RESUMEN
Nicosulfuron and Cd are common pollutants that pose significant threats to the environment and human health, particularly under combined stress. This study is the first to remediate environmental nicosulfuron and Cd under combined stress using microbiological techniques. Enterobacter ludwigii ES2 was isolated, characterized, and demonstrated to degrade 93.80 % of nicosulfuron and remove 59.64 % of Cd within 4 d. Potential functional genes, including nicosulfuron degradation genes gstA, gstB, glnQ, glnP, mreB, and sixA, and Cd tolerance/removal-related genes mntA, mntB, mntH, dnaK, znuA, and zupt, were predicted by sequencing the whole genome of strain ES2, and their expression was verified by qRT-PCR. Strain ES2 managed oxidative stress induced by Cd through superoxide dismutase, glutathione, catalase, peroxidase, and malondialdehyde. Furthermore, to repair compound stress, up to 90.48 % of nicosulfuron and 67.74 % of Cd were removed. The community structure analysis indicated that Enterobacteriaceae, Sphingomonadaceae, and Gemmatimonadaceae were dominant populations, with ES2 stably colonizing and becoming the dominant bacterium. In summary, ES2 demonstrated significant potential in remediating nicosulfuron and Cd pollution from various perspectives, providing a solid theoretical foundation.
Asunto(s)
Biodegradación Ambiental , Cadmio , Enterobacter , Enterobacter/genética , Enterobacter/metabolismo , Cadmio/metabolismo , Cadmio/toxicidad , Compuestos de Sulfonilurea/metabolismo , Contaminantes del Suelo/metabolismo , Genoma Bacteriano , Microbiota , PiridinasRESUMEN
AIMS: To construct an efficient bacterial complex to degrade nicosulfuron and clarify its degradative characteristics, promote the growth of maize (Zea mays), and provide a theoretical foundation for the efficient remediation of soil contaminated with nicosulfuron. METHODS AND RESULTS: Biocompatibility was determined by the filter paper sheet method by mixing Serratia marcescens A1 and Bacillus cereus A2 in a 1:1 ratio, yielding A12. The optimum culture conditions for the bacterial composite were obtained based on a three-factor, three-level analysis using response surface methodology, with 29.25 g l-1 for maltodextrin, 10.04 g l-1 for yeast extract, and 19.93 g l-1 for NaCl, which resulted in 92.42% degradation at 4 d. The degradation characteristics of A12 were clarified as follows: temperature 30°C, pH 7, initial concentration of nicosulfuron 20 mg l-1, and 4% inoculum. The ability to promote growth was determined by measuring the ratio of the lysosphere diameter (D) to the colony diameter (d), and the ability of the complex A12 to promote growth was higher than that of the two single strains. CONCLUSIONS: Nicosulfuron degradation in sterilized and unsterilized soils reached 85.4% and 91.2% within 28 d, respectively. The ability of the strains to colonize the soil was determined by extraction of total soil DNA, primer design, and gel electrophoresis. The bioremediation effect of A12 was confirmed by the maximum recovery of fresh weight (124.35%) of nicosulfuron-sensitive crop plants and the significant recovery of soil enzyme activities, as measured by the physiological indices in the sensitive plants.
Asunto(s)
Bacillus cereus , Biodegradación Ambiental , Piridinas , Microbiología del Suelo , Contaminantes del Suelo , Compuestos de Sulfonilurea , Compuestos de Sulfonilurea/metabolismo , Contaminantes del Suelo/metabolismo , Piridinas/metabolismo , Bacillus cereus/metabolismo , Bacillus cereus/crecimiento & desarrollo , Serratia marcescens/metabolismo , Serratia marcescens/crecimiento & desarrollo , Zea mays/metabolismo , Zea mays/microbiología , Suelo/química , Herbicidas/metabolismoRESUMEN
The extensive use of sulfonylurea herbicides has raised major concerns regarding their long-term soil residues and agroecological risks despite their role in agricultural protection. Microbial degradation is an important approach to remove sulfonylureas, whereas understanding the associated biodegradation mechanisms, enzymes, and physiological responses remains incomplete. Based on the rapid biodegradation of nicosulfuron by typical fungal isolate Talaromyces flavus LZM1, the dependency on cellular accumulation and environmental conditions, e.g. pH and nutrient supplies, was shown in the study. The biodegradation of nicosulfuron occurred intracellularly and followed the cascade of reactions including hydrolysis, Smile contraction rearrangement, hydroxylation, and opening of the pyrimidine ring. Besides 2-amino-4,6-dimethoxypyrimidine (ADMP) and 2-aminosulfonyl-N,N-dimethylnicotinamide (ASDM), numerous products and intermediates were newly identified and the structural forms of methoxypyrimidine and sulfonylurea bridge contraction rearrangement are predicted to be more toxic than nicosulfuron. The biodegradation should be enzymatically regulated by glycosylphosphatidylinositol transaminase (GPI-T) and P450s, which were manifested with the significant upregulation in proteomics. It is the first time that the hydrolysis of nicosulfuron into ADMP and ASDM have been associated with GPI-T. The integrated pathways of biodegradation were further elucidated through the involvement of various active enzymes. Except for the enzymatic catalysis, the physiological responses verified by metabolo-proteomics were critical not only to regulate material synthesis, uptake, utilization, and energy transfer but also to maintain antioxidant homeostasis, biodegradability, and tolerance of nicosulfuron by the differentially expressed metabolites, such as acetolactate synthase and 3-isopropylmalate dehydratase. The obtained results would help understand the biodegradation mechanism of sulfonylurea from chemicobiology and enzymology and promote the use of fungal biodegradation in pollution rehabilitation.
Asunto(s)
Biodegradación Ambiental , Herbicidas , Compuestos de Sulfonilurea , Herbicidas/metabolismo , Herbicidas/toxicidad , Compuestos de Sulfonilurea/metabolismo , Talaromyces/metabolismo , Proteómica , Piridinas/metabolismo , Contaminantes del Suelo/metabolismo , Contaminantes del Suelo/toxicidad , MultiómicaRESUMEN
Mesotrione is a herbicide used in agricultural production; however, its stability and long-term residues pose ecological risks to soil health and subsequent crops. In this research, the strain Amycolatopsis nivea La24 was identified as capable of completely degrading 50 mgâL-1 mesotrione within 48 h. It exhibited a broad adaptability to various environment and could degrade three sulfonylurea herbicides (nicosulfuron, chlorimuron-methyl, and cinosulfuron). Non-target metabonomic and mass spectrometry demonstrated that La24 strain broke down the mesotrione parent molecule by targeting the ß-diketone bond and nitro group, resulting in the production of five possible degradation products. The differentially expressed genes were significantly enriched in fatty acid degradation, amino acid metabolism, and other pathways, and the differentially metabolites in glutathione metabolism, arginine/proline metabolism, cysteine/methionine metabolism, and other pathways. Additionally, it was confirmed by heterologous expression that nitroreductase was directly involved in the mesotrione degradation, and NDMA-dependent methanol dehydrogenase would increase the resistance to mesotrione. Finally, the intracellular response of La24 during mesotrione degradation was proposed. This work provides insight for a comprehensive understanding of the mesotrione biodegradation mechanism, significantly expands the resources for pollutant degradation, and offers the potential for a more sustainable solution to address herbicide pollution in soil.
Asunto(s)
Amycolatopsis , Biodegradación Ambiental , Ciclohexanonas , Herbicidas , Herbicidas/metabolismo , Herbicidas/química , Ciclohexanonas/metabolismo , Amycolatopsis/metabolismo , Amycolatopsis/genética , Metabolómica , Compuestos de Sulfonilurea/metabolismo , Contaminantes del Suelo/metabolismo , MultiómicaRESUMEN
Sulfonylurea (SU) herbicides are widely used and often detected in environmental matrices and have toxic effects on ecosystems and plant development. However, the interaction between SU and soil-plant metabolism during the whole wheat growth cycle remains poorly investigated. Field trials demonstrated that bensulfuron methyl exposure reduced wheat height and a thousand grains' weight, disrupting the critical metabolic pathways, including linoleic acid and amino acid metabolism in the maturity stage. During different growth processes, bensulfuron methyl exposure decreases wheat soil and plants' defense-related indole alkaloid compounds, such as benzoxazinoids and melatonin. Microbial sequencing results showed that bensulfuron methyl treated decreased the abundance of beneficial microorganisms (Gammaproteobacteria, Bacteroidia, and Blastocatella) in the rhizosphere soil, which positively correlated with the inhibition of soil enzyme activity and the secretion of allelopathic substances (benzoxazinoids and melatonin). Molecular docking further confirmed that bensulfuron methyl affects protein molecular structure by establishing hydrogen bonds, which disequilibrate wheat benzoxazinoids and melatonin metabolism. Therefore, bensulfuron methyl exposure disrupted the interaction between soil microorganisms and indole alkaloid metabolism, hindering plant development. This study provides constructive insights into the environmental risks of herbicides and agricultural product safety throughout wheat development.
Asunto(s)
Herbicidas , Microbiología del Suelo , Compuestos de Sulfonilurea , Triticum , Triticum/efectos de los fármacos , Triticum/metabolismo , Triticum/crecimiento & desarrollo , Compuestos de Sulfonilurea/toxicidad , Compuestos de Sulfonilurea/metabolismo , Herbicidas/toxicidad , Herbicidas/metabolismo , Microbiota/efectos de los fármacos , Contaminantes del Suelo/toxicidad , Contaminantes del Suelo/metabolismo , Estrés Fisiológico/efectos de los fármacos , Redes y Vías Metabólicas/efectos de los fármacos , Simulación del Acoplamiento Molecular , RizosferaRESUMEN
Microbial herbicide degradation is an efficient bioremediation method. In this study, a strain of Streptomyces nigra, LM01, which efficiently degrades atrazine and nicosulfuron, was isolated from a corn field using a direct isolation method. The degradation effects of the identified strain on two herbicides were investigated and optimized using an artificial neural network. The maximum degradation rates of S. nigra LM01 were 58.09 % and 42.97 % for atrazine and nicosulfuron, respectively. The degradation rate of atrazine in the soil reached 67.94 % when the concentration was 108 CFU/g after 5 d and was less effective than that of nicosulfuron. Whole genome sequencing of strain LM01 helped elucidate the possible degradation pathways of atrazine and nicosulfuron. The protein sequences of strain LM01 were aligned with the sequences of the degraded proteins of the two herbicides by using the National Center for Biotechnology Information platform. The sequence (GE005358, GE001556, GE004212, GE005218, GE004846, GE002487) with the highest query cover was retained and docked with the small-molecule ligands of the herbicides. The results revealed a binding energy of - 6.23 kcal/mol between GE005358 and the atrazine ligand and - 6.66 kcal/mol between GE002487 and the nicosulfuron ligand.
Asunto(s)
Atrazina , Biodegradación Ambiental , Herbicidas , Piridinas , Streptomyces , Compuestos de Sulfonilurea , Atrazina/metabolismo , Atrazina/química , Streptomyces/metabolismo , Streptomyces/genética , Herbicidas/metabolismo , Herbicidas/química , Compuestos de Sulfonilurea/metabolismo , Compuestos de Sulfonilurea/química , Piridinas/metabolismo , Piridinas/química , Contaminantes del Suelo/metabolismo , Genes Bacterianos , Redes Neurales de la ComputaciónRESUMEN
Microbial degradation is a highly efficient and reliable approach for mitigating the contamination of sulfonylurea herbicides, such as chlorimuron-ethyl, in soil and water. In this study, we aimed to assess whether Kj-mhpC plays a pivotal role in the degradation of chlorimuron-ethyl. Kj-mhpC enzyme purified via prokaryotic expression exhibited the highest catalytic activity for chlorimuron-ethyl at 35 °C and pH 7. Bioinformatic analysis and three-dimensional homologous modeling of Kj-mhpC were conducted. Additionally, the presence of Mg+ and Cu2+ ions partially inhibited but Pb2+ ions completely inhibited the enzymatic activity of Kj-mhpC. LC/MS revealed that Kj-mhpC hydrolyzes the ester bond of chlorimuron-ethyl, resulting in the formation of 2-(4-chloro-6-methoxypyrimidine-2-amidoformamidesulfonyl) benzoic acid. Furthermore, the point mutation of serine at position 67 (Ser67) confirmed that it is the key amino acid at the active site for degrading chlorimuron-ethyl. This study enhanced the understanding of how chlorimuron-ethyl is degraded by microorganisms and provided a reference for bioremediation of the environment polluted with chlorimuron-ethyl.
Asunto(s)
Herbicidas , Pirimidinas , Contaminantes del Suelo , Klebsiella/genética , Klebsiella/metabolismo , Esterificación , Contaminantes del Suelo/metabolismo , Herbicidas/metabolismo , Compuestos de Sulfonilurea/metabolismo , IonesRESUMEN
Sulfonylurea herbicides have been widely used worldwide and play a significant role in modern agricultural production. However, these herbicides have adverse biological effects that can damage the ecosystems and harm human health. As such, rapid and effective techniques that remove sulfonylurea residues from the environment are urgently required. Attempts have been made to remove sulfonylurea residues from environment using various techniques such as incineration, adsorption, photolysis, ozonation, and microbial degradation. Among them, biodegradation is regarded as a practical and environmentally responsible way to eliminate pesticide residues. Microbial strains such as Talaromyces flavus LZM1, Methylopila sp. SD-1, Ochrobactrum sp. ZWS16, Staphylococcus cohnii ZWS13, Enterobacter ludwigii sp. CE-1, Phlebia sp. 606, and Bacillus subtilis LXL-7 can almost completely degrade sulfonylureas. The degradation mechanism of the strains is such that sulfonylureas can be catalyzed by bridge hydrolysis to produce sulfonamides and heterocyclic compounds, which deactivate sulfonylureas. The molecular mechanisms associated with microbial degradation of sulfonylureas are relatively poorly studied, with hydrolase, oxidase, dehydrogenase and esterase currently known to play a pivotal role in the catabolic pathways of sulfonylureas. Till date, there are no reports specifically on the microbial degrading species and biochemical mechanisms of sulfonylureas. Hence, in this article, the degradation strains, metabolic pathways, and biochemical mechanisms of sulfonylurea biodegradation, along with its toxic effects on aquatic and terrestrial animals, are discussed in depth in order to provide new ideas for remediation of soil and sediments polluted by sulfonylurea herbicides.
Asunto(s)
Herbicidas , Humanos , Herbicidas/análisis , Ecosistema , Compuestos de Sulfonilurea/toxicidad , Compuestos de Sulfonilurea/química , Compuestos de Sulfonilurea/metabolismo , Sulfonamidas , Agricultura , Biodegradación AmbientalRESUMEN
The widespread use of the herbicide chlorimuron-methyl is hazard to rotational crops and causes soil degradation problems. Biodegradation is considered a promising way for removing herbicide residues from the environment. Here, a new isolated strain, Cedecea sp. LAM2020, enabled complete degradation of 100 mg/L chlorimuron-methyl within five days. Transcriptome analysis revealed that ABC transporters, atrazine degradation and purine metabolism were enriched in the KEGG pathway. Integrating GO and KEGG classification with related reports, we predict that carboxylesterases are involved in the biodegradation of chlorimuron-methyl by LAM2020. Heterologous expression of the carboxylesterase gene carH showed 26.67% degradation of 50 mg/L chlorimuron-methyl within 6 h. The intracellular potential biological response and extracellular degradation process of chlorimuron-ethyl were analyzed by the nontarget metabolomic and mass spectrometry respectively, and the biodegradation characteristics and complete mineralization pathway was revealed. The cleavage of the sulfonylurea bridge and the ester bond achieved the first step in the degradation of chlorimuron-methyl. Together, these results reveal the presence of acidolysis and enzymatic degradation of chlorimuron-methyl by strain LAM2020. Hydroponic corn experiment showed that the addition of strain LAM2020 alleviated the toxic effects of chlorimuron-ethyl on the plants. Collectively, strain LAM2020 may be a promising microbial agent for plants chlorimuron-ethyl detoxification and soil biofertilizer.
Asunto(s)
Herbicidas , Contaminantes del Suelo , Contaminantes del Suelo/metabolismo , Biodegradación Ambiental , Compuestos de Sulfonilurea/metabolismo , Herbicidas/metabolismo , Enterobacteriaceae/metabolismo , SueloRESUMEN
Herbicide residues in the environment threaten high-quality agriculture and human health. Consequently, in situ remediation of herbicide contamination is vital. We synthesized a novel self-catalyzed nanozyme, ultrasmall (2-3 nm) copper peroxide nanodots modified by citric acid (CP@CA) for this purpose, which can break down into H2O2 and Cu2+ in water or soil. Ubiquitous glutathione reduces Cu2+ into Cu+, which promotes the decomposition of H2O2 into â¢OH through a Fenton-like reaction under mild acid conditions created by the presence of citric acid. The generated â¢OH efficiently degrade nicosulfuron in water and soil, and the maximum degradation efficiency could be achieved at 97.58% in water at 56 min. The possible degradation mechanisms of nicosulfuron were proposed through the 25 intermediates detected. The overall ecotoxicity of the nicosulfuron system was significantly reduced after CP@CA treatment. Furthermore, CP@CA had little impact on active components of soil bacterial community. Moreover, CP@CA nanozyme could effectively remove seven other sulfonylurea herbicides from the water. In this paper, a high-efficiency method for herbicide degradation was proposed, which provides a new reference for the in situ remediation of herbicide pollution.
Asunto(s)
Herbicidas , Humanos , Herbicidas/metabolismo , Cobre/toxicidad , Ácido Cítrico , Peróxido de Hidrógeno/metabolismo , Peróxidos , Compuestos de Sulfonilurea/toxicidad , Compuestos de Sulfonilurea/metabolismo , Suelo/química , AguaRESUMEN
Chlorimuron-ethyl is a widely used herbicide in agriculture. However, uncontrolled chlorimuron-ethyl application causes serious environmental problems. Chlorimuron-ethyl can be effectively degraded by microbes, but the underlying molecular mechanisms are not fully understood. In this study, we identified the possible pathways and key genes involved in chlorimuron-ethyl degradation by the Chenggangzhangella methanolivorans strain CHL1, a Methylocystaceae strain with the ability to degrade sulfonylurea herbicides. Using a metabolomics method, eight intermediate degradation products were identified, and three pathways, including a novel pyrimidine-ring-opening pathway, were found to be involved in chlorimuron-ethyl degradation by strain CHL1. Transcriptome sequencing indicated that three genes (atzF, atzD, and cysJ) are involved in chlorimuron-ethyl degradation by strain CHL1. The gene knock-out and complementation techniques allowed for the functions of the three genes to be identified, and the enzymes involved in the different steps of chlorimuron-ethyl degradation pathways were preliminary predicted. The results reveal a previously unreported pathway and the key genes of chlorimuron-ethyl degradation by strain CHL1, which have implications for attempts to enrich the biodegradation mechanism of sulfonylurea herbicides and to construct engineered bacteria in order to remove sulfonylurea herbicide residues from environmental media.
Asunto(s)
Herbicidas , Methylocystaceae , Contaminantes del Suelo , Biodegradación Ambiental , Herbicidas/metabolismo , Methylocystaceae/metabolismo , Pirimidinas/metabolismo , Contaminantes del Suelo/metabolismo , Compuestos de Sulfonilurea/metabolismoRESUMEN
The overuse of the herbicide nicosulfuron has become a global environmental concern. As a potential bioremediation technology, the microbial degradation of nicosulfuron shows much promise; however, the mechanism by which microorganisms respond to nicosulfuron exposure requires further study. An isolated soil-borne bacteria Pseudomonas nicosulfuronedens LAM1902 displaying nicosulfuron, chlorimuron-ethyl, and cinosulfuron degradabilities in the presence of glucose, was used to determine the transcriptional responses to nicosulfuron exposure. RNA-Seq results indicated that 1102 differentially expressed genes (DEGs) were up-regulated and 702 down-regulated under nicosulfuron stress. DEGs were significantly enriched in "ABC transporters", "sulfur metabolism", and "ribosome" pathways (p ≤ 0.05). Several pathways (glycolysis and pentose phosphate pathways, a two-component regulation system, as well as in bacterial chemotaxis metabolisms) were affected by nicosulfuron exposure. Surprisingly, nicosulfuron exposure showed positive effects on the production of oxalic acid that is synthesized by genes encoding glycolate oxidase through the glyoxylate cycle pathway. The results suggest that P. nicosulfuronedens LAM1902 adopt acid metabolites production strategies in response to nicosulfuron, with concomitant nicosulfuron degradation. Data indicates that glucose metabolism is required during the degradation and adaptation of strain LAM1902 to nicosulfuron stress. The present studies provide a glimpse at the molecular response of microorganisms to sulfonylurea pesticide toxicity and a potential framework for future mechanistic studies.
Asunto(s)
Herbicidas , Herbicidas/toxicidad , Pseudomonas/metabolismo , Piridinas/toxicidad , Compuestos de Sulfonilurea/metabolismo , TranscriptomaRESUMEN
This study aims to investigate the effects of long-term nicosulfuron residue on an herbicide factory ecosystem. High-throughput sequencing was used to investigate the environmental microbial community structure and interactions. The results showed that the main contributor to the differences in the microbial community structure was the sample type, followed by oxygen content, pH and nicosulfuron residue concentration. Regardless of the presence or absence of nicosulfuron, soil, sludge, and sewage were dominated by groups of Bacteroidetes, Actinobacteria, and Proteobacteria. Long-term exposure to nicosulfuron increased alpha diversity of bacteria and archaea but significantly decreased the abundance of Bacteroidetes and Acidobateria compared to soils without nicosulfuron residue. A total of 81 possible nicosulfuron-degrading bacterial genera, e.g., Rhodococcus, Chryseobacterium, Thermomonas, Stenotrophomonas, and Bacillus, were isolated from the nicosulfuron factory environmental samples through culturomics. The co-occurrence network analysis indicated that the keystone taxa were Rhodococcus, Stenotrophomonas, Nitrospira, Terrimonas, and Nitrosomonadaceae_MND1. The strong ecological relationship between microorganisms with the same network module was related to anaerobic respiration, the carbon and nitrogen cycle, and the degradation of environmental contaminants. Synthetic community (SynCom), which provides an effective top-down approach for the critical degradation strains obtained, enhanced the degradation efficiency of nicosulfuron. The results indicated that Rhodococcus sp. was the key genus in the environment of long-term nicosulfuron exposure.
Asunto(s)
Herbicidas , Microbiota , Bacterias/metabolismo , Bacteroidetes/metabolismo , Herbicidas/metabolismo , Herbicidas/toxicidad , Piridinas , Aguas del Alcantarillado , Suelo/química , Microbiología del Suelo , Compuestos de Sulfonilurea/metabolismo , Compuestos de Sulfonilurea/toxicidadRESUMEN
The widespread use of agrochemicals for controlling pests and diseases of crops is recognized as a main threat to biodiversity. Sulfonylurea herbicides are being increasingly used and display low levels of degradation in water which suggest that they might affect non-target organisms. In a common garden experiment, eggs of a widespread amphibian (Bufo spinosus) were exposed to sublethal environmentally relevant concentrations of a widely used sulfonylurea herbicide, nicosulfuron, during the whole embryonic development. We assessed development-related traits (i.e., development duration, hatching success, hatchling size and occurrence of malformation) as well as antioxidant markers in response to contamination (i.e., SOD, GPx, catalase, thiols and relevant ratios thereof). We found that sublethal concentrations of nicosulfuron increased embryonic development duration, increased hatchling size and tended to increase malformations. Embryos exposed to nicosulfuron displayed decreased thiols and increased catalase activity suggesting alteration of oxidative status. We did not find any effect of nicosulfuron on SOD and GPx levels. Interestingly, higher catalase activity was linked to higher proportion of malformed individuals, suggesting that exposure to nicosulfuron induced teratogenic effects. Our results suggest that alteration of antioxidant levels might be one physiological mechanism through which nicosulfuron might cause detrimental effects on amphibian embryos. Sublethal effects of pesticides at environmentally relevant concentrations have been overlooked and require further investigations, especially in non-target taxa occurring in agricultural landscapes.
Asunto(s)
Herbicidas , Anfibios/metabolismo , Animales , Desarrollo Embrionario , Herbicidas/metabolismo , Herbicidas/toxicidad , Humanos , Estrés Oxidativo , Piridinas , Compuestos de Sulfonilurea/metabolismo , Compuestos de Sulfonilurea/toxicidadRESUMEN
Bioaugmentation is a promising alternative in soil remediation. One challenge of bioaugmentation is that exogenous pollutant-degrading microbes added to soil cannot establish enough biomass to eliminate pollutants. Considering that methanotrophs have a growth advantage in the presence of methane, we hypothesize that genetically engineered methanotrophs could degrade contaminants efficiently in soil with methane. Here, methanotroph Methylomonas sp. LW13, herbicide bensulfuron-methyl (BSM), and two kinds of soil were chosen to confirm this hypothesis. The unmarked gene knock-in method was first developed for strain LW13. Then, BSM hydrolase encoding gene sulE was inserted into the chromosome of strain LW13, conferring it BSM-degrading ability. After inoculation, the cell amount of strain LW13-sulE in soil raised considerably (over 100 fold in 9 days) with methane provision; meanwhile, >90% of BSM in soil was degraded. This study provides a proof of the concept that genetically engineered methanotroph is a potential platform for soil remediation.
Asunto(s)
Biodegradación Ambiental , Metano/metabolismo , Plaguicidas/metabolismo , Contaminantes del Suelo/metabolismo , Técnicas de Sustitución del Gen , Hidrolasas/genética , Hidrolasas/metabolismo , Metano/química , Methylomonas/genética , Methylomonas/metabolismo , Plaguicidas/química , Microbiología del Suelo , Contaminantes del Suelo/química , Compuestos de Sulfonilurea/química , Compuestos de Sulfonilurea/metabolismo , Zea mays/metabolismoRESUMEN
A Gram-stain-negative, aerobic, motile, short-rod-shaped bacterium with nicosulfuron-degrading ability, designated strain LAM1902T, was isolated from a microbial consortium enriched with nicosulfuron as a sole nitrogen and energy source. The optimal temperature and pH for growth of strain LAM1902T were 30 °C and pH 6.0, respectively. Strain LAM1902T could grow in the presence of NaCl with concentration up to 4.0â% (w/v). Comparative analysis of 16S rRNA gene sequences revealed that LAM1902T was closely related to the members of the family Pseudomonadaceae to the genus Pseudomonas, with the highest similarity to Pseudomonas nitroreducens DSM 14399T (99.6â%), Pseudomonas nitritireducens WZBFD3-5A2T (99.3â%) and Pseudomonas panipatensis Esp-1T (98.8â%). Multi-locus sequence analysis based on both concatenated sequences of the 16S rRNA gene and three housekeeping genes (gyrB, rpoB and rpoD) further confirmed the intrageneric phylogenetic position of strain LAM1902T. The genomic DNA G+C content of LAM1902T was 64.8 mol%. The low values of in silico DNA-DNA hybridization (less than 43.7â%) and average nucleotide identity (less than 90.9â%) also showed that the strain was distinctly different from known species of the genus Pseudomonas. The major fatty acids were C16â:â0, C17â:â0 cyclo and anteiso C15â:â0. Ubiquinone Q-9 was detected as the predorminant respiratory quinone. The major polar lipids were diphosphatidylglycerol, phosphatidylglycerol, phosphatidylethanolamine and aminophospholipid. Based on phylogenetic, phenotypic and chemotaxonomic analyses and genome comparisons, we conclude that strain LAM1902T represents a novel species, for which the name Pseudomonas nicosulfuronedens sp. nov. is proposed. The type strain is LAM1902T (=JCM 33860T=KCTC 72830T).
Asunto(s)
Consorcios Microbianos , Filogenia , Pseudomonas/clasificación , Piridinas/metabolismo , Compuestos de Sulfonilurea/metabolismo , Técnicas de Tipificación Bacteriana , Composición de Base , ADN Bacteriano/genética , Ácidos Grasos/química , Genes Bacterianos , Fosfolípidos/química , Pseudomonas/aislamiento & purificación , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Ubiquinona/químicaRESUMEN
Nicosulfuron is one of the main sulfonylurea herbicides that have been widely used to protect maize crops. A total of 10 nicosulfuron-degrading strains were isolated from the intestine tract of earthworm Eisenia foetida. Among them, Bacillus velezensis CF57 with the highest degradation efficiency was selected and studied in detail. The degradation characteristics of CF57 showed that it was able to effectively degrade nicosulfuron in a wide range of temperature, pH, and a low inoculation amount, and the response surface analysis revealed that the optimum degradation conditions were 30.8 °C, pH 6.31, and inoculation amount 3.04%. Meanwhile, CF57 could degrade high-concentration nicosulfuron efficiently and posed a broad degradation spectrum of other sulfonylurea herbicides. Furthermore, the localization of degradation enzyme indicated that the nicosulfuron-degrading enzyme was an extracellular fraction. By analyzing the metabolites of nicosulfuron, it could be further determined that the degradation of nicosulfuron by strain CF57 was mainly through the extracellular enzyme, and its possible degradation pathway was mainly derived from the cleavage of the C-N bond of the sulfonylurea bridge. These results may provide new insights into bioremediation of nicosulfuron-contaminated environments and enrich the resources of degrading bacteria of sulfonylurea herbicides.
Asunto(s)
Bacillus/metabolismo , Biodegradación Ambiental , Herbicidas/metabolismo , Piridinas/metabolismo , Compuestos de Sulfonilurea/metabolismo , Animales , Oligoquetos/efectos de los fármacos , Zea mays/parasitologíaRESUMEN
The herbicides diuron, hexazinone, and sulfometuron-methyl present a potential risk of environmental contamination and are widely used for weed control in sugarcane cultivation. Our objectives were to measure the tolerance of Canavalia ensiformes (L.) DC., Stilizobium aterrimum L., Raphanus sativus L., Crotalaria spectabilis Röth, Lupinus albus L., and Pennisetum glaucum (L.) R. Br. To the herbicides diuron, hexazinone, and sulfometuron-methyl to assess the capacity of these species to extract and accumulate the herbicides in their tissues. Before sowing the green manure species, the soils were individually contaminated with the three 14C-radiolabeled herbicides. 14C-diuron and 14C-sulfometuron-methyl showed higher values remaining in the soil (>90%) for all species of green manure compared to hexazinone (<80%). The green manure species analyzed showed greater potential to remedy soils contaminated with hexazinone than the other herbicides. C. ensiformes showed high phytoextraction of hexazinone when compared to the other species, removing 11.2% of the pollutant from the soil, followed by L. albus (8.6%), S. aterrimum (7.3%), R. sativus (4.8%), C. spectabilis (2.5%), and P. glaucum (1.1%). The results indicate that the phytoextraction of diuron, hexazinone and sulfometuron-methyl is dependent on the species of green manure and can be an important tool for the decontamination of areas polluted by these herbicides.
Asunto(s)
Biodegradación Ambiental , Diurona/metabolismo , Herbicidas/metabolismo , Estiércol , Contaminantes del Suelo/metabolismo , Compuestos de Sulfonilurea/metabolismo , Triazinas/metabolismo , Diurona/análisis , Herbicidas/análisis , Saccharum , SueloRESUMEN
In order to solve the problem of soil, water pollution and sensitive crop drug damage caused by chlorosulfuron residue, and to provide degradation strain resources for microbial remediation of contaminated soil, a chlorimuron-ethyl-degrading strain T9DB-01 was isolated from chlorosulfuron contaminated soil by the method of enrichment culture and step by step domestication. Strain T9DB-01 was identified as Pseudomonas sp. by morphological characteristics, physiological and biochemical analysis and 16S rDNA gene sequence analysis. The effects of temperature, pH value, substrate concentration, medium volume, and inoculation volume on the degradation of chlorsulfuron-methyl by strain T9DB-01 were investigated by single factor experiment. The degradation conditions of chlorosulfuron by strain T9DB-01 were optimized by orthogonal test and verification. Results show that 30 °C, pH 8.0, inoculum 4%, liquid volume 100 mL/250 mL, substrate concentration of 200 mg/L, cultured for 5 d, the strain degraded 93.7% chlorsulfuron-methyl. The degrading strain has certain application potential for bioremediation of chlorsulfuron-contaminated soil.
Asunto(s)
Bacterias , Biodegradación Ambiental , Pirimidinas , Microbiología del Suelo , Contaminantes del Suelo , Compuestos de Sulfonilurea , Bacterias/aislamiento & purificación , Bacterias/metabolismo , Concentración de Iones de Hidrógeno , Pirimidinas/metabolismo , Contaminantes del Suelo/metabolismo , Compuestos de Sulfonilurea/metabolismo , TemperaturaRESUMEN
In this study, glimepiride and l-arginine (GA) binary mixtures at various molar ratios were prepared to evaluate whether they could improve the poor water solubility and dissolution characteristics of glimepiride. It was shown that glimepiride and arginine form a eutectic mixture, a type of crystalline solid dispersions, at a 1:1 M ratio and eutectic temperature of 426.9 K using a phase diagram constructed using differential scanning calorimetry (DSC) and thermo-microscopy. The preserved characteristic powder X-ray diffraction (PXRD) patterns and infrared (IR) spectra of each material in those of GA binary mixtures confirmed the formation of eutectic mixture without molecular interaction in solid state. The formation of GA eutectic mixture (GAEM) resulted in the improvement of solubility through pH modification and the intermolecular interaction of glimepiride and l-arginine in aqueous mediums, thereby wettability and dissolution rate of glimepiride were also enhanced. The intermolecular interaction between glimepiride and l-arginine at a 1:1 stoichiometry of the complex in solution state was identified by phase solubility, stoichiometric determination, and solution state nuclear magnetic resonance (NMR) spectroscopy. Specific molecular interactions such as hydrogen bonding and hydrophobic interaction were suggested as main mechanisms of GA complexation in solution. Therefore, this study concludes that the GAEM could be an effective way to improve the solubility and dissolution rate of glimepiride.