, such as temperature, light intensity, and phosphorus limitation. Stimulation of N 2 fixation and growth at elevated pCO 2 has been observed in both la...

Acidification of seawater caused by anthropogenic carbon dioxide (CO) is anticipated to influence the growth of dinitrogen (N)-fixing phytoplankton, which contribute a large fraction of primary production in the tropical and subtropical ocean. We found that growth and N-fixation of the ubiquitous cyanobacterium decreased under acidified conditions, notwithstanding a beneficial effect of high CO Acidification resulted in low cytosolic pH and reduced N-fixation rates despite elevated nitrogenase concentrations. Low cytosolic pH required increased proton pumping across the thylakoid membrane and elevated adenosine triphosphate production. These requirements were not satisfied under field or experimental iron-limiting conditions, which greatly amplified the negative effect of acidification.

Nitrogen fixation is critical for the biological productivity of the ocean, but clear mechanistic controls on this process remain elusive. Here, we investigate the abundance, activity, and drivers of nitrogen-fixing diazotrophs across the tropical western North Pacific. We find a basin-scale coherence of diazotroph abundances and N 2 fixation rates with the supply ratio of iron:nitrogen to the upper ocean. Across a threshold of increasing supply ratios, the abundance of nifH genes and N 2 fixation rates increased, phosphate concentrations decreased, and bioassay experiments demonstrated evidence for N 2 fixation switching from iron to phosphate limitation. In the northern South China Sea, supply ratios were hypothesized to fall around this critical threshold and bioassay experiments suggested colimitation by both iron and phosphate. Our results provide evidence for iron:nitrogen supply ratios being the most important factor in regulating the distribution of N 2 fixation across the tropical ocean.

Phytoplankton assimilation and microbial oxidation of ammonium are two critical conversion pathways in the marine nitrogen cycle. The underlying regulatory mechanisms of these two competing processes remain unclear. Here we show that ambient nitrate acts as a key variable to bifurcate ammonium flow through assimilation or oxidation, and the depth of the nitracline represents a robust spatial boundary between ammonium assimilators and oxidizers in the stratified ocean. Profiles of ammonium utilization show that phytoplankton assemblages in nitrate-depleted regimes have higher ammonium affinity than nitrifiers. In nitrate replete conditions, by contrast, phytoplankton reduce their ammonium reliance and thus enhance the success of nitrifiers. This finding helps to explain existing discrepancies in the understanding of light inhibition of surface nitrification in the global ocean, and provides further insights into the spatial linkages between oceanic nitrification and new production.

In the high-nutrient, low-chlorophyll waters of the Gulf of Alaska, microcosm manipulation experiments were used to assess the effect of CO 2 on growth and primary production under iron-limited and iron-replete conditions. As expected, iron had a strong effect on growth and photosynthesis. A modest and variable stimulation of growth and biomass production by CO 2 (high CO 2 : 77-122 Pa; low CO 2 : 11-17 Pa) was observed under both iron-replete and iron-limited conditions, though near the limit of precision of our measurements in slow-growing low-iron experiments. Physiological acclimations responsible for the changes in growth were assessed. Under iron-limited conditions, growth stimulation at high CO 2 appeared to result from an increase in photosynthetic efficiency, which we attribute to energy savings from down-regulation of the carbon concentrating mechanisms. In some cases, iron-rich photosynthetic proteins (PsbA, PsaC, and cytochrome b 6 ) were down-regulated at elevated CO 2 in iron-limited controls. Under iron-replete conditions, there was an increase in growth rate and biomass at high CO 2 in some experiments. This increase was unexpectedly supported by reductions in cellular carbon loss, most likely decreased respiration. We speculate that this effect may be due to acclimation to decreased pH rather than high CO 2 . The variability in responses to CO 2 among experiments did not appear to be caused by differences in phytoplankton community structure and may reflect the sensitivity of the net response of phytoplankton to antagonistic effects of the several parameters that co-vary with CO 2 .

In natural samples from the New Jersey coast and the Gulf of Alaska, zinc (Zn) and cadmium (Cd) uptake rates by phytoplankton decreased on average about 30% as pH was decreased from 8.5 to 7.9 or 7.7, and the partial pressure of carbon dioxide (PCO2) increased accordingly. The underlying mechanism was explored with the model species, Thalassiosira weissflogii and Emiliania huxleyi, using ethylenediaminetetraacetic acid (EDTA), desferrioxamine B, phytochelatin, and cysteine as complexing agents. Experiments with single complexing agents did not reproduce the effect of pH seen in field samples, ruling out two possible mechanisms: a direct effect on the uptake machinery or down‐regulation of uptake at high PCO2. Zn and Cd bioavailability must thus somehow decrease at low pH in natural seawater, which is counterintuitive since the protonation of complexing agents at low pH should increase the total free concentration of metals. However, in the presence of both a strong and a weak complexing agent, metal uptake rate may decrease at low pH if formation of the weak complex decreases and the metal in the weak complex is more “available” than in the strong complex. We obtained proof of concept for such a two‐ligand mechanism for Zn uptake in the presence of EDTA + phytochelatin and EDTA + cysteine. Weak ligands that bind a small fraction of essential metals in surface seawater may thus be important in metal uptake by phytoplankton, and the dual effects of strong and weak complexing agents may control not just the magnitude but also the sign of the effect of pH‐PCO2 on metal uptake rates.