- In this study, the molecular roles of exogenously H 2 S application were explored by RNA sequencing under Cd stress in T.. - Comparative transcriptome analysis showed that the expression levels of 9152 genes changed under Cd stress (4658 upregulated and 4494 downregulated). - However, only 1359 genes were differentially expressed with NaHS treatment under Cd stress (1087 upregulated and 272 downregulated). - transcripts involved in the oxidation – reduction process, oxidoreductase activity, glutathione peroxidase activity, and cell redox homeostasis were the considerable enrichments between Cd stress and NaHS treatment under Cd stress.. - Kyoto Encyclopedia of Genes and Genomes (KEGG) indicated that the carbon metabolism, glutathione metabolism, metabolism of xenobiotics by cytochrome P450, and ABC transporters were significantly differentially expressed components between Cd stress and NaHS treatment under Cd stress in T. - Conclusion: NaHS alleviated Cd stress mainly through inhibiting Cd absorbtion, promoting CdS nanoparticles formation, increasing oxidation resistance, and regulation of transport in free-living unicellular T. - These findings will expand our understanding for H 2 S functions in the freshwater protozoa.. - Exogenous H 2 S acts as a potent antioxidant under Cd stress by enhancing anti- oxidant enzymes activities in wheat seedlings [10], and alleviates Cd toxicity through regulations of Cd transport across the plasma and vacuolar membranes in Populus euphratica cells [11]. - thermophila stressed under Cd was evalu- ated by phenotypic observation, enzyme and metabolites analysis, and high throughput transcriptome sequencing technology. - thermophila under Cd stress. - Cd caused a dose-dependent decline in the viability of T. - thermophila under Cd and NaHS treatments was investigated. - The results showed exogenous NaHS play a protective role on Cd stress in T. - These physio- logical processes are interrelated under Cd stress [26].. - Exogenous cysteine treatment enhanced H 2 S level and maintained H 2 S at high level under Cd stress (Fig.. - H 2 S alleviates lipid peroxidation and improves antioxidant capacity under Cd stress. - thermophila under Cd and H 2 S treatments. - c The effects of cysteine on H 2 S content under Cd stress. - accumulation (decrease by 26%) in the T. - Cd stress caused lipid peroxidation and induced mal- ondialdehyde (MDA) production of T. - thermophila cells under Cd stress (Fig. - Under Cd stress, GSH content increased by 51%. - Further- more, SOD activity increased by 92% and CAT activity increased by 29% under Cd stress. - thermophila, spherical CdS nanoparticles in yellow colour with an average par- ticle diameter of nm were observed under Cd treatment, and the nanoparticles amount increased by adding NaHS (Fig. - The Ag nanoparticles stored intracellularly in the food vacuoles of T. - 3 Effects of NaHS on Cd accumulation, H 2 S content, lipid peroxidation, and antioxidant system under Cd stress. - To further explore the mechanisms of H 2 S alleviating Cd stress in T. - Differentially expressed genes (DEGs) in response to NaHS treatment, Cd stress, and NaHS treatment under Cd stress. - DEGs were hierarchically clustered to obtain a compre- hensive view of the differential gene expression under NaHS treatment, Cd stress, and NaHS treatment with Cd stress (Fig. - Under Cd stress, the expression level of 9152 genes sig- nificantly changed, including 4658 upregulated genes and 4494 downregulated genes. - A total of 1087 genes were upregulated and 272 genes were downregulated with NaHS treatment under Cd stress. - The expression levels of most genes recovered under Cd stress with NaHS treatment. - and 3738 genes were downregulated between Cd stress and NaHS treatment under Cd stress (Fig. - indicating that H 2 S-responsive tran- scripts under normal condition were far fewer than those under Cd stress. - The significantly upregulated genes under Cd stress (log2FC. - Compared with Cd stress, the unigenes significantly downregulated in the NaHS treatment under Cd stress (log2FC <. - The results indicated that the redox system is sensitive for NaHS treatment and Cd stress in T. - To obtain the functional annotations of the DEGs for Cd stress and H 2 S treatment under Cd stress, GO cat- egory enrichment analysis was performed. - In the. - Under Cd stress, 283 DEGs were in- cluded in oxidoreductase activity and 231 DEGs partici- pated in oxidation-reduction process. - These data indicated that H 2 S responds to Cd stress mainly through the adjustment of the redox balance.. - Under Cd stress, the 1116 DEGs were mapped to the 252 KEGG pathways. - For NaHS treatment under Cd stress compared with Cd stress, 966 DEGs were mapped to 247 KEGG pathways.. - Under Cd stress, 54 DEGs (4.84%) were dis- tributed in the carbon metabolism, and 48 DEGs (4.97%) also enriched in this pathway with adding NaHS. - 52 DEGs (4.66%) were enriched at the GSH metabolism under Cd stress, and 54 DEGs (5.59%) were also enriched with adding NaHS. - Under Cd stress, 39 DEGs (3.49%) or 40 DEGs (3.58%) were distributed in drug metabolism–cytochrome P450 or metabolism of. - 5 Responses of DEGs to H 2 S treatment, Cd stress, and H 2 S treatment under Cd stress. - Besides, under Cd stress, 67 DEGs (6.00%) were distributed in ABC transporters (Fig.. - C, indicating the important detox effects through H 2 S under Cd stress.. - Regulation of cytochrome P450 and GSH metabolism under Cd stress and with NaHS treatment. - Cytochrome P450 represents an important participant in regulatory networks of organism responses to Cd stress.. - In the wolf spider Pardosa pseudoannulata, cytochrome P450 genes were found to respond to Cd stress [36]. - Under Cd stress, metabolism of xenobiotics by cytochrome P450 was significantly enriched by KEGG pathway analysis on DEGs, of which 32 DEGs were markedly upregulated (Fig. - Under Cd stress, GSH metabolism was sig- nificantly enriched by KEGG pathway analysis on upreg- ulated genes (Fig. - Specifically, the 8 upregulated GPXs and 29 GSTs induced by Cd stress returned to normal level with adding NaHS. - Given the role of GSH in the detoxification of heavy metals, regulation of GSH, GPXs, and GSTs by H 2 S play an important role in ameliorating Cd stress.. - Regulation of ABC transporters under Cd stress and with NaHS treatment. - Under Cd stress, 49 DEGs involved in the enrichment of ABC transporters were markedly upregulated, while 18 DEGs were downregulated. - The ABCA subfamily comprises of a total of 32 mem- bers, which included 9 upregulated genes and 3 down- regulated genes under Cd stress. - The ABCG subfamily includes 39 trans- porters, and 9 genes were upregulated under Cd stress and 3 genes were inhibited. - 00034920) was induced up to 357 folds under Cd stress (Table S2), which suggested it could play an important role in detoxifying Cd toxicity. - Most of the expression of ABC transporters under Cd stress was restored and some of them even hyper activated by exogenous NaHS addition, which is in relation to H 2 S lowering the cellular concen- tration of Cd (Fig. - Under Cd stress, the expression levels of the genes were signifi- cantly upregulated. - The log2FC of the CdMT genes were ranked as MTT5≈MTT3>MTT1 under Cd stress (Table S2). - On the contrary, the expression level of MTT1 further increased and MTT5 had no obvious change with NaHS treatment under Cd stress (Fig. - patterns between Cd stress and NaHS treatment under Cd stress, indicating their functional diversification. - In the present study, IC50 of Cd for T. - Among them, H 2 S participates in suppressing Cd stress. - The GSH content was notable up- regulated under Cd stress, and with the addition of NaHS, the GSH content further increased (Fig. - Cd stress leads to the significant molecular and physiological changes in different organisms. - The whole transcriptional reprogramming is regarded as a vital mo- lecular response to Cd stress. - Under Cd stress, 27 oxidoreduc- tases and 9 glutathione peroxidase family proteins were upregulated in the oxidation–reduction process enrichment term in T. - MTT5 is the strongest induced, while MTT3 is the least induced by Cd stress at 1 h or 24 h [40, 47]. - thermophila was stressed by Cd stress at 6 h (Fig. - Inter- estingly, the upregulated expression of MTT1 under Cd stress further increased with NaHS treatment. - On the contrary, the upregulated expression of MTT3 under Cd stress decreased with NaHS treatment. - Meanwhile, the upregulated expression of MTT5 under Cd stress has no significant change with NaHS treatment. - 8 Relative expression levels of six DEGs with NaHS treatment and under Cd stress. - In the present study, three HSPs (TTHERM TTHERM_00351140, TTHERM_00171850) were found to activate by Cd and the expression level returns to normal by NaHS addition (Fig. - It seems that H 2 S involved in the detoxification of Cd through the antioxidative stress sig- nal pathway.. - H 2 S alleviates Cd stress by regulating cytochrome P450, GST and ABC transporters. - GST plays important roles through GSH me- tabolism in the protection against oxidative damage resulting from the high levels of ROS under Cd stress in Chironomus riparius [52]. - The expression level of 31 GSTs was upregulated under Cd stress. - gene, has an important role in the detoxification of Cd [58]. - We do not detect the change of ABCB15 at tran- scriptional level under Cd stress in T. - As a whole, a total of 67 DEGs were enriched in the ABC transporter pathway under Cd stress, and the ABC transporter pathway was significantly altered with NaHS addition. - H 2 S physiologically alleviated growth inhibition under Cd stress and accumulation of Cd in T. - H 2 S affected various metabolic processes and molecular func- tions under Cd stress, including glutathione metabolism, metabolism of xenobiotics by cytochrome P450, GPXs, Cytochrome P450s, GSTs, and ABC transporters. - thermophila from Cd stress by inhibiting Cd absorbtion, promoting CdS nanoparticles formation, increasing oxidation resistance, and regula- tion of transport.. - To analyze the effects on cell proliferation of H 2 S under Cd stress, cells were uniformly grown in the SPP medium with 30 μM Cd (IC50) in the presence or ab- sence of 70 μM NaHS (optimum positive effect concen- tration), and their starting concentration was 0.7× 10 5 mL − 1 . - The cells in the logarithmic phase were divided into four groups with different treatments: (1) CK, without chemical treatments (the standard SPP solution). - CS represents the group with NaHS treatment under Cd stress. - C: Control under Cd stress. - CS: Control with H 2 S treatment under Cd stress. - Genome-wide identification and characterization of cytochrome P450 monooxygenase genes in the ciliate Tetrahymena thermophila. - Expression and functional analysis of cytochrome P450 genes in the wolf spider Pardosa pseudoannulata under cadmium stress. - Arsenate toxicity and stress responses in the freshwater ciliate Tetrahymena pyriformis. - Cadmium- induced oxidative stress, histopathology, and transcriptome changes in the hepatopancreas of freshwater crayfish (Procambarus clarkii). - Proteomic analysis in the lichen Physcia adscendens exposed to cadmium stress. - Effect of heavy metals on the antioxidant enzymes in the marine ciliate Euplotes crassus
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