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Temperature is one of the environmental factors that most strongly influence the life of microalgae, from the enzymatic level up to the modification of biosynthetic and degradation potential and, t...

Recent studies indicate that aluminum (Al) could play an important role in the ocean carbon cycle by increasing phytoplankton carbon fixation and reducing organic carbon decomposition. However, how Al may influence the decomposition of organic carbon has not yet been explicitly examined. Here we report the effects of Al on carbon fixation by marine diatoms and their subsequent decomposition. By using radiocarbon as a tracer, the carbon fixation and decomposition of three model marine diatoms were examined in Aquil* media at different concentrations (0, 40, 200, and 2000 nM) of dissolved Al. Addition of Al enhanced net carbon fixation by the diatoms in the declining growth phase (by 9%–29% for Thalassiosira pseudonana, 15%–20% for T. oceanica, 15%–23% for T. weissflogii). Under axenic conditions the decomposition rates (d⁻¹) of the diatom-produced particulate organic carbon (POC) significantly decreased (by 21%–57% for T. pseudonana, 0%–41% for T. oceanica, 29%–58% for T. weissflogii) in the Al-enriched treatments. In the presence of bacteria, the decomposition rates of T. weissflogii-produced POC were still 37%–38% lower in Al-enriched treatments compared to the control. Significant increases in cell size, cellular carbon content (pmol C/cell) and cellular carbon density (pmol C/μm³) of T. weissflogii were also observed in the Al-enriched treatments compared to the control. The Al-related increase in net carbon fixation and cell size, and the decrease in POC decomposition rate may facilitate carbon export to ocean depths. The study provides new evidence for the iron–aluminum hypothesis, which suggests that Al could increase phytoplankton uptake of atmospheric CO₂ and influence climate change.

Recent development of DNA sequencing technologies has enabled successful construction of thousands of mitochondrial genomes. Nevertheless, only 33 mitochondrial genomes have been reported for species in Bacillariophyta, which includes harmful algal bloom (HAB) species. In this study, we determined the complete mitogenome of the Bacillariophyta HAB species Odontella regia. For comparison, we also constructed the mitogenome of Lithodesmium undulatum, another Bacillariophyta species. Both strains were isolated in the Jiaozhou Bay, an epitome of China’s coastal ecosystem and an ideal site for HAB research. These two mitogenomes were characterized as 37,617 bp and 37,057 bp circular-mapping molecules with AT content of 73.4% and 75.3%, respectively. The phylogenetic tree based on 32 protein-coding genes of the mitogenome encoded in 35 Bacillariophyta species revealed that the class Mediophyceae consisted of multi-phyletic clades. While O. regia formed an independent clade, L. undulatum was closely related to Thalassiosira pseudonana and Skeletonema marinoi. Synteny comparison of O. regia and L. undulatum mitogenomes and mitogenomes of three closely related species displayed substantial differences among lineages in Mediophyceae by a series of gene translocations and/or inversion with many conserved gene blocks such as rpl2-rps19-rps3-rpl16-atp9. These analyses suggested a complex evolutionary relationship in Mediophyceae in which the mitogenome of O. regia was the least conservative compared with the mitogenomes of four other Bacillariophyta species. Further studies are needed to clarify detailed phylogenetic relationships in Bacillariophyta.

© 2020, Springer Nature Switzerland AG. Temperature plays a fundamental role in determining phytoplankton community structure, distribution, and abundance. With climate models predicting increases in ocean surface temperatures of up to 3.2°C by 2100, there is a genuine need to acquire data on the phenotypic plasticity, and thus performance, of phytoplankton in relation to temperature. We investigated the effects of temperature (14–28°C) on the growth, morphology, productivity, silicification and macromolecular composition of the marine diatom Thalassiosira pseudonana. Optimum growth rate and maximum P:R ratio were obtained around 21°C. Cell volume and chlorophyll a increased with temperature, as did lipids and proteins. One of the strongest temperature-induced shifts was the higher silicification rates at low temperature. Our results reveal temperature-driven responses in physiological, morphological and biochemical traits in T. pseudonana; whereby at supra-optimal temperatures cells grew slower, were larger, had higher chlorophyll and protein content but reduced silicification, while those exposed to sub-optimal temperatures were smaller, heavily silicified with lower lipid and chlorophyll content. If these conserved across species, our findings indicate that as oceans warm, we may see shifts in diatom phenotypes and community structure, with potential biogeochemical consequences of higher remineralisation and declines in carbon and silicon export to the ocean interior.

The marine diatom Thalassiosira pseudonana was grown in continuous culture systems to study the interactive effects of temperature, irradiance, nutrient limitation, and the partial pressure of CO2 (pCO2 ) on its growth and physiological characteristics. The cells were able to grow at all combinations of low and high irradiance (50 and 300 μmol photons · m-2 · s-1 , respectively, of visible light), low and high pCO2 (400 and 1,000 μatm, respectively), nutrient limitation (nitrate-limited and nutrient-replete conditions), and temperatures of 10-32°C. Under nutrient-replete conditions, there was no adverse effect of high pCO2 on growth rates at temperatures of 10-25°C. The response of the cells to high pCO2 was similar at low and high irradiance. At supraoptimal temperatures of 30°C or higher, high pCO2 depressed growth rates at both low and high irradiance. Under nitrate-limited conditions, cells were grown at 38 ± 2.4% of their nutrient-saturated rates at the same temperature, irradiance, and pCO2 . Dark respiration rates consistently removed a higher percentage of production under nitrate-limited versus nutrient-replete conditions. The percentages of production lost to dark respiration were positively correlated with temperature under nitrate-limited conditions, but there was no analogous correlation under nutrient-replete conditions. The results suggest that warmer temperatures and associated more intense thermal stratification of ocean surface waters could lower net photosynthetic rates if the stratification leads to a reduction in the relative growth rates of marine phytoplankton, and at truly supraoptimal temperatures there would likely be a synergistic interaction between the stresses from temperature and high pCO2 (lower pH).

Global warming is altering both phytoplankton-experienced temperature and light-exposure duration through shifting their niches from low to high latitudes. We explored the growth, physiology, and compositions of a smaller Thalassiosira pseudonana and a larger Thalassiosira punctigera, temperate marine centric diatoms, in responses to a matrix of temperatures (12, 15, 18, and 21 °C) and photoperiods (light:dark cycles of 4:20, 8:16, 16:8, and 24:0). Both T. pseudonana and T. punctigera grew faster under medium temperature and longer photoperiod, under the expected optimal instantaneous light intensity. The biovolume-based pigments content of T. pseudonana responded largely to temperature, while that of T. punctigera responded more to photoperiod duration than to temperature. In T. pseudonana, shortest photoperiod enhanced cellular protein content and alleviated their temperature dependency. Continuous growth light reduced the photosynthetic capacity of T. pseudonana at the lowest temperature and reduced that of T. punctigera across temperatures. Moreover, we found the increasing temperature linearly increased the dark respiration rate (Rd) and molar ratio of carbon to nitrogen (C:N) of T. pseudonana but decreased that of T. punctigera, with the scattered effects of photoperiod. Our results demonstrated that responses of diatoms Thalassiosira across photoperiods and temperatures vary with species and possibly with cell size, suggesting that the poleward shift of the niches of phytoplankton in nature might cause a change in community structure.

International audience; Inorganic carbon availability can limit primary productivity and control species composition of freshwater phytoplankton. This is despite the presence of CO2-concentrating mechanisms (CCMs) in some species that maximize inorganic carbon uptake. We investigated the effects of inorganic carbon on the seasonal distribution, growth rates and photosynthesis of a freshwater diatom, Asterionella formosa, and the nature of its CCM using genomics. In a productive lake, the frequency of A. formosa declined with CO2 concentration below air-equilibrium. In contrast, CO2 concentrations at 2.5-times air-equilibrium did not increase growth rate, cell C-quota or the ability to remove inorganic carbon. A pH-drift experiment strongly suggested that HCO3− as well as CO2 could be used. Calculations combining hourly inorganic carbon concentrations in a lake with known CO2 and HCO3− uptake kinetics suggested that rates of photosynthesis of A. formosa would be approximately carbon saturated and largely dependent on CO2 uptake when CO2 was at or above air-equilibrium. However, during summer carbon depletion, HCO3− would be the major form of carbon taken up and carbon saturation will fall to around 30%. Genes encoding proteins involved in CCMs were identified in the nuclear genome of A. formosa. We found carbonic anhydrases from subclasses α, β, γ and θ, as well as solute carriers from families 4 and 26 involved in HCO3− transport, but no periplasmic carbonic anhydrase. A model of the components of the CCM and their location in A. formosa showed that they are more similar to Phaeodactylum tricornutum than to Thalassiosira pseudonana, two marine diatoms.

Estuarine Pettaquamscutt River is a unique habitat 10 km in length with physical and biogeochemical characteristics analogous to a miniature fjord. Its meromictic upper basins are characterized by a permanent oxic–anoxic vertical gradient in which a well-oxygenated upper layer overlies a deeper, anoxic reservoir, with persistent blooms of phototrophic anoxygenic bacteria (Chromatium, Chlorobium) at the oxic–anoxic transition layer. A diverse assemblage of nanoplanktonic, centric diatoms (Cyclotella caspia, Thalassiosira pseudonana, cf. Cyclotella cryptica) dominated the seasonal phytoplankton cycle in the aerobic layer, similar to comparable meromictic habitats elsewhere. This assemblage of nano-centric diatoms appears to be trait-separated from other species clusters and is potentially useful as a functional group flora with ecophysiology diagnostic of marine estuarine rivers and meromictic habitat niche structure. The most conspicuous phytoplankton feature, however, was the year-round occurrence of the photoautotrophic euglenid Euglenaformis (Euglena) proxima, restricted in its upper water column distribution to the O2–H2S boundary layer where it formed a consortium with the photosynthetic green and purple sulfur bacteria community. The association of E. proxima with meromixis, H2S, and phototrophic anoxygenic bacteria is similar to that reported previously in a brackish Norwegian oyster poll and a brackish loch in Scotland.

Although increasing the pCO2 for diatoms will presumably down-regulate the CO2 -concentrating mechanism (CCM) to save energy for growth, different species have been reported to respond differently to ocean acidification (OA). To better understand their growth responses to OA, we acclimated the diatoms Thalassiosira pseudonana, Phaeodactylum tricornutum, and Chaetoceros muelleri to ambient (pCO2 400 μatm, pH 8.1), carbonated (pCO2 800 μatm, pH 8.1), acidified (pCO2 400 μatm, pH 7.8), and OA (pCO2 800 μatm, pH 7.8) conditions and investigated how seawater pCO2 and pH affect their CCMs, photosynthesis, and respiration both individually and jointly. In all three diatoms, carbonation down-regulated the CCMs, while acidification increased both the photosynthetic carbon fixation rate and the fraction of CO2 as the inorganic carbon source. The positive OA effect on photosynthetic carbon fixation was more pronounced in C. muelleri, which had a relatively lower photosynthetic affinity for CO2 , than in either T. pseudonana or P. tricornutum. In response to OA, T. pseudonana increased respiration for active disposal of H+ to maintain its intracellular pH, whereas P. tricornutum and C. muelleri retained their respiration rate but lowered the intracellular pH to maintain the cross-membrane electrochemical gradient for H+ efflux. As the net result of changes in photosynthesis and respiration, growth enhancement to OA of the three diatoms followed the order of C. muelleri > P. tricornutum > T. pseudonana. This study demonstrates that elucidating the separate and joint impacts of increased pCO2 and decreased pH aids the mechanistic understanding of OA effects on diatoms in the future, acidified oceans.

Acidification of the ocean due to high atmospheric CO2 levels may increase the resilience of diatoms causing dramatic shifts in abiotic and biotic cycles with lasting implications on marine ecosystems. Here, we report a potential bioindicator of a shift in the resilience of a coastal and centric model diatom Thalassiosira pseudonana under elevated CO2. Specifically, we have discovered, through EGFP-tagging, a plastid membrane localized putative Na+(K+)/H+ antiporter that is significantly upregulated at >800 ppm CO2, with a potentially important role in maintaining pH homeostasis. Notably, transcript abundance of this antiporter gene was relatively low and constant over the diel cycle under contemporary CO2 conditions. In future acidified oceanic conditions, dramatic oscillation with >10-fold change between nighttime (high) and daytime (low) transcript abundances of the antiporter was associated with increased resilience of T. pseudonana. By analyzing metatranscriptomic data from the Tara Oceans project, we demonstrate that phylogenetically diverse diatoms express homologs of this antiporter across the globe. We propose that the differential between night- and daytime transcript levels of the antiporter could serve as a bioindicator of a shift in the resilience of diatoms in response to high CO2 conditions in marine environments.

As oceans warm, the depth of the upper mixed layer is predicted to decrease, resulting in insufficient nutrient supply and higher solar radiation for phytoplankton. In order to understand the photophysiological responses of the key eukaryotic phytoplankton diatoms to high light and nutrient limitation, we grew two diatoms, Thalassiosira weissflogii and Thalassiosira pseudonana under N starvation conditions and exposed them to high visible light. It showed that the large-sized diatom T. weissflogii can maintain photosynthetic activity for a longer period of time under nitrogen starvation as compared with the small-sized diatom T. pseudonana. The electron transfer reaction was inhibited in both diatoms and the fast closing of reaction centers promoted the development of QB non-reducing PSII centers, thus facilitated the rapid induction of NPQ, however, the induction of NPQ depended on the degree of N starvation. N starvation exacerbated the photoinhibition caused by high light. The smaller-sized T. pseudonana had a higher σi value and was more sensitive to high-light, but its PSII repair rate was also higher. In contrast, T. weissflogii was more tolerant to high light with a lower σi value, but the tolerance was severely reduced under N-starvation. This study provides helpful insight into how climate change variables impact diatom's photosynthetic physiology.

Stressful environmental conditions can induce many different acclimation mechanisms in marine phytoplankton, resulting in a range of changes in their photophysiology. Here we characterize the common photophysiological stress response of the model diatom Thalassiosira pseudonana to ten environmental stressors and identify diagnostic responses to particular stressors. We quantify the magnitude and temporal trajectory of physiological parameters including the functional absorption cross-section of PSII (σPSII ), quantum efficiency of PSII, non-photochemical quenching (NPQ), cell volume, Chl a, and carotenoid (Car) content in response to nutrient starvation (nitrogen (N), phosphorus (P), silicon (Si), and iron (Fe)), changes in temperature, irradiance, pH, and reactive oxygen species (ROS) over 5 time points (0, 2, 6, 24, 72 h). We find changes in conditions: temperature, irradiance, and ROS, often result in the most rapid changes in photophysiological parameters (

Diatom cells utilize a variety of metabolic pathways to cope with internal energy imbalances caused by stressful environmental conditions. In this study, the model diatom species, Thalassiosira pseudonana, was grown in nutrient replete and nitrate (NO3−)- and dissolved silicate (Si)-depleted media at three growth temperatures (4, 17, 28 °C) to determine how nutrient enrichment and temperature affects diatom growth, photosynthetic efficiency, nitrate reductase (NR) enzyme activity, biogenic silica (bSiO2) deposition, and NR gene expression. Growth rates for nutrient-replete cultures were highest at 17 °C. Across all nutrient treatments, the cells grown at 17 °C had an average Fv/Fm of 0.44 ± 0.006, while the cells were grown at 4 °C and 28 °C had an average Fv/Fm of 0.37 ± 0.004 and 0.38 ± 0.01, respectively. Activity of NR was variable across treatments with no significant effect of temperature. The relative expression of the targeted NR gene was, on average, ~ 10 times higher in the 4 °C cultures and ~ 4 times higher in the 28 °C than in the 17 °C cultures, while the activity of the NR enzyme was generally highest in the cultures grown at 17 °C that were enriched with NO3−. Cells grown under nutrient-replete conditions had significantly higher bSiO2 deposition rates at 4 °C than cells grown at 17 and 28 °C. These data support the notion that cold, nutrient-replete conditions lead to increases in diatom silicification and that NR activity may be regulated downstream of mRNA transcription under specific environmental conditions.

International audience; Intertidal benthic diatoms experience a highly variable light regime, which especially challenges these organisms to cope with excess light energy during low tide. Non-photochemical quenching of chlorophyll fluorescence (NPQ) is one of the most rapid mechanisms diatoms possess to dissipate excess energy. Its capacity is mainly defined by the xanthophyll cycle (XC) and Light-Harvesting Complex X (LHCX) proteins. Whereas the XC and its relation to NPQ have been relatively well-studied in both planktonic and benthic diatoms, our current knowledge about LHCX proteins and their potential involvement in NPQ regulation is largely restricted to planktonic diatoms. While recent studies using immuno-blotting have revealed the presence of light regulated LHCX proteins in benthic diatom communities and isolates, nothing is as yet known about the diversity, identity and transcriptional regulation or function of these proteins. We identified LHCX genes in the draft genome of the model benthic diatom Seminavis robusta and followed their transcriptional regulation during a day/night cycle and during exposure to high light conditions. The S. robusta genome contains 17 LHCX sequences, which is much more than in the sequenced planktonic model diatoms (4-5), but similar to the number of LHCX genes in the sea ice associated diatom Fragilariopsis cylindrus. LHCX diversification in both species, however, appears to have occurred independently. Interestingly, the S. robusta genome contains LHCX genes that are related to the LHCX6 of the model centric diatom Thalassiosira pseudonana, which are lacking in the well-studied pennate model diatom Phaeodactylum tricornutum. All investigated LHCX genes, with exception of SrLHCX6, were upregulated during the daily dark-light transition. Exposure to 2,000 µmol photons m −2 s −1 , furthermore, increased transcription of all investigated LHCX genes. Our data suggest that the diversification and involvement of several light regulated LHCX genes in the photophysiology of S. robusta may represent an adaptation to the complex and highly variable light environment this benthic diatom species can be exposed to.

This study investigated the effects of microalgae addition on the water quality, accumulation of Vibrios in the water and sediment, and growth performance of whiteleg shrimp (Litopenaeus vannamei) in the intensive culture system. To evaluate the effects of two monospecific microalgae in whiteleg shrimp culture, an 84-day experiment was performed in concrete tanks seeded with Nannochloropsis oculata and Thalassiosira pseudonana at a density of 10 × 104~80 × 104 cells/mL. The results showed that adding T. pseudonana treatment significantly decreased the levels of nitrite-N, nitrate-N, orthophosphate-P, total ammonia nitrogen and suspended solids during the intermediate and the later culture tank. Additionally, the two added microalgae kept pH steady and increased dissolved oxygen (at daytime) in the culture water. We also found that the concentration of Vibrios in water and sediment from adding microalgae treatments was significantly reduced compared to control groups. Moreover, the highest mean weight gain (1.49 ± 0.056 g/week), the growth rate (1.50 ± 0.067 g/week) and the lowest feed conversion rate (1.42 ± 0.023) were observed in T. pseudonana treatment, which were significantly better than that of the control group (F (2, 6) = 13.77, P = 0.006). Productivity in the treatment of T. pseudonana was 3.65 ± 0.095 kg/m3, which was 25% higher than in the control group (2.92 ± 0.065 kg/m3, F (1, 4) = 143.58, P

The neurotoxins β-N-methylamino-L-alanine (BMAA) and 2,4-diaminobutyric acid (DAB) are produced by cyanobacteria, diatoms and dinoflagellates and have been detected in seafood worldwide. Ou...

Phytoplankton cells living in the surface waters of oceans are experiencing alterations in environmental conditions associated with global change. Given their importance in global primary productivity, it is of considerable concern to know how these organisms will perform physiologically under the changing levels of pH, temperatures, and nutrients predicted for future oceanic ecosystems. Here we show that the model diatom, Thalassiosira pseudonana, when grown at different temperatures (20 or 24 °C), pCO2 (400 or 1000 µatm), and nitrate concentrations (2.5 or 102.5 µmol l−1), displayed contrasting performance in its physiology. Elevated pCO2 (and hence seawater acidification) under the nitrate-limited conditions led to decreases in specific growth rate, cell size, pigment content, photochemical quantum yield of PSII, and photosynthetic carbon fixation. Furthermore, increasing the temperature exacerbated the negative effects of the seawater acidification associated with elevated pCO2 on specific growth rate and chlorophyll content under the N-limited conditions. These results imply that a reduced upward transport of nutrients due to enhanced stratification associated with ocean warming might act synergistically to reduce growth and carbon fixation by diatoms under progressive ocean acidification, with important ramifications for ocean productivity and the strength of the biological CO2 pump.

Triphenyltin (TPT) often coexists with tributyltin (TBT) and Cu in coastal waters worldwide. The combined toxic effect of TPT and TBT has always been assumed to be additive without any scientific proof, and the combined effect of Cu and TPT on marine organisms has not been vigorously studied. This study, therefore, investigated the acute toxicity of binary mixture of TPT/Cu and TPT/TBT to five selected marine species including Thalassiosira pseudonana, Skeletonema costatum, Tigriopus japonicus, Brachionus koreanus and Oryzias melastigma. The interaction between TPT and TBT or Cu was modeled antagonistic based on concentration addition (CA) model, while it was synergistic according to response addition (RA) model. Both model well predicted the toxicity of binary mixtures to the five organisms. As for the environmental risk assessment, CA overestimated the toxicity in most cases and thus is a more conservative model than RA model for assessing the toxicity of these chemical mixtures.

Benthic diatoms are important primary producers in intertidal marine sediments and form the basis of the food web in these ecosystems. In order to investigate the carbon flow within diatom mats, we performed in situ 13C pulse-chase labeling experiments and followed in detail the biochemical fate of carbon fixed by the diatoms for five consecutive days. These labeling experiments were done at approximately 2-monthly intervals during 1 yr in order to cover seasonal variations. The fixed carbon was recovered in individual carbohydrates including extracellular polymeric substances (EPS), amino acids, fatty acids, and nucleic acid bases. In addition, we assessed a variety of environmental parameters and photosynthetic characteristics. The fixed carbon was initially mainly stored as carbohydrate (glucose) while nitrogen-rich compounds (e.g., amino acids and RNA/DNA) were produced more slowly. During the year, the diatoms distributed the photosynthetically fixed carbon differently among the various carbon pools that were measured. In summer, the diatoms decreased carbon fixation and accumulated relatively more lipid as a storage compound (27% ± 2% vs. 12% ± 5% in other seasons). The percentage of fixed carbon that was excreted as EPS was lower in summer compared to other seasons, amounting 9% ± 4% and 21% ± 6%, respectively. Hence, it seemed that the physiology of the microphytobenthos was different during summer and caused by higher light intensity and a shift in nitrogen source.

Acetaldehyde, methanol, acetone, and isoprene are important reactive volatile organic compounds (VOCs) in the oceans that partition to the atmosphere in significant amounts. Reports of potentially high rates of VOC turnover in the North Atlantic suggested that both biotic and abiotic processes are involved. The biological basis for VOC cycling by ocean plankton is unknown, but is potentially important because of VOC contributions to carbon cycle budgets and atmospheric chemistry. We designed dynamic stripping chambers that coupled to a proton transfer reaction mass spectrometer to measure VOC production and consumption by cultured phytoplankton and bacterioplankton. The diatom, Thalassiosira pseudonana, produced acetaldehyde in a light-dependent manner. Acetaldehyde was oxidized by the chemoheterotrophic bacterium, Pelagibacter, at rates that suggest that most acetaldehyde is recycled in the ocean before escaping to the atmosphere. These results show that field observations of acetaldehyde turnover reported previously could be explained by biological activity. Rates of production by phytoplankton cultures of methanol, acetone, and isoprene were also measured. These findings support the conclusion that VOCs are a conduit for carbon transfer directly from phytoplankton to bacterioplankton, with the remainder available for escape to the atmosphere.

We investigated the effects of iron (Fe) limitation on the elemental stoichiometry, silicic acid (Si(OH)4) uptake kinetics and cell morphology in two Southern Ocean diatoms Eucampia antarctica and Proboscia inermis and the temperate diatom Thalassiosira pseudonana. An increase in Fe-stress resulted in reductions in specific growth rate and decreases in cellular nitrogen (N) and carbon (C) content relative to cellular biogenic silica (BSi) in both Southern Ocean diatoms and a reduction in growth rate only for T. pseudonana. Both E. antarctica and P. inermis exhibited an increase in cell volume in response to Fe-limitation resulting in a decrease in the cell surface to volume ratio, while normalization of BSi content to cell surface area suggests these diatoms do not become more heavily silicified under Fe limitation. Kinetic Si(OH)4 uptake experiments performed on all three diatom species show that Si(OH)4 uptake is reduced under Fe-limited conditions. For Southern Ocean diatoms, this was manifested through a decrease in the maximum specific uptake rate of Si(OH)4 (VSi-max), along with a decrease in the half-saturation constant for Si(OH)4 uptake (KSi) under Fe stress for E. antarctica. Our data also show that when normalized to cell surface area, VSi-max of the three diatoms species exhibited a linear relationship with cellular growth rate, and was independent of cell morphological variations. Our results suggest that the morphological adaptations of Southern Ocean diatoms in response to Fe-stress have the potential to affect phytoplankton community dynamics and Si(OH)4 : NO3 uptake and export ratios in Southern Ocean waters.

Phytoplankton in the upper oceans are exposed to changing light levels due to mixing, diurnal solar cycles and weather conditions. Consequently, effects of ocean acidification are superimposed upon responses to variable light levels. We therefore grew a model diatom Thalassiosira pseudonana under either constant or variable light but at the same daily photon dose, with current low (400 μatm, LC) and future high CO2 (1000 μatm, HC) treatments. Variable light, compared with the constant light regime, decreased the growth rate, Chl a, Chl c, and carotenoid contents under both LC and HC conditions. Cells grown under variable light appeared more tolerant of high light as indicated by higher maximum relative electron transport rate and saturation light. Light variation interacted with high CO2/lowered pH to decrease the carbon fixation rate, but increased particulate organic carbon (POC) and particularly nitrogen (PON) per cell, which drove a decrease in C/N ratio, reflecting changes in the efficiency of energy transfer from photo-chemistry to net biomass production. Our results imply that elevated pCO2 under varying light conditions can lead to less primary productivity but more PON per biomass of the diatom, which might improve the food quality of diatoms and thereby influence biogeochemical nitrogen cycles.

Authentic biomarker standards were obtained from algal cultures in an attempt to accurately determine blank C added during sample processing for compound-specific radiocarbon analysis. Emiliania huxleyi and Thalassiosira pseudonana were grown under manipulated Δ14C dissolved inorganic carbon (DIC) levels and chlorophyll a and either alkenones (E. huxleyi) or low molecular weight (LMW) alkanoic acids (T. pseudonana) were isolated from the respective biomass using preparative liquid chromatography (LC), wet chemical techniques or preparative gas chromatography, respectively. DI14C in the seawater medium was determined pre- and post-growth. Biomarker Δ14C values mostly agree within 1σ or 2σ analytical uncertainties. In those cases where biomarker Δ14C values differ significantly, chlorophyll a is up to 104‰ more 14C-depleted than alkenones or LMW alkanoic acids, consistent with a larger LC blank compared to the other purification methods. However, in the majority of experimental setups pre- and post-growth DIC Δ14C values seem to be compromised by an unknown and variable blank C contribution. DIC Δ14C values deviate strongly from the anticipated Δ14C values (by up to ca. 560‰), pre- and post-growth Δ14C values differ significantly (by up to ca. 460‰), and changes are not unidirectional. Accordingly, since the substrate Δ14C value cannot unequivocally be constrained, blank C contributions for the different biomarker purification methods cannot be accurately calculated. This study illustrates the challenges and problems of producing authentic standards that are not readily commercially available and exemplifies how a laborious and time-consuming culturing approach may enter a vicious cycle of blank C contamination hampering accurate blank C determination.

Polybrominated diphenyl ethers (PBDEs), a series of highly persistent organic pollutants (POPs), are ubiquitous in marine ecosystems. As key primary producers, microalgae are of great importance on evaluating the environmental outcome of PBDEs pollution. In this study, the toxic mechanisms of BDE-47 on the marine diatom Thalassiosira pseudonana were evaluated by measuring multiple physiological processes. Three concentrations of BDE-47 (25, 15 and 5 μg L−1) were used along with two controls (blank: no BDE-47 or DMSO; negative control: only DMSO). Experiments lasted 144 h (6 days), in which the actual BDE-47 concentrations, cell densities, nutrient (nitrate and phosphate) uptake, pigment compositions, photosynthetic physiology, cell morphology and cellular contents (organic carbon and nitrogen) were measured at 12–48 h intervals. The toxic mechanisms of BDE-47 on T. pseudonana cells were evaluated by measuring multiple physiological processes including photosynthesis, nutrient uptake, cellular material synthesis and cell cycle progressions. The cell divisions of T. pseudonana were severely inhibited by the stress of BDE-47, but the photosynthetic parameters were much less declined and recovered earlier than the cell divisions in the same BDE-47 treatments. The unsuppressed uptake rates of nutrients, increased cell volume and cellular contents indicated the cellular material synthesis proceeded normally. Finally, we found that the cell cycle was arrested in G2/M phase under the stress of BDE-47, we thus concluded that the inhibition of cell divisions by BDE-47 was not due to the lack of energy or cellular materials, where the cell cycle arrest happened; this might be the most important toxicological outcome.

Light drives phytoplankton productivity, so phytoplankton must exploit variable intensities and durations of light exposure, depending upon season, latitude, and depth. We analyzed the growth, photophysiology and composition of small, Thalassiosira pseudonana, and large, Thalassiosira punctigera, centric diatoms from temperate, coastal marine habitats, responding to a matrix of photoperiods and growth light intensities. T. pseudonana showed fastest growth rates under long photoperiods and low to moderate light intensities, while the larger T. punctigera showed fastest growth rates under short photoperiods and higher light intensities. Photosystem II function and content responded primarily to instantaneous growth light intensities during the photoperiod, while diel carbon fixation and RUBISCO content responded more to photoperiod duration than to instantaneous light intensity. Changing photoperiods caused species-specific changes in the responses of photochemical yield (e- /photon) to growth light intensity. These photophysiological variables showed complex responses to photoperiod and to growth light intensity. Growth rate also showed complex responses to photoperiod and growth light intensity. But these complex responses resolved into a close relation between growth rate and the cumulative daily generation of reductant, across the matrix of photoperiods and light intensities.

Ocean acidification due to atmospheric CO2 rise is expected to influence marine phytoplankton. Diatoms are responsible for about 40% of the total primary production in the ocean. In order to investigate the physiological response of marine diatom Thalassiosira pseudonana to ocean acidification, we grew the cells under ambient CO2 level (380 µatm) versus the elevated CO2 level (800 µatm) at a light level of 180 µmol m−2 s−1 for 30 generations. Our results showed that the elevated CO2 concentration caused a decrease of the effective photochemical efficiency of PSII $$\left( {{{F_{{\text{v}}}^{\prime } } \mathord{\left/ {\vphantom {{F_{{\text{v}}}^{\prime } } {F_{{\text{m}}}^{\prime } }}} \right. \kern-\nulldelimiterspace} {F_{{\text{m}}}^{\prime } }}} \right)$$ and increase of the dark respiration in T. pseudonana. The intracellular carbonic anhydrase activity was suppressed and the photosynthetic affinity for CO2 was lowered in the high CO2-grown cells, reflecting a downregulation of the CO2 concentrating mechanism (CCM). PSI activity was enhanced to support an increase in ATP synthesis by cyclic electron transfer as required for transport of inorganic carbon and regulation of intracellular pH. The energetic benefit from the downregulation of CCM to growth as reported in other diatom species was not observed here in T. pseudonana.

The diurnal solar cycle is the single most common influential variable for algae growing in a natural environment. In spite of its importance, relatively few laboratory studies have been directed towards photophysiological responses to dynamic irradiance regimes. Further, most of these studies have utilized optically thin cultures. However, phycological technologies utilize optically dense cultures grown in depths measured by centimeters. In this case, algae are harvested under self-shading conditions (all usable incident irradiance being absorbed), and the response of the entire water column to the dynamic irradiance regime is of interest. Thus, the area integrated under some modeled curve for a dynamic irradiance regime becomes the independent variable for the photobiological response being measured of a water column in laboratory studies, and this is important. But what curve should be used to model a dynamic irradiance regime? We suggest that a Gaussian (normal) distribution be used to model this dynamic irradiance because of its well-known statistical attributes. In this study, Thalassiosira pseudonana (marine diatom) was grown under shade-limited growth conditions in a 20-cm deep water column with a dynamic irradiance regime using our SolarStat™. A Gaussian distribution modeled this regime having an 11-h day length (standard deviation ±1.85 h) with an irradiance maximum of 1900 μmol photons m−2 s−1. The daily productivity of batch culture in the linear phase of the light-controlled growth dynamic is compared with those of semi-continuous cultures maintained at 1.0, 0.5, and 0.25 doublings day−1.

Marine dissolved organic matter (DOM) is a key source of carbon and nutrients to microbial life in the oceans, but rapid biological utilization of labile DOM confounds its compositional characterization. In order to characterize potentially bioavailable DOM produced by phytoplankton, DOM from axenic cultures of Thalassiosira pseudonana cultivated in phosphorus (P) replete and low P conditions was extracted using high-recovery electrodialysis (ED) techniques, which resulted in an average dissolved organic carbon (DOC) recovery of 76% ± 7% from all cultures. Low P concentrations resulted in greater cell-normalized production of DOC relative to P replete culture controls at the same growth phase. Despite the different nutrient conditions, DOC composition and DOM molar ratios of carbon to nitrogen (C : N) were similar in all cultures. In contrast, low P concentrations influenced DOM molar carbon to phosphorus (C : P) ratios and dissolved organic phosphorus (DOP) composition. Under P replete and low P conditions, DOM C : P ratios were 130 (± 22) and 2446 (± 519), respectively. 31P Nuclear Magnetic Resonance (NMR) spectroscopy identified P esters (> 90% of DOP) as the dominant P species in DOM produced under P replete conditions, with small or negligible contributions from phosphonates or glycerol P and polyphosphates. However, based on direct fluorometric analysis, DOP from low P cultures was greater than 8 times enriched in dissolved polyphosphate compared to DOP from replete cultures, which is consistent with the growing evidence that polyphosphate is a dynamic component of total P in low P ocean regions.

There is relatively little choice in cultivation methods for growing algae outdoors, either in open pond systems or closed photobioreactors—as batch, continuous, or semi-continuous culture. Algal batch culture grown in a nutrient replete environment with adequate sunlight will become self-shaded with sufficient cell density and enter a stage in the growth dynamic known as the “phase of linear growth.” It is during this phase of linear growth that primary production is at maximum and that the highest biomass is harvested. The inherent problem with batch culture is that the exponential (and possibly lag) phases necessary to achieve densities required prior to the phase of linear growth consume time and waste surface area, and thereby make this an inefficient method to grow algae. Semi-continuous culture can be forced into shade-limiting conditions by reducing growth rate from maximum through dilution, whereby phases of lag and exponential growth are skipped, and culture growth is put into a state similar to a perpetual phase of linear growth with an appropriate culture harvest/dilution cycle. Importantly, semi-continuous culture can increase net growth efficiency over batch culture when compared by shade-limited growth rate. However, scientific study and theory covering shade-limited algal growth under semi-continuous culture conditions are nearly non-existent, which currently makes its application to phycological technologies impractical through “hit and miss” strategies. This laboratory study compares shade-limited growth dynamics for batch and semi-continuous cultures of Thalassiosira pseudonana (small-sized, marine diatom). Theory for optimizing production of mass algal culture with semi-continuous culture technique through cycle period and harvest volume is developed, and guidelines to practical industrial applications are provided.

Diatoms are important primary producers in the ocean that thrive in diverse and dynamic environments. Their survival and success over changing conditions depend on the complex coordination of gene regulatory processes. Here we present an integrated analysis of all publicly available microarray data for the diatoms Thalassiosira pseudonana and Phaeodactylum tricornutum. This resource includes shared expression patterns, gene functions, and cis-regulatory DNA sequence motifs in each species that are statistically coordinated over many experiments. These data illustrate the coordination of transcriptional responses in diatoms over changing environmental conditions. Responses to silicic acid depletion segregate into multiple distinctly regulated groups of genes, regulation by heat shock transcription factors (HSFs) is implicated in the response to nitrate stress, and distinctly coordinated carbon concentrating, CO2 and pH-related responses are apparent. Fundamental features of diatom physiology are similarly coordinated between two distantly related diatom species, including the regulation of photosynthesis, cellular growth functions and lipid metabolism. These integrated data and analyses can be explored publicly (http://networks.systemsbiology.net/diatom-portal/).

Previous field observations suggest that the composition of intracellular polyamine pools in phytoplankton determines the profile of polyamines released to the surrounding environ- ment; thus, knowing how these pools vary among species and in response to factors affecting phytoplankton growth provides a basis for understanding fluctuations in dissolved polyamines. Our analyses of the polyamine content of axenic marine phytoplankton cultures show that intra- cellular polyamine pools are large (100s to 1000s µmol l �1 of biovolume) and that putrescine and spermidine are the major compounds present; however, composition varied with species. Norsper- midine and norspermine were the dominant compounds found in the diatom Thalassiosira pseudonana and the dinoflagellate Amphidinium carterae, respectively. We explored the effects of temperature, salinity and nutrient limitation on polyamine pools in T. pseudonana, and found that either increasing temperature or decreasing salinity increased polyamine concentrations in cells and in the growth medium. Nutrient (N, P or Si) limitation caused reductions of intracellular polyamine concentrations, but release was enhanced under N or Si limitation. Polyamine ratios in the media were not the same as in intracellular pools, suggesting selective release or uptake of polyamines by phytoplankton. Thus, the composition of the dissolved polyamine pool in seawater can vary significantly and on short time scales, depending on phytoplankton community composi- tion and environmental factors affecting phytoplankton physiology. Our work provides experi- mental verification that biological mechanisms support inferences derived from environmental correlations about the factors controlling polyamine distributions in the sea.

The nutritional value of eight species of microalgae for larvae and early post-set juveniles of the Pacific geoduck clam, Panopea generosa , was evaluated. The microalgae tested included two prymnesiophytes, Pavlova lutheri (PL) and Tisochrysis lutea (TL); two chlorophytes, Dunaliella tertiolecta (DT) and Tetraselmis suecica (TS); and four diatoms, Chaetoceros calcitrans (CC), Chaetoceros muelleri (CM), Phaeodactylum tricornutum (PT), and Thalassiosira pseudonana (TP). These microalgae were first tested as mono-species diets on both larvae and early post-set juveniles. The resultant best-performing mono-species diet was then tested as the major component in mixed- (bi-, tri-, and tetra-) species diets. Further experiments examined the possibility that the best-performing mixed-species diet could be improved by adding another algal species. The optimal diet, with the fewest number of algal species to support the best growth, was the bi-species diets of CC + TL for larvae and CM + TL for early post-set juveniles. The fatty-acid profiles of juveniles were influenced by the diets. Although no clear pattern between the level of any particular nutrient and growth and development of juveniles was evident, it appears that a balanced mixture of various dietary nutrients is important. Attention was given to the ratios between n − 3 and n − 6 fatty acids and between eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA). The best-performing diets, including the optimal one, had a n − 3/n − 6 ratio of 2.17–3.03 and an EPA/DHA ratio of 1.28–3.25 for larvae and 1.84–2.58 and 1.84–2.63, respectively, for early post-set juveniles. Results of this study can be applied to improve the commercial hatchery production of geoduck seed. Statement of relevance Nutritional value of microalgae for young geoduck clams.

Polybrominated diphenyl ethers (PBDEs) are ubiquitously distributed persistent organic pollutants (POPs) in marine environments. Phytoplankton are the entrance of PBDEs entering to biotic environments from abiotic environments, while the responding mechanisms of phytoplankton to PBDEs have not been full established. Therefore, we chose the model diatom Thalassiosira pseudonana in this study, by integrating whole transcriptome analysis with physiological-biochemical data, to reveal the molecular responding mechanisms of T. pseudonana to the toxicity of BDE-47. Our results indicated the changes of genes expressions correlated to the physiological-biochemical changes, and there were multiple molecular mechanisms of T. pseudonana responding to the toxicity of BDE-47: Gene expressions evidence explained the suppression of light reaction and proved the occurrence of cellular oxidative stress; In the meanwhile, up-regulations of genes in pathways involving carbon metabolisms happened, including the Calvin cycle, glycolysis, TCA cycle, fatty acid synthesis, and triacylglycerol synthesis; Lastly, DNA damage was found and three outcome including DNA repair, cell cycle arrest and programmed cell death (PCD) happened, which could finally inhibit the cell division and population growth of T. pseudonana. This study presented the most complete molecular responding mechanisms of phytoplankton cells to PBDEs, and provided valuable information of various PBDEs-sensitive genes with multiple functions for further research involving organic pollutants and phytoplankton.

An in vitro urease enzyme assay was developed for the marine diatoms Thalassiosira pseudonana Hasle et Heimdal (clone 3H) and T. weissflogii (Grunow) Fryxell et Hasle (clone Actin). This assay involves the colorimetric measurement of ammonium following the hydrolysis of urea in crude cell homogenates and it is the first assay to account for the rate of nitrogen assimilation in both species grown on urea as the sole nitrogen source. Urease activity was found to be present regardless of nitrogen source, although activities showed distinctly different patterns depending on the species examined and form of nitrogen supplied. Under nitrogen-replete conditions, urease activity in T. pseudonana was present constitutively when grown on NH 4 1 and upregulated when grown on NO 3 2 or urea. In nitrogen-replete T. weissflogii , urease activity was present at high constitutive levels regardless of the nitrogen source and showed no upregulation. Nitrogen starvation did not upregulate activity in either species.

Secondary metabolites produced by bacteria, fungi, algae and plants could affect the growth and development of biological and agricultural systems. This natural process that occurs worldwide is known as allelopathy. The main goal of this work was to investigate the influence of metabolites obtained from phytoplankton monocultures on the growth of green algae Chlorella vulgaris. We selected 6 species occurring in the Baltic Sea from 3 different taxonomic groups: cyanobacteria (Aphanizomenon flos-aquae; Planktothrix agardhii), diatoms (Thalassiosira pseudonana; Chaetoceros wighamii) and dinoflagellates (Alexandrium ostenfeldii; Prorocentrum minimum). In this study we have demonstrated that some of selected organisms caused allelopathic effects against microalgae. Both the negative and positive effects of collected cell-free filtrates on C. vulgaris growth, chlorophyll a concentration and fluorescence parameters (OJIP, QY, NPQ) have been observed. No evidence has been found for the impact on morphology and viability of C. vulgaris cells.

Information on the nutritional requirements and preferred diets of sea cucumber (Holothuriidae) larvae is extremely limited and this has hindered development of hatchery culture methods. This study assessed ingestion and digestion of two live (TISO and Chaetoceros muelleri ) and six concentrated microalgae (Instant Algae®, Reed Mariculture Inc.) by sandfish ( Holothuria scabra ) auricularia larvae of different ages using epifluorescence microscopy. This is the first study to report the use of epifluorescence microscopy with larval echinoderms and experiments were conducted using 2, 6 and 10 day old auricularia larvae. Seven of the eight microalgae tested were ingested and digested by the larvae with digestion occurring more rapidly in older larvae. C. muelleri was rapidly digested by 6-day and 10-day old larvae but our results indicate that C. muelleri is unsuitable as a food for 2-day old sandfish larvae. TISO was well ingested by sandfish larvae in both live and concentrated forms and live TISO was the most suitable of the microalgae tested in terms of ingestion and digestibility. All commercially available microalgae concentrates tested were readily ingested and digested by H. scabra larvae with the exception of Thalassiosira pseudonana (3H 1800®) which was not ingested by larvae of any of the three ages tested. Our results show potential for using microalgae concentrates as alternatives to live microalgae in hatchery culture of sandfish. However, further research should be conducted to assess the relative nutritional values of digestible microalgae as a basis for optimising a diet for hatchery culture of sandfish, and to provide further information on the nutritional requirements of sandfish larvae.

Blooms of centric diatoms are a common feature in the Bilbao estuary during summer when river flow is at its lowest and water temperature is above 20ºC. To gain insight into the specific composition of these diatom blooms, net samples and cultures of estuarine isolates were analysed under the scanning electron microscope (SEM) and by molecular analyses of the Internal Transcribed Spacers 1 and 2 plus the coding region 5.8S (ITS region) and the 28S rRNA gene. Seven species of solitary centric diatoms belonging to four genera were found in the estuary including: Conticribra weissflogii, Cyclotella atomus var. atomus, Cyclotella cryptica, Cyclotella marina, Cyclotella meneghiniana, Discostella pseudostelligera and Thalassiosira pseudonana. Dominant species during blooms were C. meneghiniana and Co. weissflogii in the upper estuary and D. pseudostelligera and T. pseudonana in the middle estuary. The morphological traits used to differentiate between species pairs of similar morphology (C. meneghiniana/C. cryptica or D. pseudostelligera/D. woltereckii) were observed to vary with environmental conditions, denoting a great deal of phenotypic plasticity which would hinder accurate identification of the species when using morphological approaches alone.

Carbon dioxide (CO2) is the primary substrate for photosynthesis by the phytoplankton that form the base of the marine food web and mediate biogeochemical cycling of C and nutrient elements. Specific growth rate and elemental composition (C:N:P) were characterized for 7 cosmopolitan coastal and oceanic phytoplankton species (5 diatoms and 2 chlorophytes) using low density, nutrient-replete, semi-continuous culture experiments in which CO2 was manipulated to 4 levels ranging from post-bloom/glacial maxima (2900 ppm). Specific growth rates at high CO2 were from 19 to 60% higher than in low CO2 treatments in 4 species and 44% lower in 1 species; there was no significant change in 2 species. Higher CO2 availability also resulted in elevated C:P and N:P molar ratios in Thalassiosira pseudonana (~60 to 90% higher), lower C:P and N:P molar ratios in 3 species (~20 to 50% lower), and no change in 3 species. Carbonate system-driven changes in growth rate did not necessarily result in changes in elemental composition, or vice versa. In a subset of 4 species for which fatty acid composition was examined, elevated CO2 did not affect the contribution of polyunsaturated fatty acids to total fatty acids significantly. These species show relatively little sensitivity between present day CO2 and predicted ocean acidification scenarios (year 2100). The results, however, demonstrate that CO2 availability at environmentally and geologically relevant scales can result in large changes in phytoplankton physiology, with potentially large feedbacks to ocean biogeochemical cycles and ecosystem structure.

Due to the ongoing effects of climate change, phytoplankton are likely to experience enhanced irradiance, more reduced nitrogen, and increased water acidity in the future ocean. Here, we used Thalassiosira pseudonana as a model organism to examine how phytoplankton adjust energy production and expenditure to cope with these multiple, interrelated environmental factors. Following acclimation to a matrix of irradiance, nitrogen source, and CO2 levels, the diatom's energy production and expenditures were quantified and incorporated into an energetic budget to predict how photosynthesis was affected by growth conditions. Increased light intensity and a shift from NO3− to NH4+ led to increased energy generation, through higher rates of light capture at high light and greater investment in photosynthetic proteins when grown on NH4+. Secondary energetic expenditures were adjusted modestly at different culture conditions, except that NO3− utilization was systematically reduced by increasing pCO2. The subsequent changes in element stoichiometry, biochemical composition, and release of dissolved organic compounds may have important implications for marine biogeochemical cycles. The predicted effects of changing environmental conditions on photosynthesis, made using an energetic budget, were in good agreement with observations at low light, when energy is clearly limiting, but the energetic budget over-predicts the response to NH4+ at high light, which might be due to relief of energetic limitations and/or increased percentage of inactive photosystem II at high light. Taken together, our study demonstrates that energetic budgets offered significant insight into the response of phytoplankton energy metabolism to the changing environment and did a reasonable job predicting them.

The increase in dimethylsulfoniopropionate (DMSP) biosynthesis measured in several nitrogen-limited marine algae may partially replace other less dense organic solutes such as glycine betaine (GBT). This raises the possibility that phytoplankton could synthesize the denser organic solute DMSP in low nutrient oceanic surface waters facilitating sinking into zones of the euphotic zone richer in nutrients and maximizing their growth, hereafter referred to the “DMSP-ballast” hypothesis. The objective of this study was to test the DMSP-ballast hypothesis by modeling the sinking rates of the diatom Thalassiosira pseudonana and two strains of the coccolithophore Emiliania huxleyi as a function of DMSP synthesis in nitrogen-limited conditions. We also explored the potential ballast effect of DMSP in the positively buoyant non-motile algal species Ethmodiscus rex and Pyrocystis noctiluca. The model results suggest that replacement of trimethylammonium and GBT by DMSP in the naked E. huxleyi strain and T. pseudonana grown under nitrogen limitation could increase the sinking rate by 1 – 22%; while a putative increase in DMSP synthesis (in the millimolar range) in E. rex and P. noctiluca could decrease the rising rate by 43% to several orders of magnitude. The present study suggests a potential ballast role for DMSP in phytoplankton.

The health beneficial omega-3 long-chain polyunsaturated fatty acids (LC-PUFAs), eicosapentaenoic acid (EPA), and docosahexaenoic acid (DHA) are naturally synthesized by diatoms through consecutive steps of fatty acid elongase and desaturase enzymes. In Thalassiosira pseudonana, these fatty acids constitute about 10–20 % of the total fatty acids, with EPA accumulation being five to ten times higher than DHA. In order to identify the subcellular localization of enzymes in the pathway of LC-PUFA biosynthesis in T. pseudonana and to manipulate the production of EPA and DHA, we generated constructs for overexpressing each of the T. pseudonana long-chain fatty acid elongase genes. Full-length proteins were fused to GFP, and transgenic lines were generated. In addition, overexpressed native proteins with no GFP fusion were tested. The subcellular localization of each elongase protein was determined. We then examined the total amount of lipids and analyzed the fatty acid profile in each of the transgenic lines compared to wild type. Lines with overexpressed elongases showed an increase of up to 1.4-fold in EPA and up to 4.5-fold in DHA, and the type of fatty acid that was increased (EPA vs. DHA) depended on the type of elongase that was overexpressed. This data informs future metabolic engineering approaches to further improve EPA and DHA content in diatoms.

The model diatomThalassiosira pseudonanais believed to be a single species with a global distribution, but it has not been confirmed previously whether isolates from different environmental and geographic origins are genotypically and phenotypically identical. In the present study, a polyphasic approach was employed to characterize nine clonal isolates, plus an additional replicate of one of the isolates, of the diatomT. pseudonanafrom culture collections to investigate whether there was any cryptic speciation in the publicly available strains of this species. Morphological analysis using scanning electron microscopy concluded that the strains were indistinguishable. Furthermore, conventional DNA barcoding genes (SSU rDNA, ITS1 and ITS2 rDNA and rbcL), revealed no nucleotide variation among the strains tested. On employing a whole genome fingerprinting technique, Amplified Fragment Length Polymorphism (AFLP), three clusters were revealed, although the level of variation between the clusters was surprisingly low. These findings indicate a low level of diversity among these culturedT. pseudonanastrains, despite their wide spatial and temporal distribution and the salinity range of their original habitats. Based on the limited number of available strains, this suggests thatT. pseudonanais a highly conserved diatom that nevertheless has an ability to tolerate wide ranges of salinity and populate varied geographic locations.

Diatoms are one of the most important groups of phytoplankton in terms of abundance and ecological functionality in the ocean. They usually dominate the phytoplankton communities in coastal waters and experience frequent and large fluctuations in light. In order to evaluate the combined effects of ocean warming and acidification on the diatom's exploitation of variable light environments, we grew a globally abundant diatom Thalassiosira pseudonana under two levels of temperature (18, 24 °C) and pCO2 (400, 1000 μatm) to examine its physiological performance after light challenge. It showed that the higher temperature increased the photoinactivation rate in T. pseudonana at 400 μatm pCO2, while the higher pCO2 alleviated the negative effect of the higher temperature on PSII photoinactivation. Higher pCO2 stimulated much faster PsbA removal, but it still lagged behind the photoinactivation of PSII under high light. Although the sustained phase of nonphotochemical quenching (NPQs) and activity of superoxide dismutase (SOD) were provoked during the high light exposure in T. pseudonana under the combined pCO2 and temperature conditions, it could not offset the damage caused by these multiple environmental changes, leading to decreased maximum photochemical yield.

Diatoms are one of the key phytoplankton groups in the ocean, forming vast oceanic blooms and playing a significant part in global primary production. To shed light on the role of redox metabolism in diatom's acclimation to light-dark transition and its interplay with cell fate regulation, we generated transgenic lines of the diatom Thalassiosira pseudonana that express the redox-sensitive green fluorescent protein targeted to various subcellular organelles. We detected organelle-specific redox patterns in response to oxidative stress, indicating compartmentalized antioxidant capacities. Monitoring the GSH redox potential (EGSH ) in the chloroplast over diurnal cycles revealed distinct rhythmic patterns. Intriguingly, in the dark, cells exhibited reduced basal chloroplast EGSH but higher sensitivity to oxidative stress than cells in the light. This dark-dependent sensitivity to oxidative stress was a result of a depleted pool of reduced glutathione which accumulated during the light period. Interestingly, reduction in the chloroplast EGSH was observed in the light phase prior to the transition to darkness, suggesting an anticipatory phase. Rapid chloroplast EGSH re-oxidation was observed upon re-illumination, signifying an induction of an oxidative signaling during transition to light that may regulate downstream metabolic processes. Since light-dark transitions can dictate metabolic capabilities and susceptibility to a range of environmental stress conditions, deepening our understanding of the molecular components mediating the light-dependent redox signals may provide novel insights into cell fate regulation and its impact on oceanic bloom successions.

The fractionation of silicon (Si) isotopes was measured in two Southern Ocean diatoms (Proboscia inermis and Eucampia Antarctica) and a coastal diatom (Thalassiosira pseudonana) that were grown under varying iron (Fe) concentrations. Varying Fe concentrations had no effect on the Si isotope enrichment factor (ε) in T. pseudonana, whilst E. Antarctica and P. inermis exhibited significant variations in the value of ε between Fe-replete and Fe-limited conditions. Mean ε values in P. inermis and E. Antarctica decreased from (± 1SD) −1.11 ± 0.15‰ and −1.42 ± 0.41 ‰ (respectively) under Fe-replete conditions, to −1.38 ± 0.27 ‰ and −1.57 ± 0.5 ‰ (respectively) under Fe-limiting conditions. These variations likely arise from adaptations in diatoms arising from the nutrient status of their environment. T. pseudonana is a coastal clone typically accustomed to low Si but high Fe conditions whereas E. Antarctica and P. inermis are typically accustomed to High Si, High nitrate low Fe conditions. Growth induced variations in silicic acid (Si(OH)4) uptake arising from Fe-limitation is the likely mechanism leading to Si-isotope variability in E. Antarctica and P. inermis. The multiplicative effects of species diversity and resource limitation (e.g., Fe) on Si-isotope fractionation in diatoms can potentially alter the Si-isotope composition of diatom opal in diatamaceous sediments and sea surface Si(OH)4. This work highlights the need for further in vitro studies into intracellular mechanisms involved in Si(OH)4 uptake, and the associated pathways for Si-isotope fractionation in diatoms.

We hypothesize that, in algae, carbon allocation to lipids depends on the combined effects of the metabolic constraints imposed by the genotype (i.e., species-specific differences in composition) and on the acclimation responses (phenotype) to changes in the stoichiometry of available inorganic carbon (for photosynthesis) and nitrogen (primarily for amino acids, protein, and nucleic acid synthesis). We thus cultured three taxonomically distinct algae, the green alga Dunaliella salina, the diatom Thalassiosira pseudonana, and the dinoflagellate Protoceratium reticulatum, at four nitrate concentrations and constant inorganic carbon. Since energy availability also directly impacts carbon partitioning, we studied the effect of irradiance on the oil quality of P. reticulatum. We used Fourier transform infrared (FTIR) spectroscopy to study carbon allocation and biomass reduction level and gas chromatography for fatty acid analysis. The fatty acid complements of the three species were different; within each species, growth conditions substantially altered oil quality. We ranked the oils in terms of their suitability as biodiesels, using international standards as reference. We believe that this approach may help to identify the appropriate combination of taxa and culture conditions for algal biodiesel production and in general offers insight on carbon allocation to fatty acids.

With progressive future global change, marine phytoplankton in surface oceans will be subjected to ocean acidification, as well as to increased solar exposures and decreased vertical transport of nutrients from depth due to increasing stratification. We employed a simultaneous multivariate treatment approach to investigate the physiological and biochemical responses of the diatoms Thalassiosira pseudonana and Skeletonema costatum to these projected ocean changes. Diatoms were grown under different 'clustered' regimes of solar radiation, nutrients, and pCO2 (pH), reflecting present-day (2011) and potential mid-century (2050) and end-of-century (2100) scenarios. Growth rates, chlorophyll a contents and maximal photochemical quantum yield all decreased from the present to 2100 scenarios. While cellular particulate organic carbon signifi- cantly increased in both species along with increased cellular organic C:N ratios, biogenic silica content showed species-specific differences among the cluster treatments. Our results suggest that reduced thickness of the upper mixed layer with enhanced stratification may interact with ocean acidification to influence diatom-related biogeochemical processes by affecting their growth and biochemical composition in species-specific ways.