Arsenic-contaminated Paddy Soil Research Articles

Tracing the Dynamic Changes of Element Profiles by Novel Soil Porewater Samplers with Ultralow Disturbance to Soil-Water Interface.

In flooded soils, soil–water interface (SWI) is the key zone controlling biogeochemical dynamics. Chemical species and concentrations vary greatly at micro- to cm-scales. Techniques able to track these changing element profiles both in space and over time with appropriate resolution are rare. Here, we report a patent-pending technique, the Integrated Porewater Injection (IPI) sampler, which is designed for soil porewater sampling with minimum disturbance to saturated soil environment. IPI sampler employs a single hollow fiber membrane tube to passively sample porewater surrounding the tube. When working, it can be integrated into the sample introduction system, thus the sample preparation procedure is dramatically simplified. In this study, IPI samplers were coupled to ICP-MS at data-only mode. The limits of detection of IPI-ICP-MS for Ni, As, Cd, Sb, and Pb were 0.12, 0.67, 0.027, 0.029, and 0.074 μg·L–1, respectively. Furthermore, 25 IPI samplers were assembled into an SWI profiler using 3D printing in a one-dimensional array. The SWI profiler is able to analyze element profiles at high spatial resolution (∼2 mm) every ≥24 h. When deployed in arsenic-contaminated paddy soils, it depicted the distributions and dynamics of multiple elements at anoxic–oxic transition. The results show that the SWI profiler is a powerful and robust technique in monitoring dynamics of element profile in soil porewater at high spatial resolution. The method will greatly facilitate studies of elements behaviors in sediments of wetland, rivers, lakes, and oceans.

Humic Substances Facilitate Arsenic Reduction and Release in Flooded Paddy Soil.

Organic matter is important for controlling arsenic reduction and release under anoxic conditions. Humic substances (HS) represent an important fraction of natural organic matter, yet the manner in which HS affect arsenic transformation in flooded paddy soil has not been thoroughly elucidated. In this study, anaerobic microcosms were established with arsenic-contaminated paddy soil and amended with three extracted humic fractions (fulvic acid, FA; humic acid, HA; and humin, HM). The HS substantially enhanced the extent of arsenic reduction and release in the order FA > HA > HM. It was confirmed that microbially reduced HS acted as an electron shuttle to promote arsenate reduction. HS, particularly FA, provided labile carbon to stimulate microbial activity and increase the relative abundances of Azoarcus, Anaeromyxobacter, and Pseudomonas, all of which may be involved in the reduction of HS, Fe(III), and arsenate. HS also increased the abundance of transcripts for an arsenate-respiring gene ( arrA) and overall transcription in arsenate-respiring Geobacter spp. The increase in both abundances lagged behind the increases in dissolved arsenate levels. These results help to elucidate the pathways of arsenic reduction and release in the presence of HS in flooded paddy soil.

Biotransformation of arsenic-containing roxarsone by an aerobic soil bacterium Enterobacter sp. CZ-1

Roxarsone (3-nitro-4-hydroxyphenylarsonic acid, ROX) is an arsenic-containing compound widely used as a feed additive in poultry industries. ROX excreted in chicken manure can be transformed by microbes to different arsenic species in the environment. To date, most of the studies on microbial transformation of ROX have focused on anaerobic microorganisms. Here, we isolated a pure cultured aerobic ROX-transforming bacterial strain, CZ-1, from an arsenic-contaminated paddy soil. On the basis of 16S rRNA gene sequence, strain CZ-1 was classified as a member of the genus Enterobacter. During ROX biotransformation by strain CZ-1, five metabolites including arsenate (As[V]), arsenite (As[III]), N-acetyl-4-hydroxy-m-arsanilic acid (N-AHPAA), 3-amino-4-hydroxyphenylarsonic acid (3-AHPAA) and a novel sulfur-containing arsenic species (AsC9H13N2O6S) were detected and identified based on high-performance liquid chromatography-inductively coupled plasma mass spectrometry (HPLC-ICP-MS), HPLC-ICP-MS/electrospray ionization mass spectrometry (ESI-MS) and HPLC-electrospray ionization hybrid quadrupole time-of-flight mass spectrometry (ESI-qTOF-MS) analyses. N-AHPAA and 3-AHPAA were the main products, and 3-AHPAA could also be transformed to N-AHPAA. Based on the results, we propose a novel ROX biotransformation pathway by Enterobacter. sp CZ-1, in which the nitro group of ROX is first reduced to amino group (3-AHPAA) and then acetylated to N-AHPAA.

Mitigating arsenic accumulation in rice (Oryza sativa L.) from typical arsenic contaminated paddy soil of southern China using nanostructured α-MnO2: Pot experiment and field application

Manganese oxides are naturally occurring powerful oxidants and scavengers, which can control the mobility and bioavailability of arsenic (As). However, the effect of synthetic nanostructured manganese oxides on the mobilization and transportation of As at actual paddy soils are poorly understood, especially in soils with low or medium background Mn concentration. In the present study, a novel nano manganese oxide with superior reactivity and surface area has been synthesized. A 90-d soil incubation experiment combined with pot and field rice cultivation trials were designed to evaluate the effectiveness of exogenous α-MnO2 nanorods on the mobilization and transportation of As in soil-rice systems. Our results proved that the addition of α-MnO2 nanorods can effectively control the soil-to-solution partitioning of As under anaerobic conditions. After treatment with different amounts of α-MnO2 nanorods, the content of effective As decreased, offset by an increase in residual As and insoluble binding As (Ca-As and Fe-As). Enhancing the oxidation of As(III) into As(V), the α-MnO2 nanorods increased the adsorption of As onto indigenous iron (hydr)oxides which greatly reduced the soil porewater As content. In addition, pot experiments and field applications revealed that the influx of As into the aerial parts of rice plants (stems, husk and leaves) was strictly prohibited after treatments with different amount of α-MnO2 nanorods; more interestingly, significantly negative correlations have been observed between As and Mn in rice, which indicated that as Mn is increased in soil, As in brown rice decreases. Our results demonstrated that the use of α-MnO2 nanorods in As polluted paddy soil containing low levels of background Mn oxides can be a promising remediation strategy.

Decreasing arsenic accumulation in rice by overexpressing OsNIP1;1 and OsNIP3;3 through disrupting arsenite radial transport in roots.

Rice is a major dietary source of the toxic metalloid arsenic. Reducing arsenic accumulation in rice grain is important for food safety. We generated transgenic rice overexpressing two aquaporin genes, OsNIP1;1 and OsNIP3;3, under the control of a maize ubiquitin promoter or the rice OsLsi1 promoter, and tested the effect on arsenite uptake and translocation. OsNIP1;1 and OsNIP3;3 were highly permeable to arsenite in Xenopus oocyte assays. Both transporters were localized at the plasma membrane. Knockout of either gene had little effect on arsenite uptake or translocation. Overexpression of OsNIP1;1 or OsNIP3;3 in rice did not affect arsenite uptake but decreased root-to-shoot translocation of arsenite and shoot arsenic concentration markedly. The overexpressed OsNIP1;1 and OsNIP3;3 proteins were localized in all root cells without polarity. Expression of OsNIP1;1 driven by the OsLsi1 promoter produced similar effects. When grown in two arsenic-contaminated paddy soils, overexpressing lines contained significantly lower arsenic concentration in rice grain than the wild-type without compromising plant growth or the accumulation of essential nutrients. Overexpression of OsNIP1;1 or OsNIP3;3 provides a route for arsenite to leak out of the stele, thus restricting arsenite loading into the xylem. This strategy is effective in reducing arsenic accumulation in rice grain.

Transcriptional Activity of Arsenic-Reducing Bacteria and Genes Regulated by Lactate and Biochar during Arsenic Transformation in Flooded Paddy Soil.

Organic substrates and biochar are important in controlling arsenic release from sediments and soils; however, little is known about their impact on arsenic-reducing bacteria and genes during arsenic transformation in flooded paddy soils. In this study, microcosm experiments were established to profile transcriptional activity of As(V)-respiring gene (arrA) and arsenic resistance gene (arsC) as well as the associated bacteria regulated by lactate and/or biochar in anaerobic arsenic-contaminated paddy soils. Chemical analyses revealed that lactate as the organic substrate stimulated microbial reduction of As(V) and Fe(III), which was simultaneously promoted by lactate+biochar, due to biochar's electron shuttle function that facilitates electron transfer from bacteria to As(V)/Fe(III). Sequencing and phylogenetic analyses demonstrated that both arrA closely associated with Geobacter (>60%, number of identical sequences/number of the total sequences) and arsC related to Enterobacteriaceae (>99%) were selected by lactate and lactate+biochar. Compared with the lactate microcosms, transcriptions of the bacterial 16S rRNA gene, Geobacter spp., and Geobacter arrA and arsC genes were increased in the lactate+biochar microcosms, where transcript abundances of Geobacter and Geobacter arrA closely tracked with dissolved As(V) concentrations. Our findings indicated that lactate and biochar in flooded paddy soils can stimulate the active As(V)-respiring bacteria Geobacter species for arsenic reduction and release, which probably increases arsenic bioavailability to rice plants.

Roles of different active metal-reducing bacteria in arsenic release from arsenic-contaminated paddy soil amended with biochar

Although biochar has great potential for heavy metal removal from sediments or soils, its impact on arsenic biogeochemistry in contaminated paddy fields remains poorly characterized. In this study, anaerobic microcosms were established with arsenic-contaminated paddy soil to investigate arsenic transformation as well as the potentially active microbial community and their transcriptional activities in the presence of biochar. The results demonstrated that biochar can simultaneously stimulate microbial reduction of As(V) and Fe(III), releasing high levels of As(III) into the soil solution relative to the control. Total RNAs were extracted to profile the potentially active microbial communities, which suggested that biochar increased the abundance of arsenic- and iron-related bacteria, such as Geobacter, Anaeromyxobacter and Clostridium compared to the control. Reverse transcription, quantitative PCR (RT-qPCR) showed that the abundance of Geobacter transcripts were significantly stimulated by biochar throughout the incubation. Furthermore, significant positive correlations were observed between the abundance of Geobacter transcripts and As(V) concentrations, and between that of Clostridium transcripts and Fe(III) concentrations in biochar-amended microcosms. Our findings suggest that biochar can stimulate the activity of metal-reducing bacteria to promote arsenic mobility. The Geobacter may contribute to As(V) reduction in the presence of biochar, while Clostridium has a role in Fe(III) reduction.

Arsenicibacter rosenii gen. nov., sp. nov., an efficient arsenic methylating and volatilizing bacterium isolated from an arsenic-contaminated paddy soil.

A novel bacterium with strong arsenic (As) methylation and volatilization abilities, designated strain SM-1T, was isolated from an As-contaminated paddy soil. SM-1T is strictly aerobic, rod-shaped, non-motile, Gram-negative and orange-coloured. Phylogenetic analysis revealed that strain SM-1T showed low 16S rRNA gene sequence similarities (≤88 %) to members of established genera in the family Cytophagaceae. Growth of this strain was observed at 15-45 °C (optimum, 37 °C), pH 6.0-8.0 (optimum, 7.0) and 0-0.5 % (w/v) NaCl. The major cellular fatty acids were C16 : 1ω5c and iso-C15 : 0. The respiratory quinone was MK-7, and the predominant polar lipids were phosphatidylethanolamine, an unidentified lipid (L), and an unidentified aminolipid (AL2). The DNA G+C content was 51.5 mol%. On the basis of phenotypic, phylogenetic and chemotaxonomic properties, strain SM-1T represents a novel species in a new genus within the family Cytophagaceae, for which the name Arsenicibacter rosenii gen. nov., sp. nov. is proposed. The type strain of Arsenicibacter rosenii is SM-1T (=CCTCC AB 2017086T=KCTC 52624T).

Efficient Arsenic Methylation and Volatilization Mediated by a Novel Bacterium from an Arsenic-Contaminated Paddy Soil.

Microbial arsenic (As) methylation and volatilization are important processes controlling the As biogeochemical cycle in paddy soils. To further understand these processes, we isolated a novel bacterial strain, SM-1, from an As-contaminated paddy soil. SM-1 showed strong As methylation and volatilization abilities, converting almost all arsenite (10 μM) to dimethylarsenate and trimethylarsenic oxide in the medium and trimethylarsine gas into the headspace within 24 h, with trimethylarsine accounting for nearly half of the total As. On the basis of the 16S rRNA sequence, strain SM-1 represents a new species in a new genus within the family Cytophagaceae. Strain SM-1 is abundant in the paddy soil and inoculation of SM-1 greatly enhanced As methylation and volatilization in the soil. An arsenite methyltransferase gene (ArarsM) was cloned from SM-1. When expressed in Escherichia coli, ArArsM conferred the As methylation and volatilization abilities to E. coli and increased its resistance to arsenite. The high As methylation and volatilization abilities of SM-1 are likely attributed to an efficient ArArsM enzyme coupled with low arsenite efflux. These results suggest that strain SM-1 plays an important role in As methylation and volatilization in the paddy soil and has a great potential for As bioremediation.

The diversity and abundance of As(III) oxidizers on root iron plaque is critical for arsenic bioavailability to rice.

Iron plaque is a strong adsorbent on rice roots, acting as a barrier to prevent metal uptake by rice. However, the role of root iron plaque microbes in governing metal redox cycling and metal bioavailability is unknown. In this study, the microbial community structure on the iron plaque of rice roots from an arsenic-contaminated paddy soil was explored using high-throughput next-generation sequencing. The microbial composition and diversity of the root iron plaque were significantly different from those of the bulk and rhizosphere soils. Using the aoxB gene as an identifying marker, we determined that the arsenite-oxidizing microbiota on the iron plaque was dominated by Acidovorax and Hydrogenophaga-affiliated bacteria. More importantly, the abundance of arsenite-oxidizing bacteria (AsOB) on the root iron plaque was significantly negatively correlated with the arsenic concentration in the rice root, straw and grain, indicating that the microbes on the iron plaque, particularly the AsOB, were actively catalyzing arsenic transformation and greatly influencing metal uptake by rice. This exploratory research represents a preliminary examination of the microbial community structure of the root iron plaque formed under arsenic pollution and emphasizes the importance of the root iron plaque environment in arsenic biogeochemical cycling compared with the soil-rhizosphere biotope.

Anaerobic arsenite oxidation by an autotrophic arsenite-oxidizing bacterium from an arsenic-contaminated paddy soil.

Microbe-mediated arsenic (As) redox reactions play an important role in the biogeochemical cycling of As. Reduction of arsenate [As(V)] generally leads to As mobilization in paddy soils and increased As availability to rice plants, whereas oxidation of arsenite [As(III)] results in As immobilization. A novel chemoautotrophic As(III)-oxidizing bacterium, designated strain SY, was isolated from an As-contaminated paddy soil. The isolate was able to derive energy from the oxidation of As(III) to As(V) under both aerobic and anaerobic conditions using O2 or NO3(-) as the respective electron acceptor. Inoculation of the washed SY cells into a flooded soil greatly enhanced As(III) oxidation to As(V) both in the solution and adsorbed phases of the soil. Strain SY is phylogenetically closely related to Paracoccus niistensis with a 16S rRNA gene similarity of 96.79%. The isolate contains both the denitrification and ribulose 1,5-bisphosphate carboxylase/oxygenase gene clusters, underscoring its ability to denitrify and to fix CO2 while coupled to As(III) oxidation. Deletion of the aioA gene encoding the As(III) oxidase subunit A abolished the As(III) oxidation ability of strain SY and led to increased sensitivity to As(III), suggesting that As(III) oxidation is a detoxification mechanism in this bacterium under aerobic and heterotrophic growth conditions. Analysis of the aioA gene clone library revealed that the majority of the As(III)-oxidizing bacteria in the soil were closely related to the genera Paracoccus of α-Proteobacteria. Our results provide direct evidence for As(III) oxidation by Paracoccus species and suggest that these species may play an important role in As(III) oxidation in paddy soils under both aerobic and denitrifying conditions.

Phytoremediation of arsenic contaminated paddy soils with Pteris vittata markedly reduces arsenic uptake by rice

Arsenic (As) accumulation in food crops such as rice is of major concern. To investigate whether phytoremediation can reduce As uptake by rice, the As hyperaccumulator Pteris vittata was grown in five contaminated paddy soils in a pot experiment. Over a 9-month period P. vittata removed 3.5–11.4% of the total soil As, and decreased phosphate-extractable As and soil pore water As by 11–38% and 18–77%, respectively. Rice grown following P. vittata had significantly lower As concentrations in straw and grain, being 17–82% and 22–58% of those in the control, respectively. Phytoremediation also resulted in significant changes in As speciation in rice grain by greatly decreasing the concentration of dimethylarsinic acid (DMA). In two soils the concentration of inorganic As in rice grain was decreased by 50–58%. The results demonstrate an effective stripping of bioavailable As from contaminated paddy soils thus reducing As uptake by rice.