Red algae Gracilaria tenuistipitata: control vs. high copper and high cadmium concentration
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ABSTRACT: The study of macroalgae capacity to acclimate and recover in environments contaminated with Cu and Cd could prove a promising way to understand the tolerance mechanisms of these seaweeds against different pollutants. This study used as a model organism Gracilaria tenuistipitata (Rhodophyta), a macroalga with economic and ecological importance. The partial transcriptome of G. tenuistipitata was profiled using cDNA microarrays in the sixth day of exposition to EC50 metals. Genes involved in Cu and Cd chronic stress belonging to various functional categories suffered shallow modifications. This possibly indicates that G. tenuistipitata would be in the acclimatization process. In addition, the expression of nine stress genes accompanied by analysis of the photosynthetic rate of seaweed to both metals in three different time frames was analyzed. Genetic variation linked to the mechanism of resistance of the algae, determined from EC50 culture conditions established for two metals, occurred in the early hours of treatment. It was found that G. tenuistipitata was able to accumulate these two metals and to resist and acclimate to the negative effects produced by these elements. The temporal analysis from the nine specific genes showed some specific transcriptional responses of the G. tenuistipitata, exposed to Cu and Cd. Three-condition experiment, control cells cultivated in seawater enriched with von Stosch solution vs. copper and cadmium (indepedent) treated cells. Biological replicates: 4 control, 2 copper treated, 2 cadmium treated. All independently grown and harvested. Four replicates per array.
Project description:Metals, including copper (Cu) and nickel (Ni) are among the most common contaminants in soils in Europe. Although their effects are relatively well known regarding survival and reproduction of soil invertebrates, their modes of action in these organisms are still poorly studied. Enchytraeus albidus has been used in soil ecotoxicology for many years, and more recently has a gene library and an oligonucleotide microarray for this species which allowed gene expression studies. This has potentiated the means to study further in depth the mechanisms of response to stressors. The main aim of this study is to understand the mechanisms of response of E. albidus to Cu and Ni. For that we have 1) assessed and compared the transcriptomic profile of E. albidus in response to Cu and Ni and 2) compared the Cu, Ni, Cd and Zn transcriptomic profiles. For the microarray hybridizations, E. albidus were exposed to the reproduction effect concentrations EC50 and EC90 of Cu and Ni during 4 days. Results indicate that Cu and Ni have to some extent, similar mechanisms of toxicity and that have already been identified in other species, indicating cross-species conserved mechanisms. Based on hierarchical clustering, it was possible to observe a clear separation of Cd treatments from all other metals. This separation strongly correlates with the available information regarding the toxicokinetics of the tested metals, in which Ni shows properties similar to essential metals. Gene expression in E.albidus was measured 4 days after exposure to Copper, Nickel, Cadmium and Zinc at 2 concentrations of effect on reprocduction (EC50 and EC90). Three biological replicates per exposure condition and 6 biological replicates of control conditions were used.
Project description:The study of macroalgae capacity to acclimate and recover in environments contaminated with Cu and Cd could prove a promising way to understand the tolerance mechanisms of these seaweeds against different pollutants. This study used as a model organism Gracilaria tenuistipitata (Rhodophyta), a macroalga with economic and ecological importance. The partial transcriptome of G. tenuistipitata was profiled using cDNA microarrays in the sixth day of exposition to EC50 metals. Genes involved in Cu and Cd chronic stress belonging to various functional categories suffered shallow modifications. This possibly indicates that G. tenuistipitata would be in the acclimatization process. In addition, the expression of nine stress genes accompanied by analysis of the photosynthetic rate of seaweed to both metals in three different time frames was analyzed. Genetic variation linked to the mechanism of resistance of the algae, determined from EC50 culture conditions established for two metals, occurred in the early hours of treatment. It was found that G. tenuistipitata was able to accumulate these two metals and to resist and acclimate to the negative effects produced by these elements. The temporal analysis from the nine specific genes showed some specific transcriptional responses of the G. tenuistipitata, exposed to Cu and Cd.
Project description:Metals, including copper (Cu) and nickel (Ni) are among the most common contaminants in soils in Europe. Although their effects are relatively well known regarding survival and reproduction of soil invertebrates, their modes of action in these organisms are still poorly studied. Enchytraeus albidus has been used in soil ecotoxicology for many years, and more recently has a gene library and an oligonucleotide microarray for this species which allowed gene expression studies. This has potentiated the means to study further in depth the mechanisms of response to stressors. The main aim of this study is to understand the mechanisms of response of E. albidus to Cu and Ni. For that we have 1) assessed and compared the transcriptomic profile of E. albidus in response to Cu and Ni and 2) compared the Cu, Ni, Cd and Zn transcriptomic profiles. For the microarray hybridizations, E. albidus were exposed to the reproduction effect concentrations EC50 and EC90 of Cu and Ni during 4 days. Results indicate that Cu and Ni have to some extent, similar mechanisms of toxicity and that have already been identified in other species, indicating cross-species conserved mechanisms. Based on hierarchical clustering, it was possible to observe a clear separation of Cd treatments from all other metals. This separation strongly correlates with the available information regarding the toxicokinetics of the tested metals, in which Ni shows properties similar to essential metals.
Project description:Metal ecotoxicity on soil organisms, e.g. in Enchytraeids (also known as potworms), has been addressed mainly by assessing effects on survival and reproduction but very little is known about the molecular underlying mechanisms of responses. The main purpose of the present study was to assess and compare the transcriptional responses of Enchytraeus albidus to an essential (zinc) and a non-essential metal (cadmium). Exposure was performed to two concentrations of known effect on reproduction (EC50, EC90) at three time periods (2, 4 and 8 days). Results showed that transcriptional responses were influenced by exposure duration but, independently of that, the mechanisms of response to Cd and Zn were consistently different. Both metals affected pathways related with the regulation of gene expression, calcium homeostasis and cellular respiration. Mechanisms of toxicity that were exclusively associated with Cd exposures were the inhibition of DNA repair and the impairment of ubiquitin-mediated proteolysis. The microarray for E. albidus was a useful tool to detect molecular pathways affected by metal exposures. Transcriptional responses strongly correlated with known mechanisms of Cd and Zn responses in other organisms, suggesting cross-species conserved mechanisms of action. It should be highlighted that we could not only retrieve mechanistic information, but also that genes responded within 2-8 days of exposure. This represents an additional advantage of using such molecular endpoints as a complement to the traditional, more time-consuming endpoints. Gene expression in E.albidus was measured at 2, 4, and 8 days after exposure to Cadmium and Zinc at 2 concentrations of effect on reprocduction (EC50 and EC90). Three biological replicates per exposure treatment in each time point and 4 biological replicates of control conditions were used.
Project description:In this study, the metabolic adjustments performed by maize (Zea mays L) seminal roots exposed to 25 µM Cd2+ or 25 µM Cu2+ at pre-emergence are compared, focusing on the proteomic changes after metal exposure. Root width was increased, and root length was decreased after 72 h of metal treatment. Both metals induced H2O2 accumulation and lipid peroxidation in the root tip. These changes were accompanied by increases in lipoxygenase activity and 4-hydroxy-2-nonenal content. NMR spectroscopy revealed that the abundance of 38 water-soluble metabolites was significantly modified by Cd and Cu exposure; this set of metabolites comprised carboxylic acids, amino acids, carbohydrates, and unidentified phenolic compounds. Linoleic acid content significantly decreased in Cu-treated samples. The total amount of proteins detected in maize root apexes was 2,171. Gene ontology enrichment analysis of the differentially accumulated proteins was performed to detect pathways probably affected by metal additions. Both metals altered redox homeostasis, up-regulated oxylipins biosynthetic process, and shifted metabolism towards the oxidative pentose-phosphate in the root apexes. However, the methionine salvage pathway appears as a key metabolic module only under Cd stress. The integrative analysis carried out in this study suggests that most molecular features behind the reprogramming of maize root tips to cope with cadmium and copper toxicity are common, but some are not.
Project description:Iron (Fe) and copper (Cu) are essential metal micronutrients that are necessary for many redox reactions. The uptake of these metals is tightly regulated in plants. Some redox processes can alternatively use Fe-containing proteins or Cu-containing proteins, depending on nutritional status. Copper deficiency can rescue a Cucumis melo Fe uptake deficient mutant, and Fe deficiency can result in increased accumulation of Cu. However, the system responsible for Fe-deficiency-regulated Cu-uptake is unknown. To understand the genes and gene networks associated with Fe-deficiency regulated Cu uptake and Fe-Cu cross-talk, we conducted transcriptomic profiling of roots and rosettes of spl7 (a Cu uptake deficient mutant in arabidopsis) and Col-0 (WT) grown under Fe, Cu and simultaneous Fe and Cu deficiency conditions.
Project description:Anthropogenic pollution has increased the levels of heavy metals in the environment. Bacterial populations continue to thrive in highly polluted environments and bacteria must have mechanisms to counter heavy metal stress. We chose to examine the response of the environmentally-relevant organism Pseudomonas aeruginosa to two different copper treatments. A short, 45 min exposure to copper was done in the Cu shock treatment to examine the immediate transcriptional profile to Cu stress. The Cu adapted treatment was designed to view the transcriptional profile of cells that were actively growing in the presence of Cu. Keywords: stress response
Project description:The cadmium-resistant Cupriavidus sp. E324 strain has been previously shown to have a high potential for use in cadmium (Cd) remediation, due to its high capacity for cadmium bioaccumulation. According to the comparative genomic analysis, the E324 strain was most closely related to C. nantongensis X1 T , indicating that the E324 strain should be re-identified as C. nantongensis. To unravel the Cd tolerance mechanisms of C. nantongensis E324, the transcriptional response of this strain to Cd stress was assessed using RNA-seq-based transcriptome analysis, followed by validation through qRT- PCR. The results showed that the upregulated Differentially Expressed Genes (DEGs) were significantly enriched in categories related to metal binding and transport, phosphate transport, and oxidative stress response. Consistently, we observed significant increases in both the cell wall and intracellular contents of certain essential metals (Cu, Fe, Mn, and Zn) upon Cd exposure). Among these, only the Zn pretreatment resulting in high Zn accumulation in the cell walls could enhance bacterial growth under Cd stress conditions through its role in inhibiting Cd accumulation. Additionally, the promotion of catalase activity and glutathione metabolism upon Cd exposure to cope with Cd-induced oxidative stress was demonstrated. Meanwhile, the upregulation of phosphate transport-related genes upon Cd treatment seems to be the bacterial response to Cd-induced phosphate depletion. Altogether, our findings suggest that these adaptive responses are critical mechanisms contributing to increased Cd tolerance in C. nantongensis E324 strain via the enhancement of metal-chelating and antioxidant capacities of the cells.
Project description:Environmental metals are known to cause harmful effects to fish of which many molecular mechanisms still require elucidation. Particularly concentration dependence of gene expression effects is unclear. Focusing on this matter, zebrafish embryo toxicity tests were used in combination with transcriptomics. Embryos were exposed to three concentrations of copper (CuSO4), cadmium (CdCl2) and cobalt (CoSO4) from just after fertilization until the end of the 48 hpf pre- and 96 hpf post-hatch stage. The RNA was then analyzed on Agilent’s Zebrafish (V3, 4x44K) arrays. Enrichment for GO terms of biological processes illustrated for cadmium that most affected GO terms were represented in all three concentrations, while for cobalt and copper most GO terms were represented in the lowest test concentration only. This suggested a different response to the non-essential cadmium than cobalt and copper. In cobalt and copper treated embryos, many developmental and cellular processes as well as the Wnt and Notch signaling pathways were found significantly enriched. Also, different exposure concentrations affected varied functional networks. In contrast, the largest clusters of enriched GO terms for all three concentrations of cadmium included responses to cadmium ion, metal ion, xenobiotic stimulus, stress and chemicals. However, concentration dependence of mRNA levels was evident for several genes in all metal exposures. Some of these genes may be indicative of the mechanisms of action of the individual metals in zebrafish embryos.