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1.
Plant J ; 119(1): 65-83, 2024 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-38608130

RESUMEN

The determination of physiological tolerance ranges of photosynthetic species and of the biochemical mechanisms underneath are fundamental to identify target processes and metabolites that will inspire enhanced plant management and production for the future. In this context, the terrestrial green algae within the genus Prasiola represent ideal models due to their success in harsh environments (polar tundras) and their extraordinary ecological plasticity. Here we focus on the outstanding Prasiola antarctica and compare two natural populations living in very contrasting microenvironments in Antarctica: the dry sandy substrate of a beach and the rocky bed of an ephemeral freshwater stream. Specifically, we assessed their photosynthetic performance at different temperatures, reporting for the first time gnsd values in algae and changes in thylakoid metabolites in response to extreme desiccation. Stream population showed lower α-tocopherol content and thicker cell walls and thus, lower gnsd and photosynthesis. Both populations had high temperatures for optimal photosynthesis (around +20°C) and strong constitutive tolerance to freezing and desiccation. This tolerance seems to be related to the high constitutive levels of xanthophylls and of the cylindrical lipids di- and tri-galactosyldiacylglycerol in thylakoids, very likely related to the effective protection and stability of membranes. Overall, P. antarctica shows a complex battery of constitutive and plastic protective mechanisms that enable it to thrive under harsh conditions and to acclimate to very contrasting microenvironments, respectively. Some of these anatomical and biochemical adaptations may partially limit photosynthesis, but this has a great potential to rise in a context of increasing temperature.


Asunto(s)
Fotosíntesis , Tilacoides , Tilacoides/metabolismo , Regiones Antárticas , Fotosíntesis/fisiología , Chlorophyceae/fisiología , Chlorophyceae/metabolismo , Xantófilas/metabolismo , Adaptación Fisiológica/fisiología , Desecación , Aclimatación
2.
Physiol Plant ; 176(2): e14296, 2024.
Artículo en Inglés | MEDLINE | ID: mdl-38650503

RESUMEN

In Dunaliella tertiolecta, a microalga renowned for its extraordinary tolerance to high salinity levels up to 4.5 M NaCl, the mechanisms underlying its stress response have largely remained a mystery. In a groundbreaking discovery, this study identifies a choline dehydrogenase enzyme, termed DtCHDH, capable of converting choline to betaine aldehyde. Remarkably, this is the first identification of such an enzyme not just in D. tertiolecta but across the entire Chlorophyta. A 3D model of DtCHDH was constructed, and molecular docking with choline was performed, revealing a potential binding site for the substrate. The enzyme was heterologously expressed in E. coli Rosetta (DE3) and subsequently purified, achieving enzyme activity of 672.2 U/mg. To elucidate the role of DtCHDH in the salt tolerance of D. tertiolecta, RNAi was employed to knock down DtCHDH gene expression. The results indicated that the Ri-12 strain exhibited compromised growth under both high and low salt conditions, along with consistent levels of DtCHDH gene expression and betaine content. Additionally, fatty acid analysis indicated that DtCHDH might also be a FAPs enzyme, catalyzing reactions with decarboxylase activity. This study not only illuminates the role of choline metabolism in D. tertiolecta's adaptation to high salinity but also identifies a novel target for enhancing the NaCl tolerance of microalgae in biotechnological applications.


Asunto(s)
Betaína , Colina-Deshidrogenasa , Tolerancia a la Sal , Betaína/metabolismo , Tolerancia a la Sal/genética , Colina-Deshidrogenasa/metabolismo , Colina-Deshidrogenasa/genética , Colina/metabolismo , Chlorophyceae/genética , Chlorophyceae/fisiología , Chlorophyceae/enzimología , Chlorophyceae/metabolismo , Microalgas/genética , Microalgas/enzimología , Microalgas/metabolismo , Simulación del Acoplamiento Molecular , Cloruro de Sodio/farmacología
3.
Proc Natl Acad Sci U S A ; 117(29): 17438-17445, 2020 07 21.
Artículo en Inglés | MEDLINE | ID: mdl-32636259

RESUMEN

Among green plants, desiccation tolerance is common in seeds and spores but rare in leaves and other vegetative green tissues. Over the last two decades, genes have been identified whose expression is induced by desiccation in diverse, desiccation-tolerant (DT) taxa, including, e.g., late embryogenesis abundant proteins (LEA) and reactive oxygen species scavengers. This up-regulation is observed in DT resurrection plants, mosses, and green algae most closely related to these Embryophytes. Here we test whether this same suite of protective genes is up-regulated during desiccation in even more distantly related DT green algae, and, importantly, whether that up-regulation is unique to DT algae or also occurs in a desiccation-intolerant relative. We used three closely related aquatic and desert-derived green microalgae in the family Scenedesmaceae and capitalized on extraordinary desiccation tolerance in two of the species, contrasting with desiccation intolerance in the third. We found that during desiccation, all three species increased expression of common protective genes. The feature distinguishing gene expression in DT algae, however, was extensive down-regulation of gene expression associated with diverse metabolic processes during the desiccation time course, suggesting a switch from active growth to energy-saving metabolism. This widespread downshift did not occur in the desiccation-intolerant taxon. These results show that desiccation-induced up-regulation of expression of protective genes may be necessary but is not sufficient to confer desiccation tolerance. The data also suggest that desiccation tolerance may require induced protective mechanisms operating in concert with massive down-regulation of gene expression controlling numerous other aspects of metabolism.


Asunto(s)
Adaptación Fisiológica/genética , Chlorophyta/genética , Chlorophyta/fisiología , Desecación , Regulación de la Expresión Génica de las Plantas , Chlorophyceae/genética , Chlorophyceae/fisiología , Regulación hacia Abajo , Extremófilos/fisiología , Ontología de Genes , Genes de Plantas/genética , Factores de Transcripción , Transcriptoma , Regulación hacia Arriba
4.
Plant Cell ; 31(3): 579-601, 2019 03.
Artículo en Inglés | MEDLINE | ID: mdl-30787178

RESUMEN

Light and nutrients are critical regulators of photosynthesis and metabolism in plants and algae. Many algae have the metabolic flexibility to grow photoautotrophically, heterotrophically, or mixotrophically. Here, we describe reversible Glc-dependent repression/activation of oxygenic photosynthesis in the unicellular green alga Chromochloris zofingiensis. We observed rapid and reversible changes in photosynthesis, in the photosynthetic apparatus, in thylakoid ultrastructure, and in energy stores including lipids and starch. Following Glc addition in the light, C. zofingiensis shuts off photosynthesis within days and accumulates large amounts of commercially relevant bioproducts, including triacylglycerols and the high-value nutraceutical ketocarotenoid astaxanthin, while increasing culture biomass. RNA sequencing reveals reversible changes in the transcriptome that form the basis of this metabolic regulation. Functional enrichment analyses show that Glc represses photosynthetic pathways while ketocarotenoid biosynthesis and heterotrophic carbon metabolism are upregulated. Because sugars play fundamental regulatory roles in gene expression, physiology, metabolism, and growth in both plants and animals, we have developed a simple algal model system to investigate conserved eukaryotic sugar responses as well as mechanisms of thylakoid breakdown and biogenesis in chloroplasts. Understanding regulation of photosynthesis and metabolism in algae could enable bioengineering to reroute metabolism toward beneficial bioproducts for energy, food, pharmaceuticals, and human health.


Asunto(s)
Chlorophyceae/fisiología , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Glucosa/farmacología , Oxígeno/metabolismo , Fotosíntesis/efectos de los fármacos , Transcriptoma/efectos de los fármacos , Antioxidantes/metabolismo , Bioingeniería , Carbono/metabolismo , Chlorophyceae/genética , Chlorophyceae/efectos de la radiación , Chlorophyceae/ultraestructura , Regulación de la Expresión Génica de las Plantas/efectos de la radiación , Fotosíntesis/efectos de la radiación , Tilacoides/metabolismo , Tilacoides/ultraestructura , Transcriptoma/efectos de la radiación , Xantófilas/metabolismo
5.
Ecotoxicol Environ Saf ; 207: 111153, 2021 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-32896819

RESUMEN

Micro/nano-plastics (MNPs) have been found within many environments and organisms including humans, making them a significant and growing concern. Initial research into the potential detrimental effects these MNPs both from acute and chronic exposure has been ongoing but still requires substantially more data to clarify. This research presents the response of nano-polystyrene (NPS) on Raphidocelis subcapitata, a freshwater alga, under an existing acute toxicity test along with additional analytical techniques to try identifying possible sources of toxicity. R. subcapitata cells were exposed for 72 h to a concentration range of 0-100 mg/l NPS. Growth Inhibition (GI) testing showed the R. subcapitata demonstrated statistically distinct reductions in growth over 72 h at all NPS exposure concentrations while not suffering culture collapse. By the 100 mg/l NPS exposure the R. subcapitata has suffered almost a 33.7% reduction in cell concentration after 72 h compared to control samples. Confocal imaging showed the NPS wasn't permeating into the algal plasma membrane or individual organelles but agglomerated onto the algal cell wall. The agglomeration was irregular but increased in total surface area covered as NPS concentration increases. UV-Vis fluorimetry testing produced a linear response of emission intensities to algae exposed to the 0-100 mg/l range of NPS. However, comparisons of emission intensity values of algae exposed to NPS to emission intensities of pure NPS at identical concentrations showed consistent intensity reduction. This response further indicated NPS agglomerating within the media and onto the alga cells seen from confocal imaging. Finally, Raman spectroscopy on R. subcapitata attempted to distinguish the key 1001 cm-1 peak or other crucial identifier peaks of polystyrene from overall Raman spectra. This was not successful as emissions from algal component (e.g. phenylaniline) completely suppressed the signal region.


Asunto(s)
Chlorophyceae/fisiología , Poliestirenos/toxicidad , Contaminantes Químicos del Agua/toxicidad , Agua Dulce/análisis , Humanos
6.
Ecotoxicol Environ Saf ; 207: 111559, 2021 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-33254413

RESUMEN

The current study was performed to investigate the protective effects of dietary Haematococcus pluvialis (H. pluvialis) on the oxidative stress induced by arsenic in rainbow trout (Oncorhynchus mykiss). The fish (20.70 ± 0.09 g) were fed with H. pluvialis at the levels of 0.28, 0.56, and 1.12 g 100 g-1 diet for 60 days. Then, each group was divided into two subgroups. In one of the subgroups, fish were exposed to arsenic challenge at a level of 9.1 mg/L. The other subset was used as the negative control. After the 96 h of toxicity test, protein and lipid oxidative levels, antioxidant-relevant gene expression as well as several chemical factors, including pH and peroxide value and moisture content, were evaluated in the fillet samples. Results showed that feeding with H. pluvialis decreased the levels of pH, peroxide value, as well as protein and lipid oxidation levels in treatment groups. Besides, the expression of antioxidant genes was significantly increased in the groups administrated with H. pluvialis. Based on the results of this study, feeding H. pluvialis attenuated the oxidative stress induced by arsenic in rainbow trout fillet through improving the antioxidant defense system.


Asunto(s)
Arsénico/toxicidad , Chlorophyceae/fisiología , Oncorhynchus mykiss/fisiología , Contaminantes Químicos del Agua/toxicidad , Animales , Antioxidantes/metabolismo , Arsénico/metabolismo , Chlorophyceae/metabolismo , Dieta , Oncorhynchus mykiss/metabolismo , Oxidación-Reducción , Estrés Oxidativo/efectos de los fármacos
7.
Ecotoxicol Environ Saf ; 208: 111628, 2021 Jan 15.
Artículo en Inglés | MEDLINE | ID: mdl-33396148

RESUMEN

Metals may cause damage to the biota of contaminated environments. Moreover, using multiple endpoints in ecotoxicological studies is useful to better elucidate the mechanisms of toxicity of these compounds. Therefore, this study aimed to evaluate the effects of cadmium (Cd) and cobalt (Co) on growth, biochemical and photosynthetic parameters of the microalgae Raphidocelis subcapitata, through quantification of lipid classes composition, chlorophyll a (Chl a) content, maximum (ΦM) and effective (Φ'M) quantum yields and efficiency of the oxygen-evolving complex (OEC). Both metals affected the algal population growth, with an IC50-96h of 0.67 and 1.53 µM of Cd and Co, respectively. Moreover, the metals led to an increase in the total lipid content and reduced efficiency of OEC and ΦM. Cell density was the most sensitive endpoint to detect Cd toxicity after 96 h of treatment. Regarding Co, the photosynthetic parameters were the most affected and the total lipid content was the most sensitive endpoint as it was altered by the exposure to this metal in all concentrations. Cd led to increased contents of the lipid class wax esters (0.89 µM) and phospholipids (PL - at 0.89 and 1.11 µM) and decreased values of triglycerides (at 0.22 µM) and acetone-mobile polar lipids (AMPL - at 0.44 and 1.11 µM). The percentage of free fatty acids (FFA) and PL of microalgae exposed to Co increased, whereas AMPL decreased in all concentrations tested. We were able to detect differences between the toxicity mechanisms of each metal, especially how Co interferes in the microalgae at a biochemical level. Furthermore, to the best of our knowledge, this is the first study reporting Co effects in lipid classes of a freshwater Chlorophyceae. The damage caused by Cd and Co may reach higher trophic levels, causing potential damage to the aquatic communities as microalgae are primary producers and the base of the food chain.


Asunto(s)
Cadmio/toxicidad , Chlorophyceae/fisiología , Cobalto/toxicidad , Contaminantes Químicos del Agua/toxicidad , Chlorophyceae/efectos de los fármacos , Clorofila A , Ecotoxicología , Agua Dulce/química , Metales/farmacología , Microalgas/efectos de los fármacos , Fotosíntesis/efectos de los fármacos , Complejo de Proteína del Fotosistema II
8.
Ecotoxicol Environ Saf ; 207: 111264, 2021 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-32911184

RESUMEN

This study investigated the effect of the herbicide metolachlor (MET) on the redox homeostasis of the freshwater green alga Pseudokirchneriella subcapitata. At low MET concentrations (≤40 µg L-1), no effects on algal cells were detected. The exposure of P. subcapitata to 45-235 µg L-1 MET induced a significant increase of reactive oxygen species (ROS). The intracellular levels of ROS were particularly increased at high (115 and 235 µg L-1) but environmentally relevant MET concentrations. The exposure of algal cells to 115 and 235 µg L-1 MET originated a decrease in the levels of antioxidants molecules (reduced glutathione and carotenoids) as well as a reduction of the activity of scavenging enzymes (superoxide dismutase and catalase). These results suggest that antioxidant (non-enzymatic and enzymatic) defenses were affected by the excess of MET. As consequence of this imbalance (ROS overproduction and decline of the antioxidant system), ROS inflicted oxidative injury with lipid peroxidation and damage of cell membrane integrity. The results provide further insights about the toxic modes of action of MET on a non-target organism and emphasize the relevance of toxicological studies in the assessment of the impact of herbicides in freshwater environments.


Asunto(s)
Acetamidas/toxicidad , Chlorophyceae/efectos de los fármacos , Herbicidas/toxicidad , Contaminantes Químicos del Agua/toxicidad , Antioxidantes/metabolismo , Catalasa/metabolismo , Chlorophyceae/fisiología , Agua Dulce , Glutatión/metabolismo , Homeostasis/efectos de los fármacos , Peroxidación de Lípido/efectos de los fármacos , Oxidación-Reducción , Estrés Oxidativo/efectos de los fármacos , Especies Reactivas de Oxígeno/metabolismo , Superóxido Dismutasa/metabolismo
9.
BMC Biol ; 18(1): 126, 2020 09 16.
Artículo en Inglés | MEDLINE | ID: mdl-32938439

RESUMEN

BACKGROUND: Plastid electron transport systems are essential not only for photosynthesis but also for dissipating excess reducing power and sinking excess electrons generated by various redox reactions. Although numerous organisms with plastids have lost their photoautotrophic lifestyles, there is a spectrum of known functions of remnant plastids in non-photosynthetic algal/plant lineages; some of non-photosynthetic plastids still retain diverse metabolic pathways involving redox reactions while others, such as apicoplasts of apicomplexan parasites, possess highly reduced sets of functions. However, little is known about underlying mechanisms for redox homeostasis in functionally versatile non-photosynthetic plastids and thus about the reductive evolution of plastid electron transport systems. RESULTS: Here we demonstrated that the central component for plastid electron transport systems, plastoquinone/plastoquinol pool, is still retained in a novel strain of an obligate heterotrophic green alga lacking the photosynthesis-related thylakoid membrane complexes. Microscopic and genome analyses revealed that the Volvocales green alga, chlamydomonad sp. strain NrCl902, has non-photosynthetic plastids and a plastid DNA that carries no genes for the photosynthetic electron transport system. Transcriptome-based in silico prediction of the metabolic map followed by liquid chromatography analyses demonstrated carotenoid and plastoquinol synthesis, but no trace of chlorophyll pigments in the non-photosynthetic green alga. Transient RNA interference knockdown leads to suppression of plastoquinone/plastoquinol synthesis. The alga appears to possess genes for an electron sink system mediated by plastid terminal oxidase, plastoquinone/plastoquinol, and type II NADH dehydrogenase. Other non-photosynthetic algae/land plants also possess key genes for this system, suggesting a broad distribution of an electron sink system in non-photosynthetic plastids. CONCLUSION: The plastoquinone/plastoquinol pool and thus the involved electron transport systems reported herein might be retained for redox homeostasis and might represent an intermediate step towards a more reduced set of the electron transport system in many non-photosynthetic plastids. Our findings illuminate a broadly distributed but previously hidden step of reductive evolution of plastid electron transport systems after the loss of photosynthesis.


Asunto(s)
Chlorophyceae/fisiología , Transporte de Electrón/fisiología , Evolución Molecular , Plastidios/fisiología , Fotosíntesis
10.
Int J Mol Sci ; 22(3)2021 Jan 28.
Artículo en Inglés | MEDLINE | ID: mdl-33525606

RESUMEN

The oleaginous microalgae species Chlorococcum sphacosum GD is a promising feedstock for biodiesel production from soil. However, its metabolic mechanism of lipid production remains unclear. In this study, the lipid accumulation and metabolism mechanisms of Chlorococcum sphacosum GD were analyzed under salt stress based on transcriptome sequencing. The biomass and lipid content of the alga strain were determined under different NaCl concentrations, and total RNA from fresh cells were isolated and sequenced by HiSeq 2000 high throughput sequencing technology. As the salt concentration increased in culture medium, the algal lipid content increased but the biomass decreased. Following transcriptome sequencing by assembly and splicing, 24,128 unigenes were annotated, with read lengths mostly distributed in the 200-300 bp interval. Statistically significant differentially expressed unigenes were observed in different experimental groups, with 2051 up-regulated genes and 1835 down-regulated genes. The lipid metabolism pathway analysis showed that, under salt stress, gene-related fatty acid biosynthesis (ACCase, KASII, KAR, HAD, FATA) was significantly up-regulated, but some gene-related fatty acid degradation was significantly down-regulated. The comprehensive results showed that salt concentration can affect the lipid accumulation and metabolism of C. sphacosum GD, and the lipid accumulation is closely related to the fatty acid synthesis pathway.


Asunto(s)
Proteínas Algáceas/genética , Chlorophyceae/fisiología , Perfilación de la Expresión Génica/métodos , Suelo/química , Chlorophyceae/genética , Regulación de la Expresión Génica/efectos de los fármacos , Redes Reguladoras de Genes , Secuenciación de Nucleótidos de Alto Rendimiento , Lípidos/biosíntesis , Anotación de Secuencia Molecular , Estrés Salino , Análisis de Secuencia de ARN , Cloruro de Sodio/farmacología
11.
BMC Genomics ; 21(1): 477, 2020 Jul 11.
Artículo en Inglés | MEDLINE | ID: mdl-32652928

RESUMEN

BACKGROUND: Picophytoplankton are abundant and can contribute greatly to primary production in eutrophic lakes. Mychonastes species are among the common eukaryotic picophytoplankton in eutrophic lakes. We used third-generation sequencing technology to sequence the whole genome of Mychonastes homosphaera isolated from Lake Chaohu, a eutrophic freshwater lake in China. RESULT: The 24.23 Mbp nuclear genome of M.homosphaera, harboring 6649 protein-coding genes, is more compact than the genomes of the closely related Sphaeropleales species. This genome streamlining may be caused by a reduction in gene family number, intergenic size and introns. The genome sequence of M.homosphaera reveals the strategies adopted by this organism for environmental adaptation in the eutrophic lake. Analysis of cultures and the protein complement highlight the metabolic flexibility of M.homosphaera, the genome of which encodes genes involved in light harvesting, carbohydrate metabolism, and nitrogen and microelement metabolism, many of which form functional gene clusters. Reconstruction of the bioenergetic metabolic pathways of M.homosphaera, such as the lipid, starch and isoprenoid pathways, reveals characteristics that make this species suitable for biofuel production. CONCLUSION: The analysis of the whole genome of M. homosphaera provides insights into the genome streamlining, the high lipid yield, the environmental adaptation and phytoplankton evolution.


Asunto(s)
Adaptación Fisiológica/genética , Chlorophyceae/clasificación , Chlorophyceae/fisiología , Evolución Molecular , Fitoplancton/clasificación , Fitoplancton/fisiología , Biocombustibles , China , Chlorophyceae/genética , Eutrofización , Genoma del Cloroplasto , Genoma Mitocondrial , Lagos , Sistemas de Lectura Abierta/genética , Filogenia , Fitoplancton/genética
12.
Photosynth Res ; 144(3): 317-326, 2020 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-32323065

RESUMEN

The violaxanthin (V)-antheraxanthin (A)-zeaxanthin (Z) (VAZ) cycle was deemed a non-second-scale process of photoprotection in higher plants and microalgae, but the validity of this view has not been confirmed. To test this view, we explored responses of the VAZ cycle and the relationship between the VAZ cycle and non-photochemical quenching (NPQ) under highlight at second and minute scales in Heterosigma akashiwo and Platymonas sp. Both A and Z were generated in H. akashiwo during 15 s of light exposure, whereas only A rapidly accumulated within 15 s of exposure in Platymonas sp. The above results, together with a time-dependent sigmoidal relationship between the VAZ cycle (de-epoxidation state, A/Chl a, and Z/Chl a) and NPQ, proved that the VAZ cycle was a second-scale process related to NPQ. In addition, we found that not all NPQ was dependent on the VAZ cycle and suggested that NPQ model should be carefully modified due to the species-specific proportions of de-epoxidation-dependent NPQ.


Asunto(s)
Chlorophyceae/fisiología , Xantófilas/metabolismo , Zeaxantinas/metabolismo , Chlorophyceae/efectos de la radiación , Luz , Microalgas
13.
Plant Cell Environ ; 43(5): 1212-1229, 2020 05.
Artículo en Inglés | MEDLINE | ID: mdl-31994740

RESUMEN

VIPP proteins aid thylakoid biogenesis and membrane maintenance in cyanobacteria, algae, and plants. Some members of the Chlorophyceae contain two VIPP paralogs termed VIPP1 and VIPP2, which originate from an early gene duplication event during the evolution of green algae. VIPP2 is barely expressed under nonstress conditions but accumulates in cells exposed to high light intensities or H2 O2 , during recovery from heat stress, and in mutants with defective integration (alb3.1) or translocation (secA) of thylakoid membrane proteins. Recombinant VIPP2 forms rod-like structures in vitro and shows a strong affinity for phosphatidylinositol phosphate. Under stress conditions, >70% of VIPP2 is present in membrane fractions and localizes to chloroplast membranes. A vipp2 knock-out mutant displays no growth phenotypes and no defects in the biogenesis or repair of photosystem II. However, after exposure to high light intensities, the vipp2 mutant accumulates less HSP22E/F and more LHCSR3 protein and transcript. This suggests that VIPP2 modulates a retrograde signal for the expression of nuclear genes HSP22E/F and LHCSR3. Immunoprecipitation of VIPP2 from solubilized cells and membrane-enriched fractions revealed major interactions with VIPP1 and minor interactions with HSP22E/F. Our data support a distinct role of VIPP2 in sensing and coping with chloroplast membrane stress.


Asunto(s)
Chlorophyceae/metabolismo , Cloroplastos/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de la Membrana/fisiología , Proteínas de Plantas/fisiología , Chlamydomonas reinhardtii/genética , Chlamydomonas reinhardtii/metabolismo , Chlamydomonas reinhardtii/fisiología , Chlamydomonas reinhardtii/ultraestructura , Chlorophyceae/genética , Chlorophyceae/fisiología , Chlorophyceae/ultraestructura , Cloroplastos/fisiología , Cloroplastos/ultraestructura , Clonación Molecular , Inmunoprecipitación , Espectrometría de Masas , Proteínas de la Membrana/metabolismo , Microscopía Electrónica de Transmisión , Microscopía Fluorescente , Filogenia , Proteínas de Plantas/metabolismo , Proteínas Recombinantes , Tilacoides/metabolismo
14.
Biotechnol Bioeng ; 117(4): 970-980, 2020 04.
Artículo en Inglés | MEDLINE | ID: mdl-31956983

RESUMEN

Microalgae have been shown as a potential bioresource for food, biofuel, and pharmaceutical products. During the growth phases with corresponding environmental conditions, microalgae accumulate different amounts of various metabolites. We quantified the neutral lipids accumulation and analyzed the swimming signatures (speed and trajectories) of the motile green alga, Dunaliella primolecta, during the lag-exponential-stationary growth cycle at different nutrient concentrations. We discovered significant changes in the neutral lipid content and swimming signatures of microalgae across growth phases. The timing of the maximum swimming speed coincided with the maximum neutral lipid content and both maxima occurred under nutrient stress at the stationary growth phase. Furthermore, the swimming trajectories suggested statistically significant changes in swimming modes at the stationary growth phase when the maximum intracellular neutral lipid content was observed. Our results provide the potential exploitation of microalgal swimming signatures as possible indicators of the cultivation conditions and the timing of microalgal harvest to maximize the lipid yield for biofuel production. The findings can also be implemented to explore the production of food and antibiotics from other microalgal metabolites with low energy costs.


Asunto(s)
Biocombustibles , Movimiento Celular/fisiología , Chlorophyceae , Lípidos , Microalgas , Biomasa , Chlorophyceae/química , Chlorophyceae/metabolismo , Chlorophyceae/fisiología , Metabolismo de los Lípidos , Lípidos/análisis , Lípidos/química , Microalgas/química , Microalgas/metabolismo , Microalgas/fisiología , Fotobiorreactores
16.
J Fish Dis ; 43(7): 775-778, 2020 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-32323326

RESUMEN

In August 2019, visual inspection of intertidal zones of the Gulf of Maine (ME, USA) revealed young and adult wild blue mussels, Mytilus spp., in Alley Bay (Jonesport area) with the distinctive L-shaped shell deformity (LSSD) and green spots (GS) in the mantle and adductor muscle. LSSD is a characteristic sign of current or previous mussel infection by photosynthetic unicellular alga from the group Coccomyxa, while GS are algal colonies. Based on these findings, this study represents the first report of infection signs by pathogenic Coccomyxa-like algae in mussels from the coastal waters of the Northeastern United States, providing a base for future large scale monitoring of the alga in the region.


Asunto(s)
Chlorophyceae/fisiología , Mytilus/microbiología , Animales , Maine
17.
Ecotoxicol Environ Saf ; 206: 111180, 2020 Dec 15.
Artículo en Inglés | MEDLINE | ID: mdl-32861006

RESUMEN

The occurrence of pesticides and their mixtures in the environment can alter the ecological relationships between aquatic food chains. Since fipronil and 2,4-dichlorophenoxyacetic acid (2,4-D) are commonly found together in Brazilian water bodies, the present study aimed to investigate through an integrative approach the toxicity mechanisms of environmentally relevant concentrations of pesticides Regent® 800 WG (active ingredient - a.i. fipronil), DMA® 806 BR (a.i. 2,4-D) isolated and in mixtures on the green alga Raphidocelis subcapitata using multiple parameters: physiological (growth rate and chlorophyll a fluorescence), morphological (cell complexity and size), biochemical (composition of lipid classes) and related to the photosynthetic activity (variable fluorescence, the maximum quantum yield of the photosystem II - PSII - and the efficiency of the oxygen evolving complex - OEC - of PSII). The results indicated that fipronil significantly inhibited algal population growth, increased the chlorophyll a content (observed by fluorescence), cell size and lipid class content of triacylglycerol (TAG), free fatty acid (FFA) and acetone mobile polar lipid (AMPL) and, on the other hand, decreased variable fluorescence of algae. The tested concentrations of 2,4-D increased the chlorophyll a fluorescence, the cell size and the lipid classes TAG and FFA. The pesticide mixtures have had more effects on algae than isolated compounds, causing alterations in all parameters analyzed, including photosynthetic activity (maximum quantum yield and efficiency of the oxygen evolving complex of the PSII), in which no alterations were observed for the toxicity of the single pesticides. The results suggest that these analyses are important to evaluate pesticide toxicity mechanisms in ecological risk assessments of tropical regions. Thus, here we demonstrate the importance of using multiple parameters in ecotoxicological studies to obtain a better understanding of the toxicity of these compounds for phytoplankton.


Asunto(s)
Ácido 2,4-Diclorofenoxiacético/toxicidad , Chlorophyceae/fisiología , Pirazoles/toxicidad , Contaminantes Químicos del Agua/toxicidad , Brasil , Clorofila/metabolismo , Clorofila A , Ecotoxicología , Herbicidas/análisis , Plaguicidas/toxicidad , Fotosíntesis/efectos de los fármacos , Complejo de Proteína del Fotosistema II , Fitoplancton/metabolismo
18.
Sensors (Basel) ; 20(19)2020 Oct 02.
Artículo en Inglés | MEDLINE | ID: mdl-33023245

RESUMEN

We report on the observation of the detachment in situ and in vivo of Dunaliella tertiolecta microalgae cells from a glass surface using a 1064 nm wavelength trapping laser beam. The principal bends of both flagella of Dunaliella were seen self-adhered to either the top or bottom coverslip surfaces of a 50 µm thick chamber. When a selected attached Dunaliella was placed in the trapping site, it photoresponded to the laser beam by moving its body and flagellar tips, which eventually resulted in its detachment. The dependence of the time required for detachment on the trapping power was measured. No significant difference was found in the detachment time for cells detached from the top or bottom coverslip, indicating that the induced detachment was not due solely to the optical forces applied to the cells. After detachment, the cells remained within the optical trap. Dunaliella detached from the bottom were seen rotating about their long axis in a counterclockwise direction, while those detached from the top did not rotate. The rotation frequency and the minimal force required to escape from the trap were also measured. The average rotation frequency was found to be independent of the trapping power, and the swimming force of a cell escaping the laser trap ranged from 4 to 10 picoNewtons. Our observations provide insight into the photostimulus produced when a near-infrared trapping beam encounters a Dunaliella. The microalgae frequently absorb more light than they can actually use in photosynthesis, which could cause genetic and molecular changes. Our findings may open new research directions into the study of photomovement in species of Dunaliella and other swimming microorganisms that could eventually help to solve technological problems currently confronting biomass production. In future work, studies of the response to excess light may uncover unrecognized mechanisms of photoprotection and photoacclimation.


Asunto(s)
Chlorophyceae/fisiología , Microalgas/fisiología , Pinzas Ópticas , Vidrio , Rayos Láser , Luz , Fotosíntesis
19.
Planta ; 250(4): 1379-1385, 2019 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-31359139

RESUMEN

MAIN CONCLUSION: L-Arginine supports growth and resulted in increased PII signaling protein levels and lipid droplet accumulation in the colorless green alga Polytomella parva. Polytomella parva, a model system for nonphotosynthetic green algae, utilizes ammonium and several carbon sources, including ethanol and acetate. We previously reported that P. parva accumulates high amounts of arginine with the key enzyme of the ornithine/arginine biosynthesis pathway, N-acetyl-L-glutamate kinase, exhibiting high activity. Here we demonstrate that L-arginine can be used by this alga as a nitrogen source. Externally supplied arginine directly influenced the levels of PII signaling protein and formation of triacylglycerol (TAG)-filled lipid bodies (LBs). Our results suggest that the nitrogen source, but not nitrogen starvation, may be critical for the accumulation of LBs in a PII-independent manner in P. parva.


Asunto(s)
Arginina/farmacología , Chlorophyceae/fisiología , Gotas Lipídicas/metabolismo , Nitrógeno/metabolismo , Proteínas PII Reguladoras del Nitrógeno/metabolismo , Fosfotransferasas (aceptor de Grupo Carboxilo)/metabolismo , Proteínas Algáceas/genética , Proteínas Algáceas/metabolismo , Chlorophyceae/crecimiento & desarrollo , Gotas Lipídicas/efectos de los fármacos , Proteínas PII Reguladoras del Nitrógeno/genética , Fosfotransferasas (aceptor de Grupo Carboxilo)/genética , Transducción de Señal/efectos de los fármacos , Triglicéridos/metabolismo
20.
Eur Biophys J ; 48(3): 231-248, 2019 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-30806730

RESUMEN

We examined the response of algal cells to laboratory-induced cadmium stress in terms of physiological activity, autonomous features (motility and fluorescence), adhesion dynamics, nanomechanical properties, and protein expression by employing a multimethod approach. We develop a methodology based on the generalized mathematical model to predict free cadmium concentrations in culture. We used algal cells of Dunaliella tertiolecta, which are widespread in marine and freshwater systems, as a model organism. Cell adaptation to cadmium stress is manifested through cell shape deterioration, slower motility, and an increase of physiological activity. No significant change in growth dynamics showed how cells adapt to stress by increasing active surface area against toxic cadmium in the culture. It was accompanied by an increase in green fluorescence (most likely associated with cadmium vesicular transport and/or beta-carotene production), while no change was observed in the red endogenous fluorescence (associated with chlorophyll). To maintain the same rate of chlorophyll emission, the cell adaptation response was manifested through increased expression of the identified chlorophyll-binding protein(s) that are important for photosynthesis. Since production of these proteins represents cell defence mechanisms, they may also signal the presence of toxic metal in seawater. Protein expression affects the cell surface properties and, therefore, the dynamics of the adhesion process. Cells behave stiffer under stress with cadmium, and thus, the initial attachment and deformation are slower. Physicochemical and structural characterizations of algal cell surfaces are of key importance to interpret, rationalize, and predict the behaviour and fate of the cell under stress in vivo.


Asunto(s)
Cadmio/farmacología , Chlorophyceae/citología , Chlorophyceae/fisiología , Laboratorios , Estrés Fisiológico/efectos de los fármacos , Disponibilidad Biológica , Cadmio/metabolismo , Adhesión Celular/efectos de los fármacos , Movimiento Celular/efectos de los fármacos , Proliferación Celular/efectos de los fármacos , Chlorophyceae/efectos de los fármacos , Chlorophyceae/metabolismo , Electroquímica , Regulación de la Expresión Génica de las Plantas/efectos de los fármacos , Cinética , Modelos Biológicos
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