Browsing by Author "Marques, J. C."
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- Disentangling the photochemical salinity tolerance in Aster tripolium L.: connecting biophysical traits with changes in fatty acid compositionPublication . Duarte, Bernardo; Cabrita, M. T.; Gameiro, Carla; Matos, A. R.; Godinho, R.; Marques, J. C.; Caçador, IsabelA profound analysis of A. tripolium photochemical traits under salinity exposure is lacking in the literature, with very few references focusing on its fatty acid profile role in photophysiology. To address this, the deep photochemical processes were evaluated by Pulse Amplitude Modulated (PAM) Fluorometry coupled with a discrimination of its leaf fatty acid profile. Plants exposed to 125-250 mm NaCl showed higher photochemical light harvesting efficiencies and lower energy dissipation rates. under higher NaCl exposure, there is evident damage of the oxygen evolving complexes (OECs). On the other hand, Reaction Centre (RC) closure net rate and density increased, improving the energy fluxes entering the PS II, in spite of the high amounts of energy dissipated and the loss of PS II antennae connectivity. Energy dissipation was mainly achieved through the auroxanthin pathway. Total fatty acid content displayed a similar trend, being also higher under 125-250 mm NaCl with high levels of omega-3 and omega-6 fatty acids. The increase in oleic acid and palmitic acid allows the maintenance of the good functioning of the PS II. Also relevant was the high concentration of chloroplastic C16:1t in the individuals subjected to 125-250 mm NaCl, related with a higher electron transport activity and with the organization of the Light Harvesting Complexes (LHC) and thus reducing the activation of energy dissipation mechanisms. All these new insights shed some light not only on the photophysiology of this potential cash-crop, but also highlight its important saline agriculture applications of this species as forage and potential source of essential fatty acids.
- DNA Sequencing as a Tool to Monitor Marine Ecological StatusPublication . Goodwin, Kelly D.; Thompson, Luke R.; Duarte, Bernardo; Kahlke, Tim; Thompson, Andrew R.; Marques, J. C.; Caçador, IsabelMany ocean policies mandate integrated, ecosystem-based approaches to marine monitoring, driving a global need for efficient, low-cost bioindicators of marine ecological quality. Most traditional methods to assess biological quality rely on specialized expertise to provide visual identification of a limited set of specific taxonomic groups, a time-consuming process that can provide a narrow view of ecological status. In addition, microbial assemblages drive food webs but are not amenable to visual inspection and thus are largely excluded from detailed inventory. Molecular-based assessments of biodiversity and ecosystem function offer advantages over traditional methods and are increasingly being generated for a suite of taxa using a “microbes to mammals” or “barcodes to biomes” approach. Progress in these efforts coupled with continued improvements in high-throughput sequencing and bioinformatics pave the way for sequence data to be employed in formal integrated ecosystem evaluation, including food web assessments, as called for in the European Union Marine Strategy Framework Directive. DNA sequencing of bioindicators, both traditional (e.g., benthic macroinvertebrates, ichthyoplankton) and emerging (e.g., microbial assemblages, fish via eDNA), promises to improve assessment of marine biological quality by increasing the breadth, depth, and throughput of information and by reducing costs and reliance on specialized taxonomic expertise.
- Ecophysiological constraints of Aster tripolium under extreme thermal events impacts: Merging biophysical, biochemical and genetic insightsPublication . Duarte, Bernardo; Goessling, J.W.; Marques, J. C.; Caçador, IsabelCold and heat waves are phenomenon that occurs in higher frequency and intensity due to global climate changes. Commonly cultivated crop species are crucially affected by extreme weather events, and therefore alternative crops - such as halophytes - gain in agricultural interest. While halophytes are potentially able to cope with temperature extremes on the long term exposure, effects of temporary events such as cold and heat waves are not yet described. In order to unveil the effects of these altered thermal environments, Aster tripolium plants were subjected to cold (9/5 °C) and heat (42/38 °C) waves regimes during 3 days and its photochemical and biochemical traits evaluated. In the potential cash crop A. tripolium cold waves induced the gene expression of dehydrins in order to counteract desiccation and thus to prevent oxidative stress. Regulatory proteins on the RNA maturation level (Maturase K) were highly expressed. Heat stress induced the gene expression of the cystein protease gene; most likely to degrade misfolded proteins temporary. Both thermal treatments decreased the photosynthetic efficiency and capacity, driven by a loss in the connectivity between PSII antennae. Nevertheless the light absorption capacity was unaffected due to an increased RC closure net rate. Cold wave-treated individuals showed a decrease in the carotenoid pigmentation, except auroxanthin. In cold wave treated individuals the overall peroxidase activity was significantly increased. Data suggest that exposure to both, cold and heat wave treatment decreased the ecophysiological capacity of A. tripolium.
- Ecophysiological constraints of two invasive plant species under a saline gradient: Halophytes versus glycophytesPublication . Duarte, Bernardo; Santos, D.; Marques, J. C.; Caçador, IsabelSalt marsh environments are harsh environments where salinity comprises one of the most important species distribution shaping factor, presenting sediment salinities from 0 to 855 mM (0–50 ppt). Invasive species have often a high colonizing potential, due to its high plasticity and adaptation ability. Spartina patens is an invasive species already spread along several Mediterranean countries, like France and Spain. Cyperus longus is typically a freshwater species that has been spreading across the Mediterranean. In order to evaluate the ecophysiological fitness of these species, mesocosmos trials were performed subjecting both species to increasing realistic salinity levels and their photochemical and biochemical feedback was evaluated. Both species presented very different behaviours. S. patens appears to be insensitive to salt stress, mostly due to elevated proline concentrations in its leaves allowing it to maintain its osmotic balance, and thus preventing the damaging of its photochemical mechanisms. C. longus, on the other hand, was highly affected by elevated salt levels mostly due to the lack of osmotic balance driven by an incapacity to counteract the elevated ionic strength of the external medium by osmocompatible solutes. S. patens is physiologically highly adapted to saline environments and thus is capable to colonize all the marsh saline environments, while C. longus appears to be an opportunistic invader colonizing the marsh during periods of lower salinities typical from rainy seasons.
- Ecophysiological response of native and invasive Spartina species to extreme temperature events in Mediterranean marshesPublication . Duarte, Bernardo; Marques, J. C.; Caçador, IsabelThe recent IPCC WG2 5th Assessment Report (IPCC 2014), notes an increase in the frequency and duration of extreme climatic events, especially for the Mediterranean region. Together with climate change, the invasion of natural communities by non-indigenous species (NIS) constitutes a serious threat to biodiversity. One of these NIS is the American Spartina patens, now present in Western European marshes. The present study aims to understand the biochemical and photochemical responses of S. patens compared with S. maritima under extreme temperature events. Under normal and extreme heat conditions, S. patens had a higher photosynthetic efficiency (α), compared with cold wave events, where the native S. maritima was far more efficient. This reduced photosynthetic efficiency was mostly due to a decrease in the connectivity between photosystem II (PSII) antennae. This was accompanied by severe damage to the oxygen-evolving complex of PSII. On the other hand, S. patens oxygen evolving complexes (OECs) seem to be temperature insensitive. The light absorption capacity was maintained due to a higher net rate of reaction centre (RC) closure as a counteractive measure of the reduced number of RC, especially in S. maritima. The loss of connectivity between PSII antennae and damage in OECs under heat stress leads to a severe reduction in the maximum yield for photochemistry enhanced by the low probability of each absorbed quanta to produce electronic work. However, while S. maritima presents high energy losses under heat stress, S. patens developed efficient quenching mechanisms under thermal stress, through auroxanthin. In S. patens, cold wave-treated individuals also displayed a very active line of enzymatic defences for reactive oxygen species scavenging. In fact, only cold treated individuals of this species presented higher activities of anti-oxidant enzymes, revealing some degree of adaptation to this new environment. In contrast, in S. maritima the exposure to extreme heat periods led, in most cases, to a decrease in the enzymatic defences, leaving the cell prone to oxidative damage. In summary, S. patens appears to have a higher fitness for the incoming climatic scenarios, being more tolerant to heat stress, while S. maritima will have its photobiological fitness decreased. This will impose a shift in the salt marsh biodiversity, favouring the non-indigenous S. patens expansion.
- Ecotoxicity of the lipid-lowering drug bezafibrate on the bioenergetics and lipid metabolism of the diatom Phaeodactylum tricornutumPublication . Duarte, Bernardo; Prata, Diogo; Matos, Ana Rita; Cabrita, Maria Teresa; Caçador, Isabel; Marques, J. C.; Cabral, Henrique; Reis Santos, Patrick; Fonseca, VanessaPharmaceutical residues impose a new and emerging threat to the marine environment and its biota. In most countries, ecotoxicity tests are not required for all pharmaceutical residues classes and, even when mandatory, these tests are not performed using marine primary producers such as diatoms. These microalgae are among the most abundant class of primary producers in the marine realm and key players in the marine trophic web. Blood-lipid-lowering agents such as bezafibrate and its derivatives are among the most prescribed drugs and most frequently found human pharmaceuticals in aquatic environments. The present study aims to investigate the bezafibrate ecotoxicity and its effects on primary productivity and lipid metabolism, at environmentally relevant concentrations, using the model diatom Phaeodactylum tricornutum. Under controlled conditions, diatom cultures were exposed to bezafibrate at 0, 3, 6, 30 and 60 μg L-1, representing concentrations that can be found in the vicinity of discharges of wastewater treatment plants. High bezafibrate concentrations increased cell density and are suggested to promote a shift from autotrophic to mixotrophic metabolism, with diatoms using light energy generated redox potential to breakdown bezafibrate as carbon source. This was supported by an evident increase in cell density coupled with an impairment of the thylakoid electron transport and consequent photosynthetic activity reduction. In agreement, the concentrations of plastidial marker fatty acids showed negative correlations and Canonical Analysis of Principal coordinates of the relative abundances of fatty acid and photochemical data allowed the separation of controls and cells exposed to bezafibrate with high classification efficiency, namely for photochemical traits, suggesting their validity as suitable biomarkers of bezafibrate exposure. Further evaluations of the occurrence of a metabolic shift in diatoms due to exposure to bezafibrate is paramount, as ultimately it may reduce O2 generation and CO2 fixation in aquatic ecosystems with ensuing consequences for neighboring heterotrophic organisms.
- Estuarine intertidal meiofauna and nematode communities as indicator of ecosystem's recovery following mitigation measuresPublication . Alves, Ana; Caetano, A.; Costa, José Lino; Costa, Maria José; Marques, J. C.The Mondego estuary (Portugal) has been under environmental pressure since the early 1990s due to different anthropogenic stresses. The system has been studied following benthic communities’ features from an impacted situation until the recovery phase, focusing mostly on macrobenthos. Following the application of mitigation measures in the estuary, this study is focused on the variability of the intertidal meiobenthic and nematode communities in a system that has recovered after different anthropogenic stresses. While at the spatial level (among areas along the eutrophication gradient) no significant differences were observed regarding the structure and function of the nematode communities, at the seasonal level some differences stood out. These results broadly suggest that the system has recovered from the early situations of pressures being, to the best of our knowledge, the first attempt to investigate the variability of intertidal meiobenthic and nematode communities in the scope of a system's recovery along an estuarine gradient of eutrophication. Even if performed in a short timeline, this study provides a good baseline analysis of conditions, being important for future comparisons.
- Functional and ecophysiological traits of Halimione portulacoides and Sarcocornia perennis ecotypes in Mediterranean salt marshes under different tidal exposuresPublication . Duarte, Bernardo; Silva, Helena; Dias, João Miguel; Sleimi, Noomene; Marques, J. C.; Caçador, IsabelThe present paper aims to discuss the functional and ecophysiological traits of two abundant succulent halophytes in Mediterranean salt marsh systems, considering two ecotypes differing on their tidal exposure. In Halimione portulacoides the higher root/shoot (R/S) biomass ratio verified in low inundation frequency (LF) marshes, as compared with that in high inundation frequency (HF), indicates an optimal partitioning strategy, inducing differential allocation patterns along with varying environmental factors. Conversely, Sarcocornia perennis appear to have an isometric allocation strategy, based on intrinsic biomass allocation mechanisms independent from the surrounding environmental conditions. Similarly, the photosynthetic traits appear to reflect the intrinsic ecology of the species. Sarcocornia perennis habitat adaptation to HF conditions is clearly revealed by its high photosynthetic efficiency under HF conditions. On the other hand, the cosmopolitan H. portulacoides shows no differences between ecotypes, in which concerns this trait, mostly due to the high carotenoid content of the HF ecotype, which allows this specie to counteract the stress generated during submersion. Additionally, anti-oxidant enzymatic activities revealed that S. perennis presents a high degree of stress tolerance under HF conditions, while H. portulacoides is highly adapted to LF conditions. These functional (biomass-related) and physiological traits determine the optimal habitats for H. portulacoides and S. perennis. While H. portulacoides optimal biomass partitioning, along with its efficient anti-oxidant defence system, makes this species fitter to LF conditions, S. perennis isometric allocation strategy, along with a higher photosynthetic efficiency under frequent flooding conditions, provides this species the mechanisms to efficiently colonize the intertidal habitat.
- Halophyte fatty acids as biomarkers of anthropogenic-driven contamination in Mediterranean marshes: Sentinel species survey and development of an integrated biomarker response (IBR) indexPublication . Duarte, Bernardo; Carreiras, João; Pérez-Romero, Jesús Alberto; Mateos-Naranjo, Enrique; Redondo-Gómez, Susana; Matos, Ana Rita; Marques, J. C.; Caçador, IsabelEstuarine environments are extremely affected by anthropogenic-driven contamination, namely heavy metals. In the recent years, several organisms have been studied to be used as sentinel species providing a wide range of biomarkers for estuarine contamination. Only recently non-traditional biomarkers, such as fatty acids, were included in animal ecotoxicology and impact assessment studies. Nevertheless, there is a wide lack of knowledge regarding to its application in estuarine plants. Considering this the present work aimed to evaluate the application of fatty acid profiling in Mediterranean halophytic species chronically exposed to different degrees of metal contamination, while incorporating these fatty acid biomarkers into an unifying index. From the fatty acids analysis two different types of sentinel halophytes could be identified: the contamination sensitive Halimione portulacoides (sea purslane), Sarcocornia fruticosa (glasswort) and Spartina patens (saltmeadow cordgrass), and the contamination-tolerant Spartina maritima (small cordgrass). In sensitive species the most evident changes in the fatty acid profiles were the decrease in the 18:3 and the increase in the 16:0 fatty acid relative concentrations, while the inverse trend was recorded in the tolerant S. maritima under chronic contamination. Beyond the evident physiological importance, these shifts in the halophyte fatty acid profiles provide some understanding on their use as biomarkers of metal contamination. After application of an integrated biomarker response (IBR) index it was possible to conclude that in sensitive halophytes (H. portulacoides, S. fruticosa and S. patens) the samples collected from the contaminated site produce had higher IBR values than the tolerant species S. maritima being these lower values linked with an adaptation mechanism towards contamination. Furthermore, the fatty acid-based IBR index was intrinsically connected with the bioavailable metals concentrations in the rhizosediments, which make that it could be used in future impact assessment and/or ecotoxicology studies. In summary, the data showed that two of the most abundant halophytes in the Mediterranean eco-region (S. fruticose and H. portulacoides) are potential sentinel species of metal contamination, whereas its fatty acid profile is an efficient biomarker of the degree of environmental contamination.
- Impact of heat and cold events on the energetic metabolism of the C3 halophyte Halimione portulacoidesPublication . Duarte, Bernardo; Santos, D.; Marques, J. C.; Caçador, IsabelAccording to the newest predictions, it is expected that the Mediterranean systems experience more frequent and longer heat and cold treatments events. Salt marshes will be no exception. Halimione portulacoides is a widely distributed halophyte highly adapted to harsh environments. Plants exposed to heat stress showed a reduction in the maximum electron transport rates and increase in the rate of RC closure, as indicated by the increase in M0. Alongside there was also a reduction in the quinone pool size while compared to the plants maintained in the control condition. In contrast plants exposed to low temperatures didn't show any signs of damage on the ETC. Heat-exposed individuals experienced a reduction of connectivity between the PS II antennae with simultaneous inhibition of the electron transport. This was more evident in the donor side of the PS II, Being this a consequence of the damages in the oxygen-evolving complex. Also if both PS I and PS II energy fluxes are observed, there are evident differences in the thermal tolerance of both photosystems. While compared to the control group, cold exposed plants showed an increased PS I efficiency (δR0) indicating a tolerance of this photosystem to low temperatures. Nevertheless, the excessive redox potential generated by light harvesting and inefficient processing was not dissipated correctly and consequently causing a oxidative stress situation. In the present study only heat exposed plants showed a significant activation of the xanthophyll cycle. Alongside with this mechanism and similarly to what was observed for cold treated plants, it could be observed an increase in auroxanthin content, an efficient energy quencher under stress conditions. The coupled activation of the xanthophyll cycle along with a higher auroxanthin synthesis suggests that heat-treated individuals had higher needs to dissipate excessive energy than the cells exposed to cold treatment. In both cases appears to exist an efficient ROS scavenging mechanism. According to our data, heat and cold treatment events can have serious impacts on H. portulacoides photobiology reducing its primary productivity. At the ecosystem level, these climatic events could pose a serious threat to the survival of this species in the new climatic reality that our planet is facing.