Genetic response analysis of Beauveria bassiana Z1 under high concentration Cd(II) stress.

Cadmium (Cd(II)) has carcinogenic and teratogenic toxicity, which can be accumulated in the human body through the food chain, endangering human health and life. In this study, a highly Cd(II)-tolerant fungus named Beauveria bassiana Z1 was studied, and its Cd(Ⅱ) removal efficiency was 71.2% when the Cd(II) concentration was 10 mM. Through bioanalysis and experimental verification of the transcriptome data, it was found that cadmium entered the cells through calcium ion channels, and then complexed with intracellular glutathione (GSH) and stored in vacuoles or excluded extracellular by ABC transporters. Cytochrome P450 was significantly upregulated in many pathways and actively participated in detoxification related reactions. The addition of cytochrome inhibitor taxifolin reduced the removal efficiency of Cd(II) by 45%. In the analysis, it demonstrated that ACOX1 gene and OPR gene of jasmonic acid (JA) synthesis pathway were significantly up-regulated, and were correlated with bZIP family transcription factors cpc-1_0 and pa p1_0. The results showed that exogenous JA could improve the removal efficiency of Cd(II) by strain Z1.

Metabolic diversity shapes vegetation-enhanced methane oxidation in landfill covers: Multi-omics study of rhizosphere microorganisms.

Vegetation root exudates have the ability to shape soil microbial community structures, thereby enhancing CH4 bio-oxidation capacity in landfill cover systems. In this study, the CH4 oxidation capacity of indigenous vegetation rhizosphere microorganisms within operational landfill covers in Chongqing, China, was investigated for the first time, with the objective of identifying suitable plant candidates for CH4 mitigation initiatives within landfill cover systems. Furthermore, a multi-omics methodology was employed to explore microbial community structures and metabolic variances within the rhizospheric environment of diverse vegetation types. The primary aim was to elucidate the fundamental factors contributing to divergent CH4 oxidation capacities observed in rhizosphere soils. The findings demonstrated that herbaceous vegetation predominated in landfill covers. Notably, Rumex acetosa exhibited the highest CH4 oxidation capacity in the rhizosphere soil, approximately 20 times greater than that in non-rhizosphere soil. Root exudates played a crucial role in inducing the colonization of CH4-oxidizing functional microorganisms in the rhizosphere, subsequently prompting the development of specific metabolic pathways. This process, in turn, enhanced the functional activity of the microorganisms while concurrently bolstering their tolerance to microbial pollutants. Consequently, the addition of substances like Limonexic acid strengthened the CH4 bio-oxidation process, thereby underscoring the suitability of Rumex acetosa and similar vegetation species as preferred choices for landfill cover vegetation restoration.

The performance, mechanism and greenhouse gas emission potential of nitrogen removal technology for low carbon source wastewater.

Excessive nitrogen can lead to eutrophication of water bodies. However, the removal of nitrogen from low carbon source wastewater has always been challenging due to the limited availability of carbon sources as electron donors. Biological nitrogen removal technology can be classified into three categories: heterotrophic biological technology (HBT) that utilizes organic matter as electron donors, autotrophic biological technology (ABT) that relies on inorganic electrons as electron donors, and heterotrophic-autotrophic coupling technology (CBT) that combines multiple electron donors. This work reviews the research progress, microbial mechanism, greenhouse gas emission potential, and challenges of the three technologies. In summary, compared to HBT and ABT, CBT shows greater application potential, although pilot-scale implementation is yet to be achieved. The composition of nitrogen removal microorganisms is different, mainly driven by electron donors. ABT and CBT exhibit the lowest potential for greenhouse gas emissions compared to HBT. N2O, CH4, and CO2 emissions can be controlled by optimizing conditions and adding constructed wetlands. Furthermore, these technologies need further improvement to meet increasingly stringent emission standards and address emerging pollutants. Common measures include bioaugmentation in HBT, the development of novel materials to promote mass transfer efficiency of ABT, and the construction of BES-enhanced multi-electron donor systems to achieve pollutant prevention and removal. This work serves as a valuable reference for the development of clean and sustainable low carbon source wastewater treatment technology, as well as for addressing the challenges posed by global warming.

Quantitative analysis of TCE biodegradation pathway in landfill cover utilizing continuous monitoring, droplet digital PCR and multi-omics sequencing technology.

The remediation of volatile chlorinated hydrocarbons in the quasi-vadose zone has become a significant challenge. We applied an integrated approach to assess the biodegradability of trichloroethylene to identify the biotransformation mechanism. The formation of the functional zone biochemical layer was assessed by analyzing the distribution of landfill gas, physical and chemical properties of cover soil, spatial-temporal variations of micro-ecology, biodegradability of landfill cover soil and distributional difference metabolic pathway. Real-time online monitoring showed that trichloroethylene continuously undergoes anaerobic dichlorination and simultaneous aerobic/anaerobic conversion-aerobic co-metabolic degradation on the vertical gradient of the landfill cover system and reduction in trans-1,2-dichloroethylene in the anoxic zone but not 1,1-dichloroethylene. PCR and diversity sequencing revealed the abundance and spatial distribution of known dichlorination-related genes within the landfill cover, with 6.61 ± 0.25 × 104-6.78 ± 0.09 × 106 and 1.17 ± 0.78 × 103-7.82 ± 0.07 × 105 copies per g/soil of pmoA and tceA, respectively. In addition, dominant bacteria and diversity were significantly linked with physicochemical factors, and Mesorhizobium, Pseudoxanthomonas and Gemmatimonas were responsible for biodegradation in the aerobic, anoxic and anaerobic zones. Metagenome sequencing identified 6 degradation pathways of trichloroethylene that may occur in the landfill cover; the main pathway was incomplete dechlorination accompanied by cometabolic degradation. These results indicate that the anoxic zone is important for trichloroethylene degradation.

Microecological health assessment of water environment and sediment based on metagenomics: a case study of Guixi River in Chongqing, China.

River sediment is vital in containing water pollution and strengthening water remediation. This paper has conducted a study on the microecological health assessment of the sediment and water body of Guixi River in Dianjiang, Chongqing, China, using metagenomics sequencing and microbial biological integrity index (M-IBI) technology. The analysis of physical and chemical characteristics shows that the concentration of TN varies from 2.62 to 9.76 mg/L in each sampling section, and the eutrophication of the water body is relatively severe. The proportion of Cyanobacteria in the sampling section at the sink entrance is higher than that of other sites, where there are outbreaks of water blooms and potential hazards to human health. The dominant functions of each site include carbon metabolism, TCA cycle, and pyruvate metabolism. In addition, the main virulence factors and antibiotic resistance genes in sediment are Type IV pili (VF0082), LOS (CVF494), MymA operon (CVF649), and macrolide resistance genes macB, tetracyclic tetA (58), and novA. Correlation analysis of environmental factors and microorganisms was also performed, and it was discovered that Thiothrix and Acidovorax had obvious gene expression in the nitrogen metabolism pathway, and the Guixi River Basin had a self-purification capacity. Finally, based on the microecological composition of sediment and physical and chemical characteristics of the water body, the health assessment was carried out, indicating that the main pollution area was Dianjiang Middle School and the watershed near the sewage treatment plant. The findings should theoretically support an in-depth assessment of the water environment's microecological health.

Domestication and pilot-scale culture of mixed bacteria HY-1 capable of heterotrophic nitrification-aerobic denitrification.

To further investigate the potential of heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria for practical applications, the HN-AD mixed bacteria HY-1 were enriched and domesticated in this study. After five generations of domestication, the mixture was able to remove 98% of ammonia nitrogen (400 mg/L) and 81.9% of mixed nitrogen source (nitrate, nitrite). Changes in community structure in the domestication process of mixed microorganisms were studied using 16S rDNA-seq. The results indicated an increase in the abundance of Acinetobacter from 16.9% to 80%. The conditions for the expanded culture of the HY-1 were also optimized. Moreover, A pilot-scale expanded reactor with a capacity of 1000L was constructed, and the HY-1 was successfully expanded from 0.1L to 800L. The community structures of the HY-1 remained stable after the expanded culture, with Acinetobacter as the dominant species. Moreover, the HY-1 demonstrated adaptability to actual high ammonia nitrogen wastewater and showed potential for practical application.

The potential and sustainable strategy for swine wastewater treatment: Resource recovery.

Swine wastewater is highly polluted with complex and harmful substances that require effective treatment to minimize environmental damage. There are three commonly used biological technologies for treating swine wastewater: conventional biological technology (CBT), microbial electrochemical technology (MET), and microalgae technology (MT). However, there is a lack of comparison among these technologies and a lack of understanding of their unique advantages and efficient operation strategies. This review aims to compare and contrast the characteristics, influencing factors, improvement methods, and microbial mechanisms of each technology. CBT is cost-effective but has low resource recovery efficiency, while MET and MT have the highest potential for resource recovery. However, all three technologies are affected by various factors and toxic substances such as heavy metals and antibiotics. Improved methods include exogenous/endogenous enhancement, series reactor operation, algal-bacterial symbiosis system construction, etc. Though MET is limited by construction costs, CBT and MT have practical applications. While swine wastewater treatment processes have developed automatic control systems, the application need further promotion. Furthermore, key functional microorganisms involved in CBT's pollutant removal or transformation have been detected, as have related genes. The unique electroactive microbial cooperation mode and symbiotic mode of MET and MT were also revealed, respectively. Importantly, the future research should focus on broadening the scope and scale of engineering applications, preventing and controlling emerging pollutants, improving automated management level, focusing on microbial synergistic metabolism, enhancing resource recovery performance, and building a circular economy based on low-cost and resource utilization.

Water microecology is affected by seasons but not sediments: A spatiotemporal dynamics survey of bacterial community composition in Lake Changshou-The largest artificial lake in southwest China.

This study aimed to evaluate the correlation between microecology of sediments and water as well as their spatial-temporal variations in Changshou Lake. The results demonstrated that microecology in the lake exhibits spatiotemporal heterogeneity, and microbial diversity of sediments was significantly higher than that of water body. Further, it was found that there was statistically insignificant positive correlation between microecology of sediments and that of water body. PCoA and community structure analysis revealed that the predominant phyla which exhibited significant spatial differences in sediments were Proteobacteria, Actinobacteria and Planctomycetes. While, the distribution of dominant bacteria Actinobacteria and Verrucomicrobia in water body showed significant seasonal differences. Microbial networks analysis indicated that there was a cooperative symbiotic relationship between lake microbial communities. Notably, the same bacterial genus had no significant positive correlation in sediment and water, which suggested that bacteria transport between sediment-water interface does not influence the microecological functions of lake water.

Effect mechanism of individual and combined salinity on the nitrogen removal yield of heterotrophic nitrification-aerobic denitrification bacteria.

One of the biggest challenges of applying heterotrophic nitrification-aerobic denitrification (HN-AD) bacteria to treat high salt organic wastewater lies in the inhibitory effect exerted by salinity. To study the inhibition effect and underlying mechanism induced by different ion types and ion composition, the individual and combined effects of NaCl, KCl and Na2SO4 on HN-AD bacteria Acinetobacter sp. TAC-1 were systematically investigated by batch experiments. Results indicated that the ammonia nitrogen removal yield and TAC-1 activity decreased with increased salt concentration. NaCl, KCl and Na2SO4 exerted different degrees of inhibition on TAC-1, with half concentration inhibition constant values of 0.205, 0.238 and 0.110 M, respectively. A synergistic effect on TAC-1 was found with the combinations of NaCl + KCl, NaCl + Na2SO4 and NaCl + KCl + Na2SO4. The whole RNA resequencing suggested that transcripts of denitrification genes (nirB and nasA) were significantly downregulated with increased Na2SO4 concentration. Simultaneously, Na2SO4 stress disrupted cell respiration, DNA replication, transcription, translation, and induced oxidative stress. Finally, we proposed a conceptual model to summarize the inhibition mechanisms and possible response strategies of TAC-1 bacteria under Na2SO4 stress.

Characterization of a novel salt-tolerant strain Sphingopyxis sp. CY-10 capable of heterotrophic nitrification and aerobic denitrification.

A novel heterotrophic nitrification and aerobic denitrification (HN-AD) strain CY-10 was isolated and identified as Sphingopyxis sp. When ammonium, nitrate or nitrite was used as the sole nitrogen source (300 mg/L), the maximum nitrogen removal efficiency of strain CY-10 were 100%, 91.1% and 68.5%, respectively. The optimal salinity for ammonia nitrogen removal by strain CY-10 was in the range of 0-5%. At the salinity of 5%, a maximum nitrogen removal rate of 6.25 mg/(L·h) was realized. Metabonomics data showed that the metabolic levels of sucrose and D-tagatose increased significantly at 5% salinity condition, enabling the strain to regulate osmotic pressure and survive in high-salt environments. Functional genes were successfully amplified by quantitative PCR, and HN-AD pathway of strain CY-10 followed NH4+-N â†’ NH2OH â†’ NO2--N â†’ NO â†’ N2O â†’ N2. These findings show that strain CY-10 has great potential in nitrogen removal treatment of saline wastewater.

[Transformation mechanism of carbon tetrachloride and the associated micro-ecology in landfill cover, a typical functional layer zone].

Landfill is one of the important sources of carbon tetrachloride (CT) pollution, and it is important to understand the degradation mechanism of CT in landfill cover for better control. In this study, a simulated landfill cover system was set up, and the biotransformation mechanism of CT and the associated micro-ecology were investigated. The results showed that three stable functional zones along the depth, i.e., aerobic zone (0-15 cm), anoxic zone (15-45 cm) and anaerobic zone (> 45 cm), were generated because of long-term biological oxidation in landfill cover. There were significant differences in redox condition and microbial community structure in each zone, which provided microbial resources and favorable conditions for CT degradation. The results of biodegradation indicated that dechlorination of CT produced chloroform (CF), dichloromethane (DCM) and Cl- in anaerobic and anoxic zones. The highest concentration of dechlorination products occurred at 30 cm, which were degraded rapidly in aerobic zone. In addition, CT degradation rate was 13.2-103.6 µg/(m2·d), which decreased with the increase of landfill gas flux. The analysis of diversity sequencing revealed that Mesorhizobium, Thiobacillus and Intrasporangium were potential CT-degraders in aerobic, anaerobic and anoxic zone, respectively. Moreover, six species of dechlorination bacteria and eighteen species of methanotrophs were also responsible for anaerobic transformation of CT and aerobic degradation of CF and DCM, respectively. Interestingly, anaerobic dechlorination and aerobic transformation occurred simultaneously in the anoxic zone in landfill cover. Furthermore, analysis of degradation mechanism suggested that generation of stable anaerobic-anoxic-aerobic zone by regulation was very important for the harmless removal of full halogenated hydrocarbon in vadose zone, and the increase of anoxic zone scale enhanced their removal. These results provide theoretical guidance for the removal of chlorinated pollutants in landfills.

Direct aerobic oxidation (DAO) of chlorinated aliphatic hydrocarbons: A review of key DAO bacteria, biometabolic pathways and in-situ bioremediation potential.

Contamination of aquifers and vadose zones with chlorinated aliphatic hydrocarbons (CAH) is a world-wide issue. Unlike other reactions, direct aerobic oxidation (DAO) of CAHs does not require growth substrates and avoids the generation of toxic by-products. Here, we critically review the current understanding of chlorinated aliphatic hydrocarbons-DAO and its application in bioreactors and at the field scale. According to reports on chlorinated aliphatic hydrocarbons-DAO bacteria, isolates mainly consisted of Methylobacterium and Proteobacterium. Chlorinated aliphatic hydrocarbons-DAO bacteria are characterized by tolerance to a high concentration of CAHs and highly efficient removal of CAHs. Trans-1,2-dichloroethylene (t-DCE) is easily transformed biomass for bacteria, followed by 1,2-dichloroethane (1,2-DCA), dichloromethane (DCM), vinyl chloride (VC) and cis-1,2-dichloroethylene (c-DCE). Significant differences in the maximum specific growth rates were observed with different CAHs and biometabolic pathways for DCM, 1,2-DCA, VC and c-DCE degradation have been successfully parsed. Detection of the functional genes etnC and etnE is useful for the determination of active VC DAO bacteria. Additionally, DAO bacteria have been successfully applied to CAHs in new types of bioreactors with satisfactory results. To the best of the authors' knowledge, only one study on DAO-CAHs was conducted in-situ and resulted in 99% CAH removal. Lastly, we put forward future development prospect of chlorinated aliphatic hydrocarbons-DAO.

Cometabolic degradation mechanism and microbial network response of methanotrophic consortia to chlorinated hydrocarbon solvents.

The cometabolism mechanism of chlorinated hydrocarbon solvents (CHSs) in mixed consortia remains largely unknown. CHS biodegradation characteristics and microbial networks in methanotrophic consortia were studied for the first time. The results showed that all CHSs can efficiently be degraded via cometabolism with a maximum degradation rate of 4.8 mg/(h·gcell). Chloroalkane and chloroethylene were more easily degraded than chlorobenzenes by methanotrophic consortia, especially nonfully chlorinated aliphatic hydrocarbons, which were converted to Cl- with a production rate of 0.29-0.36 mg/(h·gcell). In addition, the microecological response results indicated that Methylocystaceae (49.0%), Methylomonas (65.3%) and Methylosarcina (41.9%) may be the major functional degraders in methanotrophic consortia. Furthermore, the results of the microbial correlation network suggested that interactive relationships constructed by type I methanotrophs and heterotrophs determined biodegradability. Additionally, PICRUSt analysis showed that CHSs could increase the relative abundance of CHS degradation genes and reduce the relative abundance of methane oxidation genes, which was in good agreement with the experimental results.

[Advances in microbial degradation of chlorinated hydrocarbons].

Chlorinated hydrocarbons (CAHs) threaten human health and the ecological environment due to their strong carcinogenic, teratogenic, mutagenic and heritable properties. Heterotrophic assimilation degradation can completely and effectively degrade CAHs, without secondary pollution. However, it is crucial to comprehensively understand the heterotrophic assimilation process of CAHs for its application. Therefore, we review here the characteristics and advantages of heterotrophic assimilation degradation of CAHs. Moreover, we systematically summarize current research status of heterotrophic assimilation of CAHs. Furthermore, we analyze bacterial genera and metabolism, key enzymes and characteristic genes involved in the metabolic process. Finally, we indicate existing problems of heterotrophic assimilation research and future research needs.

Temporal and spatial variation in the mechanisms used by microorganisms to form methylmercury in the water column of Changshou Lake.

The microbiome in artificial lake water and its impact on mercury (Hg) methylation remain largely unknown. We selected the largest artificial lake in southeastern china, Changshou Lake (CSL), which has high background levels of Hg, for our investigation of Hg transformation microorganisms. Five different sections of the water column of CSL were sampled during four seasons. The water samples were subjected to analysis of geochemical parameters, various Hg species and microbiome information. High concentrations of total mercury (THg) were detected in CSL in comparison with those found in natural lakes. Significant differences in microbial community structure and Hg species abundance existed among seasons. High dissolved methyl mercury (DMeHg) formation and high bacterial richness and diversity occurred in the fall. The microbiome was dominated by Proteobacteria, Actinobacteria, Bacteroidetes, Deinococcus-Thermus and many unclassified bacteria. Significant correlations were found between seasonal bacterial communities and Hg levels. Hg methylation was strongly linked to the abundance of Cyanobacteria. Methylators, including Syntrophus, Desulfovibrio and Desulfomonile species, were detected only in samples collected in the fall. The results of enzyme functional analyses revealed that many unknown types of bacteria could also be responsible for Hg transformation. This study was the first to investigate the impact of various Hg species on the microbiome of artificial lake water. The findings of this study illuminate the role of seasonal bacteria in Hg transformation.

[Advances in biotic and abiotic mutual promoting mechanism for chlorinated aliphatic hydrocarbons degradation].

Chlorinated aliphatic hydrocarbons (CAHs) with characteristics of high toxicity, biological accumulation and recalcitrance to degradation as well as carcinogenicity, teratogenesis and mutagenicity, are seriously harmful to human health and ecological environment. CAHs degradation depends on biotic and abiotic responses that exist diversified interactive effects, so it is important to clarify the mechanism of CAHs degradation via biotic and abiotic mutual promoting to significantly enhance the CAHs-contaminated site restoration. In this work, a series of pathways for CAHs degradation was first introduced and summarized as three means on reductive dechlorination, aerobic cometabolism and direct oxidation, and biotic and abiotic typical factors affecting CAHs degradation were concluded from these. Then, mechanisms of induced degradation and synergistic degradation were indicated from the perspective of mutual promoting degradation both with biotic and abiotic responses, and furthermore, the application and technical limitations of CAHs degradation enhanced via biotic and abiotic mutual promoting were reviewed and analyzed. Finally, the development of CAHs degradation technology in future was prospected.

Effects of copper on expression of methane monooxygenases, trichloroethylene degradation, and community structure in methanotrophic consortia.

Copper plays a key role in regulating the expression of enzymes that promote biodegradation of contaminants in methanotrophic consortia (MC). Here, we utilized MC isolated from landfill cover to investigate cometabolic degradation of trichloroethylene (TCE) at nine different copper (Cu2+) concentrations. The results demonstrated that an increase in Cu2+ concentration from 0 to 15 µM altered the specific first-order rate constant k 1,TCE, the expression levels of methane monooxygenase (pmoA and mmoX) genes, and the specific activity of soluble methane monooxygenase (sMMO). High efficiency TCE degradation (95%) and the expression levels of methane monooxygenase (MMO) were detected at a Cu2+ concentration of 0.03 µM. Notably, sMMO-specific activity ranged from 74.41 nmol/(mgcell h) in 15 µM Cu2+ to 654.99 nmol/(mgcell h) in 0.03 µM Cu2+, which contrasts with cultures of pure methanotrophs in which sMMO activity is depressed at high Cu2+ concentrations, indicating a special regulatory role for Cu2+ in MC. The results of MiSeq pyrosequencing indicated that higher Cu2+ concentrations stimulated the growth of methanotrophic microorganisms in MC. These findings have important implications for the elucidation of copper-mediated regulatory mechanisms in MC.

Real-time monitoring of methane oxidation in a simulated landfill cover soil and MiSeq pyrosequencing analysis of the related bacterial community structure.

Real-time CH4 oxidation in a landfill cover soil was studied using automated gas sampling that determined biogas (CH4 and CO2) and O2 concentrations at various depths in a simulated landfill cover soil (SLCS) column reactor. The real-time monitoring system obtained more than 10,000 biogas (CH4 and CO2) and O2 data points covering 32 steady states of CH4 oxidation with 32 different CH4 fluxes (0.2-125mol·m-2·d-1). The kinetics of CH4 oxidation at different depths (0-20cm, 20-40cm, and 40-60cm) of SLCS were well fit by a CH4-O2 dual-substrate model based on 32 values (averaged, n=5-15) of equilibrated CH4 concentrations. The quality of the fit (R2 ranged from 0.90 to 0.96) was higher than those reported in previous studies, which suggests that real time monitoring is beneficial for CH4 oxidation simulations. MiSeq pyrosequencing indicated that CH4 flux events changed the bacterial community structure (e.g., increased the abundance of Bacteroidetes and Methanotrophs) and resulted in a relative increase in the amount of type I methanotrophs (Methylobacter and Methylococcales) and a decrease in the amount of type II methanotrophs (Methylocystis).

[Advances in transformation and regulation biodegradation of chorinated hydrocarbons in landfill].

Understanding the biotransformation mechanism of chlorinated hydrocarbons in contaminated site is of great significance to the in-situ bioremediation. Therefore, we summed up the overlapping composition of chlorinated hydrocarbons and analyzed statistically the concentration variations and degradation rate of chlorinated hydrocarbons in various landfill which were regarded as one of the most typical compound pollution sites. The statistical data indicated that chloralkane and chloroalkene concentration ranged 0.20 to 32.45 and 0.50 to 32.45 µg/m3, respectively, which were the main components. We also found that biodegradation rates of chlorinated hydrocarbons decreased with the number of attached chlorine atoms in landfill cover. Then, we summarized the biodegradation mechanism of chlorinated hydrocarbons under different environmental conditions. The results implied that chlorinated hydrocarbons biodegradation incorporated aerobic co-metabolism, halorespiration and anaerobic reductive dechlorination involved in a wide range of substrates and a variety of functional microbes. Based on of these analyses, we constructed biodegradation models of chlorinated hydrocarbons in landfill cover. Finally, the possible development of chlorinated hydrocarbons biological removal in the future was predicated.

[Effects of copper on biodegradation mechanism of trichloroethylene by mixed microorganisms].

We isolated and enriched mixed microorganisms SWA1 from landfill cover soils supplemented with trichloroethylene (TCE). The microbial mixture could degrade TCE effectively under aerobic conditions. Then, we investigated the effect of copper ion (0 to 15 µmol/L) on TCE biodegradation. Results show that the maximum TCE degradation speed was 29.60 nmol/min with 95.75% degradation when copper ion was at 0.03 µmol/L. In addition, genes encoding key enzymes during biodegradation were analyzed by Real-time quantitative reverse transcription PCR (RT-qPCR). The relative expression abundance of pmoA gene (4.22E-03) and mmoX gene (9.30E-06) was the highest when copper ion was at 0.03 µmol/L. Finally, we also used MiSeq pyrosequencing to investigate the diversity of microbial community. Methylocystaceae that can co-metabolic degrade TCE were the dominant microorganisms; other microorganisms with the function of direct oxidation of TCE were also included in SWA1 and the microbial diversity decreased significantly along with increasing of copper ion concentration. Based on the above results, variation of copper ion concentration affected the composition of SWA1 and degradation mechanism of TCE. The degradation mechanism of TCE included co-metabolism degradation of methanotrophs and oxidation metabolism directly at copper ion of 0.03 µmol/L. When copper ion at 5 µmol/L (biodegradation was 84.75%), the degradation mechanism of TCE included direct-degradation and co-metabolism degradation of methanotrophs and microorganisms containing phenol hydroxylase. Therefore, biodegradation of TCE by microorganisms was a complicated process, the degradation mechanism included co-metabolism degradation of methanotrophs and bio-oxidation of non-methanotrophs.