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Organic carbon fixed in chloroplasts through the Calvin-Benson-Bassham Cycle can be diverted toward different metabolic fates, including cytoplasmic and mitochondrial respiration, gluconeogenesis, and synthesis of diverse plastid metabolites via the pyruvate hub. In plants, pyruvate is principally produced via cytoplasmic glycolysis, although a plastid-targeted lower glycolytic pathway is known to exist in non-photosynthetic tissue. Here, we characterized a lower plastid glycolysis-gluconeogenesis pathway enabling the direct interconversion of glyceraldehyde-3-phosphate and phospho-enol-pyruvate in diatoms, ecologically important marine algae distantly related to plants. We show that two reversible enzymes required to complete diatom plastid glycolysis-gluconeogenesis, Enolase and bis-phosphoglycerate mutase (PGAM), originated through duplications of mitochondria-targeted respiratory isoforms. Through CRISPR-Cas9 mutagenesis, integrative 'omic analyses, and measured kinetics of expressed enzymes in the diatom Phaeodactylum tricornutum, we present evidence that this pathway diverts plastid glyceraldehyde-3-phosphate into the pyruvate hub, and may also function in the gluconeogenic direction. Considering experimental data, we show that this pathway has different roles dependent in particular on day length and environmental temperature, and show that the cpEnolase and cpPGAM genes are expressed at elevated levels in high-latitude oceans where diatoms are abundant. Our data provide evolutionary, meta-genomic, and functional insights into a poorly understood yet evolutionarily recurrent plastid metabolic pathway.
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Diatomáceas , Gluconeogênese , Glicólise , Plastídeos , Diatomáceas/metabolismo , Diatomáceas/genética , Plastídeos/metabolismo , Plastídeos/genética , Glicólise/genética , Gluconeogênese/genética , FilogeniaRESUMO
Urban greenhouse gas emissions monitoring is essential to assessing the impact of climate mitigation actions. Using atmospheric continuous measurements of air quality and carbon dioxide (CO2), we developed a gradient-descent optimization system to estimate emissions of the city of Paris. We evaluated our joint CO2-CO-NOx optimization over the first SARS-CoV-2 related lockdown period, resulting in a decrease in emissions by 40% for NOx and 30% for CO2, in agreement with preliminary estimates using bottom-up activity data yet lower than the decrease estimates from Bayesian atmospheric inversions (50%). Before evaluating the model, we first provide an in-depth analysis of three emission data sets. A general agreement in the totals is observed over the region surrounding Paris (known as Île-de-France) since all the data sets are constrained by the reported national and regional totals. However, the data sets show disagreements in their sector distributions as well as in the interspecies ratios. The seasonality also shows disagreements among emission products related to nonindustrial stationary combustion (residential and tertiary combustion). The results presented in this paper show that a multispecies approach has the potential to provide sectoral information to monitor CO2 emissions over urban areas enabled by the deployment of collocated atmospheric greenhouse gases and air quality monitoring stations.
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Poluentes Atmosféricos , COVID-19 , Gases de Efeito Estufa , Humanos , Poluentes Atmosféricos/análise , Dióxido de Carbono/análise , SARS-CoV-2 , Teorema de Bayes , Controle de Doenças Transmissíveis , Gases de Efeito Estufa/análiseRESUMO
The wide latitudinal distribution of marine Synechococcus cyanobacteria partly relies on the differentiation of lineages adapted to distinct thermal environments. Membranes are highly thermosensitive cell components, and the ability to modulate their fluidity can be critical for the fitness of an ecotype in a particular thermal niche. We compared the thermophysiology of Synechococcus strains representative of major temperature ecotypes in the field. We measured growth, photosynthetic capacities and membrane lipidome variations. We carried out a metagenomic analysis of stations of the Tara Oceans expedition to describe the latitudinal distribution of the lipid desaturase genes in the oceans. All strains maintained efficient photosynthetic capacities over their different temperature growth ranges. Subpolar and cold temperate strains showed enhanced capacities for lipid monodesaturation at low temperature thanks to an additional, poorly regiospecific Δ9-desaturase. By contrast, tropical and warm temperate strains displayed moderate monodesaturation capacities but high proportions of double unsaturations in response to cold, thanks to regiospecific Δ12-desaturases. The desaturase genes displayed specific distributions directly related to latitudinal variations in ocean surface temperature. This study highlights the critical importance of membrane fluidity modulation by desaturases in the adaptive strategies of Synechococcus cyanobacteria during the colonization of novel thermal niches.
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Água do Mar , Synechococcus , Regulação da Temperatura Corporal , Oceanos e Mares , Filogenia , Synechococcus/genéticaRESUMO
Microalgae are a promising feedstock for the production of triacylglycerol (TAG) for a variety of potential applications, ranging from food and human health to biofuels and green chemistry. However, obtaining high TAG yields is challenging. A phenotypic assay for the accumulation of oil droplets was developed to screen a library of 1,200 drugs, annotated with pharmacology information, to select compounds that trigger TAG accumulation in the diatom Phaeodactylum tricornutum Using this screen, we identified 34 molecules acting in a dose-dependent manner. Previously characterized targets of these compounds include cell division and cell signaling effectors, membrane receptors and transporters, and sterol metabolism. Among the five compounds possibly acting on sterol metabolism, we focused our study on ethynylestradiol, a synthetic form of estrogen that is used in contraceptive pills and known for its ecological impact as an endocrine disruptor. Ethynylestradiol impaired the production of very-long-chain polyunsaturated fatty acids, destabilized the galactolipid versus phospholipid balance, and triggered the recycling of fatty acids from membrane lipids to TAG. The P. tricornutum transcriptomic response to treatment with ethynylestradiol was consistent with the reallocation of carbon from sterols to acetyl-coenzyme A and TAG. The mode of action and catabolism of ethynylestradiol are unknown but might involve several up-regulated cytochrome P450 proteins. A fatty acid elongase, Δ6-ELO-B1, might be involved in the impairment of very-long-chain polyunsaturated fatty acids and fatty acid turnover. This phenotypic screen opens new perspectives for the exploration of novel bioactive molecules, potential target genes, and pathways controlling TAG biosynthesis. It also unraveled the sensitivity of diatoms to endocrine disruptors, highlighting an impact of anthropogenic pollution on phytoplankton.
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Produtos Biológicos/farmacologia , Diatomáceas/efeitos dos fármacos , Diatomáceas/metabolismo , Avaliação Pré-Clínica de Medicamentos/métodos , Triglicerídeos/metabolismo , Produtos Biológicos/administração & dosagem , Diatomáceas/genética , Relação Dose-Resposta a Droga , Avaliação Pré-Clínica de Medicamentos/estatística & dados numéricos , Estrona/farmacologia , Etinilestradiol/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacosRESUMO
RATIONALE: Secondary organic aerosols (SOAs) represent a significant portion of total atmospheric aerosols. They are generated by the oxidation of volatile organic compounds (VOCs), and particularly biogenic VOCs (BVOCs). The analysis of such samples is usually performed by targeted methods that often require time-consuming preparation steps that can induce loss of compounds and/or sample contaminations. METHODS: Recently, untargeted methods using high-resolution mass spectrometry (HRMS) have been successfully employed for a broad characterization of chemicals in SOAs. Herein we propose a new application of the direct analysis in real time (DART) ionization method combined with HRMS to quickly detect several hundred chemicals in SOAs collected on a quartz filter without sample preparation or separation techniques. RESULTS: The reproducibility of measurements was good, with several hundred elemental compositions common to three different replicates. The relative standard deviations of the intensities of the chemical families ranged from 6% to 35%, with sufficient sensitivity to allow the unambiguous detection of 4 ng/mm2 of pinic acid. The presence of oligomers and specific tracers was highlighted by MSn (n ≤ 4) experiments, an achievement that is difficult to attain with other ultrahigh-resolution mass spectrometers. Contributions of this untargeted DART-HRMS method were illustrated by the analysis of fresh and aged SOAs from different gaseous precursors such as limonene, a ß-pinene/limonene mixture or scots pines emissions. CONCLUSIONS: The results show that it is possible to use DART-HRMS for the identification of tracers of specific aging reactions, or for the identification of aerosols from specific biogenic precursors.
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Aerossóis/análise , Espectrometria de Massas/métodos , Monoterpenos/análise , Compostos Orgânicos Voláteis/análise , Aerossóis/química , Poluentes Atmosféricos/análise , Poluentes Atmosféricos/química , Monoterpenos/química , Oxirredução , Compostos Orgânicos Voláteis/químicaRESUMO
Trends of long-term observations and emission inventories suggest that traffic emissions will no longer dominate the concentrations of monoaromatic compounds (i.e., TEX - toluene, xylenes, and ethylbenzene) in European urban areas. But the split limit between traffic and other emission sector contributions such as solvent use remains tenuous. Here long-term observations of an extensive set of hydrocarbons, including TEX, at traffic and urban background sites in London, Paris and Strasbourg were combined to estimate the relative importance of traffic emissions on TEX in every city. When analyzing the urban enhancement emission ratios of TEX-to-benzene on a seasonal basis, two potential source signatures other than traffic could be differentiated in all cities (1) summertime evaporation from fuel and/or solvent and (2) wintertime domestic heating. However, traffic emissions still unambiguously dominate the concentration levels of TEX in every city despite the reduction of their emissions at exhaust pipe over the last two decades. Traffic explains between 60% and 96% (at ±20%) of TEX levels while it is less clear for xylenes at some locations. Our results provide a basis to evaluate regional emission inventories. The method is applicable at any urban area where speciated hydrocarbon monitoring is available.
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Poluentes Atmosféricos , Cidades , Monitoramento Ambiental , Londres , Emissões de VeículosRESUMO
Angstrom exponent measurements of equivalent black carbon (BCeq) have recently been introduced as a novel tool to apportion the contribution of biomass burning sources to the BCeq mass. The BCeq is the mass of ideal BC with defined optical properties that, upon deposition on the aethalometer filter tape, would cause equal optical attenuation of light to the actual PM2.5 aerosol deposited. The BCeq mass hence is identical to the mass of the total light-absorbing carbon deposited on the filter tape. Here, we use simultaneously collected data from a seven-wavelength aethalometer and a high-sensitivity proton-transfer reaction mass spectrometer installed at a suburban site in Mohali (Punjab), India, to identify a number of biomass combustion plumes. The identified types of biomass combustion include paddy- and wheat-residue burning, leaf litter, and garbage burning. Traffic plumes were selected for comparison. We find that the combustion efficiency, rather than the fuel used, determines αabs, and consequently, the αabs can be â¼1 for flaming biomass combustion and >1 for older vehicles that operate with poorly optimized engines. Thus, the absorption angstrom exponent is not representative of the fuel used and, therefore, cannot be used as a generic tracer to constrain source contributions.
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Fuligem/análise , Aerossóis/análise , Biomassa , Carbono , Incêndios , Índia , Luz , Emissões de Veículos/análiseRESUMO
Aspergillus fumigatus is a major airborne nosocomial pathogen that is responsible for severe mycosis in immunocompromised patients. We studied the efficacy of an innovative mobile air-treatment device in eliminating A. fumigatus from the air following experimental massive contamination in a high-security room. Viable mycological particles were isolated from sequential air samples in order to evaluate the device's effectiveness in removing the fungus. The concentration of airborne conidia was reduced by 95% in 18 min. Contamination was reduced below the detection threshold in 29 min, even when the machine was at the lowest airflow setting. In contrast, during spontaneous settling with no air treatment, conidia remained airborne for more than 1 h. This indoor air contamination model provided consistent and reproducible results. Because the air purifier proved to be effective at eliminating a major contaminant, it may prove useful in preventing air-transmitted disease agents. In an experimental space mimicking a hospital room, the AirLyse air purifier, which uses a combination of germicidal ultraviolet C irradiation and titanium photocatalysis, effectively eliminated Aspergillus conidia. Such a mobile device may be useful in routine practice for lowering microbiological air contamination in the rooms of patients at risk.
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Filtros de Ar , Microbiologia do Ar , Aspergillus fumigatus/isolamento & purificação , Desinfecção/métodos , HumanosRESUMO
In plants and algae, photosynthetic membranes have a unique lipid composition. They differ from all other cellular membranes by their very low amount of phospholipids, besides some phosphatidylglycerol (PG), and high proportion of glycolipids. These glycolipids are the uncharged galactolipids, that is, mono- and digalactosyldiacylglycerol (MGDG and DGDG), and an anionic sulfolipid, that is, sulfoquinovosyldiacylglycerol (SQDG). In all photosynthetic membranes analyzed to date, from cyanobacteria to algae, protists, and plants, the lipid quartet constituted by MGDG, DGDG, SQDG, and PG has been highly conserved, but the composition in fatty acids of these lipids can vary a lot from an organism to another. To better understand the chloroplast biogenesis, it is therefore essential to know their lipid content. Establishing chloroplast lipidome requires first to purify chloroplast from plant or algae tissue. Here we describe the methods to extract the lipid, quantify the lipid amount of the chloroplast, and qualify and quantify the different lipid classes that might be present in these fractions.
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Cloroplastos , Lipidômica , Glicolipídeos , Galactolipídeos , Ácidos Graxos , Fosfolipídeos , Membrana Celular , PlantasRESUMO
In plants and algae, the glycerolipidome changes in response to environmental modifications. For instance, in phosphate starvation, phospholipids are degraded and replaced by non-phosphorus lipids, and in nitrogen starvation, storage lipids accumulate. In addition to the well-known applications of oil crops for food, algae lipids are becoming a model for potential applications in health, biofuel, and green chemistry and are used as a platform for genetic engineering. It is therefore important to measure accurately and quickly the glycerolipid content in plants and algae. Here we describe the methods to extract the lipid and quantify the fatty acid amount of the lipid extract and the different lipid classes that are present in these samples.
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Ácidos Graxos , Plantas , Plantas/metabolismo , Ácidos Graxos/metabolismo , FosfolipídeosRESUMO
Urban trees are often not considered in air-quality models although they can significantly impact the concentrations of pollutants. Gas and particles can deposit on leaf surfaces, lowering their concentrations, but the tree crown aerodynamic effect is antagonist, limiting the dispersion of pollutants in streets. Furthermore, trees emit Biogenic Volatile Organic Compounds (BVOCs) that react with other compounds to form ozone and secondary organic aerosols. This study aims to quantify the impacts of these three tree effects (dry deposition, aerodynamic effect and BVOC emissions) on air quality from the regional to the street scale over Paris city. Each tree effect is added in the model chain CHIMERE/MUNICH/SSH-aerosol. The tree location and characteristics are determined using the Paris tree inventory, combined with allometric equations. The air-quality simulations are performed over June and July 2022. The results show that the aerodynamic tree effect increases the concentrations of gas and particles emitted in streets, such as NOx (+4.6 % on average in streets with trees and up to +37 % for NO2). This effect increases with the tree Leaf Area Index and it is more important in streets with high traffic, suggesting to limit the planting of trees with large crowns on high-traffic streets. The effect of dry deposition of gas and particles on leaves is very limited, reducing the concentrations of O3 concentrations by -0.6 % on average and at most -2.5 %. Tree biogenic emissions largely increase the isoprene and monoterpene concentrations, bringing the simulated concentrations closer to observations. Over the two-week sensitivity analysis, biogenic emissions induce an increase of O3, organic particles and PM2.5 street concentrations by respectively +1.1, +2.4 and + 0.5 % on average over all streets. This concentration increase may reach locally +3.5, +12.3 and + 2.9 % respectively for O3, organic particles and PM2.5, suggesting to prefer the plantation of low-emitting VOC species in cities.
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Arachidonic acid (C20: 4n-6, AA) plays a fundamental role in fish physiology, influencing growth, survival and stress resistance. However, imbalances in dietary AA can have detrimental effects on fish health and performance. Optimal AA requirements for rainbow trout have not been established. This study aimed to elucidate the effects of varying dietary AA levels on survival, growth, long-chain polyunsaturated fatty acid (LC-PUFA) biosynthetic capacity, oxylipin profiles, lipid peroxidation, and stress resistance of rainbow trout fry. Over a period of eight weeks, 4000 female rainbow trout fry at the resorptive stage (0.12 g) from their first feeding were fed diets with varying levels of AA (0.6%, 1.1% or 2.5% of total fatty acids) while survival and growth metrics were closely monitored. The dietary trial was followed by an acute confinement stress test. Notably, while the fatty acid profiles of the fish reflected dietary intake, those fed an AA-0.6% diet showed increased expression of elongase5, highlighting their inherent ability to produce LC-PUFAs from C18 PUFAs and suggesting potential AA or docosapentaenoic acidn-6 (DPAn-6) biosynthesis. However, even with this biosynthetic capacity, the trout fed reduced dietary AA had higher mortality rates. The diet had no effect on final weight (3.38 g on average for the three diets). Conversely, increased dietary AA enhanced eicosanoid production from AA, suggesting potential inflammatory and oxidative consequences. This was further evidenced by an increase in non-enzymatic lipid oxidation metabolites, particularly in the AA-2.5% diet group, which had higher levels of phytoprostanes and isoprostanes, markers of cellular oxidative damage. Importantly, the AA-1.1% diet proved to be particularly beneficial for stress resilience. This was evidenced by higher post-stress turnover rates of serotonin and dopamine, neurotransmitters central to the fish's stress response. In conclusion, a dietary AA intake of 1.1% of total fatty acids appears to promote overall resilience in rainbow trout fry.
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Ácido Araquidônico , Ácidos Graxos Insaturados , Oncorhynchus mykiss , Oxilipinas , Estresse Fisiológico , Animais , Oncorhynchus mykiss/metabolismo , Oxilipinas/metabolismo , Ácido Araquidônico/metabolismo , Ácidos Graxos Insaturados/metabolismo , Feminino , Ração Animal/análise , Dieta/veterinária , Peroxidação de Lipídeos/efeitos dos fármacos , Estresse Oxidativo/efeitos dos fármacosRESUMO
New Particle Formation (NPF) is a major source of ultrafine particles that affect both air quality and climate. Despite emissions from agricultural activities having a strong potential to lead to NPF, little is known about NPF within agricultural environments. The aim of the present study was to investigate the occurrence of NPF events at an agricultural site, and any potential relationship between agricultural emissions and NPF events. A field campaign was conducted for 3 months at the FR-Gri-ICOS site (France), at an experimental farm 25 km west of Paris city centre. 16 NPF events have been identified from the analysis of particle number size distributions; 8 during the daytime, and 8 during the night-time. High solar radiation and ozone mixing ratios were observed during the days NPF occurred, suggesting photochemistry plays a key role in daytime NPF. These events were also associated with higher levels of VOCs such as isoprene, methanol, or toluene compared to non-event days. However, ammonia levels were lower during daytime NPF events, contributing to the hypothesis that daytime NPF events were not related to agricultural activities. On the other hand, temperature and ozone were lower during the nights when NPF events were observed, whereas relative humidity was higher. During these nights, higher concentrations of NO2 and ammonia were observed. As a result, agricultural activities, in particular the spreading of fertiliser on surrounding crops, are suspected to contribute to night-time NPF events. Finally, all the identified NPF events were also observed at SIRTA monitoring station 20 km from the FR-Gri ICOS site, showing that both night-time and daytime NPF events were regional processes. We hypothesise that night-time NPF may be related to fertiliser spreading over a regional scale, as opposed to the local activities at the farm. To our knowledge, this is the first time night-time NPF has been observed in the agricultural context.
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Poluentes Atmosféricos , Ozônio , Poluentes Atmosféricos/análise , Aerossóis/análise , Monitoramento Ambiental , Amônia/análise , Tamanho da Partícula , Fertilizantes/análise , Ozônio/análiseRESUMO
The Polluscope project aims to better understand the personal exposure to air pollutants in the Paris region. This article is based on one campaign from the project, which was conducted in the autumn of 2019 and involved 63 participants equipped with portable sensors (i.e., NO2, BC and PM) for one week. After a phase of data curation, analyses were performed on the results from all participants, as well as on individual participants' data for case studies. A machine learning algorithm was used to allocate the data to different environments (e.g., transportation, indoor, home, office, and outdoor). The results of the campaign showed that the participants' exposure to air pollutants depended very much on their lifestyle and the sources of pollution that may be present in the vicinity. Individuals' use of transportation was found to be associated with higher levels of pollutants, even when the time spent on transport was relatively short. In contrast, homes and offices were environments with the lowest concentrations of pollutants. However, some activities performed in indoor air (e.g., cooking) also showed a high levels of pollution over a relatively short period.
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Portable sensors have emerged as a promising solution for personal exposure (PE) measurement. For the first time in Île-de-France, PE to black carbon (BC), particulate matter (PM), and nitrogen dioxide (NO2) was quantified based on three field campaigns involving 37 volunteers from the general public wearing the sensors all day long for a week. This successful deployment demonstrated its ability to quantify PE on a large scale, in various environments (from dense urban to suburban, indoor and outdoor) and in all seasons. The impact of the visited environments was investigated. The proximity to road traffic (for BC and NO2), as well as cooking activities and tobacco smoke (for PM), made significant contributions to total exposure (up to 34%, 26%, and 44%, respectively), even though the time spent in these environments was short. Finally, even if ambient outdoor levels played a role in PE, the prominent impact of the different environments suggests that traditional ambient monitoring stations is not a proper surrogate for PE quantification.
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Levels and sources of non-Methane Hydrocarbons (NMHCs) were investigated at the urban background Thissio station, close to the historical center of Athens (Greece) from March 2016 to February 2017 (12 months), by means of an automated GC-FID. Alkanes dominated over aromatics and alkenes, with hourly mean levels ranging from detection limit up to 60 µg m-3 for i-pentane and 90 µg m-3 for toluene. Higher levels were recorded in the cold period relative to the warmer one. In addition, NMHCs seasonal diurnal cycles were characterized by a bimodal pattern, following the trend of tracers of anthropogenic sources. The Positive Matrix Factorization (PMF) was used for the allocation of NMHC to their sources. Five factors were identified and quantified, with traffic-related sources being the main one contributing up to 60% to total NMHCs, while biomass burning contributes up to 19%. A supplementary PMF assimilation was applied on a seasonal basis further including α-pinene, C6-C16 alkanes and aromatics. This PMF resulted to a seven-factor solution that allowed the examination of two additional sources, in addition to five already identified, highlighting the main contribution of anthropogenic sources (70%) to α-pinene.
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Poluentes Atmosféricos , Poluentes Atmosféricos/análise , Atmosfera , Biomassa , Hidrocarbonetos/análise , MetanoRESUMO
Trace gas measurements were performed during the LANDEX (the LANDes EXperiment) Episode 1 field campaign in the summer 2017, in one of the largest European maritime pine forests (> 95% Pinus pinaster) located in southwestern France. Efforts have been focused on obtaining a good speciation of 20 major biogenic volatile organic compounds (BVOCs, including pinenes, carenes, terpinenes, linalool, camphene, etc.). This was made possible by the development of a new and specific chromatographic method. In order to assess the role of BVOCs in the local gas phase chemistry budget, their reactivity with the main atmospheric oxidants (hydroxyl radicals (OH), ozone (O3) and nitrate radicals (NO3)) and the corresponding consumption rates were determined. When considering the OH reactivity with BVOCs, isoprene and linalool accounted for 10-47% of the OH depletion during daytime, and monoterpenes for 50-65%, whereas monoterpenes were the main contributors during the night (70-85%). Sesquiterpenes and monoterpenes were the main contributors to the ozone reactivity, especially ß-caryophyllene (30-70%), with a maximum contribution during nighttime. Nighttime nitrate reactivity was predominantly due to monoterpenes (i.e. 90-95%). Five specific groups have been proposed to classify the 19 BVOCs measured in the forest, according to their reactivity with atmospheric oxidants and their concentrations. The total amount of BVOCs consumed under and above the forest canopy was evaluated for 7 BVOCs (i.e. isoprene, α-pinene, ß-pinene, myrcene, limonene + cis-ocimene and Δ3-carene). The reactivity of atmospheric oxidants and BVOCs at a local level are discussed in order to highlight the compounds (BVOCs, other VOCs), the atmospheric oxidants and the main associated reactive processes observed under the canopy of a maritime pine forest.
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Algae belonging to the Microchloropsis genus are promising organisms for biotech purposes, being able to accumulate large amounts of lipid reserves. These organisms adapt to different trophic conditions, thriving in strict photoautotrophic conditions, as well as in the concomitant presence of light plus reduced external carbon as energy sources (mixotrophy). In this work, we investigated the mixotrophic responses of Microchloropsis gaditana (formerly Nannochloropsis gaditana). Using the Biolog growth test, in which cells are loaded into multiwell plates coated with different organic compounds, we could not find a suitable substrate for Microchloropsis mixotrophy. By contrast, addition of the Lysogeny broth (LB) to the inorganic growth medium had a benefit on growth, enhancing respiratory activity at the expense of photosynthetic performances. To further dissect the role of respiration in Microchloropsis mixotrophy, we focused on the mitochondrial alternative oxidase (AOX), a protein involved in energy management in other algae prospering in mixotrophy. Knocking-out the AOX1 gene by transcription activator-like effector nuclease (TALE-N) led to the loss of capacity to implement growth upon addition of LB supporting the hypothesis that the effect of this medium was related to a provision of reduced carbon. We conclude that mixotrophic growth in Microchloropsis is dominated by respiratory rather than by photosynthetic energetic metabolism and discuss the possible reasons for this behavior in relationship with fatty acid breakdown via ß-oxidation in this oleaginous alga.
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Biogenesis of photosynthetic membranes depends on galactolipid synthesis, which relies on several cell compartments, notably the endoplasmic reticulum (ER) and the chloroplast envelope. Galactolipid synthesis involves lipid trafficking between both membrane compartments. In Arabidopsis, ALA10, a phospholipid flippase of the P4 type-ATPase family, counteracts the limitation of monogalactosyldiacylglycerol (MGDG) production and has a positive effect on leaf development. ALA10 locates in distinct domains of the ER depending on the ALIS (ALA interacting subunit) subunit it interacts with: close to the plasma membrane with ALIS1, or next to chloroplasts with ALIS5. It interacts with FAD2 (Fatty acid desaturase 2) and prevents accumulation of linolenic (18:3) containing phosphatidylcholine (PC) stimulating an increase of MGDG synthesis. Here we report that ALA10 interacts with PUB11 (plant U-box type 11), an E3 protein ubiquitin ligase, in vitro and in vivo. ALA10 is however ubiquitinated and degraded by the 26S proteasome in a PUB11-independent process. In pub11 null mutant, the proteasome-dependent degradation of ALA10 is retained and ALA10 is still subject to ubiquitination although its ubiquitination profile appears different. In the absence of PUB11, ALA10 is constrained to the ER close to chloroplasts, which is the usual location when ALA10 is overexpressed. Additionally, in this condition, the decrease of 18:3 containing PC is no longer observed. Taken together these results suggest, that ALA10 contributes in chloroplast-distal ER interacting domains, to reduce the 18:3 desaturation of PC and that PUB11 is involved in reconditioning of ALA10 from chloroplast-proximal to chloroplast-distal ER interacting domains.
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Brown carbon (BrC) is known to absorb light at subvisible wavelengths but its optical properties and sources are still poorly documented, leading to large uncertainties in climate studies. Here, we show its major wintertime contribution to total aerosol absorption at 370 nm (18-42%) at 9 different French sites. Moreover, an excellent correlation with levoglucosan (r2 = 0.9 and slope = 22.2 at 370 nm), suggesting important contribution of wood burning emissions to ambient BrC aerosols in France. At all sites, BrC peaks were mainly observed during late evening, linking to local intense residential wood burning during this time period. Furthermore, the geographic origin analysis also highlighted the high potential contribution of local and/or small-regional emissions to BrC. Focusing on the Paris region, twice higher BrC mass absorption efficiency value was obtained for less oxidized biomass burning organic aerosols (BBOA) compared to more oxidized BBOA (e.g., about 4.9 ± 0.2 vs. 2.0 ± 0.1 m2 g-1, respectively, at 370 nm). Finally, the BBOA direct radiative effect was found to be 40% higher when these two BBOA fractions are treated as light-absorbing species, compared to the non-absorbing BBOA scenario.