Plasmid-encoded resistance to arsenic and antimony.

Resistance determinants to the toxic oxyanionic salts of arsenic and antimony are found on plasmids of both gram-negative and gram-positive organisms. In most cases these provide resistance to both the oxyanions of +III oxidation state, antimonite and arsenite, and the +V oxidation state, arsenate. In both gram-positive and -negative bacteria, resistance is correlated with efflux of the anions from cells. The determinant from the plasmid R773, isolated from a gram-negative organism, has been studied in detail. It encodes an oxyanion-translocating ATPase with three subunits, a catalytic subunit, the ArsA protein, a membrane subunit, the ArsB subunit, and a specificity factor, the ArsC protein. The first two form a membrane-bound complex with arsenite-stimulated ATPase activity. The determinants from gram-positive bacteria have only the arsB and arsC genes and encode an efflux system without the participation of an ArsA homologue.     

Families of arsenic transporters.

Bacterial arsenic resistance (ars) operons encode an arsenite-efflux system that can be a secondary carrier protein (ArsB) or an anion-translocating ATPase (ArsAB). Yeasts extrude arsenite using Acr3p, a plasma membrane carrier protein, or sequester it in vacuoles as the glutathione conjugate using Ycf1p, an ABC transporter.     

Distributions of arsenic and essential elements in pinna of arsenic hyperaccumulator Pteris vittata L.

The distributions of arsenic and 6 essential elements in the pinna of As hyperaccumulator, Pteris vittata L., were studied using synchrotron radiation X-ray fluorescence (SRXRF). Significant correlation between the distribution and mobility of the elements revealed that SRXRF study on the elemental distribution was feasible to inspect the transportations of elements in plants. The distribution of As in the pinna showed that As had great abilities to be transported in xylem vessels and from xylem to mesophyll. The distribution of K, one of the most mobile elements in plants, was similar to that of As, whereas the distributions of Fe and Ca with less mobility in plants were almost opposite to that of As in the pinna.     

Phytoremediation of arsenic contaminated soil by Pteris vittata L. II. Effect on arsenic uptake and rice yield

A greenhouse experiment evaluated the effect of phytoextraction of arsenic from a contaminated soil by Chinese Brake Fern (Pteris vittata L.) and its subsequent effects on growth and uptake of arsenic by rice (Oryza sativa L.) crop. Pteris vittata was grown for one or two growing cycles of four months each with two phosphate sources, using single super phosphate (SSP) and di-ammonium phosphate (DAP). Rice was grown on phytoextracted soils followed by measurements of biomass yield (grain, straw, and root), arsenic concentration and, uptake by individual plant parts. The biomass yield (grain, straw and rice) of rice was highest in soil phytoextracted with Pteris vittata grown for two cycles and fertilized with diammonium phosphate (DAP). Total arsenic uptake in contaminated soil ranged from 8.2 to 16.9 mg pot(-1) in first growing cycle and 5.5 to 12.0 mg pot(-1) in second growing cycle of Pteris vittata. There was thus a mean reduction of 52% in arsenic content of rice grain after two growing cycle of Pteris vittata and 29% after the one growing cycle. The phytoextraction of arsenic contaminated soil by Pteris vittata was beneficial for growing rice resulted in decreased arsenic content in rice grain of <1 ppm. There was a mean improvement in rice grain yield 14% after two growing cycle and 8% after the one growing cycle of brake fern.     

Systematic engineering of phytochelatin synthesis and arsenic transport for enhanced arsenic accumulation in E. coli

Phytochelatin (PC) is a naturally occurring peptide with high affinity towards arsenic (As). In this article, we demonstrated the systematic engineering of PC‐producing E. coli for As accumulation by addressing different bottlenecks in PC synthesis as well as As transport. Phytochelatin synthase from Schizosaccharomyces pombe (SpPCS) was expressed in E. coli resulting in 18 times higher As accumulation. PC production was further increased by co‐expressing a feedback desensitized γ‐glutamylcysteine synthetase (GshI*), resulting in 30‐fold higher PC levels and additional 2‐fold higher As accumulation. The significantly increased PC levels were exploited further by co‐expressing an arsenic transporter GlpF, leading to an additional 1.5‐fold higher As accumulation. These engineering steps were finally combined in an arsenic efflux deletion E. coli strain to achieve an arsenic accumulation level of 16.8 µmol/g DCW, a 80‐fold improvement when compared to a control strain not producing phytochelatins. Biotechnol. Bioeng. 2010. 105: 780–785. © 2009 Wiley Periodicals, Inc.     

Effects of nutrients on arsenic accumulation by arsenic hyperaccumulator Pteris vittata L.

This research investigated the effects of various nutrients on arsenic (As) removal by arsenic hyperaccumulator Pteris vittata L. in a Hoagland nutrient solution (HNS). The treatments included different concentrations of Ca and K in 20% strength of HNS, different strengths of HNS (10, 20 and 30%), different strengths of HNS (10 and 20%) with and without CaCO₃, and different concentrations of Ca, K, NO₃, NH₄, and P in 20% strength of HNS. The plants were grown in nutrient solution containing 1 mg As L^?1 for 4 weeks except the Ca/K experiment where the plants were grown in nutrient solution containing 10 or 50 mg As L^?1 for 1 week. Adding up to 4 mM Ca or 3 mM K to 20% strength HNS significantly (P < 0.05) increased plant arsenic accumulation when the solution contained 10 mg As L^?1. Plant arsenic removal was reduced with increasing Ca and K concentrations at 50 mg As L^?1. Lower strength of HNS (10%) resulted in the greatest plant arsenic removal (79%) due to lower competition of P with As for plant uptake. Addition of CaCO₃ to 20% strength of HNS significantly increased arsenic removal by P. vittata. Among the nutrients tested, NO₃ and CaCO₃ were beneficial to plant arsenic removal while NH₄, P and Cl had adverse effects. This experiment demonstrated that it is possible to optimize plant arsenic removal by adjusting nutrients in the growth medium.     

Mechanisms of arsenic hyperaccumulation in Pteris vittata. Uptake kinetics,interactions with phosphate,and arsenic speciation

The mechanisms of arsenic (As) hyperaccumulation in Pteris vittata, the first identified As hyperaccumulator, are unknown. We investigated the interactions of arsenate and phosphate on the uptake and distribution of As and phosphorus (P), and As speciation in P. vittata. In an 18-d hydroponic experiment with varying concentrations of arsenate and phosphate, P. vittata accumulated As in the fronds up to 27,000 mg As kg(-1) dry weight, and the frond As to root As concentration ratio varied between 1.3 and 6.7. Increasing phosphate supply decreased As uptake markedly, with the effect being greater on root As concentration than on shoot As concentration. Increasing arsenate supply decreased the P concentration in the roots, but not in the fronds. Presence of phosphate in the uptake solution decreased arsenate influx markedly, whereas P starvation for 8 d increased the maximum net influx by 2.5-fold. The rate of arsenite uptake was 10% of that for arsenate in the absence of phosphate. Neither P starvation nor the presence of phosphate affected arsenite uptake. Within 8 h, 50% to 78% of the As taken up was distributed to the fronds, with a higher translocation efficiency for arsenite than for arsenate. In fronds, 49% to 94% of the As was extracted with a phosphate buffer (pH 5.6). Speciation analysis using high-performance liquid chromatography-inductively coupled plasma mass spectroscopy showed that >85% of the extracted As was in the form of arsenite, and the remaining mostly as arsenate. We conclude that arsenate is taken up by P. vittata via the phosphate transporters, reduced to arsenite, and sequestered in the fronds primarily as As(III).     

EXAFS study on arsenic species and transformation in arsenic hyperaccumulator

As a cost-effective, efficient and environmental friendly method for the remediation of contaminated soils and waters, phytoremediation of arsenic-con- taminated soils has drawn more and more attention[1]. The plants with the special ability to accumulate arse-nic (hyperaccumulators) are a prerequisite for phy-toremediation. Cretan brake (Pteris cretica L. var nervosa Thunb) has been shown to accumulate arsenic as much as 694 mg/kg in pinna in field investigation[2], and such elevated arsenic…     

EXAFS study on arsenic species and transformation in arsenic hyperaccumulator

Synchrotron radiation extended X-ray absorption fine structure (SR EXAFS) was employed to study the transformation of coordination environment and the redox speciation of arsenic in a newly discovered arsenic hyperaccumulator, Cretan brake (Pteris cretica L. var nervosa Thunb). It showed that the arsenic in the plant mainly coordinated with oxygen, except that some arsenic coordinated with S as As-GSH in root. The complexation of arsenic with GSH might not be the predominant detoxification mechanism in Cretan brake. Although some arsenic in root presented as As(V) in Na₂HAsO₄ treatments, most of arsenic in plant presented as As(III)-O in both treatments, indicating that As(V) tended to be reduced to As(III) after it was taken up into the root, and arsenic was kept as As(III) when it was transported to the above-ground tissues. The reduction of As(V) primarily proceeded in the root.     

络合型含铁材料对砷胁迫下水稻生理生化特性及砷累积的影响

土壤重金属与农产品质量息息相关.本研究通过设置砷胁迫条件下水稻种植盆栽试验,分析络合型含铁材料不同施用时期和不同施用量对水稻SPAD值、抗氧化酶(SOD、POD、CAT)活性以及水稻砷吸收、累积的影响.结果表明,砷胁迫对水稻SOD、POD、CAT活性有着不同程度的抑制作用,添加络合型含铁材料可以降低水稻MDA含量,缓解砷对水稻光合作用和SOD、POD、CAT活性的抑制;同时能减少水稻对土壤砷的吸收和累积.与移栽前添加络合型含铁材料相比,选择水稻孕穗期前添加络合型含铁材料,水稻砷吸收的抑制效果较好;水稻植株地上部砷含量与络合型含铁材料的添加浓度呈显著负相关,孕穗期前添加0.2 g·kg-1络合型含铁材料对水稻植株砷的吸收抑制效果最好,水稻植株地上部砷含量降低59.22%.     

土壤中砷的形态转化及其分析方法

在分析土壤中砷的来源及存在形态的基础上,简要探讨了不同形态砷之间相互转化的过程.对土壤砷形态的提取和检测方法进行了较系统的比较,认为高效液相色谱-氢化物发生-原子荧光光谱（HPLC-HG-AFS）联用技术具有灵敏度高、检测限低、选择性强、运行成本低、不同形态砷之间相互转化较少的优点,可以作为砷形态检测的首选方法.结合相关研究结果,对土壤中砷形态转化及相关研究的发展方向进行了讨论.     

紫色土中砷、磷的吸附-解吸和竞争吸附

采用批培养法研究了As、P在三峡库区典型土壤紫色土中的吸附-解吸特点及竞争吸附对As、P迁移活化的影响.结果表明：3种紫色土中As、P的吸附-解吸特点相似,等温吸附均符合Langmuir和Freundlich方程,As在酸性、中性和石灰性紫色土中的最大吸附量分别为1428.6、1250.0和1111.1 mg·kg^-1;P在酸性、中性和石灰性紫色土中的最大吸附量分别为322.6、357.1和434.8 mg·kg^-1;As、P吸附动力学过程为先快后慢,均符合一级动力学方程与Elovich方程,快速吸附段符合一级动力学方程,为交换吸附;慢速吸附阶段满足Elovich方程,可能属于深层吸附或专性吸附.As、P竞争吸附试验表明,As、P共存时P的吸附速度和吸附量均增强,而As的吸附速度和吸附量均降低,表明As的存在能明显增强紫色土对P的吸附作用,P的存在则明显抑制紫色土对As的吸附.     

Root exudates and arsenic accumulation in arsenic hyperaccumulating Pteris vittata and non-hyperaccumulating Nephrolepis exaltata

This study compared the roles of root exudates collected from two fern species, the As hyperaccumulating Chinese Brake fern (Pteris vittata L.) and the As-sensitive Boston fern (Nephrolepis exaltata L.), on As-mobilization of two As minerals (aluminum arsenate and iron arsenate) and a CCA (chromated copper arsenate)-contaminated soil as well as plant As accumulation. Chinese Brake fern exuded 2 times more dissolved organic carbon (DOC) than Boston fern and the difference was more pronounced under As stress. The composition of organic acids in the root exudates for both ferns consisted mainly of phytic acid and oxalic acid. However, Chinese Brake fern produced 0.46 to 1.06 times more phytic acid than Boston fern under As stress, and exuded 3–5 times more oxalic acid than Boston fern in all treatments. Consequently, root exudates from Chinese Brake fern mobilized more As from aluminum arsenate (3–4 times), iron arsenate (4–6 times) and CCA-contaminated soil (6–18 times) than Boston fern. Chinese Brake fern took up more As and translocated more As to the fronds than Boston fern. The molar ratio of P/As in the roots of Chinese Brake fern was greater than in the fronds whereas the reverse was observed in Boston fern. These results suggested that As-mobilization from the soil by the root exudates (enhancing plant uptake), coupled with efficient As translocation to the fronds (keeping a high molar ratio of P/As in the roots), are both important for As hyperaccumulation by Chinese Brake fern.     

Microbial methylation of metalloids: arsenic, antimony, and bismuth.

A significant 19th century public health problem was that the inhabitants of many houses containing wallpaper decorated with green arsenical pigments experienced illness and death. The problem was caused by certain fungi that grew in the presence of inorganic arsenic to form a toxic, garlic-odored gas. The garlic odor was actually put to use in a very delicate microbiological test for arsenic. In 1933, the gas was shown to be trimethylarsine. It was not until 1971 that arsenic methylation by bacteria was demonstrated. Further research in biomethylation has been facilitated by the development of delicate techniques for the determination of arsenic species. As described in this review, many microorganisms (bacteria, fungi, and yeasts) and animals are now known to biomethylate arsenic, forming both volatile (e.g., methylarsines) and nonvolatile (e.g., methylarsonic acid and dimethylarsinic acid) compounds. The enzymatic mechanisms for this biomethylation are discussed. The microbial conversion of sodium arsenate to trimethylarsine proceeds by alternate reduction and methylation steps, with S-adenosylmethionine as the usual methyl donor. Thiols have important roles in the reductions. In anaerobic bacteria, methylcobalamin may be the donor. The other metalloid elements of the periodic table group 15, antimony and bismuth, also undergo biomethylation to some extent. Trimethylstibine formation by microorganisms is now well established, but this process apparently does not occur in animals. Formation of trimethylbismuth by microorganisms has been reported in a few cases. Microbial methylation plays important roles in the biogeochemical cycling of these metalloid elements and possibly in their detoxification. The wheel has come full circle, and public health considerations are again important.     

Porphyrins as early biomarkers for arsenic exposure in animals and humans.

We studied the effect of arsenic exposure on the haem biosynthetic pathway in the rat and humans. Significant increases in protoporphyrin IX, coproporphyrin III, coproporphyrin I were observed in the blood, liver and kidney, and in the urine of rats after a single dose of arsenic. The level of increase was dependent on the arsenic species present. Most of porphyrin concentrations in the tissues increased within 24 hr and urinary excretion elevated within 48 hr. In the human study, we collected urine samples from 113 people who live in Xing Ren of Guizhou Province, a coal-borne arsenicosis endemic area in southwest of PR China and from 30 people who live in Xing Yi (about 80 km southwest of Xing Ren) where arsenicosis is not prevalent. We analyzed the urinary porphyrins using HPLC. Results indicate that all urinary porphyrins were higher in the arsenic exposed group than those in the control group. Women, children and older age people spend much of their time indoors, they had greater increases of urinary arsenic and porphyrins. They were the higher risk groups among the study subjects. A positive correlation between the urinary arsenic levels and porphyrin concentrations demonstrated the effect of arsenic on haem biosynthesis. Significant alteration in the porphyrin excretion profiles of the younger age (<20 y) arsenic exposed group suggested that porphyrins could be used as early warning biomarkers for chronic exposure to arsenic.