Environmental stability affects phenotypic evolution in a globally distributed marine picoplankton

Marine phytoplankton can evolve rapidly when confronted with aspects of climate change because of their large population sizes and fast generation times. Despite this, the importance of environment fluctuations, a key feature of climate change, has received little attention-selection experiments with marine phytoplankton are usually carried out in stable environments and use single or few representatives of a species, genus or functional group. Here we investigate whether and by how much environmental fluctuations contribute to changes in ecologically important phytoplankton traits such as C:N ratios and cell size, and test the variability of changes in these traits within the globally distributed species Ostreococcus. We have evolved 16 physiologically distinct lineages of Ostreococcus at stable high CO2 (1031±87?µatm CO2, SH) and fluctuating high CO2 (1012±244?µatm CO2, FH) for 400 generations. We find that although both fluctuation and high CO2 drive evolution, FH-evolved lineages are smaller, have reduced C:N ratios and respond more strongly to further increases in CO2 than do SH-evolved lineages. This indicates that environmental fluctuations are an important factor to consider when predicting how the characteristics of future phytoplankton populations will have an impact on biogeochemical cycles and higher trophic levels in marine food webs.

Adaptive evolution of a key phytoplankton species to ocean acidification

Ocean acidification, the drop in seawater pH associated with the ongoing enrichment of marine waters with carbon dioxide from fossil fuel burning, may seriously impair marine calcifying organisms. Our present understanding of the sensitivity of marine life to ocean acidification is based primarily on short-term experiments, in which organisms are exposed to increased concentrations of CO2. However, phytoplankton species with short generation times, in particular, may be able to respond to environmental alterations through adaptive evolution. Here, we examine the ability of the world's single most important calcifying organism, the coccolithophore Emiliania huxleyi, to evolve in response to ocean acidification in two 500-generation selection experiments. Specifically, we exposed E. huxleyi populations founded by single or multiple clones to increased concentrations of CO2. Around 500 asexual generations later we assessed their fitness. Compared with populations kept at ambient CO2 partial pressure, those selected at increased partial pressure exhibited higher growth rates, in both the single- and multiclone experiment, when tested under ocean acidification conditions. Calcification was partly restored: rates were lower under increased CO2 conditions in all cultures, but were up to 50% higher in adapted compared with non-adapted cultures. We suggest that contemporary evolution could help to maintain the functionality of microbial processes at the base of marine food webs in the face of global change.

SeaWiFS (version R2010.0) Monthly Global 9 km Microplankton Fraction, links to NetCDF files

Phytoplankton cell size is important to biogeochemical and food web processes. The goal of this study is to estimate phytoplankton cell size distribution from satellite imagery of spectral remote sensing reflectance (Rrs(lambda)). Previous studies have indicated phytoplankton size classes have distinctive absorption spectra despite the physiological and taxonomic variability within an assemblage. For this study, the chlorophyll specific absorption spectra for phytoplankton size class extremes, pico- and microphytoplankton, are weighted by the percent microplankton (Sfm) and are the basis of phytoplankton size retrieval from SeaWiFS imagery. Satellite retrievals of Sfm are done through implementation of a forward optical model look-up table (LUT) that incorporates the range of absorption and scattering variability due to phytoplankton size, chlorophyll concentration ([Chl]) and dissolved and detrital matter (acdm(443)) in the global ocean from which Rrs(lambda) is calculated by the radiative transfer software, Hydrolight. The Hydrolight modeled Rrs(lambda) options for a given combination of [Chl] and acdm(443) within the LUT vary only due to Sfm. For a given pixel, the LUT search space was limited by satellite imagery of [Chl] and acdm(443). Within the narrowed search space, SeaWiFS Rrs(lambda) was matched with the closest LUT Rrs(lambda) option and the associated Sfm was assigned. Thresholds at which changes in Rrs(lambda) due to Sfm could be discerned were established in terms of [Chl] and acdm(443). In situ high-precision liquid chromatography-derived estimates of cell size are used in conjunction with matched daily satellite estimates of Sfm for validation and agree well. A single month is displayed as an example of the Sfm retrieval.

Interactive effects of ocean acidification and nitrogen limitation on the diatom Phaeodactylum tricornutum

Climate change is expected to bring about alterations in the marine physical and chemical environment that will induce changes in the concentration of dissolved CO2 and in nutrient availability. These in turn are expected to affect the physiological performance of phytoplankton. In order to learn how phytoplankton respond to the predicted scenario of increased CO2 and decreased nitrogen in the surface mixed layer, we investigated the diatom Phaeodactylum tricornutum as a model organism. The cells were cultured in both low CO2 (390 µatm) and high CO2 (1000 µatm) conditions at limiting (10 µmol/L) or enriched (110 µmol/L) nitrate concentrations. Our study shows that nitrogen limitation resulted in significant decreases in cell size, pigmentation, growth rate and effective quantum yield of Phaeodactylum tricornutum, but these parameters were not affected by enhanced dissolved CO2 and lowered pH. However, increased CO2 concentration induced higher rETRmax and higher dark respiration rates and decreased the CO2 or dissolved inorganic carbon (DIC) affinity for electron transfer (shown by higher values for K1/2 DIC or K1/2 CO2). Furthermore, the elemental stoichiometry (carbon to nitrogen ratio) was raised under high CO2 conditions in both nitrogen limited and nitrogen replete conditions, with the ratio in the high CO2 and low nitrate grown cells being higher by 45% compared to that in the low CO2 and nitrate replete grown ones. Our results suggest that while nitrogen limitation had a greater effect than ocean acidification, the combined effects of both factors could act synergistically to affect marine diatoms and related biogeochemical cycles in future oceans.

Seawater carbonate chemistry and Emiliania huxleyi biological processes during experiments, 2010

The calcifying phytoplankton species, coccolithophores, have their calcified coccoliths around the cells, however, their physiological roles are still unknown. Here, we hypothesized that the coccoliths may play a certain role in reducing solar UV radiation (UVR, 280-400 nm) and protect the cells from being harmed. Cells of Emiliania huxleyi with different thicknesses of the coccoliths were obtained by culturing them at different levels of dissolved inorganic carbon and their photophysiological responses to UVR were investigated. Although increased dissolved inorganic carbon decreased the specific growth rate, the increased coccolith thickness significantly ameliorated the photoinhibition of PSII photochemical efficiency caused by UVR. Increase by 91% in the coccolith thickness led to 35% increase of the PSII yield and 22% decrease of the photoinhibition of the effective quantum yield by UVR. The coccolith cover reduced more UVA (320-400 nm) than UVB (280-315 nm), leading to less inhibition per energy at the UV-A band.

Coccosphere geometry measurements from culture experiments on the coccolithophore species Calcidiscus leptoporus, Calcidiscus quadriperforatus and Helicosphaera carteri

Coccolithophores are a key phytoplankton group that exhibit remarkable diversity in their biology, ecology, and calcitic exoskeletons (coccospheres). An understanding of the physiological processes that underpin coccosphere architecture is essential for maximizing the information that can be retrieved from their extensive fossil record. Using culturing experiments on four modern species from three long-lived families, we investigate how coccosphere architecture responds to population shifts from rapid (exponential) to slowed (stationary) growth phases as nutrients become depleted. These experiments reveal statistical differences in cell size and the number of coccoliths per cell between these two growth phases, specifically that cells in exponential-phase growth are typically smaller with fewer coccoliths, whereas cells experiencing growth-limiting nutrient depletion have larger coccosphere sizes and greater numbers of coccoliths per cell. Although the exact numbers are species-specific, these growth-phase shifts in coccosphere geometry are common to four different coccolithophore families (Calcidiscaceae, Coccolithaceae, Isochrysidaceae, Helicosphaeraceae), demonstrating that this is a core physiological response to nutrient depletion across a representative diversity of this phytoplankton group. Polarised light microscopy was used for all coccosphere geometry measurements.

Mesocosm experiment on warming and acidification effects in 2012: Microzooplankton data

Global warming and ocean acidification are among the most important stressors for aquatic ecosystems in the future. To investigate their direct and indirect effects on a near-natural plankton community, a multiple-stressor approach is needed. Hence, we set up mesocosms in a full-factorial design to study the effects of both warming and high CO2 on a Baltic Sea autumn plankton community, concentrating on the impacts on microzooplankton (MZP). MZP abundance, biomass, and species composition were analysed over the course of the experiment. We observed that warming led to a reduced time-lag between the phytoplankton bloom and an MZP biomass maximum. MZP showed a significantly higher growth rate and an earlier biomass peak in the warm treatments while the biomass maximum was not affected. Increased pCO2 did not result in any significant effects on MZP biomass, growth rate, or species composition irrespective of the temperature, nor did we observe any significant interactions between CO2 and temperature. We attribute this to the high tolerance of this estuarine plankton community to fluctuations in pCO2, often resulting in CO2 concentrations higher than the predicted end-of-century concentration for open oceans. In contrast, warming can be expected to directly affect MZP and strengthen its coupling with phytoplankton by enhancing its grazing pressure.

Seawater carbonate chemistry and biological processes of coccolithophore (Gephyrocapsa oceanica and Coccolithus pelagicus ssp. braarudii) during experiments, 2011

All species of coccolithophore appear to respond to perturbations of carbonate chemistry in a different way. Here, we show that the degree of malformation, growth rate and stable isotopic composition of organic matter and carbonate produced by two contrasting species of coccolithophore (Gephyrocapsa oceanica and Coccolithus pelagicus ssp. braarudii) are indicative of differences between their photosynthetic and calcification response to changing DIC levels (ranging from ~1100 to ~7800 µmol/kg) at constant pH (8.13 ± 0.02). Gephyrocapsa oceanica thrived under all conditions of DIC, showing evidence of increased growth rates at higher DIC, but C. braarudii was detrimentally affected at high DIC showing signs of malformation, and decreased growth rates. The carbon isotopic fractionation into organic matter and the coccoliths suggests that C. braarudii utilises a common internal pool of carbon for calcification and photosynthesis but G. oceanica relies on independent supplies for each process. All coccolithophores appear to utilize bicarbonate as their ultimate source of carbon for calcification resulting in the release of a proton. But, we suggest that this proton can be harnessed to enhance the supply of CO2(aq) for photosynthesis either from a large internal HCO3- pool which acts as a pH buffer (C. braarudii), or pumped externally to aid the diffusive supply of CO2 across the membrane from the abundant HCO3- (G. oceanica), likely mediated by an internal and external carbonic anhydrase respectively. Our simplified hypothetical spectrum of physiologies may provide a context to understand different species response to changing pH and DIC, the species-specific delta p and calcite "vital effects", as well as accounting for geological trends in coccolithophore cell size.

Age model and grain size distribution of the fine fraction of ODP Site 114-704

The physical properties of sediments beneath an upwelling area in the southern part of the Atlantic Ocean (ODP Hole 704A) were investigated. Highly significant correlations characterize the relationship of carbonate content to bulk density (R = 0.85), carbonate content to porosity (R = 0.84), and carbonate content to impedance (R = 0.84). No relationship exists between carbonate content and compressional-wave velocity (R = 0.24), indicating that amplitude variations in impedance are primarily controlled by variations in bulk density, which, in turn, are controlled by climatically driven biogenic opal and carbonate deposition. In general, maxima in impedance correspond to maxima in carbonate content (minima in opal content). The impedance record exhibits its most drastic change at about 2.4 Ma, marking dramatic increases in the average content of biogenic opal and the beginning of large-amplitude fluctuations. Between 0.7 and 0.4 Ma carbonate content, bulk density, and grain density decrease while opal content drastically increases. Similar changes have been observed in sediments beneath an upwelling cell off northwest Africa, indicating an oceanwide enhancement in upwelling or in the calcite corrosiveness of bottom water that appears to be isochronous.

Data base on heterotrophic dinoflagellates grazing and growth rate as a function of size, prey size and type compiled from the literature

Microzooplankton (the 20 to 200 µm size class of zooplankton) is recognised as an important part of marine pelagic ecosystems. In terms of biomass and abundance heterotrophic dinoflagellates are one of the important groups of organism in microzooplankton. However, their rates - grazing and growth - , feeding behaviour and prey preferences are poorly known and understood. A set of data was assembled in order to derive a better understanding of heterotrophic dinoflagellates rates, in response to parameters such as prey concentration, prey type (size and species), temperature and their own size. With these objectives, literature was searched for laboratory experiments with information on one or more of these parameters effect studied. The criteria for selection and inclusion in the database included: (i) controlled laboratory experiment with a known dinoflagellate feeding on a known prey; (ii) presence of ancillary information about experimental conditions, used organisms - cell volume, cell dimensions, and carbon content. Rates and ancillary information were measured in units that meet the experimenter need, creating a need to harmonize the data units after collection. In addition different units can link to different mechanisms (carbon to nutritive quality of the prey, volume to size limits). As a result, grazing rates are thus available as pg C dinoflagellate-1 h-1, µm3 dinoflagellate-1 h-1 and prey cell dinoflagellate-1 h-1; clearance rate was calculated if not given and growth rate is expressed as the growth rate per day.

(Table 1) Statistics of size measurements of Lagenoeca antarctica sp. n. and Salpingoeca abyssalis sp. n. (floating form) sampled during POLARSTERN cruise ANT-XXII/2 and METEOR cruise M63/2

Despite their high abundance and their high importance for the oceanic matter flux, heterotrophic nanoflagellates are only poorly studied in the deep-sea regions. Studies on the choanoflagellate distribution during two deep-sea expeditions, to the South Atlantic (5038 m) and Antarctica (Weddell Sea, 2551 m), revealed the deepest records of choanoflagellates so far. A new species, (Lagenoeca antarctica) with a conspicuous spike structure on the theca is described from deep Antarctic waters. Lagenoeca antarctica sp. n. is a solitary unstalked free living salpingoecid-like choanoflagellate. The protoplast is surrounded by a typical theca with unique spikes only visible in SEM micrographs. The ovoid cell nearly fills the whole theca and ranges in size from 4 to 6 µm. The collar measures 2-3 µm and the flagellum 3-5 µm. A second species, Salpingoeca abyssalis sp. n., was isolated from the abyssal plain of the South Atlantic (5038 m depth). Floating and attached forms were observed. The protoplast ranges from to 2 to 4 µm in length and 1 to 2 µm in width. The collar is about the same length as the protoplast and the flagellum has 2 to 2.5 × the length of the protoplast. Phylogenetic analyses based on a fragment of SSU rDNA revealed Salpingoeca abyssalis to cluster together with a marine isolate of Salpingoeca infusionum while Lagenoeca antarctica clusters separately from the other codonosigid and salpingoecid taxa. Salpingoeca abyssalis and an undetermined Monosiga species seems to be the first choanoflagellate species recorded from the abyssal plain.