Complete genome sequence of Pseudomonas moorei strain m318 isolated from rhizosphere of Pteris multifida in high-arsenic-content soil.

Pseudomonas moorei strain m318 is an arsenite-oxidizing rhizobacterium isolated from the rhizosphere of an arsenic hyperaccumulator plant, beneficial for arsenic phytoremediation. Here, we report the complete genome sequence of this strain, which consists of a circular chromosome assembled using long reads sequenced on Nanopore and polished with Illumina paired-end reads.

Effective selenate removal using pH modulated synthesis of biogenic jarosite: Comparative insight with non-biogenic jarosite and biogenic schwertmannite.

Selenium, a crucial trace element for many organisms, including prokaryotes and humans, is toxic at high concentrations, necessitating its removal from wastewater. This study investigates the use of jarosite, a naturally occurring iron sulfate mineral with excellent heavy metal attenuation properties, for selenate (Se(VI)) removal for the first time. Biogenic jarosite was synthesized through Fe(II) oxidation by Acidithiobacillus ferrooxidans at an initial pH ranging from 1.5 to 4.0 (J-1.5 to J-4.0). This resulted in the formation of morphologically diverse particles of biogenic jarosite owing to varying Fe(II) oxidation and precipitation rates. For comparative analysis, non-biogenic jarosite (J-90C) and biogenic schwertmannite (S-2.5) were also synthesized. At 0.2 mM initial Se(VI) concentration, J-2.5 demonstrated superior Se(VI) removal compared to J-3.5 and J-90C. At 2.0 mM Se(VI), J-2.5 still outperformed J-3.5 and J-90C although its overall removal efficiency decreased. Notably, at 0.2 mM concentration, Se(VI) removal by J-2.5 was 63 %, which is comparable to 77 % removal by S-2.5. Furthermore, sulfate release from J-2.5 was significantly lower than that from S-2.5 in both Se-free and Se-containing solutions. This study provides critical insights into the synthesis and application of biogenic jarosite as a replacement for metastable schwertmannite, emphasizing its potential as an excellent Se sink for wastewater treatment.

Effects of temperature on plant growth and arsenic removal efficiency of Pteris vittata in purifying arsenic-contaminated water in winter: A two-year year-round field study.

Phytoremediation is a cost-effective and eco-friendly alternative method for arsenic (As) contaminated water treatment. This study conducted a two-year year-round field study (cycle1 and cycle2) in a temperate area (Sendai, Japan) using small As-hyperaccumulator Pteris vittata seedlings to reduce pre-cultivation time and associated costs. The number of seedlings was changed from 256 in the cycle1 period to 165 in the cycle2 period to evaluate the As removal efficiency of P. vittata for As-contaminated water in field conditions with different plant densities. Before the winter season, with continuously increasing fronds, rhizomes, and roots growth, this reduction did not affect the plant's As removal efficiency for As-contaminated water to decrease the As concentration from 30 µg/L to the environmental quality standard for As in water, set at 10 µg/L in Japan. During the winter season, we found that cold weather caused P. vittata to wither and release the accumulated As into water without a greenhouse (cycle1). In the meantime, the bioaccumulation factor (BAF) and the translocation factor (TF) values for fronds of P. vittata decreased (BAF for fronds: from 66,089 to 8,460; TF for fronds: from 13.4 to 3.4). On the other hand, with greenhouse protection (cycle2), P. vittata did not severely wither and kept accumulating As. Moreover, BAF and TF values for fronds of P. vittata increased (BAF for fronds: from 24,372 to 36,740; TF for fronds: from 5.2 to 17.2). Maintaining the air temperature inside the greenhouse, particularly around the rhizomes, above 0 °C may be the reason why P. vittata remained alive and functional during the cold winter. These results indicate that a single-layer polyethylene greenhouse was sufficient for the tropical-subtropical As-hyperaccumulator fern P. vittata to survive the cold winter and snow in the temperate area, enabling year-round phytoremediation treatment of As-contaminated water in the open field.

Bayesian network highlights the contributing factors for efficient arsenic phytoextraction by Pteris vittata in a contaminated field.

Phytoextraction is a low-cost and eco-friendly method for removing pollutants, such as arsenic (As), from contaminated soil. One of the most studied As hyperaccumulators for soil remediation include Pteris vittata. Although phytoextraction using plant-assisted microbes has been considered a promising soil remediation method, microbial harnessing has not been achieved due to the complex and difficult to understand interactions between microbes and plants. This problem can possibly be addressed with a multi-omics approach using a Bayesian network. However, limited studies have used Bayesian networks to analyze plant-microbe interactions. Therefore, to understand this complex interaction and to facilitate efficient As phytoextraction using microbial inoculants, we conducted field cultivation experiments at two sites with different total As contents (62 and 8.9 mg/kg). Metabolome and microbiome data were obtained from rhizosphere soil samples using nuclear magnetic resonance and high-throughput sequencing, respectively, and a Bayesian network was applied to the obtained multi-omics data. In a highly As-contaminated site, inoculation with Pseudomonas sp. strain m307, which is an arsenite-oxidizing microbe having multiple copies of the arsenite oxidase gene, increased As concentration in the shoots of P. vittata to 157.5 mg/kg under this treatment; this was 1.5-fold higher than that of the other treatments. Bayesian network demonstrated that strain m307 contributed to As accumulation in P. vittata. Furthermore, the network showed that microbes belonging to the MND1 order positively contributed to As accumulation in P. vittata. Based on the ecological characteristics of MND1, it was suggested that the rhizosphere of P. vittata inoculated with strain m307 was under low-nitrogen conditions. Strain m307 may have induced low-nitrogen conditions via arsenite oxidation accompanied by nitrate reduction, potentially resulting in microbial iron reduction or the prevention of microbial iron oxidation. These conditions may have enhanced the bioavailability of arsenate, leading to increased As accumulation in P. vittata.

A microbial consortium led by a novel Pseudomonas species enables degradation of carbon tetrachloride under aerobic conditions.

Carbon tetrachloride (CT) is a recalcitrant and high priority pollutant known for its toxicity, environmental prevalence, and inhibitory activities. Although much is known about anaerobic CT biodegradation, microbial degradation of CT under aerobic conditions has not yet been reported. This study reports for the first time the enrichment of a stable aerobic CT-degrading bacterial consortium, from a CT-contaminated groundwater sample, capable of co-metabolically degrading 30 µM of CT within a week. A Pseudomonas strain (designated as Stari2) that is the predominant bacterium in this consortium was isolated, and further characterization showed that this bacterium can tolerate and co-metabolically degrade up to 5 mM of CT under aerobic conditions in the presence of different carbon/energy sources. The CT biodegradation profiles of strain Stari2 and the consortium were found to be identical, while no significant positive correlation between strain Stari2 and other bacteria was observed in the consortium during the period of higher CT biodegradation. These results confirmed that the isolated Pseudomonas strain Stari2 is the key player in the consortium catalyzing the biodegradation of CT. No chloroform (CF) or other chlorinated compound was detected during the cometabolism of CT. The whole genome sequencing of strain Stari2 showed that it is a novel Pseudomonas species. The findings demonstrated that biodegradation of CT under aerobic conditions is feasible, and the isolated CT-degrader Pseudomonas sp. strain Stari2 has a great potential for in-situ bioremediation of CT-contaminated environments.

Temperature Dependence of Metals Accumulation and Removal Kinetics by Arabidopsis halleri ssp. gemmifera.

Cadmium (Cd), which is present in zinc (Zn) ore, is a toxic metal and causes contamination globally. Phytoremediation is a promising technology for the remediation of sites with low and moderate contamination. Temperature is an important factor in phytoremediation because it has an impact on both plant biomass and the accumulation of heavy metals. However, little is known about the influence of temperature on heavy metal accumulation by the Cd and Zn hyperaccumulator Arabidopsis halleri ssp. gemmifera. The effect of temperature on the distribution of Cd and Zn in A. halleri ssp. gemmifera and the mechanism of metal removal from solution were investigated in this study. Our results showed that the temperature dependence of the distribution of Cd and Zn in the plant was different, which may suggest that the mechanisms of xylem loading were different between Cd and Zn. Although Cd and Zn have partially similar transport pathways, the removal kinetics based on the first-order reaction rate constant revealed that the temperature which maximized rate of absorption was different between Cd and Zn. This study suggests a potential for efficient Cd phytoextraction using A. halleri ssp gemmifera in Cd and Zn co-existing environments.

Efficient biodegradation of 1,4-dioxane commingled with additional organic compound: Role of interspecies interactions within consortia.

Microbial consortia-mediated biodegradation of 1,4-dioxane (1,4-D), an emerging water contaminant, is always a superior choice over axenic cultures. Thus, better understanding of the functions of coexisting microbes and their interspecies interactions within the consortia is crucial for predicting biodegradation efficiency and designing efficient 1,4-D-degrading microbial consortia. This study evaluated how microbial community compositions and interspecies interactions govern the microbial consortia-mediated 1,4-D biodegradation by investigating the biodegradability and microbial community dynamics of both enriched (N112) and synthetic (SCDs and SCDNs) microbial consortia in the absence or presence of additional organic compound (AOC). In the absence of AOC, N112 exhibited 100% 1,4-D biodegradation efficiency at a rate of 12.5 mg/L/d, whereas the co-occurrence of AOC resulted in substrate-dependent biodegradation inhibition and thereby reduced the biodegradation efficiency and activity (2.0-10.0 mg/L/d). The coexistence and negative influence of certain low-abundant non-degraders on both 1,4-D-degraders and key non-degraders in N112 was identified as the prime cause behind such biodegradation inhibition. Comparing with N112, SCDN-1 composed of 1,4-D-degraders and key non-degraders significantly improved the 1,4-D biodegradation efficiency in the presence of AOC, confirming the absence of negative influence of low-abundant non-degraders and cooperative interactions between 1,4-D-degraders and key non-degraders in SCDN-1. On the contrary, both two-species and three-species SCDs comprised of only 1,4-D-degraders resulted in lower 1,4-D biodegradation efficiency as compared to SCDN-1 under all treatment conditions, while max. 91% 1,4-D biodegradation occurred by SCDs in the absence of AOC. These results were attributed to the negative interaction among 1,4-D-degraders and the absence of complementary roles of key non-degraders in SCDs. The findings improve our understanding of how interspecies interactions can regulate the intrinsic abilities and functions of coexisting microbes during biodegradation in complex environments and provide valuable guidelines for designing highly efficient and robust microbial consortia for practical bioremediation of 1,4-D like emerging organic contaminants.

Rhizospheric plant-microbe synergistic interactions achieve efficient arsenic phytoextraction by Pteris vittata.

Phytoextraction is a cost-effective and eco-friendly technology to remove arsenic (As) from contaminated soil using plants and associated microorganisms. Pteris vittata is the most studied As hyperaccumulator, which effectively takes up inorganic arsenate via roots. Arsenic solubilization and speciation occur prior to plant absorption in the rhizosphere, which play a key role in As phytoextraction by P. vittata. This study investigated the metabolomic correlation of P. vittata and associated rhizospheric microorganisms during As phytoextraction. Three-month pot cultivation of P. vittata in As polluted soil was conducted. In rhizosphere, an increase of water-soluble As concentration and a decrease of pH was observed in the second month, suggesting acidic metabolites as a possible cause of As solubilization. A correlation network was built to elucidate the interactions among metabolites, bacteria and fungi in the rhizosphere of P. vittata. Our results demonstrate that the plant is the major driving force of rhizospheric microbiota generation, and both microbial community and metabolites in rhizosphere of P. vittata correlate to increased bioavailable As. Multi-omics analysis revealed that pterosins enrich microbes that potentially promote As phytoextraction. This study extends the current view of rhizospheric plant-microbes synergistic effects of hyperaccumulators on phytoextraction, which provides clues for developing efficient As phytoremediation approaches.

Arsenic uptake by Pteris vittata in a subarctic arsenic-contaminated agricultural field in Japan: An 8-year study.

In this study, the phytoremediation potential of tropical and subtropical arsenic (As) hyperaccumulating fern Pteris vittata in an As contaminated farmland field near an abandoned goldmine was investigated. The tested field is located in a subarctic area of northeast Japan. This study was aimed at decreasing the risk of As in the soil (water-soluble As) with nurturing the soil and respecting the plant life cycle for the sustainable phytoremediation for 8 years. The field was tilled and planted with new seedlings of the fern every spring and the grown fern was harvested every autumn. The biomass and As concentration in fronds, rhizomes and roots of the fern were analyzed separately after harvesting each year. The biomass of the fronds of P. vittata was significantly affected by the yearly change of the weather condition, but As concentration in fronds was kept at 100-150 mg/kg dry weight. The accumulated As in P. vittata was higher than that of As-hyperaccumulator fern Pteris cretica, the native fern in the field trial area. Harvested biomass of P. vittata per plant was also higher than that of P. cretica. More than 43.5 g As/154 m2 (convertible to 2.82 kg of As per hectare) was removed from the farmland field by P. vittata phytoremediation at the end of the 8-year experiment. Because of the short-term plant growth period and soil tilling process, total As in soil did not show significant depletion. However, the water-soluble As in the surface and deeper soil, which is phytoavailable and easily taken in cultivated plants, decreased to 10 µg/L (Japan Environmental Quality Standard for water-soluble As in soil) by the 8-year phytoremediation using P. vittata. These research data elucidate that the tropical and subtropical As hyperaccumulating fern, P. vittata, is applicable for As phytoremediation in the subarctic climate area.

Empirical Evidence of Arsenite Oxidase Gene as an Indicator Accounting for Arsenic Phytoextraction by Pteris vittata.

Arsenic (As) is a toxic semi-metallic element that is ubiquitous in the environment and poses serious human health risks. Phytoextraction by Pteris vittata is considered a low-cost and environmentally friendly approach to treat As-contaminated soil. P. vittata mainly absorbs arsenate thus the bioavailability of As to P. vittata depends on the chemical form of As. Microbial redox of As contributes to the biogeochemical cycling of As, and rhizobacterium-assisted phytoextraction by P. vittata was proposed. In this study, this microbe-assisted phytoextraction was applied to two fields, and the effectiveness of phytoextraction was evaluated. The results revealed that P. vittata was able to grow in temperate and subarctic climate zones. The biomass was influenced by the weather, and the As concentration in plants was dependent on the As content in the soil. The ratio of arsenite oxidase genes (aioA-like genes) to 16S rRNA genes was employed to evaluate the effect of As phytoextraction, and the results exhibited that the ratio was related to the As concentration in P. vittata. Our results showed that arsenite oxidation in the rhizosphere might not be achieved by single-strain inoculation, while this study provided empirical evidence that the rhizospheric aioA-like genes could be an indicator for evaluating the effectiveness of As phytoextraction.

Separation of microplastic from soil by centrifugation and its application to agricultural soil.

The increasing contamination of the environment with microplastic requires efficient methods for the separation and detection of these plastic particles. In this work, we present a protocol that uses Fenton oxidation to remove biological material, centrifugation to separate microplastics from soil, and Nile Red staining, fluorescence microscopy, and image processing to detect and quantify of microplastic. The main component of this work was the separation process using centrifugation. All the main polymers used in this work, polyethylene, polypropylene, polystyrene, poly (vinyl chloride), and poly (ethylene terephthalate), were efficiently recovered at more than 94 wt% from heat-altered soil using CaCl2 solution with a density of 1.4 g ml-1. The hydrophilicity of the polymer had a greater effect on the recovery than density. The protocol was then tested on agricultural soil sampled near a contaminated site. The number of microplastic particles was quantified, and the weight of microplastic in the soil was estimated. The highest contamination was observed near the hotspot at a distance of 1 m with 75✕103 particles kg-1, corresponding to a weight between 20 and 6 mg kg-1. The number of particles decreased logarithmically to 30✕103 particles kg-1 or 5 to 2 mg kg-1.

Influence of low temperature on comparative arsenic accumulation and release by three Pteris hyperaccumulators.

Arsenic (As) hyperaccumulator Pteris ferns are renowned for their capacity to accumulate As and have been used to remediate As-contaminated environmental. However, there is less information on how they perform under low temperature though it is important for practical phytoremediation. The purpose of this study was to identify the effect of temperature on As accumulation by three As hyperaccumulators, Pteris multifida, Pteris cretica and Pteris vittata. Ferns were cultured with 5 mg/L As addition under 25 °C to 5 °C for 15 days. The results showed that dropping of temperatures reduced As accumulation by P. vittata moderately but not P. multifida and P. cretica until 10 °C. At 5 °C, all ferns discontinued As accumulation, and the morphology showed necrosis in P. vittata, wherein P. multifida and P. cretica kept healthy. The As distribution showed that As was mainly accumulated in fronds, while P. multifida stored partial As in its root. Both translocation factor and As efflux showed that temperate zone ferns manage As more strictly as compared to P. vittata. Our findings demonstrated that temperature should be considered when applying Pteris ferns for As phytoremediation, and P. multifida could be the most suitable fern for treating As-contaminated water in temperate zone area.

New evidence of arsenic translocation and accumulation in Pteris vittata from real-time imaging using positron-emitting 74As tracer.

Pteris vittata is an arsenic (As) hyperaccumulator plant that accumulates a large amount of As into fronds and rhizomes (around 16,000 mg/kg in both after 16 weeks hydroponic cultivation with 30 mg/L arsenate). However, the sequence of long-distance transport of As in this hyperaccumulator plant is unclear. In this study, we used a positron-emitting tracer imaging system (PETIS) for the first time to obtain noninvasive serial images of As behavior in living plants with positron-emitting 74As-labeled tracer. We found that As kept accumulating in rhizomes as in fronds of P. vittata, whereas As was retained in roots of a non-accumulator plant Arabidopsis thaliana. Autoradiograph results of As distribution in P. vittata showed that with low As exposure, As was predominantly accumulated in young fronds and the midrib and rachis of mature fronds. Under high As exposure, As accumulation shifted from young fronds to mature fronds, especially in the margin of pinna, which resulted in necrotic symptoms, turning the marginal color to gray and then brown. Our results indicated that the function of rhizomes in P. vittata was As accumulation and the regulation of As translocation to the mature fronds to protect the young fronds under high As exposure.

Isolation and Characterization of Novel Bacteria Capable of Degrading 1,4-Dioxane in the Presence of Diverse Co-Occurring Compounds.

Biodegradation is found to be a promising, cost-effective and eco-friendly option for the treatment of industrial wastewater contaminated by 1,4-dioxane (1,4-D), a highly stable synthetic chemical and probable human carcinogen. This study aimed to isolate, identify, and characterize metabolic 1,4-D-degrading bacteria from a stable 1,4-D-degrading microbial consortium. Three bacterial strains (designated as strains TS28, TS32, and TS43) capable of degrading 1,4-D as a sole carbon and energy source were isolated and identified as Gram-positive Pseudonocardia sp. (TS28) and Gram-negative Dokdonella sp. (TS32) and Afipia sp. (TS43). This study, for the first time, confirmed that the genus Dokdonella is involved in the biodegradation of 1,4-D. The results reveal that all of the isolated strains possess inducible 1,4-D-degrading enzymes and also confirm the presence of a gene encoding tetrahydrofuran/dioxane monooxygenase (thmA/dxmA) belonging to group 5 soluble di-iron monooxygenases (SDIMOs) in both genomic and plasmid DNA of each of the strains, which is possibly responsible for the initial oxidation of 1,4-D. Moreover, the isolated strains showed a broad substrate range and are capable of degrading 1,4-D in the presence of additional substrates, including easy-to-degrade compounds, 1,4-D biodegradation intermediates, structural analogs, and co-contaminants of 1,4-D. This indicates the potential of the isolated strains, especially strain TS32, in removing 1,4-D from contaminated industrial wastewater containing additional organic load. Additionally, the results will help to improve our understanding of how multiple 1,4-D-degraders stably co-exist and interact in the consortium, relying on a single carbon source (1,4-D) in order to develop an efficient biological 1,4-D treatment system.

Long-term effectiveness of microbe-assisted arsenic phytoremediation by Pteris vittata in field trials.

Phytoremediation is a promising inexpensive method of detoxifying arsenic (As) contaminated soils using plants and associated soil microorganisms. The potential of Pteris vittata to hyperaccumulate As contamination has been investigated widely. Since As(V) is efficiently taken up by P. vittata than As(III), As speciation by associated rhizobacteria could offer enormous possibility to enhance As phytoremediation. Specifically, increased rhizobacteria mediated As(III) to As(V) conversion appeared to be a crucial step in As mobilization and translocation. In this study, Pseudomonasvancouverensis strain m318 with the potential to improve As phytoremediation was inoculated to P. vittata in a field trial for three years to evaluate its long-term efficacy and stability for enhancing As phytoextraction. The biomass, As concentration, and As accumulation of ferns showed to be increased by inoculation treatment. Although this trend occasionally declined which may be accounted to lower As concentration in soil and amount of precipitation during experiments, the potential of inoculation was observed in increased enrichment coefficients. Further, the arsenite oxidase (aioA-like) genes in the rhizosphere were detected to evaluate the influence of inoculation on As phytoremediation. The findings of this study suggested the potential application of rhizosphere regulation to improve phytoremediation technologies for As contaminated soils. However, the conditions which set the efficacy of this method could be further optimized.

Hydroponic approach to assess rhizodegradation by sudangrass (Sorghum x drummondii) reveals pH- and plant age-dependent variability in bacterial degradation of polycyclic aromatic hydrocarbons (PAHs).

Rhizodegradation of polycyclic aromatic hydrocarbons (PAHs) is a product of complex interactions between plant and bacteria. In this study, hydroponic culture of sudangrass was established in order to investigate the effects of the plant on PAHs degradation and vice versa through changes in rhizosphere bacterial community. Results showed a plant-induced variability in PAHs degradation dependent on a characteristic shift in bacterial community, with pH and plant age as driving factors. Moreover, bacterial communities with high diversity seemed to abate the phytotoxic effects of PAHs degradation as observed in the plant's gross health. Polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and next-generation sequencing revealed that regardless of plant age and culture conditions, the increase or decrease of Sphingobium sp. could dictate the PAHs degradation potential of the bacterial consortium. Overall, this study utilized hydroponic culture of sudangrass to show that plant even of same species can suppress, support, or enhance PAHs degradation of bacteria depending on specific factors.

A multifunctional rhizobacterial strain with wide application in different ferns facilitates arsenic phytoremediation.

Pteris vittata and Pteris multifida are widely studied As hyperaccumulators that absorb As mainly via roots. Hence, rhizobacteria exhibit promising potential in phytoextraction owing to their immense microbial diversity and interactions with plants. Pseudomonas vancouverensis strain m318 that contains aioA-like genes was screened from P. multifida's rhizosphere through the high As resistance (minimum inhibitory concentrations (MICs) against As(III): 16 mM; MICs against As(V): 320 mM), rapid As oxidation (98% oxidation by bacterial cultures (OD600nm = 1) from 200 µL of 0.1 mM As(III) within 24 h), predominant secretion of IAA (12.45 mg L-1) and siderophores (siderophore unit: 88%). Strain m318 showed significant chemotactic response and high colonization efficiency to P. vittata roots, which suggested its wide host affinity. Interestingly, inoculation with strain m318 enhanced the proportion of aioA-like genes in the rhizosphere. And in field trials, inoculation with strain m318 increased As accumulation in P. vittata by 48-146% and in P. multifida by 42-233%. Post-transplantation inoculations also increased As accumulation in both ferns. The abilities of the isolated multifunctional strain m318 and the increase in the rhizosphere microbial aioA-like genes are thus speculated to be involved in As transformation in the rhizospheres and roots of P. vittata and P. multifida.

Cupriavidus basilensis strain r507, a toxic arsenic phytoextraction facilitator, potentiates the arsenic accumulation by Pteris vittata.

As a toxic and carcinogenic metalloid, arsenic has posed serious threat to human health. Phytoremediation has emerged as a promising approach to circumvent this problem. Arsenic uptake by Pteris vittata is largely determined by arsenic speciation and mainly occurs via roots; thus, rhizospheric microbial activities may play a key role in arsenic accumulation. The aim of this study was to investigate the potential of arsenic resistant rhizobacteria to enhance arsenic phytoextraction. A total of 49 cultivable rhizobacteria were isolated from the arsenic hyperaccumulating fern, Pteris vittata, and subjected to an initial analysis to identify potentially useful traits for arsenic phytoextraction, such as arsenic resistance and the presence of aioA(aroA)-like (arsenite oxidase-like) gene. Isolated strain r507, named as Cupriavidus basilensis strain r507, was a selected candidate for its outstanding arsenic tolerance, rapid arsenite oxidation ability, and strong colonization to P. vittata. Strain r507 was used in co-cultivation trials with P. vittata in the field for six months. Results showed that the inoculation with strain r507 potentiated As accumulation of P. vittata up to 171%. Molecular analysis confirmed that the inoculation increased the abundance of aioA-like genes in the rhizosphere, which might have facilitated arsenite oxidation and absorption. The findings of this study suggested the feasibility of co-cultivating hyperaccumulators with facilitator bacteria for practical arsenic phytoremediation.

Enrichment and Analysis of Stable 1,4-dioxane-Degrading Microbial Consortia Consisting of Novel Dioxane-Degraders.

Biodegradation of 1,4-dioxane, a water contaminant of emerging concern, has drawn substantial attention over the last two decades. A number of dioxane-degraders have been identified, though many of them are unable to metabolically utilize 1,4-dioxane. Moreover, it is considered more preferable to use microbial consortia rather than the pure strains, especially in conventional bioreactors for industrial wastewater treatment. In the present study, a stable 1,4-dioxane-degrading microbial consortium was enriched, namely 112, from industrial wastewater by nitrate mineral salt medium (NMSM). The consortium 112 is capable of utilizing 1,4-dioxane as a sole carbon and energy source, and can completely degrade 1,4-dioxane up to 100 mg/l. From the consortium 112, two 1,4-dioxane-degrading bacterial strains were isolated and identified, in which the Variovorax sp. TS13 was found to be a novel 1,4-dioxane-degrader that can utilize 100 mg/l of 1,4-dioxane. The efficacy of the consortium 112 was increased significantly when we cultured the consortium with mineral salt medium (MSM). The new consortium, N112, could utilize 1,4-dioxane at a rate of 1.67 mg/l·h. The results of the ribosomal RNA intergenic spacer analysis (RISA) depicted that changes in the microbial community structure of consortium 112 was the reason behind the improved degradation efficiency of consortium N112, which was exhibited as a stable and effective microbial consortium with a high potential for bioremediation of the dioxane-impacted sites and contaminated industrial wastewater.

Identification of A Novel Arsenic Resistance Transposon Nested in A Mercury Resistance Transposon of Bacillus sp. MB24.

A novel TnMERI1-like transposon designated as TnMARS1 was identified from mercury resistant Bacilli isolated from Minamata Bay sediment. Two adjacent ars operon-like gene clusters, ars1 and ars2, flanked by a pair of 78-bp inverted repeat sequences, which resulted in a 13.8-kbp transposon-like fragment, were found to be sandwiched between two transposable genes of the TnMERI1-like transposon of a mercury resistant bacterium, Bacillus sp. MB24. The presence of a single transcription start site in each cluster determined by 5'-RACE suggested that both are operons. Quantitative real time RT-PCR showed that the transcription of the arsR genes contained in each operon was induced by arsenite, while arsR2 responded to arsenite more sensitively and strikingly than arsR1 did. Further, arsenic resistance complementary experiments showed that the ars2 operon conferred arsenate and arsenite resistance to an arsB-knocked out Bacillus host, while the ars1 operon only raised arsenite resistance slightly. This transposon nested in TnMARS1 was designated as TnARS1. Multi-gene cluster blast against bacteria and Bacilli whole genome sequence databases suggested that TnMARS1 is the first case of a TnMERI1-like transposon combined with an arsenic resistance transposon. The findings of this study suggested that TnMERI1-like transposons could recruit other mobile elements into its genetic structure, and subsequently cause horizontal dissemination of both mercury and arsenic resistances among Bacilli in Minamata Bay.