Arsenic-contaminated soils phytotoxicity studies with sunflower and sorghum

Background, Aim and Scope  Environmental pollution caused by arsenic (As) is a major ecological problem. There has been intense worldwide effort to find As-hyperaccumulating plants that can be used in phytoremediation—the green-plant-assisted removal of chemical pollutants from soils. For phytoremediation, it is natural to prefer cultivated rather than wild plants, because their agriculture is well known. This study was conducted to evaluate the tolerance of common sunflower(Helianthus annuus L.) and sugar sorghum(Sorghum saccharatum Pers.) for soil-As contents of 10–100 mg As kg^-1 soil, with sodium arsenite as a model contaminant. Methods  Plants were grown in a growth chamber for 30 days. Microfield experiments were conducted on experimental plots. To study the phytoremediation effect of the auxins indole-3-acetic acid (IAA) and 2,4-dichlorophenoxyacetic acid (2,4-D), we treated 1- and 3-day-old plant seedlings with water solutions of the auxins (concentrations 10^-5, 10^-7, and 10^-9 g l^-1). The soil and plant-biomass samples were analyzed for total As by using the color reaction of ammonium molybdate with As. Results and Discussion  Phytotoxicity studies showed that 100 mg As kg^-1 soil poisoned sunflower and sorghum growth by 50%. There was a linear correlation between soil-As content and As accumulation in the plants. Laboratory experiments showed that the soil-As content was reduced two- to threefold after sunflower had been grown with 10–100 mg As kg^-1 soil for 30 days. Treatment of sunflower and sorghum seedlings with IAA and 2,4-D at a concentration of 10^-5 g l^-1 in microfield experiments enhanced the phytoremediation two- to fivefold as compared with untreated control plants. The best results were obtained with 3-day-old seedlings. Conclusion, Recommendation and Outlook  (a) Sunflower and sorghum are good candidates to remediate As-polluted soils. (b) Phytoremediation can be improved with IAA or 2,4-D. (c) Mixed cropping of sorghum and sunflower may be another way of improving phytoremediation.     

亚砷酸氧化菌Sinorhizobium sp. GW3的鉴定与亚砷酸氧化酶基因的分离

亚砷酸氧化细菌能够将毒性大的As(Ⅲ)氧化成毒性小的As(Ⅴ),在生物修复砷污染方面具有应用价值。对一株新型亚砷酸氧化菌Sinorhizobium sp.GW3进行了较全面的鉴定,并分离及分析了亚砷酸氧化酶基因aoxAB。形态学、生理生化鉴定和16S rRNA基因等分析结果表明该菌为Sinorhizobium属。该菌对As(III)抗性的MIC(Minimum inhibitory concentration)为9 mmol/L。首次在Sinorhizobium属中分离了包括编码小亚基aoxA和大亚基aoxB在内的亚砷酸氧化酶基因,其编码的大小亚基与已发现的Agrobacterium tumefaciens(ABB51929)的大小亚基在氨基酸水平上分别有86%、80%的同源性。     

Arsenic accumulation by rice grown in soil treated with roxarsone

Poultry litter is widely used as a fertilizer for lowland rice in Taiwan and China. However, the organic‐arsenic compound roxarsone (additive of poultry feed) in poultry litter can be absorbed by the plants and the resulting arsenic (As) contamination may pose a serious threat to human health. This study used various amounts of poultry litter contaminated with roxarsone in pot experiments to evaluate the effect of roxarsone on rice agronomic parameters and the bioaccumulation of total and inorganic As in rice‐plant tissues. Rice‐grain yield decreased significantly with increasing As content of the soil, and the critical threshold that killed rice was 200 mg roxarsone (kg soil)^–1. The As concentrations in root, straw, leaf, husk, and grain increased with increasing soil As (p < 1%). At 100 mg roxarsone per kg of soil, the As concentration in the rice grain exceeded the statutory permissible limit of 1.0 mg As (kg dry weight)^–1 and at 25 mg roxarsone (kg soil)^–1, the inorganic As concentrations in grains exceeded the statutory limit of 0.15 mg of inorganic As kg^–1 in China. For all treatments, the As concentrations in various plant tissues at maturity follow the order: root > stem > leaf > husk > grain. Arsenite was the predominant species in root, straw, and grain, while arsenate was the predominant species in leaf and husk. No significant difference existed between the amounts of arsenite and arsenate when various amounts of poultry litter were applied. This result illustrates that large amounts of added roxarsone are not only toxic to rice but also accumulate in grains in the inorganic As forms, potentially posing a threat to human health via the food chain.     

三价砷氧化细菌Acidovorax sp.GW2中As(Ⅲ)氧化酶基因和调控序列的克隆鉴定

通过反向PCR和细菌Fosmid文库筛选,克隆得到1株二三价砷[As(Ⅲ)]氧化细菌Acidovorax sp.GW2的As(Ⅲ)氧化酶Aox基因簇,包括aoxRSXABCD 7个基因,分别预测编码双组分信号传导系统转录调控子AoxR(同源性68%),周质感应组氨酸激酶AoxS(同源性55%),周质结合蛋白AoxX(同...     

Effects of p38 MAPK on SETD4 expression and location in p38 +/+ and p38 -/- cells treated with sodium arsenite

AIM: To investigate the expression and location of SET domain-containing 4(SETD4) protein in p38 ^+/+ and p38 ^-/- cells treated with sodium arsenite(NaAsO₂). METHODS: The expression and location of SETD4 were detected in different cells with or without NaAsO₂ treatment by Western blotting and immunofluorescence technique. RESULTS: The expression of SETD4 was detectable in both murine- and human-derived cells. Its distribution was found to be located in the whole cell, mainly in the cytoplasm. Further investigation also suggested that the protein expression of SETD4 was reduced in both p38 ^+/+ and p38 ^-/- cells 6 h after NaAsO₂ treatment. Moreover, SETD4 protein was translocated from cytoplasm to nucleus in p38 ^+/+ cells treated with NaAsO₂, which was unobvious in p38 ^-/- cells. CONCLUSION: SETD4 protein is expressed in various cells derived from different species and tissues, and it is mainly located in cytoplasm. NaAsO₂ treatment influences the expression of SETD4, and induces the translocation of SETD4 protein to nucleus, which might be involved in the p38 MAPK signal pathway.     

Effects of JNK signaling pathway on hnRNP K protein aggregates in NIH3T3 cells treated with sodium arsenite

AIM: To explore the role of JNK signaling pathway in the formation of heterogeneous nuclear ribonucleoprotein (hnRNP) K protein aggregates by observing the expression and localization of hnRNP K in NIH3T3 cells induced by sodium arsenite(NaAsO2).METHODS: The recombinant vector pcDNA3-hnRNP K-HA was constructed and transfected into NIH3T3 cells. The expression and localization of endogenous hnRNP K and distribution of the protein aggregates stimulated with NaAsO2 at different time points were observed under fluorescence microscope. The level of reactive oxygen species (ROS) in the cells was detected by a ROS assay kit. After pretreated with the inhibitors of 5 signaling pathways (JNK, MEK, PI3K/Akt, NF-κB and nuclear transport), the changes of the protein aggregates were observed.RESULTS: The recombinant plasmid was correctly constructed. The hnRNP K was mainly distributed in the nucleus. The intracellular fluorescent aggregates increased with the prolonging stimulation of NaAsO2 in the cells transfected with the recombinant plasmid, and the overexpression of hnRNP K inhibited ROS generation in the cells induced by NaAsO2. SP600125, an inhibitor of JNK signaling pathway, significantly inhibited the formation of protein aggregates. CONCLUSION: The hnRNP K is primarily located in the nucleus of NIH3T3 cells, with a small amount in the cytoplasm. The formation of protein aggregates is significant after NaAsO2 stimulation, which can restrain the level of intracellular ROS, and the process is involved in JNK signaling pathway.     

亚砷酸盐提高藻与蚤培养基下纳米二氧化钛的稳定性

进入水环境的纳米二氧化钛(n Ti O2)会影响水体中其他污染物的环境行为,进而影响其生态风险,这种影响机制受其在水环境中分散、团聚及沉降等稳定性的影响。为更好地认识和预测淡水环境中n Ti O2的生态风险,以超纯水和常见的培养基(绿藻培养基BG11、大型水蚤培养基SM7)作为分散介质,分析了亚砷酸盐[As(Ⅲ)]影响下n Ti O2在分散介质中的稳定性。结果表明:n Ti O2稳定性与其初始浓度、分散介质离子强度显著负相关;As(Ⅲ)能够影响分散介质的pH值、Zeta电位,从而影响n Ti O2的稳定性,n Ti O2稳定性与As(Ⅲ)浓度显著正相关。这说明水环境中的As(Ⅲ)能增强n Ti O2的稳定性和迁移能力,使绿藻和大型蚤暴露于稳定的n Ti O2-As(Ⅲ)体系,增大两者的生态风险。     

水稻砷污染健康风险与砷代谢机制的研究

水稻是人类主要的粮食作物之一，然而，目前稻米主要生产区（东南亚地区）的土壤和灌溉水砷污染严重，导致稻米中砷的积累；砷在稻米中的积累通过食物链传递，对人体健康构成严重威胁。所以，稻米砷污染问题已成为东南亚地区比较突出且急需解决的环境问题之一，而减少水稻对砷的吸收、控制水稻体内砷向籽粒转移、降低籽粒中砷的生物有效性是解决这一问题的关键途径。因此，深入理解水稻对砷的吸收、体内转运和转化等代谢机制非常有必要。近年来有关水稻对砷的吸收和体内砷代谢机制的研究取得了很大进展。研究表明，水稻根系通过磷吸收通道吸收五价砷（As（Ⅴ）），水通道吸收三价砷（As（Ⅲ））。进入水稻体内的砷，一部分由地下部转运到地上部，其中一部分进一步被转运到水稻籽粒中。根表铁膜能抑制水稻对As（Ⅴ）的吸收和向地上部的转运。在水稻的地下部和（或）地上部还存在着As（Ⅴ）还原、As（Ⅲ）甲基化等砷形态的转化过程。最近，水稻砷酸盐还原酶基因已经被克隆和表征。     

Silicon decreases the arsenic level in rice grain by limiting arsenite transport

Silicon (Si) reduces arsenic (As) levels in rice shoot and grain. However, the underlying mechanisms remain unclear. In this study, we examined the effect of Si application to three rice paddy soils on the dynamics of Si, iron (Fe), phosphorus (P), and As in the soil solution, As accumulation in rice straw, flag leaf, husk, brown rice, and polished rice, and on As speciation in polished rice. Silicon application to soil increased the concentrations of Si, Fe, As, and P in the soil solution, while the redox potential was unaffected. Arsenic concentrations of straw, flag leaf, and husk were reduced by half by Si application, while As concentrations of brown and polished rice were decreased by 22%. The main As species in polished rice was arsenite, As(III), with a fraction of 70%, followed by dimethylarsinic acid (DMA) and arsenate, As(V), with 24% and 6%, respectively. Silicon application to the soil did not affect DMA or As(V) concentration of polished rice, while the As(III) concentration was reduced by 33%. These results confirm that Si reduces As(III) uptake and translocation into the shoot. Furthermore, data indicate that decrease of As concentration of polished rice is due to decreased As(III) transport into grain. Possible underlying mechanisms are discussed.     

一个水稻铜锌SOD酶基因在应答亚砷酸盐胁迫中的作用

【目的】水稻Os08g44770.1基因编码一个铜锌SOD酶,但其在响应亚砷酸盐[As(Ⅲ)]胁迫中的生物学功能未知。本研究旨在深入揭示由该基因调控水稻砷耐性改变的分子机理并为水稻抗逆育种提供理论参考。【方法】以野生型日本晴(WT)和2个Os08g44770.1过表达转基因株系为试材,通过胁迫处理、生理指标测定和基因表达分析等,系统探究了转基因植株对As(Ⅲ)的耐受性表现,并初步揭示了其调控水稻砷耐性的生理和分子机理。【结果】与WT相比,过表达转基因株系对As(Ⅲ)更加敏感;转基因植株在砷胁迫下的根系相对伸长量、生物量(干质量)、叶绿素含量、根系细胞质膜完整性、叶片抗氧化程度等均显著低于WT;Os08g44770.1在砷处理后的WT和转基因植株叶片中的表达模式略有不同,但均表现为处理24 h时被显著诱导表达。【结论】过度表达Os08g44770.1基因可导致水稻的砷耐受性极显著下降。