Hydrogen isotope fractionation is controlled by CO2 in coccolithophore lipids.

Hydrogen isotope ratios (δ2H) represent an important natural tracer of metabolic processes, but quantitative models of processes controlling H-fractionation in aquatic photosynthetic organisms are lacking. Here, we elucidate the underlying physiological controls of 2H/1H fractionation in algal lipids by systematically manipulating temperature, light, and CO2(aq) in continuous cultures of the haptophyte Gephyrocapsa oceanica. We analyze the hydrogen isotope fractionation in alkenones (αalkenone), a class of acyl lipids specific to this species and other haptophyte algae. We find a strong decrease in the αalkenone with increasing CO2(aq) and confirm αalkenone correlates with temperature and light. Based on the known biosynthesis pathways, we develop a cellular model of the δ2H of algal acyl lipids to evaluate processes contributing to these controls on fractionation. Simulations show that longer residence times of NADPH in the chloroplast favor a greater exchange of NADPH with 2H-richer intracellular water, increasing αalkenone. Higher chloroplast CO2(aq) and temperature shorten NADPH residence time by enhancing the carbon fixation and lipid synthesis rates. The inverse correlation of αalkenone to CO2(aq) in our cultures suggests that carbon concentrating mechanisms (CCM) do not achieve a constant saturation of CO2 at the Rubisco site, but rather that chloroplast CO2 varies with external CO2(aq). The pervasive inverse correlation of αalkenone with CO2(aq) in the modern and preindustrial ocean also suggests that natural populations may not attain a constant saturation of Rubisco with the CCM. Rather than reconstructing growth water, αalkenone may be a powerful tool to elucidate the carbon limitation of photosynthesis.

Influence of Temperature and CO2 On Plasma-membrane Permeability to CO2 and HCO3- in the Marine Haptophytes Emiliania huxleyi and Calcidiscus leptoporus (Prymnesiophyceae).

Membrane permeabilities to CO2 and HCO3- constrain the function of CO2 concentrating mechanisms that algae use to supply inorganic carbon for photosynthesis. In diatoms and green algae, plasma membranes are moderately to highly permeable to CO2 but effectively impermeable to HCO3- . Here, CO2 and HCO3- membrane permeabilities were measured using an 18 O-exchange technique on two species of haptophyte algae, Emiliania huxleyi and Calcidiscus leptoporus, which showed that the plasma membranes of these species are also highly permeable to CO2 (0.006-0.02 cm · s-1 ) but minimally permeable to HCO3- . Increased temperature and CO2 generally increased CO2 membrane permeabilities in both species, possibly due to changes in lipid composition or CO2 channel proteins. Changes in CO2 membrane permeabilities showed no association with the density of calcium carbonate coccoliths surrounding the cell, which could potentially impede passage of compounds. Haptophyte plasma-membrane permeabilities to CO2 were somewhat lower than those of diatoms but generally higher than membrane permeabilities of green algae. One caveat of these measurements is that the model used to interpret 18 O-exchange data assumes that carbonic anhydrase, which catalyzes 18 O-exchange, is homogeneously distributed in the cell. The implications of this assumption were tested using a two-compartment model with an inhomogeneous distribution of carbonic anhydrase to simulate 18 O-exchange data and then inferring plasma-membrane CO2 permeabilities from the simulated data. This analysis showed that the inferred plasma-membrane CO2 permeabilities are minimal estimates but should be quite accurate under most conditions.

Late Miocene threshold response of marine algae to carbon dioxide limitation.

Coccolithophores are marine algae that use carbon for calcification and photosynthesis. The long-term adaptation of these and other marine algae to decreasing carbon dioxide levels during the Cenozoic era has resulted in modern algae capable of actively enhancing carbon dioxide at the site of photosynthesis. This enhancement occurs through the transport of dissolved bicarbonate (HCO3(-)) and with the help of enzymes whose expression can be modulated by variable aqueous carbon dioxide concentration, [CO2], in laboratory cultures. Coccolithophores preserve the geological history of this adaptation because the stable carbon and oxygen isotopic compositions of their calcite plates (coccoliths), which are preserved in the fossil record, are sensitive to active carbon uptake and transport by the cell. Here we use a model of cellular carbon fluxes and show that at low [CO2] the increased demand for HCO3(-) at the site of photosynthesis results in a diminished allocation of HCO3(-) to calcification, which is most pronounced in larger cells. This results in a large divergence between the carbon isotopic compositions of small versus large coccoliths only at low [CO2]. Our evaluation of the oxygen and carbon isotope record of size-separated fossil coccoliths reveals that this isotopic divergence first arose during the late Miocene to the earliest Pliocene epoch (about 7-5 million years ago). We interpret this to be a threshold response of the cells' carbon acquisition strategies to decreasing [CO2]. The documented coccolithophore response is synchronous with a global shift in terrestrial vegetation distribution between 8 and 5 Myr ago, which has been interpreted by some studies as a floral response to decreasing partial pressures of carbon dioxide () in the atmosphere. We infer a global decrease in carbon dioxide levels for this time interval that has not yet been identified in the sparse proxy record but is synchronous with global cooling and progressive glaciations.

Hydrological change in Southern Europe responding to increasing North Atlantic overturning during Greenland Stadial 1.

Greenland Stadial 1 (GS-1) was the last of a long series of severe cooling episodes in the Northern Hemisphere during the last glacial period. Numerous North Atlantic and European records reveal the intense environmental impact of that stadial, whose origin is attributed to an intense weakening of the Atlantic Meridional Overturning Circulation in response to freshening of the North Atlantic. Recent high-resolution studies of European lakes revealed a mid-GS-1 transition in the climatic regimes. The geographical extension of such atmospheric changes and their potential coupling with ocean dynamics still remains unclear. Here we use a subdecadally resolved stalagmite record from the Northern Iberian Peninsula to further investigate the timing and forcing of this transition. A solid interpretation of the environmental changes detected in this new, accurately dated, stalagmite record is based on a parallel cave monitoring exercise. This record reveals a gradual transition from dry to wet conditions starting at 12,500 y before 2000 A.D. in parallel to a progressive warming of the subtropical Atlantic Ocean. The observed atmospheric changes are proposed to be led by a progressive resumption of the North Atlantic convection and highlight the complex regional signature of GS-1, very distinctive from previous stadial events.

Detection of a variable intracellular acid-labile carbon pool in Thalassiosira weissflogii (Heterokontophyta) and Emiliania huxleyi (Haptophyta) in response to changes in the seawater carbon system.

Accumulation of an intracellular pool of carbon (C(i) pool) is one strategy by which marine algae overcome the low abundance of dissolved CO2 (CO2 (aq) ) in modern seawater. To identify the environmental conditions under which algae accumulate an acid-labile C(i) pool, we applied a (14) C pulse-chase method, used originally in dinoflagellates, to two new classes of algae, coccolithophorids and diatoms. This method measures the carbon accumulation inside the cells without altering the medium carbon chemistry or culture cell density. We found that the diatom Thalassiosira weissflogii [(Grunow) G. Fryxell & Hasle] and a calcifying strain of the coccolithophorid Emiliania huxleyi [(Lohmann) W. W. Hay & H. P. Mohler] develop significant acid-labile C(i) pools. C(i) pools are measureable in cells cultured in media with 2-30 µmol l(-1) CO2 (aq), corresponding to a medium pH of 8.6-7.9. The absolute C(i) pool was greater for the larger celled diatoms. For both algal classes, the C(i) pool became a negligible contributor to photosynthesis once CO2 (aq) exceeded 30 µmol l(-1) . Combining the (14) C pulse-chase method and (14) C disequilibrium method enabled us to assess whether E. huxleyi and T. weissflogii exhibited thresholds for foregoing accumulation of DIC or reduced the reliance on bicarbonate uptake with increasing CO2 (aq) . We showed that the C(i) pool decreases with higher CO2 :HCO3 (-) uptake rates.

Rapid northern hemisphere ice sheet melting during the penultimate deglaciation.

The rate and consequences of future high latitude ice sheet retreat remain a major concern given ongoing anthropogenic warming. Here, new precisely dated stalagmite data from NW Iberia provide the first direct, high-resolution records of periods of rapid melting of Northern Hemisphere ice sheets during the penultimate deglaciation. These records reveal the penultimate deglaciation initiated with rapid century-scale meltwater pulses which subsequently trigger abrupt coolings of air temperature in NW Iberia consistent with freshwater-induced AMOC slowdowns. The first of these AMOC slowdowns, 600-year duration, was shorter than Heinrich 1 of the last deglaciation. Although similar insolation forcing initiated the last two deglaciations, the more rapid and sustained rate of freshening in the eastern North Atlantic penultimate deglaciation likely reflects a larger volume of ice stored in the marine-based Eurasian Ice sheet during the penultimate glacial in contrast to the land-based ice sheet on North America as during the last glacial.

Phenotypic Variability in the Coccolithophore Emiliania huxleyi.

Coccolithophores are a vital part of oceanic phytoplankton assemblages that produce organic matter and calcium carbonate (CaCO3) containing traces of other elements (i.e. Sr and Mg). Their associated carbon export from the euphotic zone to the oceans' interior plays a crucial role in CO2 feedback mechanisms and biogeochemical cycles. The coccolithophore Emiliania huxleyi has been widely studied as a model organism to understand physiological, biogeochemical, and ecological processes in marine sciences. Here, we show the inter-strain variability in physiological and biogeochemical traits in 13 strains of E. huxleyi from various biogeographical provinces obtained from culture collections commonly used in the literature. Our results demonstrate that inter-strain genetic variability has greater potential to induce larger phenotypic differences than the phenotypic plasticity of single strains cultured under a broad range of variable environmental conditions. The range of variation found in physiological parameters and calcite Sr:Ca highlights the need to reconsider phenotypic variability in paleoproxy calibrations and model parameterizations to adequately translate findings from single strain laboratory experiments to the real ocean.

Decrease in coccolithophore calcification and CO2 since the middle Miocene.

Marine algae are instrumental in carbon cycling and atmospheric carbon dioxide (CO2) regulation. One group, coccolithophores, uses carbon to photosynthesize and to calcify, covering their cells with chalk platelets (coccoliths). How ocean acidification influences coccolithophore calcification is strongly debated, and the effects of carbonate chemistry changes in the geological past are poorly understood. This paper relates degree of coccolith calcification to cellular calcification, and presents the first records of size-normalized coccolith thickness spanning the last 14 Myr from tropical oceans. Degree of calcification was highest in the low-pH, high-CO2 Miocene ocean, but decreased significantly between 6 and 4 Myr ago. Based on this and concurrent trends in a new alkenone ɛp record, we propose that decreasing CO2 partly drove the observed trend via reduced cellular bicarbonate allocation to calcification. This trend reversed in the late Pleistocene despite low CO2, suggesting an additional regulator of calcification such as alkalinity.

Micropicking of nannofossils in preparation for analysis by secondary ion mass spectrometry.

This protocol describes a technique for picking individual nannofossils for geochemical analysis. The protocol allows for preparation of both individual nannofossil specimens and monospecific populations for analysis. A micromanipulator is adapted on an inverted microscope to hold a very fine tungsten needle capable of picking up individual selected nannofossils from an uncovered glass slide. Nannofossils can then be placed in designated areas on a Tacky Dot slide. Subsequently, they can be transferred to epoxy resin and disinterred from the Tacky slide. Nannofossils prepared using this protocol are suitable for subsequent analysis by secondary ion mass spectrometry. The preparation procedure in total takes 90-120 min per sample.

Potential and limitations of Sr/Ca ratios in coccolith carbonate: new perspectives from cultures and monospecific samples from sediments.

The Sr/Ca ratio of coccoliths was recently proposed as a potential indicator of past growth rates of coccolithophorids, marine algae, which play key roles in both the global carbonate and carbon cycles. We synthesize calibrations of this proxy through laboratory culture studies and analysis of monospecific coccolith assemblages from surface sediments. Cultures of coccolithophorids Helicosphaera carteri, Syracosphaera pulchra and Algirospira robusta confirm a 1-2% increase in Sr/Ca per degrees C previously identified in Emiliania huxleyi and Gephyrocapsa oceanica. This effect is not due merely to increases in growth rate with temperature and must be considered in palaeoceanographic studies. In light-limited cultures of E. huxleyi, Calcidiscus leptoporus and G. oceanica at constant temperature, coccolith Sr/Ca ratios vary by 10% across the range of possible growth and calcification rates for a given species. Among different species under similar culture conditions, Sr/Ca ratios vary by 30%. Although the highest ratios are in the cells with highest calcification and organic carbon fixation rates, at lower rates there is much scatter, indicating that different mechanisms control interspecific and intraspecific coccolith Sr/Ca variations. In field studies in the Equatorial Pacific and Somalia coastal region, coccolith Sr/Ca correlates with upwelling intensity and productivity. A more dynamic response is observed in larger coccoliths like C. leptoporus (23-55% variation in Sr/Ca) than in smaller coccoliths of G. oceanica or Florisphaera profunda (6-15% variation in Sr/Ca). This response suggests that, despite temperature effects, coccolith Sr/Ca has potential as an indicator of coccolithophorid productivity. If the variable Sr/Ca response of different species accurately reflects their variable productivity response to upwelling (and not different slopes of Sr/Ca with productivity), coccolith Sr/Ca could provide useful data on past changes in coccolith ecology. The mechanism of coccolith Sr/Ca variations remains poorly understood but is probably more closely tied to biochemical cycles during carbon acquisition than to chemical kinetic effects on Sr incorporation in the calcite coccolith crystals.