ALBERT

Description: The HAB-forming, toxic dinoflagellate Karenia mikimotoi, previously found to benefit from ocean acidification (OA), was cultivated to investigate its transcriptional response to simulated OA for 30 generations. Batch cultures were grown under two CO2 concentrations, 450 (control) and 1100 (simulated OA) μatm, and physiological parameters [growth, pigments, catalase (CAT), glutathione reductase (GR), and superoxide dismutase (SOD) activity], as well as transcriptomes (obtained via RNA-seq), were compared. Chlorophyll a (Chl a) and carotenoid (Caro) contents, as well as CAT and GR activities, were significantly increased under OA conditions. Transcriptomic analysis revealed 2,490 differentially expressed unigenes in response to OA, which comprised 1.54% of all unigenes. A total of 1,121 unigenes were upregulated, and 1,369 unigenes were downregulated in OA compared to control conditions. The downregulated expression of bicarbonate transporter and carbonic anhydrase genes was a landmark of OA acclimation. Key genes involved in energy metabolism, e.g., photosynthesis, tricarboxylic acid cycle, oxidative phosphorylation, and nitrogen metabolism, were highly upregulated under OA, contributing to increases in the Chl a (55.05%) and Caro (28.37%). The enhanced antioxidant enzyme activities (i.e. CAT, GR) and upregulated genes (i.e. glutathione peroxidase, ascorbate peroxidase, heat shock protein, 20S proteasome, aldehyde dehydrogenase, and apolipoprotein) benefit cells against the potential lower pH stress condition under OA. In addition, the downregulation of four genes associated with motility suggested that the preserved energy could further boost growth. In conclusion, the present study suggests that K. mikimotoi exhibits efficient gene expression regulation for the utilization of energy and resistance to OA-induced stress. Taken together, K. mikimotoi appeared as a tolerant species in response to OA. Thus, more extensive algal blooms that threaten marine organisms are likely in the future. These findings expand current knowledge on the gene expression of HAB-forming species in response to future OA.

Description: 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.

Description: The effects and interactive effects of different nitrogen (N) sources (ammonium, nitrate, and urea) and carbon dioxide (CO2) concentrations were investigated on Alexandrium tamarense, a harmful marine dinoflagellate, by measuring its growth (μ), extracellular carbonic anhydrase (CA), and its toxicity to zebrafish (Danio rerio) embryo. The μ and CA were influenced more strongly by CO2 concentrations rather than by N sources; significant effects of CO2 on μ and CA were observed under low CO2 concentration (LC) conditions compared to high CO2 concentration (HC) conditions. The ammonium and nitrate media under LC conditions had the maximum μ and CA, which was inhibited under HC conditions. The embryotoxic effects were influenced more strongly by the N sources than by CO2 concentrations, thus excluding the lower deformation in urea under HC conditions. Moreover, the antioxidant enzymes superoxide dismutase (SOD), glutathione peroxidase (GPX), glutathione S-transferase (GST), and catalase (CAT) were detected in normal (untreated) zebrafish embryos, and among them, the level of SOD was the highest. In summary, this study provides a clear insight for understanding the effects and interactive effects of N sources and CO2 concentrations on the growth and toxicity of harmful dinoflagellates.

Description: This work demonstrated a 10-day batch culture experiment to test the physiology and toxicity of harmful dinoflagellate Karenia mikimotoi in response to ocean acidification (OA) under two different phosphate concentrations. Cells were previously acclimated in OA (pH = 7.8 and CO2 = 1100 μatm) condition for about three months before testing the responses of K. mikimotoi cells to a two-factorial combinations experimentation. This work measured the variation in physiological parameters (growth, rETR) and toxicity (hemolytic activity and its toxicity to zebrafish embryos) in four treatments, representing two factorial combinations of CO2 (450 and 1100 μatm) and phosphate concentration (37.75 and 4.67 umol l−1). Results: OA stimulated the faster growth, and the highest rETRmax in high phosphate (HP) treatment, low phosphate (LP) and a combination of high CO2 and low phosphate (HC*LP) inhibited the growth and Ek in comparison to low CO2*high phosphate (LCHP) treatment. The embryotoxicity of K. mikimotoi cells enhanced in all high CO2 (HC) conditions irrespective of phosphate concentration, but the EC50 of hemolytic activity increased in all high CO2 (HC) and low phosphate (LP) treatments in comparison of LCHP. Ocean acidification (high CO2 and lower pH) was probably the main factor that affected the rETRmax, hemolytic activity and embryotoxicity, but low phosphate was the main factor that affected the growth, α, and Ek. There were significant interactive effects of OA and low phosphate (LP) on growth, rETRmax, and hemolytic activity, but there were no significant effects on α, Ek, and embryotoxicity. If these results are extrapolated to the aquatic environment, it can be hypothesized that the K. mikimotoi cells were impacted significantly by future changing ocean (e.g., ocean acidification and nutrient stoichiometry).

Description: Due to global climate change, marine phytoplankton will likely experience low pH (ocean acidification), high temperatures and high irradiance in the future. Here, this work report the results of a batch culture experiment conducted to study the interactive effects of elevated CO2, increased temperature and high irradiance on the harmful dinoflagellate Akashiwo sanguinea, isolated at Dongtou Island, Eastern China Sea. The A. sanguineacells were acclimated in high CO2 condition for about three months before testing the responses of cells to a full factorial matrix experimentation during a 7-day period. This study measured the variation in physiological parameters and hemolytic activity in 8 treatments, representing full factorial combinations of 2 levels each of exposure to CO2(400 and 1000 μatm), temperature (20 and 28 °C) and irradiance (50 and 200 μmol photons /m**2/s). Sustained growth of A. sanguinea occurred in all treatments, but high CO2 (HC) stimulated faster growth than low CO2 (LC). The pigments (chlorophyll a and carotenoid) decreased in all HC treatments. The quantum yield (Fv/Fm) declined slightly in all high-temperature (HT) treatments. High irradiance (HL) induced the accumulation of ultraviolet-absorbing compounds (UVabc) irrespective of temperature and CO2. The hemolytic activity in the LC treatments, however, declined when exposed to HT and HL, but HC alleviated the adverse effects of HT and HL on hemolytic activity. All HC and HL conditions and the combinations of high temperature*high light (HTHL) and high CO2*high temperature*high light (HCHTHL) positively affected the growth in comparison to the low CO2*low temperature*low light (LCLTLL) treatment. High temperature (HT), high light (HL) and a combination of HT*HL, however, negatively impacted hemolytic activity. CO2 was the main factor that affected the growth and hemolytic activity. There were no significant interactive effects of CO2*temperature*irradiance on growth, pigment, Fv/Fm or hemolytic activity, but there were effects on Pm, α, and Ek. If these results are extrapolated to the natural environment, it can be hypothesized that A. sanguinea cells will benefit from the combination of ocean acidification, warming, and high irradiance that are likely to occur under future climate change. It is assumed that faster growth and higher hemolytic activity and UVabc of this species will occur under future conditions compared with those the current CO2 (400 μatm) and temperature (20 °C) conditions.

Description: Ocean acidification (OA) is affecting marine ecosystems through changes in carbonate chemistry that may influence consumers of phytoplankton, often via trophic pathways. Using a mesocosm approach, we investigated OA effects on a subtropical zooplankton community during oligotrophic, bloom, and post-bloom phases under a range of different pCO2 levels (from 400 to 1480 μatm). Furthermore, we simulated an upwelling event by adding 650 m-depth nutrient-rich water to the mesocosms, which initiated a phytoplankton bloom. No effects of pCO2 on the zooplankton community were visible in the oligotrophic conditions before the bloom. The zooplankton community responded to phytoplankton bloom by increased abundances in all treatments, although the response was delayed under high-pCO2 conditions. Microzooplankton was dominated by small dinoflagellates and aloricate ciliates, which were more abundant under medium- to high-pCO2 conditions. The most abundant mesozooplankters were calanoid copepods, which did not respond to CO2 treatments during the oligotrophic phase of the experiment but were found in higher abundance under medium- and high-pCO2 conditions toward the end of the experiment, most likely as a response to increased phyto- and microzooplankton standing stocks. The second most abundant mesozooplankton taxon were appendicularians, which did not show a response to the different pCO2 treatments. Overall, CO2 effects on zooplankton seemed to be primarily transmitted through significant CO2 effects on phytoplankton and therefore indirect pathways. We conclude that elevated pCO2 can change trophic cascades with significant effects on zooplankton, what might ultimately affect higher trophic levels in the future.

Description: The study aimed to unravel the interaction between ocean acidification and solar ultraviolet radiation (UVR) in Chaetoceros curvisetus. Chaetoceros curvisetus cells were acclimated to high CO2 (HC, 1000 ppmv) and low CO2 concentration (control, LC, 380 ppmv) for 14 days. Cell density, specific growth rate and chlorophyll were measured. The acclimated cells were then exposed to PAB (photosynthetically active radiation (PAR) + UV-A + UV-B), PA (PAR + UV-A) or P (PAR) for 60 min. Photochemical efficiency (phi PSII), relative electron transport rate (rETR) and the recovery of PHPSII were determined. HC induced higher cell density and specific growth rate compared with LC. However, no difference was found in chlorophyll between HC and LC. Moreover, phi PSII and rETRs were higher under HC than LC in response to solar UVR. P exposure led to faster recovery of phi PSII, both under HC and LC, than PA and PAB exposure. It appeared that harmful effects of UVR on C. curvisetus could be counteracted by ocean acidification simulated by high CO2 when the effect of climate change is not beyond the tolerance of cells.

Description: A batch culture experiment was conducted to study the interactive effects of ocean acidification (OA) and solar ultraviolet radiation (UVR, 280–400 nm) on the harmful dinoflagellate Karenia mikimotoi. Cells were incubated in 7-days trials under four treatments. Physiological (growth, pigments, UVabc) and toxicity (hemolytic activity and its toxicity to zebrafish embryos) response variables were measured in four treatments, representing two factorial combinations of CO2 (400 and 1000 μatm) and solar irradiance (with or without UVR). Toxic species K. mikimotoi showed sustained growth in all treatments, and there was not statistically significant difference among four treatments. Cell pigment content decreased, but UVabc and hemolytic activity increased in all HC treatments and PAB conditions. The toxicity to zebrafish embryos of K. mikimotoi was not significantly different among four treatments. All HC and UVR conditions and the combinations of HC*UVR (HC-PAB) positively affected the UVabc, hemolytic activity in comparison to the LC*P (LC-P) treatment, and negatively affected the pigments. Ocean acidification (OA) was probably the main factor that affected the chlorophyll-a (Chl-a) and UVabc, but UVR was the main factor that affected the carotenoid (Caro) and hemolytic activity. There were no significant interactive effects of OA*UVR on growth, toxicity to zebrafish embryos. If these results are extrapolated to the natural environment, it can be hypothesized that this strain (DP-C32) of K. mikimotoi cells have the efficient mechanisms to endure the combination of ocean acidification and solar UVR. It is assumed that this toxic strain could form harmful bloom and enlarge the threatening to coastal communities, marine animals, even human health under future conditions.