Arsenic (As) biovolatilization is an important component of the global As biogeochemical cycle. Soils can emit various methylarsine gases, but the underlying microbial processes remain unclear. Here, we show that the addition of molybdate (Mo), an inhibitor of sulfate-reducing bacteria, greatly enhanced dimethylarsine evolution from dimethylarsenate [DMAs(V)] added to two paddy soils. Molybdate addition significantly affected the microbial community structure. The aerobic enrichment cultures from both soils volatilized substantial amounts of dimethylarsine from DMAs(V) in the presence of Mo, whereas the anaerobic enrichment cultures did not. A Bacillus strain (CZ-2) capable of reducing DMAs(V) to dimethylarsine was isolated from the aerobic enrichment culture, and its volatilization ability was enhanced by Mo. RNA-seq analysis identified 10 reductase genes upregulated by Mo. Addition of the reducing agent NADH increased dimethylarsine volatilization by strain CZ-2, suggesting that DMAs(V) reductase is an NADH-dependent enzyme. The strain could not methylate arsenite or convert monomethylarsenate and DMAs(V) to trimethylarsine. Our results show that dimethylarsine evolution from DMAs(V) is independent of the As methylation pathway and that Mo enhances dimethylarsine evolution from paddy soils by shifting the microbial community structure and enhancing the reduction of DMAs(V) to dimethylarsine, possibly through upregulating the expression of DMAs(V) reductase gene(s).

Cadmium (Cd), a common toxic heavy metal in soil, has relatively high bioavailability, which seriously threatens agricultural products. In this study, 8 different soils with contrasting soil properties were collected from different regions in China to investigate the Cd transfer coefficient from soil to Chinese cabbage (Brassica chinensis L.) and the threshold levels of Cd in soils for production of Chinese cabbage according to the food safety standard for Cd. Exogenous Cd (0–4 mg kg−1) was added to the soils and equilibrated for 2 weeks before Chinese cabbage was grown under greenhouse conditions. The influence of soil properties on the relationship between soil and cabbage Cd concentrations was investigated. The results showed that Cd concentration in the edible part of Chinese cabbage increased linearly with soil Cd concentration in 5 soils, but showed a curvilinear pattern with a plateau at the highest dose of exogenous Cd in the other 3 soils. The Cd transfer coefficient from soil to plant varied significantly among the different soils and decreased with increasing soil pH from 4.7 to 7.5. However, further increase in soil pH to > 8.0 resulted in a significant decrease in the Cd transfer coefficient. According to the measured Cd transfer coefficient and by reference to the National Food Safety Standards of China, the safety threshold of Cd concentration in soil was predicted to be between 0.12 and 1.7 mg kg−1 for the tested soils. The predicted threshold values were higher than the current soil quality standard for Cd in 5 soils, but lower than the standard in the other 3 soils. Regression analysis showed a significant positive relationship between the predicted soil Cd safety threshold value and soil pH in combination with soil organic matter or clay content.

•Reductive arsenic mobilization differed by > 100 times among six paddy soils with similar total As.•Oxalate-extractable As was a good indicator of potentially mobilizable As.•Indigenous soil Mn oxide content influenced As speciation and mobilization in paddy soil.•Addition of synthetic Mn oxide decreased reductive As mobilization and As uptake by rice.Reductive mobilization of arsenic (As) in paddy soils under flooded conditions is an important reason for the relatively high accumulation of As in rice, posing a risk to food safety and human health. The extent of As mobilization varies widely among paddy soils, but the reasons are not well understood. In this study, we investigated As mobilization in six As-contaminated paddy soils (total As ranging from 73 to 122 mg kg−1) in flooded incubation and pot experiments. Arsenic speciation in the solution and solid phases were determined. The magnitude of As mobilization into the porewater varied by > 100 times among the six soils. Porewater As concentration correlated closely with the concentration of oxalate-extractable As, suggesting that As associated with amorphous iron (oxyhydr)oxides represents the potentially mobilizable pool of As under flooded conditions. Soil containing a high level of manganese oxides showed the lowest As mobilization, likely because Mn oxides retard As mobilization by slowing down the drop of redox potential upon soil flooding and maintaining a higher arsenate to arsenite ratio in the solid and solution phases. Additions of a synthetic Mn oxide (hausmannite) to two paddy soils increased arsenite oxidation, decreased As mobilization into the porewater and decreased As concentrations in rice grain and straw. Consistent with previous studies using simplified model systems or pure mineral phases, the present study shows that Mn oxides and amorphous Fe (oxyhydr)oxides are important factors controlling reductive As mobilization in As-contaminated paddy soils. In addition, this study also suggests a potential mitigation strategy using exogenous Mn oxides to decrease As uptake by rice in paddy soils containing low levels of indigenous Mn oxides, although further work is needed to verify its efficacy and possible secondary effects under field conditions.Download high-res image (467KB)Download full-size image

Rice is a major dietary source of cadmium (Cd) and arsenic (As) for populations consuming rice as the staple food. Excessive Cd and As accumulation in rice grain is of great concern worldwide, especially in South China where soil contamination with heavy metals and metalloids is widespread. It is important to reduce Cd and As accumulation in rice grain through selection and breeding of cultivars accumulating low levels of Cd or As.To assess the genetic and environmental variations in the concentrations of Cd and As in rice grains, 471 locally adapted high-yielding rice cultivars were grown at three moderately contaminated sites in South China for two years. Cadmium and As concentrations in brown rice varied by 10 – 32 and 2.5 – 4 fold, respectively. Genotype (G), environment (E) and G x E interactions were highly significant factors explaining the variations. Brown rice Cd concentration was found to correlate positively with the heading date among different cultivars, whereas As concentration and heading date correlated negatively. There was a significant and negative correlation between grain Cd and As concentrations.Eight and 6 rice cultivars were identified as stable low accumulators of Cd and As, respectively, based on the multiple site and season trials. These cultivars are likely to be compliant with the grain Cd or As limits of the Chinese Food Safety Standards when grown in moderately contaminated paddy soils in South China.

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.

Arsenic (As) contamination in soil can lead to elevated transfer of As to the food chain. One potential mitigation strategy is to genetically engineer plants to enable them to transform inorganic As to methylated and volatile As species. In this study, we genetically engineered two ecotypes of Arabidopsis thaliana with the arsenite (As(III)) S-adenosylmethyltransferase (arsM) gene from the eukaryotic alga Chlamydomonas reinhardtii. The transgenic A. thaliana plants gained a strong ability to methylate As, converting most of the inorganic As into dimethylarsenate [DMA(V)] in the shoots. Small amounts of volatile As were detected from the transgenic plants. However, the transgenic plants became more sensitive to As(III) in the medium, suggesting that DMA(V) is more phytotoxic than inorganic As. The study demonstrates a negative consequence of engineered As methylation in plants and points to a need for arsM genes with a strong ability to methylate As to volatile species.

Land applications of municipal sewage sludge may pose a risk of introducing antibiotic resistance genes (ARGs) from urban environments into agricultural systems. However, how the sewage sludge recycling and application method influence soil resistome and mobile genetic elements (MGEs) remains unclear. In the present study, high through-put quantitative PCR was conducted on the resistome of soils from a field experiment with past (between 1994 and 1997) and annual (since 1994) applications of five different sewage sludges. Total inputs of organic carbon were similar between the two modes of sludge applications. Intrinsic soil resistome, defined as the ARGs shared by the soils in the control and sludge-amended plots, consisted of genes conferring resistance to multidrug, β-lactam, Macrolide-Lincosamide-Streptogramin B (MLSB), tetracycline, vancomycin, and aminoglycoside, with multidrug resistance genes as the most abundant members. There was a strong correlation between the abundance of ARGs and MGE marker genes in soils. The composition and diversity of ARGs in the five sludges were substantially different from those in soils. Considerable proportions of ARGs and MGE marker genes in the sludges attenuated following the application, especially aminoglycoside and tetracycline resistance genes. Annual applications posed a more significant impact on the soil resistome, through both continued introduction and stimulation of the soil intrinsic ARGs. In addition, direct introduction of sludge-specific ARGs into soil was observed especially from ARG-rich sludge. These results provide a better insight into the characteristics of ARG dissemination from urban environment to the agricultural system through sewage sludge applications.

Tea is a major dietary source of aluminium (Al) to humans. There is a need to understand how environmental factors influence Al accumulation in tea leaves in order to devise strategies to lower Al intake from tea.Paired soil and tea shoots were collected from 197 micro-locations across 11 tea plantation estates in Kericho, Kenya. The concentrations of Al and other minerals in tea samples and a range of soil properties were determined. Multiple regression analysis was performed to identify environmental variables that significantly affected tea Al concentration. A comparison was made between washed and unwashed tea samples. Tea samples from a long-term lime and elemental sulphur trial were analysed.Four environmental variables were identified as having a significant positive effect on tea Al concentration, including soil dust contamination represented by tea Ti concentration, the age of plantation, the number of years after pruning and Al saturation in topsoil, whereas two variables had a significant negative effect, including the altitude of plantation and rainfall prior to tea plucking. The effect of soil dust contamination was further verified by a leaf washing experiment. Soil pH was found to have no significant effect on tea Al concentration in both the field survey and the lime and sulphur experiment.Liming would not be an effective method to decrease Al concentration in tea. Avoiding soil dust contamination, plucking tea shoots from plantations at higher altitudes or from younger plantations could lower tea Al concentration.

Tea is a strong accumulator of both aluminium (Al) and fluoride (F). We tested the hypothesis that Al helps detoxify F in tea plants by forming Al-F complexes.Tea plants were grown hydroponically with a range of Al and F concentrations and in a soil pot experiment with amendments of NaF and acids. Growth and the uptake of F and Al were determined. Chemical species of F in the nutrient solutions and the cell saps of roots and leaves were determined by 19F NMR.In hydroponic experiments, F inhibited the growth of new roots and shoot tips in the absence of Al in the nutrient solutions, whereas Al stimulated root growth and alleviated the toxicity of F. Aluminium generally increased F concentration in roots but decreased F concentration in leaves. Geochem-PC calculations and 19F NMR showed the presence of AlF2+, AlF2+ and AlF30 in the nutrient solutions when Al and F were present. Aluminium markedly decreased the NMR peak of free F in the cell saps from roots and leaves. An Al-F complex likely to be AlF2+ was detected in the leaf cell sap from the plants treated with both F and Al. In the soil pot experiment, F caused leaf necrosis when the leaf Al to F molar ratio was smaller than 1.Tea plants are sensitive to F toxicity in the absence of Al. Aluminium alleviates F toxicity in tea by forming Al-F complexes.

Plants are able to take up inorganic arsenic (As) and methylated As, but whether the mode of phytotoxicity and the detoxification mechanism differ between different As species remains unclear. This study aimed to investigate the differences in phytotoxicity and detoxification mechanism between arsenate [As(V)], monomethylarsonic acid [MMA(V)] and dimethylarsinic acid [DMA(V)].Arabidopsis thaliana was grown in agar-solidified medium, hydroponic or soil pot experiments. Root and shoot growth, seed production, As accumulation and oxidative stress indicators of wild-type plants exposed to As(V), MMA(V) and DMA(V) were compared. The role of thiols in As detoxification was investigated using a specific inhibitor of glutathione (GSH) biosynthesis and the mutants defective in GSH synthesis (cad2-1), phytochelatin (PC) synthesis (cad1-3) or tonoplast transporters for As(III)-PCs (abcc1-2).Methylated As species, especially DMA(V), were more toxic than As(V) for growth and seed production in A. thaliana. Methylated As species were more efficiently translocated from roots to shoots and from shoots to seeds than As(V). DMA(V) exposure resulted in a greater oxidative stress than other As species. As(V) and MMA(V) induced the production of non-protein thiols (NPT), but DMA(V) did not. The GSH inhibitor BSO greatly enhanced the sensitivity to As(V) and MMA(V), but decreased the sensitivity to DMA(V). The mutants cad1-3, cad2-1 and abcc1-2 were similarly hypersensitive to As(V) and MMA(V), but not to DMA(V). As(V) and MMA(V) enhanced the expression of the sulphur assimilation genes encoding ATP Sulphurylase (ATPS) and adenosine-5′-phosphosulphate reductase (APR1) more than DMA(V).DMA(V) is more toxic to A. thaliana than As(V) or MMA(V). The detoxification mechanism for MMA(V) is similar to that for As(V), involving thiol production, complexation with PCs and vacuolar sequestration. This mechanism is ineffective for the detoxification of DMA(V).

China faces great challenges in protecting its soil from contamination caused by rapid industrialization and urbanization over the last three decades. Recent nationwide surveys show that 16% of the soil samples, 19% for the agricultural soils, are contaminated based on China’s soil environmental quality limits, mainly with heavy metals and metalloids. Comparisons with other regions of the world show that the current status of soil contamination, based on the total contaminant concentrations, is not worse in China. However, the concentrations of some heavy metals in Chinese soils appear to be increasing at much greater rates. Exceedance of the contaminant limits in food crops is widespread in some areas, especially southern China, due to elevated inputs of contaminants, acidic nature of the soil and crop species or cultivars prone to heavy metal accumulation. Minimizing the transfer of contaminants from soil to the food chain is a top priority. A number of options are proposed, including identification of the sources of contaminants to agricultural systems, minimization of contaminant inputs, reduction of heavy metal phytoavailability in soil with liming or other immobilizing materials, selection and breeding of low accumulating crop cultivars, adoption of appropriate water and fertilizer management, bioremediation, and change of land use to grow nonfood crops. Implementation of these strategies requires not only technological advances, but also social-economic evaluation and effective enforcement of environmental protection law.

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.

Cereals are an important source of iron (Fe) and zinc (Zn) for humans, but their concentrations are generally low in white flour (which corresponds to the starchy endosperm) and their bioavailability is limited by high phytate content in the bran. The aim of this study was to investigate whether the phytate content of wheat grain affects the distribution of Fe and Zn to the bran and starchy endosperm.The stable isotopes 57Fe and 68Zn were applied to the flag leaves of four wheat lines differing in their phytate content. The isotopes were also applied to roots of a low phytate line. Isotopic compositions in bran, starchy endosperm and straw were quantified by inductively coupled plasma mass spectrometry.The low phytate line contained approximately half of the phytate-phosphorus concentration of the other three lines. Foliar applications of 57Fe and 68Zn increased the isotopic ratios 57Fe/56Fe and 68Zn/66Zn more in the bran than in the starchy endosperm. The low phytate line did not have a greater distribution of the isotope labels toward the starchy endosperm. The four wheat lines differed in the distribution of 57Fe and 68Zn from the flag leaves to the rest of the vegetative tissues and the grain. Little of the isotopes applied to roots at the anthesis stage reached the grain.57Fe and 68Zn were preferentially distributed to the bran of wheat grain. Low phytate content in the bran does not appear to facilitate the distribution of 57Fe and 68Zn into the starchy endosperm.

Increasing nitrogen supply can increase Fe and Zn concentrations in wheat grain, but the underlying mechanisms remain unclear. Size-exclusion chromatography coupled with inductively coupled plasma mass spectrometry was used to determine Fe and Zn speciation in the soluble extracts of grain pearling fractions of two wheat cultivars grown at two N rates (100 and 350 kg of N ha–1). Increasing N supply increased the concentrations of total Fe and Zn and the portions of Fe and Zn unextractable with a Tris–HCl buffer and decreased the concentrations of Tris–HCl-extractable (soluble) Fe and Zn. Within the soluble fraction, Fe and Zn bound to low molecular weight compounds, likely to be Fe–nicotianamine and Fe–deoxymugineic acid or Zn–nicotianamine, were decreased by 5–12% and 4–37%, respectively, by the high N treatment, whereas Fe and Zn bound to soluble high molecular weight or soluble phytate fractions were less affected. The positive effect of N on grain Fe and Zn concentrations was attributed to an increased sink in the grain, probably in the form of water-insoluble proteins.

This study aimed to determine differences among wheat cultivars in the distribution and speciation of Fe and Zn in grain milling fractions. Cultivars with higher Fe and Zn concentrations in the wholemeal flour were found to contain higher concentrations in the white flour. Soluble Fe and Zn were extracted and analyzed by size exclusion–inductively coupled plasma mass spectrometry. Fe speciation varied between milling fractions with a low molecular weight (LMW) complex likely to be Fe–deoxymugenic acid/nicotianamine being the predominant extractable Fe species in white flour, accounting for approximately 85% of the extractable Fe. Bran fractions had a lower amount of LMW-Fe form but more as soluble Fe–phytate and an unidentified high molecular weight peak. In the white flour fraction soluble Zn was found to be present mainly as a LMW peak likely to be Zn–nicotianamine complex. Soluble Fe–phytate was found in the white flour fraction of a high-Fe cultivar but not in a low-Fe cultivar.

Rice is a major source of inorganic arsenic (iAs) in the human diet because paddy rice is efficient at accumulating As. Rice As speciation is dominated by iAs and dimethylarsinic acid (DMA). Here we review the global pattern in rice As speciation and the factors causing the variation. Rice produced in Asia shows a strong linear relationship between iAs and total As concentration with a slope of 0.78. Rice produced in Europe and the United States shows a more variable, but generally hyperbolic relationship with DMA being predominant in U.S. rice. Although there is significant genotypic variation in grain As speciation, the regional variations are primarily attributed to environmental factors. Emerging evidence also indicates that methylated As species in rice are derived from the soil, while rice plants lack the As methylation ability. Soil flooding and additions of organic matter increase microbial methylation of As, although the microbial community responsible for methylation is poorly understood. Compared with iAs, methylated As species are taken up by rice roots less efficiently but are transported to the grain much more efficiently, which may be an important factor responsible for the spikelet sterility disorder (straight-head disease) in rice. DMA is a weak carcinogen, but the level of ingestion from rice consumption is much lower than that of concern. Questions that require further investigations are identified.

Methylation of arsenic in soil influences its environmental behavior and accumulation by plants, but little is known about the factors affecting As methylation. As speciation was determined in the pore waters of six soils from diverse geographical locations over 54 days of incubation under flooded conditions. The concentration of methylated As (monomethylarsonic acid, MMA, and dimethylarsinic acid, DMA) varied from 0 to 85 μg L–1 (0 – 69% of the total As in pore water). Two Bangladeshi paddy soils contaminated by irrigation of As-laden groundwater produced large concentrations of inorganic As but relatively little methylated As. Two contaminated paddy soils from China produced a transient peak of DMA during the early phase of incubation. Methylated As represented considerable proportions of the total soluble As in the two uncontaminated soils from the UK and U.S. The copy number of the microbial arsenite methyltransferase gene (arsM) correlated positively with soil pH. However, pore-water methylated As correlated negatively with pH or arsM copy number, and positively with dissolved organic C. GeoChip assay revealed considerable arsM diversity among the six soils, with 27–35 out of 66 sequences in the microarray being detected. As speciation in rice plants grown in the soils generally mirrored that in the pore water. The results suggest that methylated As species in plants originated from the soil and As methylation in soil was influenced strongly by the soil conditions.

The role of lateral roots in the acquisition of nutrients and contaminants from the soil may vary between mobile and immobile solutes. The aim of the present study was to quantify the contributions of lateral roots to growth and elemental uptake under different conditions.A lateral rootless mutant of rice (Oryza sativa) with a gain-of-function mutation in OsIAA11 was compared with its wild-type (WT) in hydroponic, pot and field conditions. Three soils varying in the P availability were used in the pot experiment. Synchrotron fast X-ray fluorescence (XRF) was used to map the distribution of trace elements in fresh hydrated roots.The Osiaa11 mutant grew smaller compared with the WT in all three experiments, especially in the field. The difference was larger in a P-sufficient soil than in P-deficient soils in the pot experiment. Elemental concentrations in the roots and shoots were affected differently by the mutation, depending on the elements and the growth media. The apparent contributions of lateral roots to elemental uptake varied from 2.7 to 82.5% in the hydroponic experiment, from −19.8 to 76.4% in the pot experiment, and from 30 to 76% in the field experiment. In general, the apparent contributions to the uptake of P, Mn, Zn, Cu and As were larger than that for the biomass, whereas for N, S and K the apparent contributions were smaller than or similar to the effect on plant biomass. Synchrotron XRF revealed strong accumulation of Mn, Zn, Cu, As and Se in the lateral roots of the WT.Lateral roots play an important role in the acquisition of less mobile elements such as P, Mn, Zn, Cu and As, but have relatively small effects on the acquisition of mobile elements such as N, S and K.