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1.
New Phytol ; 241(3): 1210-1221, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38013640

RESUMO

Marine diatoms express genes encoding potential phosphate transporter and alkaline phosphatase (APase) under phosphate-limited (-P) condition. This indicates that diatoms use high-affinity phosphate uptake system with organic phosphate hydration. The function of molecules playing roles for Pi uptake was determined in this study. Pi uptake and APase activity of two marine diatoms, Phaeodactylum tricornutum and Thalassiosira pseudonana, were monitored during acclimation to -P condition. The transcript levels of Pi transporter were analyzed, and Pi transporters were localized with GFP tagging in diatom cells. KO mutants of plasma membrane solute carrier proteins (SLC34s) or APase were established, and their phenotype was evaluated. Some Na+ /Pi transporter candidates, SLC34s in P. tricornutum and T. pseudonana, increased transcript under -P condition. Whole-cell Pi transport was specifically stimulated by sodium ion but independent of potassium, lithium, or proton. Genome-editing KO of PtSLC34-5 and APase (Pt49678) in P. tricornutum was highly inhibitory for Pi uptake, and KO of TpSLC34-2 was also highly inhibitory for Pi uptake in T. pseudonana. SLC34s and APase were co-expressed under -P conditions in marine diatoms. SLC34s play a major role in the initial acclimation stage of diatom cells to -P condition and APase plays an increasing role in the prolonged Pi-starved condition.


Assuntos
Diatomáceas , Diatomáceas/genética , Diatomáceas/metabolismo , Fosfatase Alcalina/metabolismo , Fosfatos/metabolismo , Transporte Biológico , Proteínas de Membrana Transportadoras/metabolismo
2.
Photosynth Res ; 156(2): 217-229, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36862281

RESUMO

Carbonic anhydrase (CA) is a crucial component for the operation of CO2-concentrating mechanisms (CCMs) in the majority of aquatic photoautotrophs that maintain the global primary production. In the genome of the centric marine diatom, Thalassiosira pseudonana, there are four putative gene sequences that encode θ-type CA, which was a type of CA recently identified in marine diatoms and green algae. In the present study, specific subcellular locations of four θCAs, TpθCA1, TpθCA2, TpθCA3, and TpθCA4 were determined by expressing GFP-fused proteins of these TpθCAs in T. pseudonana. As a result, C-terminal GFP fusion proteins of TpθCA1, TpθCA2, and TpθCA3 were all localized in the chloroplast; TpθCA2 was at the central chloroplast area, and the other two TpθCAs were throughout the chloroplast. Immunogold-labeling transmission electron microscopy was further performed for the transformants expressing TpθCA1:GFP and TpθCA2:GFP with anti-GFP-monoclonal antibody. TpθCA1:GFP was localized in the free stroma area, including the peripheral pyrenoid area. TpθCA2:GFP was clearly located as a lined distribution at the central part of the pyrenoid structure, which was most likely the pyrenoid-penetrating thylakoid. Considering the presence of the sequence encoding the N-terminal thylakoid-targeting domain in the TpθCA2 gene, this localization was likely the lumen of the pyrenoid-penetrating thylakoid. On the other hand, TpθCA4:GFP was localized in the cytoplasm. Transcript analysis of these TpθCAs revealed that TpθCA2 and TpθCA3 were upregulated in atmospheric CO2 (0.04% CO2, LC) levels, while TpθCA1 and TpθCA4 were highly induced under 1% CO2 (HC) condition. The genome-editing knockout (KO) of TpθCA1, by CRISPR/Cas9 nickase, gave a silent phenotype in T. pseudonana under LC-HC conditions, which was in sharp agreement with the case of the previously reported TpθCA3 KO. In sharp contrast, TpθCA2 KO is so far unsuccessful, suggesting a housekeeping role of TpθCA2. The silent phenotype of KO strains of stromal CAs suggests that TpαCA1, TpθCA1, and TpθCA3 may have functional redundancy, but different transcript regulations in response to CO2 of these stromal CAs suggest in part their independent roles.


Assuntos
Anidrases Carbônicas , Diatomáceas , Diatomáceas/genética , Diatomáceas/metabolismo , Anidrases Carbônicas/genética , Anidrases Carbônicas/metabolismo , Dióxido de Carbono/metabolismo , Cloroplastos/metabolismo , Proteínas/metabolismo
3.
Microb Ecol ; 86(1): 144-153, 2023 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35622094

RESUMO

The interaction between marine phyto- and bacterioplankton is regulated by multiple environmental and biological factors. Among them, phages as the major regulators of bacterial mortality are considered to have important impacts on algae-associated bacteria and algae-bacteria relationship. However, little is currently known about the actual impact of phages from this perspective. Here, we revealed that phage infection improved the maximum quantum efficiency of photosystem II of Phaeodactylum tricornutum by regulating the associated bacterial community. Specifically, phage infection weakened bacterial abundance and eliminated their negative effects on the diatom. Unexpectedly, the structure of the bacterial community co-cultured with the diatom was not significantly affected, likely because the shaping effect of the diatom on the bacterial community structure can far outcompete or mask the impact of phage infection. Our results established a link between algae, bacteria, and phages, suggesting that phage infection benefits the diatom by regulating the associated bacterial community.


Assuntos
Bacteriófagos , Diatomáceas , Diatomáceas/fisiologia , Bactérias , Organismos Aquáticos
4.
Mar Drugs ; 21(4)2023 Apr 03.
Artigo em Inglês | MEDLINE | ID: mdl-37103370

RESUMO

In the present investigation, 24-methylcholesta-5(6), 22-diene-3ß-ol (MCDO), a major phytosterol was isolated from the cultured marine diatom, Phaeodactylum tricornutum Bohlin, and in vitro and in vivo anti-inflammatory effects were determined. MCDO demonstrated very potent dose-dependent inhibitory effects on the production of nitric oxide (NO) and prostaglandin E2 (PGE2) against lipopolysaccharide (LPS)-induced RAW 264.7 cells with minimal cytotoxic effects. MCDO also demonstrated a strong and significant suppression of pro-inflammatory cytokines of interleukin-1ß (IL-1ß) production, but no substantial inhibitory effects were observed on the production of cytokines, including tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) at the tested concentrations against LPS treatment on RAW macrophages. Western blot assay confirmed the suppression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) protein expressions against LPS-stimulated RAW 264.7 cells. In addition, MCDO was assessed for in vivo anti-inflammatory effects using the zebrafish model. MCDO acted as a potent inhibitor for reactive oxygen species (ROS) and NO levels with a protective effect against the oxidative stress induced by LPS in inflammatory zebrafish embryos. Collectively, MCDO isolated from the cultured marine diatom P. tricornutum exhibited profound anti-inflammatory effects both in vitro and in vivo, suggesting that this major sterol might be a potential treatment for inflammatory diseases.


Assuntos
Diatomáceas , Animais , Diatomáceas/metabolismo , Peixe-Zebra/metabolismo , NF-kappa B/metabolismo , Lipopolissacarídeos/farmacologia , Anti-Inflamatórios/farmacologia , Transdução de Sinais , Citocinas/metabolismo , Interleucina-6/metabolismo , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Ciclo-Oxigenase 2/metabolismo
5.
Ecotoxicol Environ Saf ; 203: 111000, 2020 Oct 15.
Artigo em Inglês | MEDLINE | ID: mdl-32736119

RESUMO

Microplastics are identified as a great threat to marine environments. However, knowledge of their impacts on phytoplankton, especially for the diatoms is scarce. Herein, the effects of different polyvinyl chloride (PVC) microplastic concentrations and contact times (24, 48, 72 and 96 h) on the Fv/Fm and cell density of Phaeodactylum tricornutum (B255), Chaetoceros gracilis (B13) and Thalassiosira sp. (B280) were investigated to evaluate the toxic effects of microplastics on marine diatoms. The effects of PVC microplastics on the morphology of the diatoms was observed by SEM. The order of sensitivity to 1 µm PVC microplastics among three marine diatoms was B13 > B280 > B255, showing that the toxic effects varied with different microalgae species. Furthermore, the presence of a siliceous cell wall played a minimal role in protecting cells from the physical attack of PVC microplastics, with no significant difference from the common cell wall. PVC microplastics caused dose-dependent adverse effects on three marine diatoms. High PVC concentrations (200 mg/L) reduced the chlorophyll content, inhibited Fv/Fm, and affected the photosynthesis of three marine diatoms. The PVC microplastics adsorbed and caused physical damage on the structure of algal cells. Interactions between PVC microplastics and diatoms may be the probable reason for the negative effects of PVC on diatoms.


Assuntos
Diatomáceas/efeitos dos fármacos , Microplásticos/toxicidade , Cloreto de Polivinila/toxicidade , Poluentes Químicos da Água/toxicidade , Adsorção , Clorofila/metabolismo , Diatomáceas/crescimento & desenvolvimento , Diatomáceas/fisiologia , Relação Dose-Resposta a Droga , Microalgas/efeitos dos fármacos , Microalgas/crescimento & desenvolvimento , Microalgas/fisiologia , Fotossíntese/efeitos dos fármacos , Fitoplâncton/efeitos dos fármacos , Fitoplâncton/crescimento & desenvolvimento , Fitoplâncton/fisiologia , Fatores de Tempo
6.
J Enzyme Inhib Med Chem ; 34(1): 510-518, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30688123

RESUMO

Carbonic anhydrases (CAs) are ubiquitous metalloenzymes, which started to be investigated in detail in pathogenic, as well as non-pathogenic species since their pivotal role is to accelerate the physiological CO2 hydration/dehydration reaction significantly. Here, we propose the marine unicellular diatom Phaeodactylum tricornutum as a model organism for testing the membrane penetrability of CA inhibitors (CAIs). Seven inhibitors belonging to the sulphonamide type and possessing a diverse scaffold have been explored for their in vitro inhibition of the whole diatom CAs and the in vivo inhibitory effect on the growth of P. tricornutum. Interesting, inhibition of growth was observed, in vivo, demonstrating that this diatom is a good model for testing the cell wall penetrability of this class of pharmacological agents. Considering that many pathogens are difficult and dangerous to grow in the laboratory, the growth inhibition of P. tricornutum with different such CAIs may be subsequently used to design inhibition studies of CAs from pathogenic organisms.


Assuntos
Inibidores da Anidrase Carbônica/farmacologia , Anidrases Carbônicas/metabolismo , Diatomáceas/efeitos dos fármacos , Sulfonamidas/farmacologia , Inibidores da Anidrase Carbônica/síntese química , Inibidores da Anidrase Carbônica/química , Anidrases Carbônicas/isolamento & purificação , Permeabilidade da Membrana Celular/efeitos dos fármacos , Diatomáceas/enzimologia , Diatomáceas/crescimento & desenvolvimento , Relação Dose-Resposta a Droga , Modelos Moleculares , Estrutura Molecular , Relação Estrutura-Atividade , Sulfonamidas/síntese química , Sulfonamidas/química
7.
Mar Drugs ; 17(2)2019 02 09.
Artigo em Inglês | MEDLINE | ID: mdl-30744121

RESUMO

An unprecedented phosphatidylmonogalactosyldiacylglycerol pool (PGDG, 1) rich in polyunsaturated fatty acids was isolated from the marine diatoms Thalassiosira weissflogii. Here we report for the first time the NMR characterization of this rare lipid from marine organisms along with a synthetic strategy for the preparation of a PGDG analog (2). PGDG 1 exhibited immunostimulatory activity in human dendritic cells (DCs) and the synthetic PGDG 2 was prepared to explore its mechanism of action. A Toll-like receptor-4 (TLR-4) agonistic activity was evidenced in human and murine DCs underlying the antigen-specific T-cell activation of this class of molecules.


Assuntos
Adjuvantes Imunológicos/química , Adjuvantes Imunológicos/farmacologia , Diatomáceas/química , Glicolipídeos/química , Glicolipídeos/farmacologia , Fosfatidilgliceróis/química , Fosfatidilgliceróis/farmacologia , Receptor 4 Toll-Like/agonistas , Adjuvantes Imunológicos/síntese química , Adjuvantes Imunológicos/isolamento & purificação , Animais , Espectroscopia de Ressonância Magnética Nuclear de Carbono-13 , Células Cultivadas , Células Dendríticas/efeitos dos fármacos , Células Dendríticas/imunologia , Feminino , Glicolipídeos/síntese química , Glicolipídeos/isolamento & purificação , Humanos , Camundongos , Camundongos Endogâmicos C57BL , Estrutura Molecular , Ressonância Magnética Nuclear Biomolecular , Fosfatidilgliceróis/síntese química , Fosfatidilgliceróis/isolamento & purificação , Receptor 4 Toll-Like/imunologia
8.
Proc Natl Acad Sci U S A ; 113(35): 9828-33, 2016 08 30.
Artigo em Inglês | MEDLINE | ID: mdl-27531955

RESUMO

The algal pyrenoid is a large plastid body, where the majority of the CO2-fixing enzyme, ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) resides, and it is proposed to be the hub of the algal CO2-concentrating mechanism (CCM) and CO2 fixation. The thylakoid membrane is often in close proximity to or penetrates the pyrenoid itself, implying there is a functional cooperation between the pyrenoid and thylakoid. Here, GFP tagging and immunolocalization analyses revealed that a previously unidentified protein, Pt43233, is targeted to the lumen of the pyrenoid-penetrating thylakoid in the marine diatom Phaeodactylum tricornutum The recombinant Pt43233 produced in Escherichia coli cells had both carbonic anhydrase (CA) and esterase activities. Furthermore, a Pt43233:GFP-fusion protein immunoprecipitated from P. tricornutum cells displayed a greater specific CA activity than detected for the purified recombinant protein. In an RNAi-generated Pt43233 knockdown mutant grown in atmospheric CO2 levels, photosynthetic dissolved inorganic carbon (DIC) affinity was decreased and growth was constantly retarded; in contrast, overexpression of Pt43233:GFP yielded a slightly greater photosynthetic DIC affinity. The discovery of a θ-type CA localized to the thylakoid lumen, with an essential role in photosynthetic efficiency and growth, strongly suggests the existence of a common role for the thylakoid-luminal CA with respect to the function of diverse algal pyrenoids.


Assuntos
Proteínas de Algas/metabolismo , Dióxido de Carbono/metabolismo , Anidrases Carbônicas/metabolismo , Diatomáceas/enzimologia , Fotossíntese/fisiologia , Tilacoides/enzimologia , Proteínas de Algas/genética , Sequência de Aminoácidos , Ciclo do Carbono/fisiologia , Anidrases Carbônicas/genética , Clonagem Molecular , Diatomáceas/genética , Diatomáceas/crescimento & desenvolvimento , Diatomáceas/ultraestrutura , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Vetores Genéticos/química , Vetores Genéticos/metabolismo , Cinética , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Ribulose-Bifosfato Carboxilase/genética , Ribulose-Bifosfato Carboxilase/metabolismo , Alinhamento de Sequência , Tilacoides/genética , Tilacoides/ultraestrutura
9.
Ecotoxicol Environ Saf ; 169: 85-92, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30439583

RESUMO

Aluminum (Al) is widespread in the environment including the ocean. The effects of Al on marine organisms have attracted more and more attention in recent years. However, the mechanisms of uptake of Al by marine organisms and the subcellular distribution of Al once assimilated are unknown. Here we report the uptake and subcellular distribution of Al in a marine diatom Thalassiosira weissflogii. Short-term (< 120 min) uptake experiments showed that the Al uptake rate by the diatom was 0.033 ±â€¯0.013 fmol-1 cell-1 min-1 (internalization flux normalized to the exposure Al concentration of 2 µM = 0.034 ±â€¯0.013 nmol m-2 min-1 nM-1). Subcellular fractionation experiments showed that the internalized Al was partitioned to subcellular components in the following order: granules (69 ±â€¯5%) > debris (17 ±â€¯4%) > organelles (12 ±â€¯2%) > heat-stable peptides (HSP) (~2%) > heat-denaturable proteins (HDP) (< 1%), indicating that the majority of intracellular Al was detoxified and stored in inorganic forms. The subcellular distribution of Al in the diatom is different from that of Al in freshwater green algae, in which most of the internalized Al is partitioned to organelles. We also evaluated an artificial seawater-based EDTA rinse solution to remove Al adsorbed on the diatom cell surface. Overall, our study provides new information to understand the mechanisms of uptake of Al by marine diatoms, and the mechanisms responsible for the biological effects (both toxic and beneficial) of Al on the growth of marine phytoplankton, especially diatoms.


Assuntos
Alumínio/metabolismo , Diatomáceas/efeitos dos fármacos , Água do Mar/química , Poluentes Químicos da Água/metabolismo , Adsorção , Alumínio/análise , Alumínio/toxicidade , Diatomáceas/metabolismo , Fitoplâncton/efeitos dos fármacos , Fitoplâncton/metabolismo , Poluentes Químicos da Água/análise , Poluentes Químicos da Água/toxicidade
10.
J Exp Bot ; 68(14): 3763-3772, 2017 06 01.
Artigo em Inglês | MEDLINE | ID: mdl-28633304

RESUMO

Diatoms operate a CO2-concentrating mechanism (CCM) that drives upwards of 20% of annual global primary production. Recent progress in CCM research in the marine pennate diatom Phaeodactylum tricornutum revealed that this diatom directly takes up HCO3- from seawater through low-CO2-inducible plasma membrane HCO3- transporters, which belong to the solute carrier (SLC) 4 family. Apart from this, studies of carbonic anhydrases (CAs) in diatoms have revealed considerable diversity in classes and localization among species. This strongly suggests that the CA systems, which control permeability and flux of dissolved inorganic carbon (DIC) by catalysing reversible CO2 hydration, have evolved from diverse origins. Of particular interest is the occurrence of low-CO2-inducible external CAs in the centric marine diatom Thalassiosira pseudonana, offering a strategy of CA-catalysed initial CO2 entry via passive diffusion, contrasting with active DIC transport in P. tricornutum. Molecular mechanisms to transport DIC across chloroplast envelopes are likely also through specific HCO3- transporters, although details have yet to be elucidated. Furthermore, recent discovery of a luminal θ-CA in the diatom thylakoid implied a common strategy in the mechanism to supply CO2 to RubisCO in the pyrenoid, which is conserved among green algae and some heterokontophytes. These results strongly suggest an occurrence of convergent coevolution between the pyrenoid and thylakoid membrane in aquatic photosynthesis.


Assuntos
Dióxido de Carbono/metabolismo , Diatomáceas/metabolismo , Fotossíntese , Biofísica
11.
Mar Drugs ; 15(10)2017 Oct 18.
Artigo em Inglês | MEDLINE | ID: mdl-29057803

RESUMO

Antifreeze proteins (AFPs) protecting the cells against freezing are produced in response to extremely low temperatures in diverse psychrophilic organisms, and they are encoded by multiple gene families. The AFP of Antarctic marine diatom Chaetoceros neogracile is reported in our previous research, but like other microalgae, was considered to probably have additional genes coding AFPs. In this paper, we reported the cloning and characterization of additional AFP gene from C. neogracile (Cn-isoAFP). Cn-isoAFP protein is 74.6% identical to the previously reported Cn-AFP. The promoter sequence of Cn-isoAFP contains environmental stress responsive elements for cold, thermal, and high light conditions. Cn-isoAFP transcription levels increased dramatically when cells were exposed to freezing (-20 °C), thermal (10 °C), or high light (600 µmol photon m-2 s-1) stresses. The thermal hysteresis (TH) activity of recombinant Cn-isoAFP was 0.8 °C at a protein concentration of 5 mg/mL. Results from homology modeling and TH activity analysis of site-directed mutant proteins elucidated AFP mechanism to be a result of flatness of B-face maintained via hydrophobic interactions.


Assuntos
Proteínas Anticongelantes/fisiologia , Diatomáceas/fisiologia , Congelamento/efeitos adversos , Isoformas de Proteínas/fisiologia , Regiões Antárticas , Proteínas Anticongelantes/química , Clonagem Molecular , Interações Hidrofóbicas e Hidrofílicas , Modelos Moleculares , Mutagênese Sítio-Dirigida , Regiões Promotoras Genéticas/genética , Isoformas de Proteínas/química , Proteínas Recombinantes/química , Proteínas Recombinantes/genética , Elementos de Resposta/genética , Homologia de Sequência de Aminoácidos , Estresse Fisiológico/fisiologia
12.
J Exp Bot ; 67(13): 3939-51, 2016 06.
Artigo em Inglês | MEDLINE | ID: mdl-27225826

RESUMO

Diatoms are phytoplanktonic organisms that grow successfully in the ocean where light conditions are highly variable. Studies of the molecular mechanisms of light acclimation in the marine diatom Phaeodactylum tricornutum show that carotenoid de-epoxidation enzymes and LHCX1, a member of the light-harvesting protein family, both contribute to dissipate excess light energy through non-photochemical quenching (NPQ). In this study, we investigate the role of the other members of the LHCX family in diatom stress responses. Our analysis of available genomic data shows that the presence of multiple LHCX genes is a conserved feature of diatom species living in different ecological niches. Moreover, an analysis of the levels of four P. tricornutum LHCX transcripts in relation to protein expression and photosynthetic activity indicates that LHCXs are differentially regulated under different light intensities and nutrient starvation, mostly modulating NPQ capacity. We conclude that multiple abiotic stress signals converge to regulate the LHCX content of cells, providing a way to fine-tune light harvesting and photoprotection. Moreover, our data indicate that the expansion of the LHCX gene family reflects functional diversification of its members which could benefit cells responding to highly variable ocean environments.


Assuntos
Proteínas de Algas/genética , Diatomáceas/genética , Regulação da Expressão Gênica , Complexos de Proteínas Captadores de Luz/genética , Fitoplâncton/genética , Transdução de Sinais , Proteínas de Algas/metabolismo , Diatomáceas/metabolismo , Complexos de Proteínas Captadores de Luz/metabolismo , Fotossíntese , Fitoplâncton/metabolismo
13.
Mar Drugs ; 13(8): 5334-57, 2015 Aug 20.
Artigo em Inglês | MEDLINE | ID: mdl-26308005

RESUMO

Carotenoids exert beneficial effects on human health through their excellent antioxidant activity. To increase carotenoid productivity in the marine Pennales Phaeodactylum tricornutum, we genetically engineered the phytoene synthase gene (psy) to improve expression because RNA-sequencing analysis has suggested that the expression level of psy is lower than other enzyme-encoding genes that are involved in the carotenoid biosynthetic pathway. We isolated psy from P. tricornutum, and this gene was fused with the enhanced green fluorescent protein gene to detect psy expression. After transformation using the microparticle bombardment technique, we obtained several P. tricornutum transformants and confirmed psy expression in their plastids. We investigated the amounts of PSY mRNA and carotenoids, such as fucoxanthin and ß-carotene, at different growth phases. The introduction of psy increased the fucoxanthin content of a transformants by approximately 1.45-fold relative to the levels in the wild-type diatom. However, some transformants failed to show a significant increase in the carotenoid content relative to that of the wild-type diatom. We also found that the amount of PSY mRNA at log phase might contribute to the increase in carotenoids in the transformants at stationary phase.


Assuntos
Organismos Aquáticos/genética , Vias Biossintéticas/genética , Carotenoides/biossíntese , Diatomáceas/genética , Expressão Gênica/genética , Geranil-Geranildifosfato Geranil-Geraniltransferase/genética , Carotenoides/genética , Plastídeos/genética , RNA Mensageiro/genética , Xantofilas/genética , beta Caroteno/genética
14.
Sci Total Environ ; 923: 171371, 2024 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-38432364

RESUMO

The wide application of benzophenones (BPs), such as benzophenone-3 (BP3), as an ingredient in sunscreens, cosmetics, coatings, and plastics, has led to their global contamination in aquatic environments. Using the marine diatom Chaetoceros neogracilis as a model, this study assessed the toxic effects and mechanisms of BP3 and its two major metabolites (BP8 and BP1). The results showed that BP3 exhibited higher toxicity on C. neogracilis than BP8 and BP1, with their 72-h median effective concentrations being 0.4, 0.8 and 4 mg/L, respectively. Photosynthesis efficiencies were significantly reduced after exposure to environmentally relevant concentrations of the three benzophenones, while cell viability, membrane integrity, membrane potential, and metabolic activities could be further impaired at their higher concentrations. Comparative transcriptomic analysis, followed by gene ontology and KEGG pathway enrichment analyses unraveled that all the three tested benzophenones disrupted photosynthesis and nitrogen metabolism of the diatom through alteration of similar pathways. The toxic effect of BP3 was also attributable to its unique inhibitory effects on eukaryotic ribosome biosynthesis and DNA replication. Taken together, our findings underscore that benzophenones may pose a significant threat to photosynthesis, oxygen production, primary productivity, carbon fixation, and the nitrogen cycle of diatom in coastal waters worldwide.


Assuntos
Cosméticos , Diatomáceas , Diatomáceas/metabolismo , Protetores Solares/toxicidade , Protetores Solares/metabolismo , Cosméticos/metabolismo , Benzofenonas/toxicidade , Benzofenonas/metabolismo
15.
PhytoKeys ; 232: 77-88, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37732132

RESUMO

Craspedostauros E.J. Cox is a diatom genus comprising 17 taxa reported from various regions of the world. While many species of Craspedostauros are epibiontic, the taxa have variable ecological preferences. In this study we formally describe Craspedostaurosnazmiisp. nov., an epilithic species discovered along the Turkish Black Sea Coast, based on light and scanning electron microscopy. Craspedostaurosnazmiisp. nov. is characterized by valves that are lanceolate to narrowly lanceolate, slightly constricted near the apices with uniseriate, parallel throughout the whole valve, transapical striae and and the presence of an apical silica flap. The areolae are distributed over the valve face and the mantle. The differences and similarities between C.nazmiisp. nov. and established species of Craspedostauros are discussed. Based on shape and morphometrics, the most similar species is Craspedostauroscapensis, but it is easily distinguished from C.nazmiisp. nov. by its lack of an apical silica flap.

16.
Mar Environ Res ; 186: 105929, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36863076

RESUMO

Multifaceted changes in marine environments as a result of anthropogenic activities are likely to have a compounding impact on the physiology of marine phytoplankton. Most studies on the combined effects of rising pCO2, sea surface temperature, and UVB radiation on marine phytoplankton were only conducted in the short-term, which does not allow to test the adaptive capacity of phytoplankton and associated potential trade-offs. Here, we investigated populations of the diatom Phaeodactylum tricornutum that were long-term (∼3.5 years, ∼3000 generations) adapted to elevated CO2 and/or elevated temperatures, and their physiological responses to short-term (∼2 weeks) exposure of two levels of ultraviolet-B (UVB) radiation. Our results showed that while elevated UVB radiation showed predominantly negative effects on the physiological performance of P. tricornutum regardless of adaptation regimes. Elevated temperature alleviated these effects on most of the measured physiological parameters (e.g., photosynthesis). We also found that elevated CO2 can modulate these antagonistic interactions, and conclude that long-term adaptation to sea surface warming and rising CO2 may alter this diatom's sensitivity to elevated UVB radiation in the environment. Our study provides new insights into marine phytoplankton's long-term responses to the interplay of multiple environmental changes driven by climate change.


Assuntos
Diatomáceas , Temperatura , Dióxido de Carbono , Fitoplâncton/fisiologia , Aclimatação
17.
Mar Genomics ; 61: 100921, 2022 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-35030498

RESUMO

Promoters are key elements for the regulation of gene expression. Recently, we investigated the activity of promoters derived from marine diatom-infecting viruses (DIVs) in marine diatoms. Previously, we focused on potential promoter regions of the replication-associated protein gene and the capsid protein gene of the DIVs. In addition to these genes, two genes of unknown function (VP1 and VP4 genes) have been found in the DIV genomes. In this study, the promoter regions of the VP1 gene and VP4 gene derived from a Chaetoceros lorenzianus-infecting DNA virus (named ClP3 and ClP4, respectively) were newly isolated. ClP4 was found to be a constitutive promoter and displayed the highest activity. In particular, the 3' region of ClP4 (ClP4 3' region) showed a higher promoter activity than full-length ClP4. The ClP4 3' region might involve high-level promoter activity of ClP4. In addition, the ClP4 3' region may be useful for substance production and metabolic engineering of diatoms.


Assuntos
Vírus de DNA , Diatomáceas , Fases de Leitura Aberta , Regiões Promotoras Genéticas , Vírus de DNA/genética , Diatomáceas/genética , Diatomáceas/virologia
18.
J Hazard Mater ; 409: 124460, 2021 05 05.
Artigo em Inglês | MEDLINE | ID: mdl-33183842

RESUMO

The current knowledge about the ecological effects of microplastic (MP) remains limited, and to-date ecotoxicity tests often utilize standard microplastic with one or two distinct size classes and expose the organisms to unrealistically high MP concentrations. We exposed the marine diatom Phaeodactylum tricornutum to microplastic particles of a mimicked realistic size frequency distribution complemented with serial experiments with distinct size classes. To do so, we exposed this diatom to a concentration series of different sized polyethylene (PE) microbeads (sizes: 10-106 µm; 1.25 ×102-1.25 ×107 particles/L) in a 72-h growth inhibition test. No effect on the growth of P. tricornutum by virgin PE microbeads up to 1.25 × 107 particles/L (or 499 mg/L), indicating environmentally relevant concentrations and sizes of MP does not alter the growth of marine diatoms. Results of smaller sized MPs (10-20 µm) did not differ from those obtained with larger MPs (90-106 µm) and mix sized MPs (10-106 µm), i.e. no impact on the microalgae growth. As a pioneer work, our results contribute with high quality dose-response data to an improved risk assessment of microplastic under realistic present and future marine MP pollution.


Assuntos
Diatomáceas , Microalgas , Poluentes Químicos da Água , Microplásticos , Plásticos/toxicidade , Poluentes Químicos da Água/toxicidade
19.
Mar Pollut Bull ; 163: 111950, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33444995

RESUMO

A big challenge of the 21st century is to cope with the huge amounts of plastic waste on Earth. Especially the oceans are heavily polluted with plastics. To counteract this issue, biological (enzymatic) plastic decomposition is increasingly gaining attention. Recently it was shown that polyethylene terephthalate (PET) can be degraded in a saltwater-based environment using bacterial PETase produced by a marine diatom. At moderate temperatures, plastic biodegradation is slow and requires sensitive methods for detection, at least at initial stages. However, conventional methods for verifying the plastic degradation are either complex, expensive, time-consuming or they interfere with the degradation process. Here, we adapt lensless digital holographic microscopy (LDHM) as a new application for efficiently monitoring enzymatic degradation of a PET glycol copolymer (PETG). LDHM is a cost-effective, compact and sensitive optical method. We demonstrate enzymatic PETG degradation over a time course of 43 days employing numerical analysis of LDHM images.


Assuntos
Microscopia , Plásticos , Bactérias , Biodegradação Ambiental , Oceanos e Mares
20.
Plant Sci ; 296: 110475, 2020 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-32540005

RESUMO

Marine diatoms constitute a major group of unicellular photosynthetic eukaryotes. Diatoms are widely applicable for both basic studies and applied studies. Molecular tools and techniques have been developed for diatom research. Among these tools, several endogenous gene promoters (e.g., the fucoxanthin chlorophyll a/c-binding protein gene promoter) have become available for expressing transgenes in diatoms. Gene promoters that drive transgene expression at a high level are very important for the metabolic engineering of diatoms. Various marine diatom-infecting viruses (DIVs), including both DNA viruses and RNA viruses, have recently been isolated, and their genome sequences have been characterized. Promoters from viruses that infect plants and mammals are widely used as constitutive promoters to achieve high expression of transgenes. Thus, we recently investigated the activity of promoters derived from marine DIVs in the marine diatom, Phaeodactylum tricornutum. We discuss novel viral promoters that will be useful for the future metabolic engineering of diatoms.


Assuntos
Vírus de DNA/metabolismo , Diatomáceas/genética , Engenharia Genética/métodos , Regiões Promotoras Genéticas , Vírus de RNA/metabolismo , Diatomáceas/virologia , Engenharia Metabólica/métodos , Regiões Promotoras Genéticas/genética
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