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1.
Bioresour Technol ; 410: 131299, 2024 Oct.
Article in English | MEDLINE | ID: mdl-39153691

ABSTRACT

Phytohormones play a role in regulating microalgae cells tolerance to adversity. This paper examines the effects of different temperatures (20 °C, 25 °C, 30 °C and 35 °C) on the physiological characteristics and endogenous phytohormones of the Isochrysis Zhanjiangensis (IZ) and its mutagenic strain (3005). The results showed that the endogenous phytohormones indole acetic acid (IAA) and jasmonic acid (JA) exhibited significant differences (P<0.05) between the two strains. The addition of 0.5 mg·L-1 exogenous JA inhibitor ibuprofen (IBU) improved cell growth of IZ, and was extremely effective in the accumulation of polysaccharides, which accounted for 33.25 %. Transcriptomic analyses revealed that genes involved in photosynthesis, such as PetC and PsbO, exhibited significantly elevated expression of the strain IZ, while the pathways related to JA synthesis may be the factor affecting microalgae temperature tolerance. This study provides a theoretical foundation for elucidating the underlying mechanisms and potential applications for high temperature tolerance in IZ.


Subject(s)
Haptophyta , Microalgae , Oxylipins , Plant Growth Regulators , Microalgae/metabolism , Microalgae/drug effects , Plant Growth Regulators/pharmacology , Plant Growth Regulators/metabolism , Oxylipins/metabolism , Oxylipins/pharmacology , Haptophyta/metabolism , Haptophyta/drug effects , Indoleacetic Acids/metabolism , Indoleacetic Acids/pharmacology , Cyclopentanes/pharmacology , Cyclopentanes/metabolism , Cell Proliferation/drug effects , Temperature , Ibuprofen/pharmacology , Photosynthesis/drug effects , Polysaccharides/metabolism
2.
Nat Microbiol ; 9(8): 2006-2021, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38969820

ABSTRACT

The lag phase is key in resuming bacterial growth, but it remains underexplored particularly in environmental bacteria. Here we use transcriptomics and 13C-labelled metabolomics to show that the lag phase of the model marine bacterium Phaeobacter inhibens is shortened by methylated compounds produced by the microalgal partner, Emiliania huxleyi. Methylated compounds are abundantly produced and released by microalgae, and we show that their methyl groups can be collected by bacteria and assimilated through the methionine cycle. Our findings underscore the significance of methyl groups as a limiting factor during the lag phase and highlight the adjustability of this growth phase. In addition, we show that methylated compounds, typical of photosynthetic organisms, prompt diverse reductions in lag times in bacteria associated with algae and plants, potentially favouring early growth in some bacteria. These findings suggest ways to accelerate bacterial growth and underscore the significance of studying bacteria within an environmental context.


Subject(s)
Haptophyta , Microalgae , Haptophyta/metabolism , Haptophyta/growth & development , Haptophyta/genetics , Microalgae/metabolism , Microalgae/growth & development , Microalgae/genetics , Methylation , Metabolomics , Bacteroidetes/genetics , Bacteroidetes/growth & development , Bacteroidetes/metabolism , Methionine/metabolism , Photosynthesis , Rhodobacteraceae
3.
J Phycol ; 60(4): 968-979, 2024 Aug.
Article in English | MEDLINE | ID: mdl-38980982

ABSTRACT

Phaeocystis globosa is an important bloom-forming marine phytoplankton species that often accumulates to large levels in temperate and tropical waters and has significant impacts on food webs and biogeochemical cycles. It can form "giant" colonies that reach 3 cm in diameter. Microscopic observations, colony elemental composition, and pigment composition were analyzed to assess the characteristics of colonies as a function of colony size. Particulate organic carbon (POC) per unit surface area, colonial cell density, and chlorophyll a per unit surface area all increased with colony size, in contrast to results from temperate waters. Cellular chl a averaged 0.85 pg chl · cell-1. Colonies had both photosynthetic and protective pigments, with fucoxanthin being the dominant accessory pigment. Based on chl a and pigment levels, it appears colonies were acclimated to relatively low irradiances, likely due to their life cycle and the extremely turbulent environment in which they grew. Mucous carbon ranged from 16.2% to 79.2% of the total POC, and mucous carbon per unit surface area increased with colony size, suggesting that the mucous envelope did not thin as the colony grew. Based on elemental composition, nitrogen did not appear to limit growth, but phosphorus:carbon ratios were similar to those of P-limited cultures. Giant colonies represent an extreme response to the environment, but they do not appear to have greatly different characteristics than other tropical strains.


Subject(s)
Haptophyta , Vietnam , Haptophyta/growth & development , Haptophyta/metabolism , Carbon/metabolism , Carbon/analysis , Chlorophyll A/metabolism , Chlorophyll A/analysis , Phytoplankton/growth & development , Phytoplankton/metabolism , Phytoplankton/chemistry
4.
Environ Microbiol Rep ; 16(4): e13312, 2024 Aug.
Article in English | MEDLINE | ID: mdl-39049182

ABSTRACT

Biological nitrogen fixation provides fixed nitrogen for microbes living in the oligotrophic open ocean. UCYN-A2, the previously known symbiont of Braarudosphaera bigelowii, now believed to be an early-stage B. bigelowii organelle that exchanges fixed nitrogen for fixed carbon, is globally distributed. Indirect evidence suggested that B. bigelowii might be a mixotrophic (phagotrophic) phototrophic flagellate. The goal of this study was to determine if B. bigelowii can graze on bacteria using several independent approaches. The results showed that B. bigelowii grazed on co-occurring bacteria at a rate of 5-7 cells/h/B. bigelowii and that the overall grazing rate was significantly higher at nighttime than at daytime. Bacterial abundance changes, assessed with 16S rRNA gene amplicon sequencing analysis, may have indicated preferential grazing by B. bigelowii on specific bacterial genotypes. In addition, Lysotracker™ staining of B. bigelowii suggested digestive activity inside B. bigelowii. Carbon and nitrogen fixation measurements revealed that the carbon demand of B. bigelowii could not be fulfilled by photosynthesis alone, implying supplementation by heterotrophy. These independent lines of evidence together revealed that B. bigelowii engages in phagotrophy, which, beyond serving as a supplementary source of carbon and energy, may also facilitate the indirect assimilation of inorganic nutrients.


Subject(s)
Haptophyta , Nitrogen Fixation , Symbiosis , Haptophyta/metabolism , Haptophyta/growth & development , Haptophyta/physiology , Nitrogen/metabolism , Bacteria/classification , Bacteria/genetics , Bacteria/metabolism , Bacteria/isolation & purification , Carbon/metabolism , RNA, Ribosomal, 16S/genetics , Phagocytosis , Phylogeny
5.
Proc Natl Acad Sci U S A ; 121(26): e2318570121, 2024 Jun 25.
Article in English | MEDLINE | ID: mdl-38905238

ABSTRACT

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


Subject(s)
Carbon Dioxide , Haptophyta , Lipids , Photosynthesis , Carbon Dioxide/metabolism , Haptophyta/metabolism , Lipids/chemistry , Hydrogen/metabolism , Chloroplasts/metabolism , Deuterium/metabolism , NADP/metabolism , Temperature , Chemical Fractionation/methods , Lipid Metabolism
6.
Microbiol Res ; 286: 127785, 2024 Sep.
Article in English | MEDLINE | ID: mdl-38851011

ABSTRACT

Carbohydrates play a pivotal role in nutrient recycling and regulation of algal-bacterial interactions. Despite their ecological significance, the intricate molecular mechanisms governing regulation of phycosphere carbohydrates by bacterial taxa linked with natural algal bloom have yet to be fully elucidated. Here, a comprehensive temporal metagenomic analysis was conducted to explore the carbohydrate-active enzyme (CAZyme) genes in two discrete algal bloom microorganisms (Gymnodinium catenatum and Phaeocystis globosa) across three distinct bloom stages: pre-bloom, peak bloom, and post-bloom. Elevated levels of extracellular carbohydrates, primarily rhamnose, galactose, glucose, and arabinose, were observed during the initial and post-peak stages. The prominent CAZyme families identified-glycoside hydrolases (GH) and carbohydrate-binding modules (CBMs)-were present in both algal bloom occurrences. In the G. catenatum bloom, GH23/24 and CBM13/14 were prevalent during the pre-bloom and peak bloom stages, whereas GH2/3/30 and CBM12/24 exhibited increased prevalence during the post-bloom phase. In contrast, the P. globosa bloom had a dominance of GH13/23 and CBM19 in the initial phase, and this was succeeded by GH3/19/24/30 and CBM54 in the later stages. This gene pool variation-observed distinctly in specific genera-highlighted the dynamic structural shifts in functional resources driven by temporal alterations in available substrates. Additionally, ecological linkage analysis underscored a correlation between carbohydrates (or their related genes) and phycospheric bacteria, hinting at a pattern of bottom-up control. These findings contribute to understanding of the dynamic nature of CAZymes, emphasizing the substantial influence of substrate availability on the metabolic capabilities of algal symbiotic bacteria, especially in terms of carbohydrates.


Subject(s)
Bacteria , Carbohydrate Metabolism , Eutrophication , Carbohydrate Metabolism/genetics , Bacteria/genetics , Bacteria/classification , Bacteria/metabolism , Dinoflagellida/genetics , Dinoflagellida/metabolism , Metagenomics , Glycoside Hydrolases/genetics , Glycoside Hydrolases/metabolism , Plankton/genetics , Plankton/metabolism , Haptophyta/genetics , Haptophyta/metabolism , Seawater/microbiology , Metagenome , Phylogeny
7.
Appl Microbiol Biotechnol ; 108(1): 352, 2024 May 31.
Article in English | MEDLINE | ID: mdl-38819468

ABSTRACT

Fucoxanthin is a versatile substance in the food and pharmaceutical industries owing to its excellent antioxidant and anti-obesity properties. Several microalgae, including the haptophyte Pavlova spp., can produce fucoxanthin and are potential industrial fucoxanthin producers, as they lack rigid cell walls, which facilitates fucoxanthin extraction. However, the commercial application of Pavlova spp. is limited owing to insufficient biomass production. In this study, we aimed to develop a mixotrophic cultivation method to increase biomass and fucoxanthin production in Pavlova gyrans OPMS 30543X. The effects of culturing OPMS 30543X with different organic carbon sources, glycerol concentrations, mixed-nutrient conditions, and light intensities on the consumption of organic carbon sources, biomass production, and fucoxanthin accumulation were analyzed. Several organic carbon sources, such as glycerol, glucose, sucrose, and acetate, were examined, revealing that glycerol was well-consumed by the microalgae. Biomass and fucoxanthin production by OPMS 30543X increased in the presence of 10 mM glycerol compared to that observed without glycerol. Metabolomic analysis revealed higher levels of the metabolites related to the glycolytic, Calvin-Benson-Bassham, and tricarboxylic acid cycles under mixotrophic conditions than under autotrophic conditions. Cultures grown under mixotrophic conditions with a light intensity of 100 µmol photons m-2 s-1 produced more fucoxanthin than autotrophic cultures. Notably, the amount of fucoxanthin produced (18.9 mg/L) was the highest reported thus far for Pavlova species. In conclusion, the use of mixotrophic culture is a promising strategy for increasing fucoxanthin production in Pavlova species. KEY POINTS: • Glycerol enhances biomass and fucoxanthin production in Pavlova gyrans • Metabolite levels increase under mixotrophic conditions • Mixotrophic conditions and medium-light intensity are appropriate for P. gyrans.


Subject(s)
Biomass , Glycerol , Haptophyta , Xanthophylls , Xanthophylls/metabolism , Glycerol/metabolism , Haptophyta/metabolism , Haptophyta/growth & development , Haptophyta/radiation effects , Microalgae/metabolism , Microalgae/growth & development , Culture Media/chemistry , Carbon/metabolism , Light , Metabolomics
8.
ISME J ; 18(1)2024 Jan 08.
Article in English | MEDLINE | ID: mdl-38709871

ABSTRACT

Chirality, a fundamental property of matter, is often overlooked in the studies of marine organic matter cycles. Dihydroxypropanesulfonate (DHPS), a globally abundant organosulfur compound, serves as an ecologically important currency for nutrient and energy transfer from phytoplankton to bacteria in the ocean. However, the chirality of DHPS in nature and its transformation remain unclear. Here, we developed a novel approach using chiral phosphorus-reagent labeling to separate DHPS enantiomers. Our findings demonstrated that at least one enantiomer of DHPS is present in marine diatoms and coccolithophores, and that both enantiomers are widespread in marine environments. A novel chiral-selective DHPS catabolic pathway was identified in marine Roseobacteraceae strains, where HpsO and HpsP dehydrogenases at the gateway to DHPS catabolism act specifically on R-DHPS and S-DHPS, respectively. R-DHPS is also a substrate for the dehydrogenase HpsN. All three dehydrogenases generate stable hydrogen bonds between the chirality-center hydroxyls of DHPS and highly conserved residues, and HpsP also form coordinate-covalent bonds between the chirality-center hydroxyls and Zn2+, which determines the mechanistic basis of strict stereoselectivity. We further illustrated the role of enzymatic promiscuity in the evolution of DHPS metabolism in Roseobacteraceae and SAR11. This study provides the first evidence of chirality's involvement in phytoplankton-bacteria metabolic currencies, opening a new avenue for understanding the ocean organosulfur cycle.


Subject(s)
Diatoms , Phytoplankton , Rhodobacteraceae , Phytoplankton/metabolism , Stereoisomerism , Diatoms/metabolism , Rhodobacteraceae/metabolism , Rhodobacteraceae/genetics , Haptophyta/metabolism , Oxidoreductases/metabolism , Oxidoreductases/genetics , Biotransformation , Metabolic Networks and Pathways , Alkanesulfonates
9.
Environ Pollut ; 351: 124084, 2024 Jun 15.
Article in English | MEDLINE | ID: mdl-38697245

ABSTRACT

Due to the potential impacts of microplastics (MPs) and nanoplastics (NPs) on algal growth and thereby affect the climate-relevant substances, dimethylsulfoniopropionate (DMSP) and dimethyl sulfide (DMS), we studied the polystyrene (PS) MPs and NPs of 1 µm and 80 nm impacts on the growth, chlorophyll content, reactive oxygen species (ROS), antioxidant enzyme activity, and DMS/DMSP production in Emiliania huxleyi. E. huxleyi is a prominent oceanic alga that plays a key role in DMS and DMSP production. The results revealed that high concentrations of MPs and NPs inhibited the growth, carotenoid (Car), and Chl a concentrations of E. huxleyi. However, short-time exposure to low concentrations of PS MPs and NPs stimulated the growth of E. huxleyi. Furthermore, high concentrations of MPs and NPs resulted in an increase in the superoxide anion radical (O2.-) production rate and a decrease in the malondialdehyde (MDA) content compared with the low concentrations. Exposure to MPs and NPs at 5 mg L-1 induced superoxide dismutase (SOD) activity as a response to scavenging ROS. High concentrations of MPs and NPs significantly inhibited the production of DMSP and DMS. The findings of this study support the potential ecotoxicological impacts of MPs and NPs on algal growth, antioxidant system, and dimethylated sulfur compounds production, which maybe potentially impact the global climate.


Subject(s)
Antioxidants , Haptophyta , Reactive Oxygen Species , Sulfides , Sulfonium Compounds , Water Pollutants, Chemical , Antioxidants/metabolism , Sulfonium Compounds/metabolism , Haptophyta/growth & development , Haptophyta/metabolism , Haptophyta/drug effects , Reactive Oxygen Species/metabolism , Water Pollutants, Chemical/toxicity , Microplastics/toxicity , Chlorophyll/metabolism , Superoxide Dismutase/metabolism , Nanoparticles/toxicity , Polystyrenes/toxicity
10.
Sci Adv ; 10(15): eadk1954, 2024 Apr 12.
Article in English | MEDLINE | ID: mdl-38598627

ABSTRACT

The globally distributed marine alga Emiliania huxleyi has cooling effect on the Earth's climate. The population density of E. huxleyi is restricted by Nucleocytoviricota viruses, including E. huxleyi virus 201 (EhV-201). Despite the impact of E. huxleyi viruses on the climate, there is limited information about their structure and replication. Here, we show that the dsDNA genome inside the EhV-201 virion is protected by an inner membrane, capsid, and outer membrane. EhV-201 virions infect E. huxleyi by using fivefold vertices to bind to and fuse the virus' inner membrane with the cell plasma membrane. Progeny virions assemble in the cytoplasm at the surface of endoplasmic reticulum-derived membrane segments. Genome packaging initiates synchronously with the capsid assembly and completes through an aperture in the forming capsid. The genome-filled capsids acquire an outer membrane by budding into intracellular vesicles. EhV-201 infection induces a loss of surface protective layers from E. huxleyi cells, which enables the continuous release of virions by exocytosis.


Subject(s)
Haptophyta , Phycodnaviridae , Viruses , Haptophyta/metabolism , Phycodnaviridae/genetics , Virion , Climate
11.
Cell ; 187(7): 1762-1768.e9, 2024 Mar 28.
Article in English | MEDLINE | ID: mdl-38471501

ABSTRACT

Biological dinitrogen (N2) fixation is a key metabolic process exclusively performed by prokaryotes, some of which are symbiotic with eukaryotes. Species of the marine haptophyte algae Braarudosphaera bigelowii harbor the N2-fixing endosymbiotic cyanobacteria UCYN-A, which might be evolving organelle-like characteristics. We found that the size ratio between UCYN-A and their hosts is strikingly conserved across sublineages/species, which is consistent with the size relationships of organelles in this symbiosis and other species. Metabolic modeling showed that this size relationship maximizes the coordinated growth rate based on trade-offs between resource acquisition and exchange. Our findings show that the size relationships of N2-fixing endosymbionts and organelles in unicellular eukaryotes are constrained by predictable metabolic underpinnings and that UCYN-A is, in many regards, functioning like a hypothetical N2-fixing organelle (or nitroplast).


Subject(s)
Cyanobacteria , Haptophyta , Nitrogen Fixation , Cyanobacteria/metabolism , Haptophyta/cytology , Haptophyta/metabolism , Haptophyta/microbiology , Nitrogen/metabolism , Symbiosis
12.
J Sci Food Agric ; 104(7): 4354-4362, 2024 May.
Article in English | MEDLINE | ID: mdl-38318717

ABSTRACT

BACKGROUND: Alcoholic liver disease (ALD) is responsible for 3.3 million deaths per annum. Efficacious therapeutic modalities or drug treatments for ALD have not yet been found, so it is urgent to seek new agents for preventing ALD and its related disease. Many experiments have indicated that modulating the gut microbiota and regulating the toll-like receptor 4 (TLR4)/nuclear transcription factor-κB (NF-κB) inflammatory pathway can provide a new target for prevention and treatment of ALD. Marine microalgae have their natural metabolic pathways to synthesize various of bioactive compounds as promising candidates for hepatoprotection. In this study, we investigated ethanol extracts from Isochrysis zhanjiangensis (EEIZ) to evaluate their ability to alleviate acute alcoholic liver injury, regulate TLR4/NF-κB inflammatory pathway and modulate intestinal bacteria dysbiosis in mice for ALD treatment. RESULTS: In the acute ALD mouse model, EEIZ reduced levels of aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, triacylglyceride, total cholesterol and low-density lipoprotein, while increasing the level of high-density lipoprotein. Besides, TLR4, myeloid differentiation factor 88, NF-κB and tumor necrosis factor-α expression levels in liver tissue were effectively downregulated by EEIZ. Furthermore, treatment with EEIZ enhanced intestinal homeostasis and significantly alleviated the damage caused by alcohol. CONCLUSION: EEIZ showed effective hepatoprotective activity against alcohol-induced acute liver injury in mice as it could alleviate hepatocyte damage, suppress the TLR4/NF-κB inflammatory pathway and regulate the intestinal flora structure. EEIZ could be a good candidate for preventing acute alcoholic liver injury. © 2024 Society of Chemical Industry.


Subject(s)
Haptophyta , Liver Diseases, Alcoholic , Mice , Animals , Ethanol/metabolism , Toll-Like Receptor 4/genetics , Toll-Like Receptor 4/metabolism , Toll-Like Receptor 4/therapeutic use , Haptophyta/metabolism , NF-kappa B/genetics , NF-kappa B/metabolism , Dysbiosis/drug therapy , Dysbiosis/metabolism , Liver/metabolism , Liver Diseases, Alcoholic/etiology , Bacteria/metabolism , Mice, Inbred C57BL
13.
Molecules ; 29(3)2024 Feb 02.
Article in English | MEDLINE | ID: mdl-38338436

ABSTRACT

Parkinson's disease (PD) is a progressive neurodegenerative disorder of the elderly for which there is no cure or disease-modifying therapy. Mitochondrial dysfunction and oxidative stress play a central role in dopaminergic neurodegeneration in PD. Therefore, antioxidants are considered a promising neuroprotective approach. In in vivo activity studies, 6-OHDA-induced oxidative stress in SH-SY5Y cells was established as a model of PD for cellular experiments. IIAVE (Ile-Ile-Ala-Val-Glu) was derived from Isochrysis zhanjiangensis octapeptide (IIAVEAGC), which has a small molecular weight. The structure and antioxidant activity of IIAVE were tested in a previous study and proved to have good antioxidant potential. In this study, the chemical properties of IIAVE were calculated using quantum chemical methods, including frontier molecular orbital (FMO), molecular electrostatic potential (MEP), natural population analysis (NPA), and global reactivity properties. The interaction of IIAVE with Bcl-2 and DJ-1 was investigated using the molecular docking method. The results showed that IIAVE promoted the activation of the Keap1/Nrf2 pathway and up-regulated the expression of the superoxide dismutase 1 (SOD-1) protein by inhibiting the level of reactive oxygen species (ROS) in cells. In addition, IIAVE inhibits ROS production and prevents 6-OHDA-induced oxidative damage by restoring mitochondrial membrane potential. Furthermore, IIAVE inhibited cell apoptosis by increasing the Bcl-2/Bax ratio and inhibiting the activation of Caspase-9 and Caspase-3. Thus, IIAVE may become a potential drug for the treatment and prevention of PD.


Subject(s)
Haptophyta , Neuroblastoma , Neuroprotective Agents , Parkinson Disease , Humans , Aged , Neuroprotection , Reactive Oxygen Species/metabolism , Oxidopamine/pharmacology , Kelch-Like ECH-Associated Protein 1/metabolism , Haptophyta/metabolism , Molecular Docking Simulation , Neuroprotective Agents/pharmacology , NF-E2-Related Factor 2/metabolism , Cell Line, Tumor , Apoptosis , Antioxidants/pharmacology , Parkinson Disease/drug therapy , Proto-Oncogene Proteins c-bcl-2/metabolism
14.
ACS Synth Biol ; 13(1): 77-84, 2024 01 19.
Article in English | MEDLINE | ID: mdl-38147049

ABSTRACT

Coccolithophores are a group of unicellular marine phytoplankton that exhibit a prolific capacity for carbon conversion and are critical to ocean biogeochemistry. A fundamental understanding of coccolithophore biomineralization has been limited, in part, by the lack of genetic and molecular tools to investigate the organisms. In particular, it has proven to be difficult to deliver macromolecules across the coccosphere-membrane complex. To overcome this barrier, we employed cell-penetrating peptides (CPP) in the Emiliania huxleyi coccolithophores. We evaluated three established CPPs (TAT, R9, and KFF) and designed a CPP that incorporates a high proline content identified in the protein transduction domain of EhV060, an E. huxleyi virus lectin protein. To measure the delivery performance, we covalently linked CPPs to synthetic peptide nucleic acids (PNA) and attached a fluorescein marker. CPP-PNA-FITC complexes were efficiently delivered across the coccosphere-membrane complex to the cytoplasm of E. huxleyi cells. Characterization of E. huxleyi demonstrates that CPP-PNA are nontoxic and reveals specific effects of CPP-PNA on cell biology and calcification. Direct delivery and characterization of synthetic nucleic acids represent a step forward in synthetic biology to explore coccolithophore biomineralization.


Subject(s)
Cell-Penetrating Peptides , Haptophyta , Nucleic Acids , Haptophyta/genetics , Haptophyta/metabolism , Cell-Penetrating Peptides/metabolism , Nucleic Acids/metabolism , Calcification, Physiologic , Phytoplankton/genetics
15.
Nat Commun ; 14(1): 3749, 2023 06 23.
Article in English | MEDLINE | ID: mdl-37353496

ABSTRACT

Coccolithophores are globally abundant, calcifying microalgae that have profound effects on marine biogeochemical cycles, the climate, and life in the oceans. They are characterized by a cell wall of CaCO3 scales called coccoliths, which may contribute to their ecological success. The intricate morphologies of coccoliths are of interest for biomimetic materials synthesis. Despite the global impact of coccolithophore calcification, we know little about the molecular machinery underpinning coccolithophore biology. Working on the model Emiliania huxleyi, a globally distributed bloom-former, we deploy a range of proteomic strategies to identify coccolithogenesis-related proteins. These analyses are supported by a new genome, with gene models derived from long-read transcriptome sequencing, which revealed many novel proteins specific to the calcifying haptophytes. Our experiments provide insights into proteins involved in various aspects of coccolithogenesis. Our improved genome, complemented with transcriptomic and proteomic data, constitutes a new resource for investigating fundamental aspects of coccolithophore biology.


Subject(s)
Haptophyta , Proteomics , Calcification, Physiologic/genetics , Oceans and Seas , Genomics , Haptophyta/genetics , Haptophyta/metabolism
16.
Mar Biotechnol (NY) ; 25(3): 463-472, 2023 Jun.
Article in English | MEDLINE | ID: mdl-37289264

ABSTRACT

Isochrysis zhangjiangensis is an important microalgal species used as bait in aquaculture. However, its optimal cultivation temperature is around 25 °C, limiting its use in summer when temperature is higher. To overcome this limitation, we aimed to develop a consortia of I. zhangjiangensis and bacteria that are more resistant to heat stress. Here, six thermotolerance-promoting bacterial strains were isolated from the culture of a heat-tolerant mutant strain of I. zhangjiangensis (IM), and identified as Algoriphagus marincola, Nocardioides sp., Pseudidiomarina sp., Labrenzia alba, Nitratireductor sp., and Staphylococcus haemolyticus. Further, co-culturing I. zhangjiangensis with A. marincola under high temperature conditions increased cell density, chlorophyll a, PSII maximum photochemical efficiency (Fv/Fm), and soluble protein content of microalgae. The presence of A. marincola positively influenced the activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and total antioxidant capacity (T-AOC) in I. zhangjiangensis cells, while concurrently reducing the levels of reactive oxygen species (ROS). Additionally, gene expression studies confirmed that co-culturing with A. marincola upregulated the expression of antioxidant-related genes (sod and pod) and stress tolerance genes (heat shock protein genes). Our findings indicate that A. marincola effectively helps I. zhangjiangensis withstand high temperature stress, leading to improved yield of microalgae during high temperature conditions. The thermotolerance-promoting bacteria can be exploited as potential inoculants for enhancing the productivity and sustainability of bait microalgae in aquaculture.


Subject(s)
Haptophyta , Thermotolerance , Antioxidants/metabolism , Haptophyta/metabolism , Chlorophyll A/metabolism , Bacteria
17.
Mar Drugs ; 21(6)2023 Jun 09.
Article in English | MEDLINE | ID: mdl-37367680

ABSTRACT

Microalgae have enormous potential for human nutrition, yet the European Commission has authorized the consumption of only eleven species. Strains of fifteen rarely researched microalgae from two kingdoms were screened regarding their nutritional profile and value for human health in two cultivation phases. Contents of protein, fiber, lipids, fatty acids, minerals, trace elements and heavy metals were determined. In the growth phase, microalgae accumulated more arginine, histidine, ornithine, pure and crude protein, Mg, Mn, Fe and Zn and less Ni, Mo and I2 compared to the stationary phase. Higher contents of total fat, C14:0, C14:1n5, C16:1n7, C20:4n6, C20:5n3 and also As were observed in microalgae from the chromista kingdom in comparison to microalgae from the plantae kingdom (p < 0.05). Conversely, the latter had higher contents of C20:0, C20:1n9 and C18:3n3 as well as Ca and Pb (p < 0.05). More precisely, Chrysotila carterae appeared to have great potential for human nutrition because of its high nutrient contents such as fibers, carotenoids, C20:6n3, Mg, Ca, Mn, Fe, Se, Zn, Ni, Mo and I2. In summary, microalgae may contribute to a large variety of nutrients, yet the contents differ between kingdoms, cultivation phases and also species.


Subject(s)
Haptophyta , Microalgae , Trace Elements , Humans , Micronutrients , Microalgae/metabolism , Fatty Acids/metabolism , Haptophyta/metabolism
18.
Proc Natl Acad Sci U S A ; 120(27): e2217121120, 2023 07 04.
Article in English | MEDLINE | ID: mdl-37364132

ABSTRACT

Marine viruses play a key role in regulating phytoplankton populations, greatly affecting the biogeochemical cycling of major nutrients in the ocean. Resistance to viral infection has been reported for various phytoplankton species under laboratory conditions. Nevertheless, the occurrence of resistant cells in natural populations is underexplored due to the lack of sensitive tools to detect these rare phenotypes. Consequently, our current understanding of the ecological importance of resistance and its underlying mechanisms is limited. Here, we sought to identify lipid biomarkers for the resistance of the bloom-forming alga Emiliania huxleyi to its specific virus, E. huxleyi virus (EhV). By applying an untargeted lipidomics approach, we identified a group of glycosphingolipid (GSL) biomarkers that characterize resistant E. huxleyi strains and were thus termed resistance-specific GSLs (resGSLs). Further, we detected these lipid biomarkers in E. huxleyi isolates collected from induced E. huxleyi blooms and in samples collected during an open-ocean E. huxleyi bloom, indicating that resistant cells predominantly occur during the demise phase of the bloom. Last, we show that the GSL composition of E. huxleyi cultures that recover following infection and gain resistance to the virus resembles that of resistant strains. These findings highlight the metabolic plasticity and coevolution of the GSL biosynthetic pathway and underscore its central part in this host-virus arms race.


Subject(s)
Haptophyta , Virus Diseases , Viruses , Humans , Phytoplankton/metabolism , Haptophyta/metabolism , Biomarkers/metabolism , Oceans and Seas , Lipids
19.
Sci Total Environ ; 874: 162445, 2023 May 20.
Article in English | MEDLINE | ID: mdl-36848993

ABSTRACT

The phycotoxin ß-N-methylamino-l-alanine (BMAA) has attracted attention due to its risks to marine organisms and human health. In this study, approximately 85 % of synchronized cells of the marine microalga Isochrysis galbana were arrested at the cell cycle G1 phase by BMAA at 6.5 µM for a 24-h exposure. The concentration of chlorophyll a (Chl a) gradually decreased, while the maximum quantum yield of PSII (Fv/Fm), the maximum relative electron transport rate (rETRmax), light utilization efficiency (α) and half-saturated light irradiance (Ik) reduced early and recovered gradually in I. galbana exposed to BMAA in 96-h batch cultures. Transcriptional expression of I. galbana analyzed at 10, 12, and 16 h disclosed multiple mechanisms of BMAA to suppress the microalgal growth. Production of ammonia and glutamate was limited by the down-regulation of nitrate transporters, glutamate synthase, glutamine synthetase, cyanate hydrolase, and formamidase. Diverse extrinsic proteins related to PSII, PSI, cytochrome b6f complex, and ATPase were influenced by BMAA at transcriptional level. Suppression of the DNA replication and mismatch repair pathways increased the accumulation of misfolded proteins, which was reflected by the up-regulated expression of proteasome to accelerate proteolysis. This study improves our understanding of the chemical ecology impacts of BMAA in marine ecosystems.


Subject(s)
Amino Acids, Diamino , Haptophyta , Microalgae , Humans , Neurotoxins/toxicity , Haptophyta/metabolism , Microalgae/metabolism , Chlorophyll A , Ecosystem , Amino Acids, Diamino/toxicity , Cell Cycle
20.
Environ Pollut ; 320: 121008, 2023 Mar 01.
Article in English | MEDLINE | ID: mdl-36608724

ABSTRACT

Simultaneous ocean warming and acidification will alter marine ecosystem structure and directly affect marine organisms. The alga Phaeocystis globosa commonly causes harmful algal blooms in coastal areas of eastern China. P. globosa often outcompetes other species due to its heterotypic life cycle, primarily including colonies and various types of solitary cells. However, little is known about the adaptive response of P. globosa to ocean warming and acidification. This study aimed to reveal the global molecular regulatory networks implicated in the response of P. globosa to simultaneous warming and acidification. After exposure to warming and acidification, the phosphatidylinositol (PI) and mitogen-activated protein kinase (MAPK) signaling pathways of P. globosa were activated to regulate other molecular pathways in the cell, while the light harvesting complex (LHC) genes were downregulated to decrease photosynthesis. Exposure to warming and acidification also altered the intracellular energy flow, with more energy allocated to the TCA cycle rather than to the biosynthesis of fatty acids and hemolytic substances. The upregulation of genes associated with glycosaminoglycan (GAG) degradation prevented the accumulation of polysaccharides, which led to a reduction in colony formation. Finally, the upregulation of the Mre11 and Rad50 genes in response to warming and acidification implied an increase in meiosis, which may be used by P. globosa to increase the number of solitary cells. The increase in genetic diversity through sexual reproduction may be a strategy of P. globosa that supports rapid response to complex environments. Thus, the life cycle of P. globosa underwent a transition from colonies to solitary cells in response to warming and acidification, suggesting that this species may be able to rapidly adapt to future climate changes through life cycle transitions.


Subject(s)
Haptophyta , Haptophyta/metabolism , Ecosystem , Harmful Algal Bloom , Hydrogen-Ion Concentration , Oceans and Seas
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