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The widespread application of isoproturon (IPU) can cause serious pollution to the environment and threaten ecological functions. In this study, the IPU bacterial N-demethylase gene pdmAB was transferred and expressed in the chloroplast of soybean (Glycine max L. 'Zhonghuang13'). The transgenic soybeans exhibited significant tolerance to IPU and demethylated IPU to a less phytotoxic metabolite 3-(4-isopropylphenyl)-1-methylurea (MDIPU) in vivo. The transgenic soybeans removed 98% and 84% IPU from water and soil within 5 and 14 days, respectively, while accumulating less IPU in plant tissues compared with the wild-type (WT). Under IPU stress, transgenic soybeans showed a higher symbiotic nitrogen fixation performance (with higher total nodule biomass and nitrogenase activity) and a more stable rhizosphere bacterial community than the WT. This study developed a transgenic (TS) soybean capable of efficiently removing IPU from its growing environment and recovering a high-symbiotic nitrogen fixation capacity under IPU stress, and provides new insights into the interactions between rhizosphere microorganisms and TS legumes under herbicide stress.

Plastic pollution and antibiotic resistance are two emerging environmental and human health crises today. Although it was revealed that microplastics can serve as vectors for the dissemination of antibiotic resistance, it is still unclear how the nanoplastics influence the horizontal transfer of antibiotic resistance genes (ARGs). Herein, we firstly compared the effect of polystyrene (PS) micro/nanoplastics on the transformation of plasmid-borne ARG, using a transformation model consisting of plasmid pUC19 (amp

Gut microbiota and their metabolites are increasingly recognized for their crucial role in regulating the health and growth of the host. The mechanism by which the gut microbiome affects the growth rate of fish (Cyprinus carpio) in the rice-fish coculture system, however, remains unclear. In this study, the gut contents of the fast-growing and slow-growing (FG and SG) carp were collected from the rice-fish coculture system for both the fish gut microbiome and metabolome analyses. High throughput 16 S rRNA gene sequencing showed that the overall gut microbiota of FG group was distinct from that of SG group. For example, the cyanobacteria were highly enriched in the guts of SG carp (18.61%), in contrast, they only represented a minor fraction of gut microbiota for FG group (0.20%). The liquid chromatography-mass spectrometry (LC-MS)-based metabolomics analysis revealed that 191 identified metabolites mostly located in 18 KEGG pathways were differentially present between the two groups, of which more than 50% of these metabolites were involved in lipid and amino acids metabolism. Compared with the FG group, the gut microbiota of SG group significantly enriched the metabolic pathways involved in the steroid (hormone) biosynthesis, whereas reducing those associated with beta-alanine metabolism, biosynthesis of unsaturated fatty acids and bile secretion. The enrichment and depletion of these metabolic pathways resulted in an increase in steroid metabolites and a decrease in the concentration of spermidine, which may have a major impact on the growth rate of carp. The metabolome results were further supported by the predicated KEGG functions of the gut microbiomes of the two groups, pointing out that the gut microbiota could substantially affect the growth of fish via their unique metabolic functions. Together, our integrated fish gut microbiome and metabolome analysis has substantial implications for the development of engineered microbiome technologies in aquaculture.

Genetic redundancy is prevalent in organisms and plays important roles in the evolution of biodiversity and adaptation to environmental perturbation. However, selective advantages of genetic redundancy in overcoming metabolic disturbance due to structural analogues have received little attention. Here, functional divergence of the three 4-hydroxybenzoate 3-hydroxylase (PHBH) genes (phbh1~3) was found in Pigmentiphaga sp. strain H8. The genes phbh1/phbh2 were responsible for 3-bromo-4-hydroxybenzoate (3-Br-4-HB, an anthropogenic pollutant) catabolism, whereas phbh3 was primarily responsible for 4-hydroxybenzoate (4-HB, a natural intermediate of lignin) catabolism. 3-Br-4-HB inhibited 4-HB catabolism by competitively binding PHBH3 and was toxic to strain H8 cells especially at high concentrations. The existence of phbh1/phbh2 not only enabled strain H8 to utilize 3-Br-4-HB but also ensured the catabolic safety of 4-HB. Molecular docking and site-directed mutagenesis analyses revealed that Val199 and Phe384 of PHBH1/PHBH2 were required for the hydroxylation activity towards 3-Br-4-HB. Phylogenetic analysis indicated that phbh1 and phbh2 originated from a common ancestor and evolved specifically in strain H8 to adapt to 3-Br-4-HB-contaminated habitats, whereas phbh3 evolved independently. This study deepens our understanding of selective advantages of genetic redundancy in prokaryote's metabolic robustness and reveals the factors driving the divergent evolution of redundant genes in adaptation to environmental perturbation.

Ammonia oxidising archaea (AOA) are ecologically important nitrifiers in acidic agricultural soils. Two AOA phylogenetic clades, belonging to order-level lineages of Nitrososphaerales (clade C11; also classified as NS-Gamma-2.3.2) and family-level lineage of Candidatus Nitrosotaleaceae (clade C14; NT-Alpha-1.1.1), usually dominate AOA population in low pH soils. This study aimed to investigate the effect of different fertilisation histories on community composition and activity of acidophilic AOA in soils. High-throughput sequencing of ammonia monooxygenase gene (amoA) was performed on six low pH agricultural plots originating from the same soil but amended with different types of fertilisers for over 20 years and nitrification rates in those soils were measured. In these fertilised acidic soils, nitrification was likely dominated by Nitrososphaerales AOA and ammonia-oxidising bacteria, while Ca. Nitrosotaleaceae AOA activity was non-significant. Within Nitrososphaerales AOA, community composition differed based on the fertilisation history, with Nitrososphaerales C11 only representing a low proportion of the community. This study revealed that long-term soil fertilisation selects for different acidophilic nitrifier communities, potentially through soil pH change or through direct effect of nitrogen, potassium and phosphorus. Comparative community composition among the differently fertilised soils also highlighted the existence of AOA phylotypes with different levels of stability to environmental changes, contributing to the understanding of high AOA diversity maintenance in terrestrial ecosystems.

Ammonia-oxidizing archaea (AOA) are among the most abundant and ubiquitous microorganisms in the ocean, exerting primary control on nitrification and nitrogen oxides emission. Although united by a common physiology of chemoautotrophic growth on ammonia, a corresponding high genomic and habitat variability suggests tremendous adaptive capacity. Here, we compared 44 diverse AOA genomes, 37 from species cultivated from samples collected across diverse geographic locations and seven assembled from metagenomic sequences from the mesopelagic to hadopelagic zones of the deep ocean. Comparative analysis identified seven major marine AOA genotypic groups having gene content correlated with their distinctive biogeographies. Phosphorus and ammonia availabilities as well as hydrostatic pressure were identified as selective forces driving marine AOA genotypic and gene content variability in different oceanic regions. Notably, AOA methylphosphonate biosynthetic genes span diverse oceanic provinces, reinforcing their importance for methane production in the ocean. Together, our combined comparative physiological, genomic, and metagenomic analyses provide a comprehensive view of the biogeography of globally abundant AOA and their adaptive radiation into a vast range of marine and terrestrial habitats.

Long-term nitrogen (N) fertilization imposes strong selection on nitrifying communities in agricultural soil, but how a progressively changing niche affects potentially active nitrifiers in the field remains poorly understood. Using a 44-year grassland fertilization experiment, we investigated community shifts of active nitrifiers by DNA-based stable isotope probing (SIP) of field soils that received no fertilization (CK), high levels of organic cattle manure (HC), and chemical N fertilization (CF). Incubation of DNA-SIP microcosms showed significant nitrification activities in CF and HC soils, whereas no activity occurred in CK soils. The 44 years of inorganic N fertilization selected only 13C-ammonia-oxidizing bacteria (AOB), whereas cattle slurry applications created a niche in which both ammonia-oxidizing archaea (AOA) and AOB could be actively 13C-labeled. Phylogenetic analysis indicated that Nitrosospira sp. 62-like AOB dominated inorganically fertilized CF soils, while Nitrosospira sp. 41-like AOB were abundant in organically fertilized HC soils. The 13C-AOA in HC soils were affiliated with the 29i4 lineage. The 13C-nitrite-oxidizing bacteria (NOB) were dominated by both Nitrospira- and Nitrobacter-like communities in CF soils, and the latter was overwhelmingly abundant in HC soils. The 13C-labeled nitrifying communities in SIP microcosms of CF and HC soils were largely similar to those predominant under field conditions. These results provide direct evidence for a strong selection of distinctly active nitrifiers after 44 years of different fertilization regimes in the field. Our findings imply that niche differentiation of nitrifying communities could be assessed as a net result of microbial adaption over 44 years to inorganic and organic N fertilization in the field, where distinct nitrifiers have been shaped by intensified anthropogenic N input.

Anaerobic degradation is the major pathway for microbial degradation of benzene, toluene, ethylbenzene, and xylenes (BTEX) under electron acceptor lacking conditions. However, how exogenous electron acceptors modulate BTEX degradation through shaping the microbial community structure remains poorly understood. Here, we investigated the effect of various exogenous electron acceptors on BTEX degradation as well as methane production in anaerobic microbiota, which were enriched from the same contaminated soil. It was found that the BTEX degradation capacities of the anaerobic microbiota gradually increased along with the increasing redox potentials of the exogenous electron acceptors supplemented (WE: Without exogenous electron acceptors SS: Sulfate supplement FS: Ferric iron supplement NS: Nitrate supplement), while the complexity of the co-occurring networks (e.g., avgK and links) of the microbiota gradually decreased, showing that microbiota supplemented with higher redox potential electron acceptors were less dependent on the formation of complex microbial interactions to perform BTEX degradation. Microbiota NS showed the highest degrading capacity and the broadest substrate-spectrum for BTEX, and it could metabolize BTEX through multiple modules which not only contained fewer species but also different key microbial taxa (eg. Petrimonas, Achromobacter and Comamonas). Microbiota WE and FS, with the highest methanogenic capacities, shared common core species such as Sedimentibacter, Acetobacterium, Methanobacterium and Smithella/Syntrophus, which cooperated with Geobacter (microbiota WE) or Desulfoprunum (microbiota FS) to perform BTEX degradation and methane production. This study demonstrates that electron acceptors may alter microbial function by reshaping microbial community structure and regulating microbial interactions and provides guidelines for electron acceptor selection for bioremediation of aromatic pollutant-contaminated anaerobic sites.

rac-Dichlorprop, a commonly used phenoxyalkanoic acid herbicide, is frequently detected in environments and poses threats to environmental safety and human health. Microbial consortia are thought to play key roles in rac-dichlorprop degradation. However, the compositions of the microbial consortia involved in rac-dichlorprop degradation remain largely unknown. In this study, DNA stable isotope probing (SIP) and metagenomic analysis were integrated to reveal the key microbial consortium responsible for rac-dichlorprop degradation in a rac-dichlorprop-degrading enrichment. OTU340 (Sphingobium sp.) and OTU348 (Sphingopyxis sp.) were significantly enriched in the rac-[13C]dichlorprop-labeled heavy DNA fractions. A rac-dichlorprop degrader, Sphingobium sp. strain L3, was isolated from the enrichment by a traditional enrichment method but with additional supplementation of the antibiotic ciprofloxacin, which was instructed by metagenomic analysis of the associations between rac-dichlorprop degraders and antibiotic resistance genes. As revealed by functional profiling of the metagenomes of the heavy DNA, the genes rdpA and sdpA, involved in the initial degradation of the (R)- and (S)-enantiomers of dichlorprop, respectively, were mostly taxonomically assigned to Sphingobium species, indicating that Sphingopyxis species might harbor novel dichlorprop-degrading genes. In addition, taxonomically diverse bacterial genera such as Dyella, Sphingomonas, Pseudomonas, and Achromobacter were presumed to synergistically cooperate with the key degraders Sphingobium/Sphingopyxis for enhanced degradation of rac-dichlorprop. IMPORTANCE Understanding of the key microbial consortium involved in the degradation of the phenoxyalkanoic acid herbicide rac-dichlorprop is pivotal for design of synergistic consortia used for enhanced bioremediation of herbicide-contaminated sites. However, the composition of the microbial consortium and the interactions between community members during the biodegradation of rac-dichlorprop are unclear. In this study, DNA-SIP and metagenomic analysis were integrated to reveal that the metabolite 2,4-dichlorophenol degraders Dyella, Sphingomonas, Pseudomonas, and Achromobacter synergistically cooperated with the key degraders Sphingobium/Sphingopyxis for enhanced degradation of rac-dichlorprop. Our study provides new insights into the synergistic degradation of rac-dichlorprop at the community level and implies the existence of novel degrading genes for rac-dichlorprop in nature.

rac-Dichlorprop, a commonly used phenoxyalkanoic acid herbicide, is frequently detected in environments and poses threats to environmental safety and human health. Microbial consortia are thought to play key roles in rac-dichlorprop degradation. However, the compositions of the microbial consortia involved in rac-dichlorprop degradation remain largely unknown. In this study, DNA stable isotope probing (SIP) and metagenomic analysis were integrated to reveal the key microbial consortium responsible for rac-dichlorprop degradation in a rac-dichlorprop-degrading enrichment. OTU340 (Sphingobium sp.) and OTU348 (Sphingopyxis sp.) were significantly enriched in the rac-[(13)C]dichlorprop-labeled heavy DNA fractions. A rac-dichlorprop degrader, Sphingobium sp. strain L3, was isolated from the enrichment by a traditional enrichment method but with additional supplementation of the antibiotic ciprofloxacin, which was instructed by metagenomic analysis of the associations between rac-dichlorprop degraders and antibiotic resistance genes. As revealed by functional profiling of the metagenomes of the heavy DNA, the genes rdpA and sdpA, involved in the initial degradation of the (R)- and (S)-enantiomers of dichlorprop, respectively, were mostly taxonomically assigned to Sphingobium species, indicating that Sphingopyxis species might harbor novel dichlorprop-degrading genes. In addition, taxonomically diverse bacterial genera such as Dyella, Sphingomonas, Pseudomonas, and Achromobacter were presumed to synergistically cooperate with the key degraders Sphingobium/Sphingopyxis for enhanced degradation of rac-dichlorprop. IMPORTANCE Understanding of the key microbial consortium involved in the degradation of the phenoxyalkanoic acid herbicide rac-dichlorprop is pivotal for design of synergistic consortia used for enhanced bioremediation of herbicide-contaminated sites. However, the composition of the microbial consortium and the interactions between community members during the biodegradation of rac-dichlorprop are unclear. In this study, DNA-SIP and metagenomic analysis were integrated to reveal that the metabolite 2,4-dichlorophenol degraders Dyella, Sphingomonas, Pseudomonas, and Achromobacter synergistically cooperated with the key degraders Sphingobium/Sphingopyxis for enhanced degradation of rac-dichlorprop. Our study provides new insights into the synergistic degradation of rac-dichlorprop at the community level and implies the existence of novel degrading genes for rac-dichlorprop in nature.

The abilities of three phylogenetically distant ammonia oxidizers, Nitrososphaera gargensis, an ammonia-oxidizing archaeon (AOA); Nitrosomomas nitrosa Nm90, an ammonia-oxidizing bacterium (AOB); and Nitrospira inopinata, the only complete ammonia oxidizer (comammox) available as a pure culture, to biotransform seven sulfonamides (SAs) were investigated. The removals and protein-normalized biotransformation rate constants indicated that the AOA strain N. gargensis exhibited the highest SA biotransformation rates, followed by N. inopinata and N. nitrosa Nm90. The transformation products (TPs) of sulfadiazine (SDZ), sulfamethazine (SMZ) and sulfamethoxazole (SMX) and the biotransformation mechanisms were evaluated. Based on the analysis of the TP formulas and approximate structures, it was found that during biotransformation, i) the AOA strain carried out SA deamination, hydroxylation, and nitration; ii) the AOB strain mainly performed SA deamination; and iii) the comammox isolate participated only in deamination reactions. It is proposed that deamination was catalyzed by deaminases while hydroxylation and nitration were mediated by nonspecific activities of the ammonia monooxygenase (AMO). Additionally, it was demonstrated that among the three ammonia oxidizers, only AOB contributed to the formation of pterin-SA conjugates. The biotransformation of SDZ, SMZ and SMX occurred only when ammonia oxidation was active, suggesting a cometabolic transformation mechanism. Interestingly, SAs could also be transformed by hydroxylamine, an intermediate of ammonia oxidation, suggesting that in addition to enzymatic conversions, a microbially induced abiotic mechanism contributes to SA transformation during ammonia oxidation. Overall, using experiments with pure cultures, this study provides important insights into the roles played by ammonia oxidizers in SA biotransformation.

To determine bacterial communities involved, directly or indirectly, in the anaerobic degradation of cellulose, we conducted a microcosm experiment with soil treated with 13C-cellulose, 12C-cellulose, or without cellulose with analyses of DNA-based stable isotope probing (DNA-SIP), real-time quantitative PCR, and high-throughput sequencing. Firmicutes, Actinobacteria, Verrucomicrobia, and Fibrobacteres were the dominant bacterial phyla-degrading cellulose. Generally, bacteria possessing gene-encoding enzymes involved in the degradation of cellulose and hemicellulose were stimulated. Phylotypes affiliated to Geobacter were also stimulated by cellulose, probably due to their role in electron transfer. Nitrogen-fixing bacteria were also detected, probably due to the decreased N availability during cellulose degradation. High-throughput sequencing showed the presence of bacteria not incorporating 13C and probably involved in the priming effect caused by the addition of cellulose to soil. Collectively, our findings revealed that a more diverse microbial community than expected directly and indirectly participated in anaerobic cellulose degradation.

Biochar management has been proposed as a promising strategy to mitigate climate change. However, the long-term effects of biochar amendment on soil greenhouse gas (GHG) production and microbial community in forest ecosystems under projected warming remain highly uncertain. In this study, we conducted a 49-day incubation experiment to investigate the impact of biochar application on soil physico-chemical properties, GHG production rates, and microbial community at three temperature levels using a temperate forest soil amended with spruce biochar four years ago. Our results showed that temperature exerted a positive effect on soil CO2, CH4 and N2O production, leading to an increase in total global warming potential by 169% and 87% as temperature rose from 5 to 15 °C and from 15 to 25 °C, respectively, and thus a positive feedback to warming. Moreover, warming was found to reduce soil microbial biomass significantly, but at the same time promote the selection of an activated microbial community towards some phyla, e.g. Acidobacteria and Actinobacteria. We observed that biochar amendment reduced soil CH4 consumption and N2O production in the absence of litter by 106% and 94%, respectively, but did not affect soil CO2 production. While biochar had no significant influence of total global warming potential of forest soil, it could promote climate change mitigation by increasing the total soil carbon content by 26% in the presence of litter. In addition, biochar application was shown to enhance soil available phosphorus and dissolved organic carbon concentrations, as well as soil microbial biomass under a warmer environment. Our findings highlighted the potential of spruce biochar as a soil amendment in improving soil fertility and carbon sequestration in temperate forest over the long term, without creating any adverse climatic impacts associated with soil GHG production.

The membrane rotary energy-yielding ATPases represent the cornerstone of cellular bioenergetics for all three domains of life. The archaeal ATPases (A-type ATPases) are functionally similar to the eukaryotic and bacterial F-type ATPases that catalyze ATP synthesis using a PMF. However, they are structurally more similar to the vacuolar-type (V-type) ATPases of eukaryotes and some bacteria that function as proton pumps driven by ATP hydrolysis. Significant variation in subunit composition, structure, and mechanism of the archaeal ATPases is thought to confer adaptive advantage in the variety of extreme environments that archaea inhabit.The ammonia-oxidizing archaea are recognized to exert primary control of nitrification in the marine environment, are major contributors to soil nitrification, and have a habitat range extending from geothermal systems, to acidic soils and the oceanic abyss. The basis for their remarkable adaptive radiation is obscured by a relatively simple metabolism – autotrophic growth using ammonia for energy and nitrogen. In this study, we find that their adaptation to acidic habitats and the extreme pressures of the hadal zone of the ocean at depths below 6000 meters is correlated with horizontal transfer of a variant of the energy-yielding ATPase (atp) operon. Whereas the ATPase genealogy of neutrophilic soil and upper ocean pelagic AOA is congruent with their organismal genealogy inferred from concatenated conserved proteins, a common clade of V-type ATPases unites phylogenetically disparate clades of acidophilic and piezophilic species.A function of the so-acquired V-ATPases in pumping excessive cytoplasmic protons at low pH is consistent with its increased expression by acid-tolerant AOA with decreasing pH. Consistently, heterologous expression of the thaumarchaeotal V-ATPase significantly increased the growth rate of E.coli at low pH. Additional support for adaptive significance derives from our observation that horizontal transfer is also associated with the adaptive radiation of Micrarchaeota, Parvarchaeota and Marsarchaeota into acidic environments. Their ATPases are affiliated with the acidophilic lineage ATPases of Thermoplasmatales and phylogenetically divergent from the corresponding species tree.Another notable finding is that single hadopelagic AOA species contain both A- and V-type ATPases, suggesting that extensive horizontal transfer of atp operons is a highly active and ongoing process within AOA. The presence of an additional V-type ATPase in hadopelagic AOA may provide fitness advantages in the deep ocean with elevated hydrostatic pressure, as the proposed function of V-ATPase in pumping excessive cytoplasmic protons at high pressure may serve to maintain the cytosolic pH homeostasis in marine AOA.Taken together, our study provides the first clear evidence of a significant role of horizontal transfer of atp operon in the adaptive radiation of AOA, one of the most successful organisms on Earth, and other archaeal species, spanning the TACK and DPANN superphyla as well as Euryarchaeota phylum.

Thaumarchaeota are responsible for a significant fraction of ammonia oxidation in the oceans and in soils that range from alkaline to acidic. However, the adaptive mechanisms underpinning their habitat expansion remain poorly understood. Here we show that expansion into acidic soils and the high pressures of the hadopelagic zone of the oceans is tightly linked to the acquisition of a variant of the energy-yielding ATPases via horizontal transfer. Whereas the ATPase genealogy of neutrophilic Thaumarchaeota is congruent with their organismal genealogy inferred from concatenated conserved proteins, a common clade of V-type ATPases unites phylogenetically distinct clades of acidophilic/acid-tolerant and piezophilic/piezotolerant species. A presumptive function of pumping cytoplasmic protons at low pH is consistent with the experimentally observed increased expression of the V-ATPase in an acid-tolerant thaumarchaeote at low pH. Consistently, heterologous expression of the thaumarchaeotal V-ATPase significantly increased the growth rate of E. coli at low pH. Its adaptive significance to growth in ocean trenches may relate to pressure-related changes in membrane structure in which this complex molecular machine must function. Together, our findings reveal that the habitat expansion of Thaumarchaeota is tightly correlated with extensive horizontal transfer of atp operons.

Root-associated fungal communities play a key role in plant productivity and soil processes in forest ecosystems. However, how diversity and composition of root-associated fungi change with forest development and their linkage with soil enzyme activities remain largely unknown. We characterized the root-associated fungal communities, plant and soil properties, and extracellular enzyme activities along a chronosequence spanning young (15 years old) to over-mature (63 years old) Pinus massoniana forest development stages. Our results showed that P. massoniana roots harbored diverse root-associated fungal communities and they varied with forest development. Near-mature (36 years old) forest stands had the lowest alpha-diversity but higher relative abundances of ericoid mycorrhizal and activities of enzymes involved in C, N and P acquisition. The relative abundances of ectomycorrhizal fungi and endophyte were higher in middle-age (24 years old) and mature stands (45 years old), whereas the relative abundance of ericoid mycorrhizal fungi was highest in the near-mature stand. Soil pH, soil C:P and N:P ratios were important factors shaping the diversity and composition of root-associated fungal communities. Structural equation modeling indicated that changes in the community composition, but not richness of root-associated fungi, had significant correlations with soil enzymatic C:N and N:P stoichiometry. In conclusion, this study suggests that different stand development stages exhibit distinct diversity and composition of root-associated fungal communities, which affecting soil functions in terms of enzymatic activity.

The environmental fate of the recalcitrant organic chlorine insecticide lindane and its removal from contaminated soils are still of great concern. However, the key factors influencing microbial removal of lindane from paddy soils with intermittent flooding and draining remain largely unknown. Here, we conducted laboratory experiments to investigated lindane biodegradation in different layers of typical acidic paddy soils under different water managements and bioremediation strategies, together with the changes of functional bacterial consortium, key genes and metabolic pathways. It was found that under flooded conditions, lindane spiking significantly stimulated the growth of some bacterial genera with potential anaerobic catabolic functions in both top- (0–20 cm depth) and subsoil (20–40 cm depth), leading to the shortest half-life of lindane with 7.6–9.0 d in the topsoil. In contrary, lindane spiking dramatically stimulated the growth of bacterial members with aerobic catabolic functions under drained conditions, exhibiting half-lives of lindane with 85–131 d and 18–23 d in the top- and subsoil, respectively. Overall, biostimulation coupled with flooding management would be the better combination for increased lindane bioremediation. Functional genes involved in lindane degradation and retrieved from metagenomic data further supported the anaerobic and aerobic biodegradation of lindane under flooded and drained conditions, respectively. Moreover, the integrated network analysis suggested water management and organic matter were the primary factors shaped the assembly of functional bacteria in lindane degradation, among which Clostridium and Rhodanobacter were the key anaerobic and aerobic functional genera, respectively. Taken together, our study provides a comprehensive understanding of lindane biodegradation in distinct layers of acidic paddy soils that were combinedly affected by different water managements and bioremediation strategies.

The recent discovery of complete ammonia oxidation (comammox) process in a single organism challenged the division of labor between two functional groups in the classical two-step nitrification model. However, the distribution and activity of comammox bacteria in various environments remain largely unknown. This study presented a large-scale investigation of the geographical distribution, phylogenetic diversity, and activity of comammox Nitrospira in typical agricultural soils. Among the 23 samples harvested across China, comammox Nitrospira clade A was ubiquitously detected at 4.14 × 104–1.65 × 107 amoA gene copies/g dry soil, with 90% belonging to the subclade A2. The abundance of comammox Nitrospira clade B was two orders of magnitude lower than clade A. In all samples, comammox Nitrospira were 1–2 orders of magnitude less abundant than canonical nitrifiers, and soils with slightly high pH and C/N tended to enrich more comammox Nitrospira. Unlike canonical nitrifiers, comammox Nitrospira had sustained amoA gene transcription regardless of external ammonia supply, indicating their competitive advantage over other nitrifiers under low-ammonia conditions. When fed with 1 mM ammonium for 15 days, comammox Nitrospira in tested soils were enriched 2.36 times higher than those enriched by the same amount of nitrite, indicating their preference to utilizing ammonia as the substrate. DNA-SIP further confirmed the in situ nitrification activity of comammox Nitrospira. This study provided new insights into the broad distribution and diversity of comammox Nitrospira in agricultural soils, which could potentially play an important role in the microbial nitrogen cycle in soils.

Peanut yield and quality are seriously compromised by continuous monoculturing in the red soil region of southern China. Monoculturing can cause soil degradation and an increase in soil-borne diseases. This research aimed to investigate the influence of long-term peanut monocropping and different fertilization treatments on peanut growth, soil physical and chemical properties and soil microbial community. A long-term fertilization experiment established in 1996 was utilized to examine the effect of various fertilization treatments including chemical and organic fertilizers treatments. Deep 16S rRNA gene pyrosequencing highlighted changes in the abundance and structure of bacterial communities, especially of the pathogenic and beneficial bacterial communities in long term chemical fertilizer treatment in comparison to the organic manure treatment. Chemical fertilizer treatment causes a shift in bacterial community structure and decrease in diversity under the long-term monocropping in comparison to organic fertilizer. The abundance of the bacterial pathogen Ralstonia solanacearum, a causative agent of peanut wilt, was found to be associated with a loss of community diversity and loss of the peanut yield. The organic fertilizers more effectively increase microbial diversity in the soil and changed the community structure. Long-term use of the chemical fertilizer leads to a decrease in microbial diversity of the soil and an increase in R. solanacearum with associated increase of peanut wilt. The potential decrease in diversity and competition between the bacterial community and the pathogen may be a contributing factor to increased disease during long-term chemical fertilizer use.

Rice paddy fields are characterized by regular flooding and nitrogen fertilization, but the functional importance of aerobic ammonia oxidizers and nitrite oxidizers under unique agricultural management is poorly understood. In this study, we report the differential contributions of ammonia-oxidizing archaea (AOA), bacteria (AOB) and nitrite-oxidizing bacteria (NOB) to nitrification in four paddy soils from different geographic regions (Zi-Yang (ZY), Jiang-Du (JD), Lei-Zhou (LZ) and Jia-Xing (JX)) that are representative of the rice ecosystems in China. In urea-amended microcosms, nitrification activity varied greatly with 11.9, 9.46, 3.03 and 1.43 μg NO3(-)-N g(-1) dry weight of soil per day in the ZY, JD, LZ and JX soils, respectively, over the course of a 56-day incubation period. Real-time quantitative PCR of amoA genes and pyrosequencing of 16S rRNA genes revealed significant increases in the AOA population to various extents, suggesting that their relative contributions to ammonia oxidation activity decreased from ZY to JD to LZ. The opposite trend was observed for AOB, and the JX soil stimulated only the AOB populations. DNA-based stable-isotope probing further demonstrated that active AOA numerically outcompeted their bacterial counterparts by 37.0-, 10.5- and 1.91-fold in (13)C-DNA from ZY, JD and LZ soils, respectively, whereas AOB, but not AOA, were labeled in the JX soil during active nitrification. NOB were labeled to a much greater extent than AOA and AOB, and the addition of acetylene completely abolished the assimilation of (13)CO2 by nitrifying populations. Phylogenetic analysis suggested that archaeal ammonia oxidation was predominantly catalyzed by soil fosmid 29i4-related AOA within the soil group 1.1b lineage. Nitrosospira cluster 3-like AOB performed most bacterial ammonia oxidation in the ZY, LZ and JX soils, whereas the majority of the (13)C-AOB in the JD soil was affiliated with the Nitrosomona communis lineage. The (13)C-NOB was overwhelmingly dominated by Nitrospira rather than Nitrobacter. A significant correlation was observed between the active AOA/AOB ratio and the soil oxidation capacity, implying a greater advantage of AOA over AOB under microaerophilic conditions. These results suggest the important roles of soil physiochemical properties in determining the activities of ammonia oxidizers and nitrite oxidizers.

All cultivated ammonia-oxidizing archaea (AOA) within the Nitrososphaera cluster (former soil group 1.1b) are neutrophilic. Molecular surveys also indicate the existence of Nitrososphaera -like phylotypes in acidic soil, but their ecological roles are poorly understood. In this study, we present molecular evidence for the chemolithoautotrophic growth of Nitrososphaera -like AOA in an acidic soil with pH 4.92 using DNA-based stable isotope probing (SIP). Soil microcosm incubations demonstrated that nitrification was stimulated by urea fertilization and accompanied by a significant increase in the abundance of AOA rather than ammonia-oxidizing bacteria (AOB). Real-time PCR analysis of amoA genes as a function of the buoyant density of the DNA gradient following the ultracentrifugation of the total DNA extracted from SIP microcosms indicated a substantial growth of soil AOA during nitrification. Pyrosequencing of the total 16S rRNA genes in the “heavy” DNA fractions suggested that archaeal communities were labeled to a much greater extent than soil AOB. Acetylene inhibition further showed that 13 CO 2 assimilation by nitrifying communities depended solely on ammonia oxidation activity, suggesting a chemolithoautotrophic lifestyle. Phylogenetic analysis of both 13 C-labeled amoA and 16S rRNA genes revealed that most of the active AOA were phylogenetically closely related to the neutrophilic strains Nitrososphaera viennensis EN76 and JG1 within the Nitrososphaera cluster. Our results provide strong evidence for the adaptive growth of Nitrososphaera -like AOA in acidic soil, suggesting a greater metabolic versatility of soil AOA than previously appreciated.

Both bacteria and archaea potentially contribute to ammonia oxidation, but their roles in freshwater sediments are still poorly understood. Seasonal differences in the relative activities of these groups might exist, since cultivated archaeal ammonia oxidizers have higher temperature optima than their bacterial counterparts. In this study, sediment collected from eutrophic freshwater Lake Taihu (China) was incubated at different temperatures (4°C, 15°C, 25°C, and 37°C) for up to 8 weeks. We examined the active bacterial and archaeal ammonia oxidizers in these sediment microcosms by using combined stable isotope probing (SIP) and molecular community analysis. The results showed that accumulation of nitrate in microcosms correlated negatively with temperature, although ammonium depletion was the same, which might have been related to enhanced activity of other nitrogen transformation processes. Incubation at different temperatures significantly changed the microbial community composition, as revealed by 454 pyrosequencing targeting bacterial 16S rRNA genes. After 8 weeks of incubation, [ 13 C]bicarbonate labeling of bacterial amoA genes, which encode the ammonia monooxygenase subunit A, and an observed increase in copy numbers indicated the activity of ammonia-oxidizing bacteria in all microcosms. Nitrosomonas sp. strain Is79A3 and Nitrosomonas communis lineages dominated the heavy fraction of CsCl gradients at low and high temperatures, respectively, indicating a niche differentiation of active bacterial ammonia oxidizers along the temperature gradient. The 13 C labeling of ammonia-oxidizing archaea in microcosms incubated at 4 to 25°C was minor. In contrast, significant 13 C labeling of Nitrososphaera -like archaea and changes in the abundance and composition of archaeal amoA genes were observed at 37°C, implicating autotrophic growth of ammonia-oxidizing archaea under warmer conditions.

Paddy field methanogenic archaea are responsible for methane (CH4) production and contribute significantly to climate change. The information regarding the spatial variations in the abundance, the diversity and the composition of such ecologically important microbes, however, is quite limited at large scale. In this investigation, we studied the abundance, alpha diversity and geographical distribution of methanogenic archaeal communities in nine representative paddy sites, along a large latitudinal gradient in China, using pyrosequencing and real-time quantitative PCR. It is found that all paddy soils harbor constant methanogenic archaeal constituents, which is dominated by family Methanocellaceae (37.3%), Methanobacteriaceae (22.1%), Methanosaetaceae (17.2%), and Methanosarcinaceae (9.8%). Methanogenic archaeal abundance is primarily influenced by soil C (R = 0.612, P = 0.001) and N (R = 0.673, P = 0.001) contents, as well as alpha diversity by soil pH (PD: R = -0.552, P = 0.006; Chao1: R = -0.615, P = 0.002). Further exploration revealed that both spatial distance (R = 0.3469, P = 0.001, partial mental test) and soil chemical variables mainly about soil C and N (R = 0.2847, P = 0.001) are the two major factors affecting methanogenic archaeal community composition distribution in paddy soils. This finding will allow us to develop a better picture of the biogeographic ranges of these ecologically important microbes and get deeper insights into their ecology.

Soil harbors remarkably stabilise bacterial communities at the phylum level. However, no two soils have exactly the same structure of bacterial phyla. The structure of microbial community is strongly influenced by the type of land-use through changes in soil attributes. Using high-throughput pyrosequencing and quantitative polymerase chain reaction techniques, soil microbial community structures were investigated along a land-use gradient of 100- and 27-year farmlands, a 33-year Pinus forest, a 28-year poplar forest, and a 21-year shrubland, as well as a native desert from which all cultivated systems were converted. The results revealed that the dominant phylotypes in the native soil comprised primarily of Alphaproteobacteria, Actinobacteria, Bacteroidetes, and Firmicutes, accounting for > 71.4% of the total bacterial 16S rRNA sequence reads. Changes in land-use led to a significant; decrease in these dominant phylotypes down to 33.4%. In contrast, the phylotypes with low abundance, such as Acidobacteria, Chloroflexi, Nitrospira, and Gammaproteobacteria, increased sharply from 4.5%-5.9% in the native soil to 20.9%-30.2% of the total 16S rRNA gene sequences in the cultivated soils except for the soil from the shrubland. These contrasting changes in the major taxa appear to be correlated with the changes in soil attributes. For instance, bacterial and archaeal amoA genes were found to be 960- and 3800-fold more abundant in the soil from the 100-year farmland than the native soil. The changes in numerically less dominant nitrifying phylotypes are consistent with soil inorganic nitrogen dynamics. Quantification of the 16S rRNA genes demonstrated that bacteria and archaea were about two to three orders of magnitude more abundant in the cultivated soil than in the native soil. Hence, land-use type affects the soil bacterial community structure, which has profound consequences on ecosystem function.

Long-term field fertilization experiments may provide useful insight into the relative contributions of NH 3 –oxidizing bacteria (AOB) and NH 3 –oxidizing archaea (AOA) to soil nitrification. In this study, the abundance and composition of AOB and AOA were investigated in bulk soil from paddy field plots (4 by 5 m) that received no fertilization (CK), 180 kg urea N ha−1 yr −1 (NPK), or 180 kg urea N plus 4500 kg rice (Oryza sativa L.) straw ha −1 yr−1 (NPK/OM) since 1988 (each with three replicated plots). Our results clearly indicate that long-term fertilization (22 yr) significantly altered the community structure of AOB rather than AOA in the soil within fertilized plots compared with the CK plot. Quantitative polymerase chain reaction analysis of the amoA genes that encode the ammonia monooxygenase subunit A revealed that the AOB in the fertilized plots were 50 times more abundant than in the CK plot. Denaturing gradient gel electrophoresis fingerprinting of the amoA genes demonstrated a drastic shift in the AOB community structure, resulting in highly enriched AOB similar to Nitrosospira cluster 3 in the fertilized plots. In contrast, the population size and composition of the AOA community remained largely unchanged in all plots. Furthermore, a positive correlation was observed between the bacterial rather than archaeal amoA gene copy numbers and potential nitrification activity among the soils. The distinct responses of AOA and AOB to long-term field fertilization were probably related to the availability of NH 3 . Our results suggest that AOB might play major roles in the NH 3 oxidation that occurs in bulk soil under the in situ fertilized paddy field conditions tested in this study.

A synthetic pyrethroid (SP)-degrading bacterium, designated pyd-1, was isolated from SPcontaminated soil. Based on its phenotypic and genotypic properties, the strain was identified as Ochrobactrum tritici. Strain pyd-1 was able to degrade a wide range of SPs, and its degradation efficiencies were dependent on the molecular structure of the SP. Interestingly, the strain degraded cis- and trans-permethrin (cypermethrin) at nearly the same rate and possessed approximately equal hydrolysis activities toward the two enantiomers of fenpropathrin. These results suggest that different isomers of SPs are degraded with equal efficiency by strain pyd-1. We studied the metabolic pathway of fenpropathrin degradation in strain pyd-1 by metabolite identification and enzymatic analysis. Fenpropathrin is degraded by hydrolysis of the carboxylester linkage to yield 2,2,3,3-tetramethylcyclopropanecarboxylic acid and 3-phenoxybenzaldehyde, which is converted to 3-phenoxybenzoic acid (PBA). PBA is further metabolized to 4-hydroxy-3-phenoxybenzoic acid (4-hydroxy-PBA). 4-Hydroxy-PBA is oxidized to protocatechuate and p-hydroquinone. Protocatechuate is further oxidized through an ortho-cleavage pathway, and p-hydroquinone is degraded via 1,2,4-benzenetriol.

A pyrethroid-degrading bacterium strain JZ-2 was isolated from activated sludge treating pyrethroid-manufacturing wastewater. Based on the morphological, physiological and biochemical characterization, and phylogenetic analysis of the 16S rRNA gene sequence, the strain was identified as Sphingobium sp. Strain JZ-2 was capable of degrading fenpropathrin, cypermethrin, permethrin, cyhalothrin, deltamethrin, fenvalerate and bifenthrin. This strain degraded fenpropathrin by hydrolysis of the carboxylester linkage to yield 3-phenoxybenzaldehyde and 2,2,3,3-tetramethylcyclopropanecarboxylic acid. 3-Phenoxybenzaldehyde, 3-phenoxybenzoate, protocatechuate and catechol are the intermediates of fenpropathrin degradation. Protocatechuate and catechol were further oxidized by ortho -cleavage pathway. A novel pyrethroid hydrolase from cell-free extract was purified 108.5-fold to apparent homogeneity with a 10.2% overall recovery. It was a monomer with a molecular mass of 31 ± 1 kDa, a p I of 4.85. The optimal pH and temperature were 7.5 and 40 °C, respectively. No cofactors or coenzymes were required for the pyrethroid-hydrolysis activity. The enzyme was strongly inhibited by many irons (Ag + , Cu 2+ , Hg 2+ and Zn 2+ ), SDS, p-chloromercuribenzoic acid, phenylmethylsulfonyl fluoride and malathion.

The methanogenic archaea plays many ecological roles in paddy ecosystems such as the regulation of CH4 emissions. However, knowledge gaps exist about the mechanisms of their spatial shift in population.We collected 8 latitudinal paddy soil samples from 20.55 degrees N to 47.43 degrees N in China, and studied their methanogenic archaeal community compositions by PCR-DGGE fingerprinting and phylogenetic analyses. Mechanism of spatial shift in community composition was unraveled by canonical correspondence analysis and Venn diagram.Soil pH was one of the main environmental triggers to the composition of methanogenic archaea community in paddy soils. In addition, community composition shifted along latitudinal gradients.It is the first report on biogeography of methanogenic archaeal community in arable soils and its environmental driving factors. The information would contribute to understanding spatial shifts in the transformation of organic matters along Chinese latitudinal gradient.

Paddy fields represent a unique ecosystem in which regular flooding occurs, allowing for rice cultivation. However, the taxonomic identity of the microbial functional guilds that catalyze soil nitrification remains poorly understood. In this study, we provide molecular evidence for distinctly different phylotypes of nitrifying communities in a neutral paddy soil using high-throughput pyrosequencing and DNA-based stable isotope probing (SIP). Following urea addition, the levels of soil nitrate increased significantly, accompanied by an increase in the abundance of the bacterial and archaeal amoA gene in microcosms subjected to SIP (SIP microcosms) during a 56-day incubation period. High-throughput fingerprints of the total 16S rRNA genes in SIP microcosms indicated that nitrification activity positively correlated with the abundance of Nitrosospira -like ammonia-oxidizing bacteria (AOB), soil group 1.1b-like ammonia-oxidizing archaea (AOA), and Nitrospira -like nitrite-oxidizing bacteria (NOB). Pyrosequencing of 13 C-labeled DNA further revealed that 13 CO 2 was assimilated by these functional groups to a much greater extent than by marine group 1.1a-associated AOA and Nitrobacter -like NOB. Phylogenetic analysis demonstrated that active AOB communities were closely affiliated with Nitrosospira sp. strain L115 and the Nitrosospira multiformis lineage and that the 13 C-labeled AOA were related to phylogenetically distinct groups, including the moderately thermophilic “ Candidatus Nitrososphaera gargensis,” uncultured fosmid 29i4, and acidophilic “ Candidatus Nitrosotalea devanaterra” lineages. These results suggest that a wide variety of microorganisms were involved in soil nitrification, implying physiological diversification of soil nitrifying communities that are constantly exposed to environmental fluctuations in paddy fields.

Bacteria and archaea sustain subsurface cave ecosystems by dominating primary production and fueling biogeochemical cyclings, despite the permanent darkness and shortage of nutrients. However, the heterogeneity and underlying mechanism of microbial diversity in caves, in particular those well connect to surface environment are largely unexplored. In this study, we examined the bacterial abundance and composition in Jinjia Cave, a small and shallow limestone cave located on the western Loess Plateau of China, by enumerating and pyrosequencing small subunit (SSU) rRNA genes. The results clearly reveal the contrasting bacterial community compositions in relation to cave habitat types, i.e., rock wall deposit, aquatic sediment and sinkhole soil, which are differentially connected to the surface environment. The deposits on the cave walls were dominated by putative cave-specific bacterial lineages within the -Proteobacteria or Actinobacteria that are routinely found on cave rocks around the world. In addition, sequence identity with known functional groups suggests enrichments of chemolithotrophic bacteria potentially involved in autotrophic C fixation and inorganic N transformation on rock surfaces. By contrast, bacterial communities in aquatic sediments were more closely related to those in the overlying soils. This is consistent with the similarity in elemental composition between the cave sediment and the overlying soil, implicating the influence of mineral chemistry on cave microhabitat and bacterial composition. These findings provide compelling molecular evidence of the bacterial community heterogeneity in an East Asian cave, which might be controlled by both subsurface and surface environments.

To evaluate the effect of long-term fertilization on soil microbial community and soil chemical and physical properties.Using a high-throughput pyrosequencing technique, we studied microbial community in the 0-300 cm soil samples covering a 20-year field-experiment with different fertilization applications including inorganic fertilizer alone (N 300 kg/hm2, P2O5 150 kg/hm2 and K2O 60 kg/hm2) and inorganic fertilizer combined with straw (same application rate of N and P fertilizer combined with 5.4 t straw).Actinobacteria and alpha-proteobacteria were the predominant groups in the topsoil (0-20 cm). As the soil depth increased, the relative abundance of actinobacteria decreased whereas that of proteobacteria, especially gamma-proteobacteria and beta-proteobacteria increased and gradually became the dominant groups in the subsoil (20-300 cm). Long-term fertilizing applications significantly affected soil microbial communities throughout the soil profile, and increased the relative abundance of ammonia-oxidizing archaea at 0-40 cm depth. In addition, agriculture management, e. g. irrigation may be an important driving factor for the distribution of ammonia-oxidizing bacteria in soil profile. Total nitrogen and organic carbon contents were the most influential factors on microbial community in the topsoil and in the subsoil, respectively.Long-term fertilizer applications altered soil nutrient availability within the soil profile, which was likely to result in the different microbial community structure between the fertilizer treatments, especially for the subsoil.

Pure culture studies have demonstrated that methanotrophs and ammonia oxidizers can both carry out the oxidation of methane and ammonia. However, the expected interactions resulting from these similarities are poorly understood, especially in complex, natural environments. Using DNA-based stable isotope probing and pyrosequencing of 16S rRNA and functional genes, we report on biogeochemical and molecular evidence for growth stimulation of methanotrophic communities by ammonium fertilization, and that methane modulates nitrogen cycling by competitive inhibition of nitrifying communities in a rice paddy soil. Pairwise comparison between microcosms amended with CH4, CH4+Urea, and Urea indicated that urea fertilization stimulated methane oxidation activity 6-fold during a 19-day incubation period, while ammonia oxidation activity was significantly suppressed in the presence of CH4. Pyrosequencing of the total 16S rRNA genes revealed that urea amendment resulted in rapid growth of Methylosarcina-like MOB, and nitrifying communities appeared to be partially inhibited by methane. High-throughput sequencing of the 13C-labeled DNA further revealed that methane amendment resulted in clear growth of Methylosarcina-related MOB while methane plus urea led to an equal increase in Methylosarcina and Methylobacter-related type Ia MOB, indicating the differential growth requirements of representatives of these genera. An increase in 13C assimilation by microorganisms related to methanol oxidizers clearly indicated carbon transfer from methane oxidation to other soil microbes, which was enhanced by urea addition. The active growth of type Ia methanotrops was significantly stimulated by urea amendment, and the pronounced growth of methanol-oxidizing bacteria occurred in CH4-treated microcosms only upon urea amendment. Methane addition partially inhibited the growth of Nitrosospira and Nitrosomonas in urea-amended microcosms, as well as growth of nitrite-oxidizing bacteria. These results suggest that type I methanotrophs can outcompete type II methane oxidizers in nitrogen-rich environments, rendering the interactions among methane and ammonia oxidizers more complicated than previously appreciated.

The hydrolysis of urea as a source of ammonia has been proposed as a mechanism for the nitrification of ammonia-oxidizing bacteria (AOB) in acidic soil. The growth of Nitrososphaera viennensis on urea suggests that the ureolysis of ammonia-oxidizing archaea (AOA) might occur in natural environments. In this study, (15)N isotope tracing indicates that ammonia oxidation occurred upon the addition of urea at a concentration similar to the in situ ammonium content of tea orchard soil (pH 3.75) and forest soil (pH 5.4) and was inhibited by acetylene. Nitrification activity was significantly stimulated by urea fertilization and coupled well with abundance changes in archaeal amoA genes in acidic soils. Pyrosequencing of 16S rRNA genes at whole microbial community level demonstrates the active growth of AOA in urea-amended soils. Molecular fingerprinting further shows that changes in denaturing gradient gel electrophoresis fingerprint patterns of archaeal amoA genes are paralleled by nitrification activity changes. However, bacterial amoA and 16S rRNA genes of AOB were not detected. The results strongly suggest that archaeal ammonia oxidation is supported by hydrolysis of urea and that AOA, from the marine Group 1.1a-associated lineage, dominate nitrification in two acidic soils tested.

A synthetic pyrethroid (SP)-degrading bacterial strain, designated JZ-1T, was isolated from activated sludge of a SP-manufacturing wastewater treatment facility and studied using a polyphasic taxonomic approach. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain JZ-1T belonged to the genus Sphingobium, showing highest sequence similarities to Sphingobium faniae DSM 21829T (98.6 %), Sphingobium cloacae JCM 10874T (98.5 %), Sphingobium vermicomposti DSM 21299T (97.4 %) and Sphingobium ummariense CCM 7431T (96.9 %). The polar lipid pattern, the presence of spermidine and ubiquinone Q-10, the predominance of the cellular fatty acids C18 : 1ω7c, C19 : 0 cyclo ω8c, 11 methyl C18 : 1ω7c, C16 : 0 and C14 : 0 2-OH, and the G+C content of the genomic DNA also supported the affiliation of the strain with the genus Sphingobium. Strain JZ-1T showed low DNA–DNA relatedness values with S. faniae DSM 21829T (30.2 %), S. cloacae JCM 10874T (23.3 %), S. vermicomposti DSM 21299T (10.9 %) and S. ummariense CCM 7431T (7.9 %). Based on its phylogenetic position and its phenotypic and genotypic properties, strain JZ-1T represents a novel species of the genus Sphingobium, for which the name Sphingobium wenxiniae sp. nov. is proposed. The type strain is JZ-1T ( = CGMCC 1.7748T = DSM 21828T).

A novel esterase gene, pytH , encoding a pyrethroid-hydrolyzing carboxylesterase was cloned from Sphingobium sp. strain JZ-1. The gene contained an open reading frame of 840 bp. Sequence identity searches revealed that the deduced enzyme shared the highest similarity with many α/β-hydrolase fold proteins (20 to 24% identities). PytH was expressed in Escherichia coli BL21 and purified using Ni-nitrilotriacetic acid affinity chromatography. It was a monomeric structure with a molecular mass of approximately 31 kDa and a pI of 4.85. PytH was able to transform p -nitrophenyl esters of short-chain fatty acids and a wide range of pyrethroid pesticides, and isomer selectivity was not observed. No cofactors were required for enzyme activity.

A bacterial strain capable of degrading pyrethroid, designated JZ-2T, was isolated from activated sludge treating pyrethroid-contaminated wastewater. Phylogenetic analysis based on 16S rRNA gene sequences indicated that strain JZ-2Tbelongs to the genusSphingobium. It showed the highest levels of 16S rRNA gene sequence similarity toSphingobium cloacaeJCM 10874T(98.3 %) andSphingobium ummarienseCCM 7431T(97.1 %), and 94.8–96.9 % similarity to the type strains of other members of the genusSphingobium. Strain JZ-2Tcontained C18 : 1ω7cas the predominant fatty acid, C14 : 02-OH as the major 2-hydroxy fatty acid, ubiquinone Q-10 as the main respiratory quinone, diphosphatidylglycerol, phosphatidylglycerol, phosphatidylcholine, phosphatidylmonomethylethanolamine, phosphatidylethanolamine and two sphingoglycolipids as the predominant polar lipids and spermidine as the major polyamine. DNA–DNA hybridization results showed that strain JZ-2Thad low genomic relatedness withS. cloacaeJCM 10874T(34 %) andS. ummarienseCCM 7431T(23 %). Based on the phenotypic, genotypic and phylogenetic data presented, strain JZ-2Tis considered to represent a novel species of the genusSphingobium, for which the nameSphingobium faniaesp. nov. is proposed. The type strain is JZ-2T(=CGMCC 1.7749T=DSM 21829T).