Coupling of metataxonomics and culturing improves bacterial diversity characterization and identifies a novel Rhizorhapis sp. with metal resistance potential in a multi-contaminated waste sediment.

Long-term contaminated environments have been recognized as potential hotspots for bacterial discovery in taxonomic and functional terms for bioremediation purposes. Here, bacterial diversity in waste sediment collected from a former industrial dumpsite and contaminated with petroleum hydrocarbon and heavy metals was investigated through the parallel application of culture-independent (16S rRNA gene amplicon sequencing) and -dependent (plate culturing followed by colony picking and identification of isolates by 16S rRNA gene Sanger sequencing) approaches. The bacterial diversities retrieved by both approaches greatly differed. Bacteroidetes and Proteobacteria were dominant in the culture-independent community, while Firmicutes and Actinobacteria were the main culturable groups. Only 2.7% of OTUs (operational taxonomic units) in the culture-independent dataset were cultured. Most of the culturable OTUs were absent or in very low abundances in the culture-independent dataset, revealing that culturing is a useful tool to study the rare bacterial biosphere. One culturable OTUs (comprising only the isolate SPR117) was identified as a potential new species in the genus Rhizorhapis (class Alphaproteobacteria) and was selected for further characterization. Phytopathogenicity tests showed that Rhizorhapis sp. strain SPR117 (ATCC TSD-228) is not pathogenic to lettuce, despite the only described species in this genus, Rhizorhapis suberifaciens, is causal agent of the lettuce corky root disease. The genome of the strain SPR117 was sequenced, assembled in 256 contigs, with a length of 4,419,522 bp and a GC content of 59.9%, and its further annotation revealed the presence of genes related to the resistance to arsenic, copper, iron, and mercury, among other metals. Therefore, the coupling of metataxonomics and culturing is a useful tool to obtain not only an improved description of bacterial communities in contaminated environments, but also to isolate microorganisms with bioremediation potential.

Development of a Strategy for Enhancing the Biomass Growth and Lipid Accumulation of Chlorella sp. UJ-3 Using Magnetic Fe3O4 Nanoparticles.

In this study, magnetic Fe3O4 nanoparticles (NPs) were used as an effective enhancer to increase the biomass and total lipid production of Chlorella sp. UJ-3. It was found that the biomass of algal cells increased significantly when they were exposed to low concentrations of Fe3O4 NPs (20 mg/L), while the best total lipid content of algal cells was achieved when they were exposed to high concentrations of Fe3O4 NPs (100 mg/L). Therefore, we established a strategy to promote the growth and lipid accumulation of microalgae by initially exposing the algal cells to low concentrations of Fe3O4 NPs and then treating them with an increased concentration of Fe3O4 NPs after 12 days of culture. For this strategy, the biomass and total lipid production of algal cells increased by 50% and 108.7%, respectively, compared to the untreated control. The increase in lipid production and change in the fatty acid composition of Chlorella cells were found to help them to cope with the increased number of reactive oxygen species produced due to oxidative stress in alga cells after the addition of Fe3O4 NPs. This study provided a highly efficient way to improve the lipid production of microalgae using nanoparticles.

Horizontal 'gene drives' harness indigenous bacteria for bioremediation.

Engineering bacteria to clean-up oil spills is rapidly advancing but faces regulatory hurdles and environmental concerns. Here, we develop a new technology to harness indigenous soil microbial communities for bioremediation by flooding local populations with catabolic genes for petroleum hydrocarbon degradation. Overexpressing three enzymes (almA, xylE, p450cam) in Escherichia coli led to degradation of 60-99% of target hydrocarbon substrates. Mating experiments, fluorescence microscopy and TEM revealed indigenous bacteria could obtain these vectors from E. coli through several mechanisms of horizontal gene transfer (HGT), including conjugation and cytoplasmic exchange through nanotubes. Inoculating petroleum-polluted sediments with E. coli carrying the vector pSF-OXB15-p450camfusion showed that the E. coli cells died after five days but a variety of bacteria received and carried the vector for over 60 days after inoculation. Within 60 days, the total petroleum hydrocarbon content of the polluted soil was reduced by 46%. Pilot experiments show that vectors only persist in indigenous populations when under selection pressure, disappearing when this carbon source is removed. This approach to remediation could prime indigenous bacteria for degrading pollutants while providing minimal ecosystem disturbance.

Fungal Laccase Production from Lignocellulosic Agricultural Wastes by Solid-State Fermentation: A Review.

Laccases are copper-containing oxidase enzymes found in many fungi. They have received increasing research attention because of their broad substrate specificity and applicability in industrial processes, such as pulp delignification, textile bleaching, phenolic removal, and biosensors. In comparison with traditional submerged fermentation (SF), solid-state fermentation (SSF) is a simpler technique for laccase production and has many advantages, including higher productivity, efficiency, and enzyme stability as well as reduced production costs and environmental pollution. Here, we review recent advances in laccase production technology, with focus on the following areas: (i) Characteristics and advantages of lignocellulosic agricultural wastes used as SSF substrates of laccase production, including detailed suggestions for the selection of lignocellulosic agricultural wastes; (ii) Comparison of fungal laccase production from lignocellulosic substrates by either SSF or SF; (iii) Fungal performance and strain screening in laccase production from lignocellulosic agricultural wastes by SSF; (iv) Applications of laccase production under SSF; and (v) Suggestions and avenues for future studies of laccase production by fungal SSF with lignocellulosic materials and its applications.

Optimization of Polysaccharide Production from Cordyceps militaris by Solid-State Fermentation on Rice and Its Antioxidant Activities.

Polysaccharides are an important class of bioactive components of medical mushroom and herbs and are now used as natural drugs or dietary supplements on a global scale. In this paper, we aimed to increase the polysaccharide production of Cordyceps militaris and the antioxidant activities of fermented rice by solid-state fermentation. The media components and culture condition were optimized by orthogonal design and mono-factor tests using rice as the raw material. The optimal media consisted of (g/L): rice (50), fructose (7), glycerin (7), peptone (1), MgCl2 (0.11), VB1 (0.05), VB2 (0.05), CaCl2 (1.5), corn bran (6), and a water-materials ratio of 100%. The fermentation condition was as follows: inoculum volume of 5.5% (v/w), rice weight of 50 g in one bowl with a diameter of 120 mm and a depth of 90 mm, incubation temperature of 26 °C, and incubation time of seven days. Under the optimized condition, the maximal C. militaris polysaccharide content and free radical scavenging ratio were 68.3 mg/g dry substrate and 98.9%, respectively. This study provides a new strategy for the production of healthy food from traditional food.

A High-Throughput Fluorescence-Based Assay for Rapid Identification of Petroleum-Degrading Bacteria.

Over the past 100 years, oil spills and long-term waste deposition from oil refineries have significantly polluted the environment. These contaminants have widespread negative effects on human health and ecosystem functioning. Natural attenuation of long chain and polyaromatic hydrocarbons is slow and often incomplete. Bioaugmentation of polluted soils with indigenous bacteria that naturally consume petroleum hydrocarbons could speed up this process. However, the characterization of bacterial crude oil degradation efficiency - which often relies upon expensive, highly specialized gas-chromatography mass spectrometry analyses - can present a substantial bottleneck in developing and implementing these bioremediation strategies. Here, we develop a low-cost, rapid, high-throughput fluorescence-based assay for identifying wild-type bacteria that degrade crude oil using the dye Nile Red. We show that Nile Red fluoresces when in contact with crude oil and developed a robust linear model to calculate crude oil content in liquid cell cultures based on fluorescence intensity (FI). To test whether this assay could identify bacteria with enhanced metabolic capacities to break down crude oil, we screened bacteria isolated from a former Shell Oil refinery in Bay Point, CA, and identified one strain (Cupriavidus sp. OPK) with superior crude oil depletion efficiencies (up to 83%) in only 3 days. We further illustrate that this assay can be combined with fluorescence microscopy to study how bacteria interact with crude oil and the strategies they use to degrade this complex substance. We show for the first time that bacteria use three key strategies for degrading crude oil: biofilm formation, direct adherence to oil droplets, and vesicle encapsulation of oil. We propose that the quantitative and qualitative data from this assay can be used to develop new bioremediation strategies based on bioaugmentation and/or biomimetic materials that imitate the natural ability of bacteria to degrade crude oil.

Enhanced crude oil depletion by constructed bacterial consortium comprising bioemulsifier producer and petroleum hydrocarbon degraders.

The aim of this work was to study the production of bioemulsifier by Rhodococcus erythropolis OSDS1, and the improvement of crude oil depletion efficiency using a consortium of petroleum hydrocarbon degraders and OSDS1. The results showed that R. erythropolis OSDS1 produced highly stable bioemulsifier under various salinity (0-35 g/L NaCl) and pH (5.0-9.0) conditions; more than 90% of the initial emulsification activity was retained after 168 h. Emulsification capacity of the bioemulsifier on different petroleum hydrocarbons was diesel > mineral oil/crude oil > gasoline. A mixed bacterial consortium combining OSDS1 and four other petroleum hydrocarbon degraders was constructed. GC-MS results revealed that the constructed consortium achieved 85.26% depletion efficiency of crude oil in 15 days, which was significantly higher than that of individual strains. During the process, alkane hydroxylase gene (alkB) was successfully amplified from the consortium, confirming presence of crude oil degrading enzymes.

Removal of boron from wastewater: Evaluation of seven poplar clones for B accumulation and tolerance.

Boron (B) contamination of wastewater is a serious environmental and health problem, which has proved difficult to remediate. One potential approach is phytoremediation, i.e., the use of plants to extract B into the shoot systems, which can then be harvested and removed. The purpose of this study was to investigate the ability of seven hybrid poplar clones to accumulate and tolerate B at different levels of B and salinity. The clones were grown in quarter strength Hoagland's nutrient solution and exposed to four B treatments: 0.25, 25, 75 and 125 mg B L-1 in factorial combination with three salinity treatments: 0, 0.5 and 5 g L-1. Measurements were made of biomass (dry weight) and B concentrations (analyzed by inductively coupled plasma-optical emission spectroscopy) of plant tissues. The results showed that, with increasing level of B supply, the B concentrations in the shoot tissue of the poplar clones increased, while shoot biomass decreased. Comparison of the seven different poplar clones revealed that the clone designated 195-529 exhibited the greatest ability for B accumulation and tolerance. Generally, the 0.5 g L-1 level of salinity was beneficial for poplar growth and B removal, while the high salinity level of 5 g L-1 significantly inhibited poplar growth and B translocation from roots to shoots. Based on the results obtained with this hydroponic system, the clone 195-529 (P. trichocarpa × P. deltoides) is likely to be the most useful for the removal of B from B contaminated wastewater.

Development of a constructed wetland water treatment system for selenium removal: incorporation of an algal treatment component.

On the basis of the fact that algae have the ability to volatilize substantial quantities of selenium (Se), we investigated the concept of including an algal pretreatment unit into a constructed wetland system for the removal of Se from river water entering the Salton Sea. Of six different algal strains tested, the most effective in terms of Se volatilization and Se removal from the water column was a Chlorella vulgaris strain (designated Cv). Cv removed 96% of Se (supplied as selenate) from the microcosm water column within 72 h, with up to 61% being removed by volatilization to the atmosphere. X-ray absorption spectroscopy revealed that the major forms of Se likely to be accumulated in an algal-wetland system are selenomethionine, a precursor of volatile Se formation, and elemental Se. Our results suggest that the inclusion of an algal pretreatment unit within a constructed wetland water treatment system should not only enhance the efficiency of Se removal but also significantly reduce the risk of the buildup of ecotoxic forms of Se by promoting the biological volatilization of Se.

Development of a constructed wetland water treatment system for selenium removal: use of mesocosms to evaluate design parameters.

The Salton Sea in California is an important habitat for fish and waterfowl. Its ecosystem is threatened due to diminishing water supplies and increasing salinity. An alternative source of water to support species conservation habitat may be obtained from local rivers (e.g., Alamo or New Rivers), provided that a wetland treatment system can be developed to remove selenium (Se), fertilizer nutrients, and other contaminants. Here, we used mesocosms to evaluate a number of potential design options (e.g., plant species selection, sediment composition and arrangement, forced aeration, organic amendments, etc.) to improve the efficiency of Se removal using treatment wetlands. Our results show that, of five different substrate arrangements tested for Se removal, the most efficient was obtained for cattails growing in a substrate of cattail litter overlying sand and peat moss sediment (water column Se was reduced from 15 µg Se/L to <0.1 µg Se/L in 72 h). The addition of organic amendments in the form of alfalfa hay or alfalfa meal was also helpful in lowering Se levels. These results suggest that it may be possible to design constructed wetland water treatment systems capable of reducing Se concentrations in river water to values below 1 µg Se/L.

Isolation of an extremely boron-tolerant strain of Bacillus firmus.

A strain of Bacillus firmus (designated strain KC) isolated from a boron (B) mine in California exhibited extreme tolerance to B, provided it was first acclimated at intermediate B supply concentrations. Strain KC tolerated up to 1000 mmol/L B (boric acid-B) and 1800 mmol/L B (sodium tetraborate-B), and attained the greatest growth (as measured by absorbance) at 300 mmol/L B. Despite its extreme tolerance to high B, there was no evidence that it was able to remove significant quantities of B from the growth media, suggesting that strain KC is not likely to be useful for the removal of B from wastewaters in an engineered bioreactor.

A comparative transcriptomic analysis of the extremely boron tolerant plant Puccinellia distans with the moderately boron tolerant Gypsophila arrostil.

UNLABELLED: The Turkish ecotype of Puccinellia distans displays exceptional boron (B) tolerance, >1,250 mg B L⁻¹, compared to <50 mg B L⁻¹ for Gypsophila arrostil. In the present study, we compare the molecular basis for the difference in B tolerance between the two species by constructing high B-responsive suppression subtractive hybridization libraries to identify the upregulated genes. A total of 219 and 113 unique non-redundant expressed sequence tags (ESTs) were identified and functionally classified in P. distans and G. arrostil, respectively. In addition, 63 ESTs were down-regulated in P. distans in response to high B. The majority of the high B upregulated genes belong to four categories: metabolism, protein synthesis, cellular organization, and stress/defense. We hypothesize that the superior B tolerance exhibited by P. distans may be due to its ability to restrict the accumulation of B in plant tissues through the upregulated expression of efflux transporters comparable to the Bot1 transporter of barley. In addition, our results are consistent with the view that other molecular mechanisms involved in stress/defense, such as detoxification, anti-oxidative, and signaling pathways, are needed to tolerate B-toxicity stress. KEY MESSAGE: The molecular basis of boron tolerance of two plant species (Puccinellia distans and Gypsophila arrostil) that differ greatly in their boron tolerance was studied in this manuscript.

Evaluation of the boron tolerant grass, Puccinellia distans, as an initial vegetative cover for the Phytorestoration of a boron-contaminated mining site in Southern California.

Land damaged by boron (B) mining should be restored to its natural state with a zero net impact on biodiversity. In an earlier study (Environ. Sci. Technol.2010,44, 7089-7095), we characterized a Turkish ecotype of the grass, Puccinellia distans, which exhibited extreme tolerance to B. Here we evaluated the use of a US ecotype of P. distans as an initial vegetative cover for the phytorestoration of a B mine in southern California. Hydroponic studies revealed that this P. distans ecotype tolerated B concentrations >100 mg B/L and could be germinated and grown in B-contaminated soils taken from the sites to be restored. P. distans grew well in moderately B-contaminated soil (∼88 mg B/L saturated extract) amended with added organic matter (peat moss); other soil treatments such as gypsum addition or pH correction were not needed. P. distans also grew in severely B-contaminated soil (∼1506 mg B/L) provided that toxic levels of soil B were diluted by the addition of sand and/or organic matter. Our results provide evidence in support of the concept of using the US ecotype of P. distans as an initial vegetative cover for the phytorestoration of B-contaminated soil.

Mechanisms of boron tolerance and accumulation in plants: a physiological comparison of the extremely boron-tolerant plant species, Puccinellia distans, with the moderately boron-tolerant Gypsophila arrostil.

The physiological characteristics of the extremely boron (B)-tolerant plant species, Puccinellia distans, were compared with those of the moderately tolerant Gypsophila arrostil, two species collected from a B-mining area of Eskisehir, Turkey. Boron was supplied to plants hydroponically at B concentrations ranging from 0.5 to 50 mg B/L for G. arrostil, and from 0.5 to 2000 mg B/L for P. distans. The results show that P. distans has a strikingly greater tolerance to B than G. arrostil. While G. arrostil was unable to survive B supply concentrations greater than 50 mg B/L, P. distans grew at B supply concentrations exceeding 1250 mg B/L. Our research supports the conclusion that from 0.5 to 50 mg B/L, P. distans is better able to restrict the accumulation of B in the whole plant, and the transport of B from root to shoot, than G. arrostil. We propose that P. distans uses several strategies to achieve B tolerance including the ability to restrict the accumulation of B relative to its accumulation of biomass, the ability to restrict the transport of B from root to shoot, and, to a lesser extent, the ability to tolerate high concentrations of B in its shoot and root tissues.

Heavy metal tolerance and accumulation in Indian mustard (Brassica juncea L.) expressing bacterial gamma-glutamylcysteine synthetase or glutathione synthetase.

The overexpression of either gamma-glutamylcysteine synthetase (gamma-ECS) or glutathione synthetase (GS) in Brassica juncea transgenics was shown previously to result in higher accumulation of glutathione (GSH) and phytochelatins (PCs), as well as enhanced Cd tolerance and accumulation. The present study was aimed at analyzing the effects of gamma-ECS or GS overexpression on tolerance to and accumulation of other metal/loids supplied individually in agar medium (seedlings) or in hydroponics (mature plants). Also, as pollution in nature generally consists of mixtures of metals, glutamylcysteine synthetase (ECS) and GS seedlings were tested on combinations of metals. Compared to wild-type plants, ECS and GS transgenics exhibited a significantly higher capacity to tolerate and accumulate a variety of metal/loids (particularly As, Cd, and Cr) as well as mixed-metal combinations (As, Cd, Zn/As, Pb, and Zn). This enhanced metal tolerance and accumulation of the ECS and GS transgenics may be attributable to enhanced production of PCs, sustained by a greater availability of GSH as substrate, as suggested by their higher concentrations of GSH, PC2, PC3, and PC4 as compared to wild-type plants. Overexpression of GS and gamma-ECS may represent a promising strategy for the development of plants with an enhanced phytoremediation capacity for mixtures of metals.

Selenium volatiles as proxy to the metabolic pathways of selenium in genetically modified Brassica juncea.

In this study we demonstrate that the headspace selenium volatiles could be used as proxy to the metabolic pathways in the Se-accumulator plant Brassica juncea. The selenium metabolic pathways in wild type plants are compared to those of several genetically modified cultures. Complementary use of atomic and molecular mass spectrometric techniques also allowed for identification of yet unreported minor headspace Se-containing volatiles such as CH3SeSeSeCH3, CH3SeSSeCH3, and CH3SeCH2CH3. By combining the information resulting from this research with the previously known information about selenium metabolism in B. juncea, it is possible that a more efficacious phytoremediation tool can be constructed.

Transgenic Indian mustard overexpressing selenocysteine lyase or selenocysteine methyltransferase exhibit enhanced potential for selenium phytoremediation under field conditions.

Two new transgenic Indian mustard [Brassica juncea (L.) Czern.] lines were tested under field conditions for their ability to accumulate selenium (Se)from Se- and boron-contaminated saline sediment. The transgenic lines overexpress genes encoding the enzymes selenocysteine lyase (cpSL) and selenocysteine methyltransferase (SMT), respectively. In the first Spring planting, cpSL, SMT, and wildtype plants (WT) were compared, while SMT and WT were compared in a second, Fall planting. In the Spring planting, shoots of the cpSL transgenic plants accumulated 2-fold more Se (p < 0.01), had 1.8 times higher leaf Se concentrations (p < 0.01), and grew better on contaminated soil than WT. The SMT plants had a 1.7-fold higher leaf Se concentration than WT (p < 0.05). In the Fall planting, the SMT transgenic plants accumulated 1.6-fold more Se in their shoots than WT (p < 0.01) with Se concentrations being higher in both leaves and stems. These results conclusively demonstrate that cpSL and SMT transgenic lines have significantly greater Se phytoremediation potential than wildtype Indian mustard. Further, this study confirms the importance of field testing for evaluating future transgenic lines.

Characterization of a selenate-resistant Arabidopsis mutant. Root growth as a potential target for selenate toxicity.

Screening an Arabidopsis (Arabidopsis thaliana) T-DNA mutant library for selenate resistance enabled us to isolate a selenate-resistant mutant line (sel1-11). Molecular and genetic characterization showed that the mutant contained a lesion in the SULTR1;2 gene that encodes a high affinity root sulfate transporter. We showed that SULTR1;2 is the only gene among 13 mutated genes of the Arabidopsis sulfate transporter family whose mutation conferred selenate resistance to Arabidopsis. The selenate resistance phenotype of the sel1-11 mutant was mirrored by an 8-fold increase of root growth in the presence of selenate as shown by the calculated lethal concentration values. The impairment of SULTR1;2 activity in sel1-11 resulted in a reduced (35)S-sulfate uptake capacity by both roots and calli and a reduced sulfate and selenate content in root, shoot, and calli. Comparing sulfate-to-selenate ratios instead of absolute sulfate and selenate contents in roots and shoots enabled us to gain better insight into the mechanism of selenate toxicity in Arabidopsis. Roots of the sel1-11 mutant line showed a higher sulfate to selenate ratio than that of wild-type roots, while there were no significant differences in sulfate to selenate ratios in shoots of wild-type and mutant lines. These results indicated that the mechanism that confers the selenate resistance phenotype to the sel1-11 line takes place rather in the roots. It might be in part the result of a lower selenate uptake and of a protective effect of sulfate against the toxic effects of selenate on root growth. These results revealed in plants a central and specific role of the transporter SULTR1;2 in selenate sensitivity; they further suggested that root growth and potentially the root tip activity might be a specific target of selenate toxicity in Arabidopsis.

Phytoremediation of mercury and organomercurials in chloroplast transgenic plants: enhanced root uptake, translocation to shoots, and volatilization.

Transgenic tobacco plants engineered with bacterial merA and merB genes via the chloroplast genome were investigated to study the uptake, translocation of different forms of mercury (Hg) from roots to shoots, and their volatilization. Untransformed plants, regardless of the form of Hg supplied, reached a saturation point at 200 microM of phenylmercuric acetate (PMA) or HgCl2, accumulating Hg concentrations up to 500 microg g(-1) with significant reduction in growth. In contrast, chloroplast transgenic lines continued to grow well with Hg concentrations in root tissues up to 2000 microg g(-1). Chloroplasttransgenic lines accumulated both the organic and inorganic Hg forms to levels surpassing the concentrations found in the soil. The organic-Hg form was absorbed and translocated more efficiently than the inorganic-Hg form in transgenic lines, whereas no such difference was observed in untransformed plants. Chloroplast-transgenic lines showed about 100-fold increase in the efficiency of Hg accumulation in shoots compared to untransformed plants. This is the first report of such high levels of Hg accumulation in green leaves or tissues. Transgenic plants attained a maximum rate of elemental-Hg volatilization in two days when supplied with PMA and in three days when supplied with inorganic-Hg, attaining complete volatilization within a week. The combined expression of merAB via the chloroplast genome enhanced conversion of Hg2+ into Hg,0 conferred tolerance by rapid volatilization and increased uptake of different forms of mercury, surpassing the concentrations found in the soil. These investigations provide novel insights for improvement of plant tolerance and detoxification of mercury.

Constitutive expression of a high-affinity sulfate transporter in Indian mustard affects metal tolerance and accumulation.

The Stylosanthes hamata SHST1 gene encodes a high-affinity sulfate transporter located in the plasma membrane. In this study the S. hamata SHST1 gene was constitutively expressed in Indian mustard [Brassica juncea (L.) Czern.] to investigate its importance for tolerance and accumulation of various oxyanions that may be transported by SHST1 and for cadmium, which is detoxified by sulfur-rich compounds. The transgenic SHST1 lines SHST1-12C and SHST1-4C were compared with wild-type Indian mustard for tolerance and accumulation of arsenate, chromate, tungstate, vanadate, and cadmium. As seedlings the SHST1 plants accumulated significantly more Cd and W, and somewhat more Cr and V. The SHST1 seedlings were less tolerant to Cd, Mo, and V compared to wild-type plants. Mature SHST1 plants were less tolerant than wild-type plants to Cd and Cr. SHST1 plants accumulated significantly more Cd, Cr, and W in their roots than wild-type plants. In their shoots they accumulated significantly more Cr and somewhat more V and W. Shoot Cd accumulation was significantly lower than in wild-type, and As levels were somewhat reduced. Compared to wild-type plants, sulfur accumulation was enhanced in roots of SHST1 plants but not in shoots. Together these results suggest that SHST1 can facilitate uptake of other oxyanions in addition to sulfate and that SHST1 mediates uptake in roots rather than root-to-shoot translocation. Since SHST1 overexpression led to enhanced accumulation of Cr, Cd, V, and W, this approach shows some potential for phytoremediation, especially if it could be combined with the expression of a gene that confers enhanced metal translocation or tolerance.