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[Objective] The biological regulatory mechanisms of soil microorganisms in response to single heavy metals or organic pollutants have been extensively studied, while the biological mechanisms of microbial responses to combined pollution remain unclear. This study aims to reveal the biological mechanisms of Enterococcus faecalis HHT-1 in response to single and combined stress of cadmium (Cd) and aniline (AN) by transcriptomics. [Methods] Transcriptomics was employed to explore the transcriptional regulation of HHT-1 under single (Cd: 150 mg/L; AN: 2 g/L) and combined stress (150 mg/L Cd and 2 g/L AN) at half inhibitory concentrations (IC50). [Results] Under single Cd stress, HHT-1 upregulated the expression of genes encoding metal-binding proteins and transporters to promote the sequestration or efflux of Cd2+. High concentrations of Cd induced oxidative stress in cells, and HHT-1 upregulated the expression of ribosome-related genes and nucleotide-related genes to repair the protein damage and DNA damage caused by oxidative stress, and cleared intracellular reactive oxygen species (ROS) by producing catalase. Moreover, Cd stress upregulated the expression of genes related to virulence and antibiotic resistance in HHT-1. Under the single stress of AN, HHT-1 activated the expression of AN-degrading enzyme genes to reduce its toxicity and upregulated the expression of genes encoding efflux pumps to excrete AN from the cell. AN also caused intracellular oxidative stress, and HHT-1 cleared the ROS by upregulating the expression of glutathione synthase genes. Under the combined stress of Cd and AN, HHT-1 exhibited a complex array of adaptive mechanisms. Initially, HHT-1 upregulated the expression of genes encoding metal transporters and efflux pumps to transport Cd and AN out of the cell, thereby mitigating their toxic effects on the cell. At the same time, HHT-1 cleared intracellular ROS via the dual pathway of catalase and glutathione. In addition, HHT-1 upregulates the expression of virulence and antibiotic resistance genes under combined stress, which may enhance its pathogenicity and resistance. It is worth noting that HHT-1 did not significantly upregulate the expression of AN-degrading enzyme-related genes under the combined stress. [Conclusion] The transcriptional regulation mechanism of E. faecalis HHT-1 in response to the combined stress of Cd and AN is a combination of the transcriptional regulation mechanisms observed under single Cd stress and single AN stress, and it is more similar to the mechanism observed under single Cd stress. HHT-1 mainly responds to the combined stress of Cd and AN by enhancing cell wall synthesis, Cd2+ efflux, DNA repair, and ROS scavenging. Genes associated with efflux pumps and glutathione under AN stress continue to be expressed under the combined stress. Both single Cd stress and combined stress of Cd and AN may increase the potential pathogenicity and drug resistance of HHT-1.
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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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