一株新的砷还原菌分离鉴定及其对天然矿物臭葱石的溶解作用

【目的】探究江汉平原土著砷还原微生物如何介导臭葱石的溶解和释放过程,以及硝酸盐和硫酸盐对该过程的影响。【方法】采集江汉平原高砷沉积物,利用多轮传代富集方法筛选出一株兼性厌氧砷还原菌;克隆其16S rRNA基因、砷还原酶基因(arsC)、硫代硫酸盐还原酶基因(phsA)、硝酸盐还原酶基因(nar)以获得其分类地位;分析该细菌的As(V)、NO3–、Fe(III)、S_2O_3~(2–)还原功能;利用microcosm技术分析该菌株催化臭葱石中不可溶砷和铁的溶解和释放作用及硝酸盐和硫酸盐对此过程的影响;采用X-射线衍射(XRD)和扫描电镜(SEM)等方法对细菌作用前后的矿物表面形貌进行分析。【结果】16S rRNA基因测序结果表明该细菌为柠檬酸杆菌属(Citrobacter sp.),故命名为Citrobacter sp. A11;在Citrobacter sp. A11作用下,0.45 mmol/L As(V)在4 d内被还原成As(III),2.0 mmol/L S_2O_3~(2–)在6 d内被还原成S~(2–),1.0 mmol/L Fe(III)在3 d内被还原成Fe(II),140.0 mg/L NO_3~–在28 h内被还原成NO_2~–;经过28 d该细菌的催化作用使得体系中不可溶砷和铁的释放量分别为33.68μmol/L、51.93μmol/L;硫酸根的加入使得砷和铁的释放量分别增长了41.04%和34.30%,硝酸根的加入则使砷和铁释放量分别降低了35.07%和53.46%。XRD、SEM-EDS分析表明,细菌作用后的臭葱石表面形貌发生明显改变,并出现细小且分散的溶解性颗粒。【结论】本次研究从江汉平原高砷沉积物中富集分离得到一株兼性厌氧砷还原细菌Citrobacter sp. A11,能有效还原As(V)、S_2O_3~(2–)、NO_3~–、Fe(III);砷还原细菌Citrobacter sp. A11能显著促进臭葱石中砷和铁的溶解和释放,硫酸根离子的存在会促进细菌介导臭葱石中固态砷、铁的释放,而硝酸根离子的存在则对此过程起明显抑制作用。     

砷超富集植物中砷化学形态其转化的EXAFS研究

通过同步辐射扩展X射线吸收精细结构(SR EXAFS)研究砷超富集植物大叶井口边草(Pteris nervosa)中砷的化学形态及其在植物体中的转化. 结果表明, 在大叶井口边草中砷主要与O配位, 根部存在与谷胱苷肽(GSH)结合的砷, 但是在羽叶中没有发现与GSH结合的砷. 在NaAsO2和Na2HAsO4处理中, 植物根系的砷分别以As(Ⅲ)和 As(Ⅴ)为主, 但是在叶柄和羽叶中砷都以As(Ⅲ)的形态为主. 植物根系吸收的As(Ⅴ)在向上转运的过程中具有向As(Ⅲ)转化的趋势, 其转化过程主要发生在根部. 实验证明, 与GSH结合并不是大叶井口边草中砷解毒的主要机理, 超富集植物可能具有与一般耐性植物不同的重金属解毒机制.     

砷超富集植物中砷化学形态及其转化的EXAFS研究

通过同步辐射扩展X射线吸收精细结构(SR EXAFS)研究砷超富集植物大叶井口边草(Pteris nervosa)中砷的化学形态及其在植物体中的转化。结果表明,在大叶井口边草中砷主要与O配位,根部存在与谷胱苷肽(GSH)结合的砷,但是在羽叶中没有发现与GSH结合的砷,在NaAsO_2和Na_2HAsO_4处理中,植物根系的砷分别以As(Ⅲ)和As(Ⅴ)为主,但是在叶柄和羽叶中砷都以As(Ⅲ)的形态为主,植物根系吸收的As(Ⅴ)在向上转运的过程中具有向As(Ⅲ)转化的趋势,其转化过程主要发生在根部。实验证明,与GSH结合并不是大叶井口边草中砷解毒的主要机理,超富集植物可能具有与一般耐性植物不同的重金属解毒机制。     

砷超富集植物中砷化学形态及其转化的EXAFS研究

通过同步辐射扩展X射线吸收精细结构(SR EXAFS)研究砷超富集植物大叶井口边草(Pteris nervosa)中砷的化学形态及其在植物体中的转化. 结果表明, 在大叶井口边草中砷主要与O配位, 根部存在与谷胱苷肽(GSH)结合的砷, 但是在羽叶中没有发现与GSH结合的砷. 在NaAsO₂和Na₂HAsO₄处理中, 植物根系的砷分别以As(Ⅲ)和As(Ⅴ)为主, 但是在叶柄和羽叶中砷都以As(Ⅲ)的形态为主. 植物根系吸收的As(Ⅴ)在向上转运的过程中具有向As(Ⅲ)转化的趋势, 其转化过程主要发生在根部. 实验证明, 与GSH结合并不是大叶井口边草中砷解毒的主要机理, 超富集植物可能具有与一般耐性植物不同的重金属解毒机制.     

抗砷性微生物及其抗砷分子机制研究进展

砷(Arsenic, As)是一种剧毒类金属(Metalloid), 在自然环境中主要以三价亚砷酸盐[Arsenite, AsO2-, As(III)]和五价砷酸盐[Arsenate, AsO43-, As(V)]的无机形式广泛存在。许多微生物在含砷环境的长期适应过程中, 进化了多种不同的砷解毒抗性机制。目前研究发现主要存在4种类型的砷抗性机理, 包括: As(III)氧化, 细胞质As(V)还原, 呼吸性As(V)还原, As(III)甲基化, 这些机制赋予微生物砷抗性并在砷的转化和地球化学循环中起着极     

4种蚯蚓肠道微生物对砷毒性的响应差异研究

蚯蚓肠道是微生物多样性的一个潜在存储库。砷对蚯蚓肠道微生物群落的影响已被证实,但砷在不同蚯蚓肠道菌群中生物转化的差异仍不清楚。为了进一步阐述土壤中广泛存在的低浓度砷(浓度为5,15,25 mg/kg)对不同种类蚯蚓肠道微生物影响的差异,将4种典型蚯蚓暴露于砷污染土壤后,测定其肠道微生物组成变化,并分析砷对不同蚯蚓肠道内砷富集、形态和砷生物转化基因的影响。结果显示,所有蚯蚓组织内均存在明显的砷富集,其富集系数由高到低依次为:安德爱胜蚓(1.93)>加州腔蚓(0.80)>通俗腔蚓(0.78)>湖北远盲蚓(0.52),蚯蚓组织和肠道内砷形态主要以无机砷为主,其中As(III)含量比例> 80%,部分蚯蚓组织内还发现少量有机砷。4种蚯蚓肠道微生物群落在门水平上主要以变形菌、厚壁菌和放线菌为主,并与周围土壤细菌群落组成存在显著差异。同时,在土壤和肠道内共检测到17个砷转化基因,其中蚯蚓肠道内As(V)还原和砷转运相关基因相对丰度较高,而砷(去)甲基化基因丰度较低。此外,低浓度砷污染对蚯蚓生长无显著影响,却能引起蚯蚓肠道微生物群落的紊乱。蚯蚓种类和砷污染是引起蚯蚓肠道微生物...     

淡水湖泊生态系统中砷的赋存与转化行为研究进展

砷(As)是一种无处不在的元素,目前已有很多关于湖泊水体、沉积物、浮游生物、底栖动物、鱼类及水生植物中As的含量分布与赋存形态的研究报道.As在全球淡水湖泊中分布不均,区域差异性较大;湖泊沉积物中As含量水平对底栖动物的自然生境影响很大,甚至造成底栖物种生物区系的改变;水生植物普遍具有富集As的能力,一般表现为沉水植物＞浮水植物＞挺水植物;水生动物中As含量一般为底栖动物＞浮游动物＞鱼类.相对于海洋生态系统,目前对湖泊生态系统环境与生物质中砷的赋存形态及其转化的认识还很不足,今后应加强人类活动影响下我国重要湖泊As的迁移、富集与转化行为的研究.     

河套平原不同深度高砷地下水硫酸盐还原菌群落分布特征及环境意义

【目的】探究不同深度的高砷含水层中硫酸盐还原菌的丰度、群落组成和多样性的差异,并结合硫酸盐硫同位素等多种水化参数,揭示不同深度高砷地下水中硫酸盐还原菌群落分布特征及其环境意义。【方法】以我国典型高砷地下水分布区河套平原为研究区,采集不同深度含水层中的高砷地下水样品,测定水化参数,采用qPCR对样品16S rRNA基因和dsrB基因进行定量;通过dsrB基因高通量测序对硫酸盐还原菌群落进行分析,并将dsrB基因相对丰度、群落组成及多样性与水化因子结合,进行统计学分析。【结果】基于dsrB基因的定量结果表明,浅层地下水中dsrB基因相对丰度高于深层地下水。浅层地下水中,dsrB基因相对丰度与CH₄浓度呈显著正相关,且δ³⁴S-SO₄^2–与CH₄浓度显著正相关。而深层高砷地下水中,dsrB基因相对丰度与SO₄^2–浓度、DOC浓度存在显著正相关性。高通量测序结果表明,深层地下水中硫酸盐还原菌的α多样性显著高于浅层地下水。研究区内硫酸盐还原菌可...     

一株芽孢杆菌对含砷矿物中砷的还原作用

在自然环境中,砷通常吸附于铁氧化物、铝氧化物等金属氧化物矿物上,或与这些氧化物矿物形成共沉淀。厌氧条件下,微生物可能通过直接还原砷或者还原铁氧化物等载砷矿物从而影响砷的迁移转化。本研究筛选得到芽孢杆菌属的一株细菌DX-04,并研究了该菌株对不同形态砷酸盐的还原作用和还原途径。厌氧条件下,在12~24 h内菌株DX-04对溶解态砷的还原能力最强,溶解态砷对提高细菌生物量具有明显的促进作用。接种菌株DX-04的铁砷共沉淀培养基中液相As(III)浓度呈先升高后降低的趋势,砷发生还原与释放,进而发生二次沉淀再次被固持。当以载砷氧化铝矿物为载砷的模型矿物时,在DX-04菌株的还原作用下,吸附的As(V)首先从氢氧化铝矿物上释放到液相,进一步被还原为As(III)。微生物的这一作用能够引起含砷矿物中的砷向水体、沉积物环境中释放,成为人类健康的潜在威胁。     

砷还原菌群对砷的还原作用及菌群的多样性分析

从砷污染土壤中富集砷抗性细菌,在厌氧环境中进行培养,观察其对砷的还原能力。结果表明：在21 h之内,As(V)就被完全还原为As(Ⅲ);培养72 h后,培养基中出现黄色沉淀,采用X射线衍射分析(XRD)和扫描电镜-能谱分析(SEM-EDS)技术对沉淀进行分析表明,沉淀主要是以3种晶型存在的硫化砷(AsS);培养150 h后,大约有65%的As以上述沉淀的方式从溶液中移除。此外,本文还采用了构建16S rDNA文库的方式对该体系中的微生物种群进行分析,利用RFLP技术对16S rDNA片段进行分型,共得到72个操作单元类型(OTU),其中6个OTU占了库容的51%,从这6个OTU中各选取1个克隆进行测序,结果表明,富集到的砷还原细菌属于喜热菌属(Caloramator)、梭菌属(Clostridium)和杆菌属(Bacillus)。     

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.     

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…     

Effects of selenium on arsenic uptake in arsenic hyperaccumulator Pteris vittata L

Selenium (Se) is a non-metallic element, which has the capability to increase the antioxidative capacity and stress tolerance of plants to heavy metals. Plants vary considerably in their physiological response to Se. The reported research investigated the effects of Se on arsenic (As) uptake by As hyperaccumulator Pteris vittata L. and determined possible mechanisms of interaction. Pteris vittata plants were exposed hydroponically to 0, 150 or 300 microM of Na(2)HAsO(4) in the presence of 0, 5 or 10 microM of Na(2)SeO(4) for 5 or 10d. Application of 5 microM Se enhanced As concentration by P. vittata fronds by 7-45%. At 5 microM, Se acted as an antioxidant, inhibiting lipid peroxidation (reduced by 26-42% in the fronds) via increased levels of thiols and glutathione (increased by 24% in the fronds). The results suggest that Se is either an antioxidant or it activates plant protective mechanisms, thereby alleviating oxidative stress and improving arsenic uptake in P. vittata.     

Recent trend in nanoparticle research in regulating arsenic bioaccumulation and mitigating arsenic toxicity in plant species

Journal of Plant Biochemistry and Biotechnology - Atmospheric contamination by heavy metals/metalloids is a widespread global issue. Industrial discharges, along with agricultural and anthropogenic...     

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.     

Effect of arsenic on visible symptom and arsenic concentration in hydroponic Japanese mustard spinach

Responses of Japanese mustard spinach (JM-spinach; Brassica rapa L. var. pervirdis) were investigated at elevated levels of arsenic (As). Plants were grown hydroponically in the greenhouse under 0, 6.7, 33.5 and 67 μM As (equal to 0, 0.5, 2.5 and 5 mg L^?1 As, respectively) for 14 days. Arsenic was used as sodium meta-arsenite (NaAsO₂). Toxicity symptom was solely shown as shoot growth repression at 33.5 and 67 μM As exposures. Dry weight (DW) enhanced by 19.4% in shoot and 38.9% in root in the 6.7 μM As level as compared to control but decreased by 48.1% and 72.1% DW in shoot and 24.1% and 61.1% DW in root in the 33.5 and 67 μM As levels, respectively. This result indicated that As at lower concentration might have slight stimulating effect on JM-spinach growth, but toxicity increased with increasing As. Based on the regression lines between growth and As concentration in the plant tissues, the critical toxicity level (CTL) of As in JM-spinach shoot was 7.85 μg g^?1 DW considering 10% DW reduction. The CTL for the root was almost 2110 μg As g^?1 DW, indicating that shoot of JM-spinach was more sensitive to As-toxicity than that of root. Arsenic concentrations increased in plant parts with increasing As in the medium. Arsenic concentrations were also compared in DW and fresh weight (FW) basis. The JM-spinach concentrated unaccepted level of As in shoots for human consumption in the higher As levels without showing visible toxicity symptom. In spite of decreasing iron (Fe) concentration in shoot in the highest As level, chlorophyll index did not decrease accordingly. Phosphorus (P) concentration also decreased. Phosphorus concentration decreased much more than Fe concentration. Low P might help to mobilize Fe in shoots, resulting in higher chlorophyll index at 67 μM As level. Phosphorus might compete with Fe in shoot tissues of As-stressed JM-spinach.     

Evaluation of amino acid profile in contrasting arsenic accumulating rice genotypes under arsenic stress

Amino acids (AAs) play significant roles in metal binding, antioxidant defense, and signaling in plants during heavy metal stress. In the present study, the essential amino acids (EAAs), non-essential amino acids (NEAAs), as well as the enzymes of proline and cysteine biosynthetic pathways were studied in contrasting arsenic accumulating rice genotypes grown in hydroponic solutions with addition of arsenate (As^V) or arsenite (As^III). Under a mild As stress, the total AAs content significantly increased in both the rice genotypes with a greater increase in a low As accumulating rice genotype (LAARG; IET-19226) than in a high As accumulating rice genotype (HAARG; BRG-12). At the equimolar concentration (10 μM), As^III had a greater effect on EAAs than As^V. Conversely, As^V was more effective in inducing a proline accumulation than As^III. Among NEAAs, As significantly induced the accumulation of histidine, aspartic acid, and serine. In contrast, a higher As concentration (50 μM) reduced the content of most AAs, the effect being more prominent during As^III exposure. The inhibition of glutamate kinase activity was noticed in HAARG, conversely, serine acetyltransferase and cysteine synthase activities were increased which was positively correlated with the cysteine synthesis.     

Resistance to arsenic compounds in microorganisms

Abstract: Arsenic ions, frequently present as environmental pollutants, are very toxic for most microorganisms. Some microbial strains possess genetic determinants that confer resistance. In bacteria, these determinants are often found on plasmids, which has facilitated their study at the molecular level. Bacterial plasmids conferring arsenic resistance encode specific efflux pumps able to extrude arsenic from the cell cytoplasm thus lowering the intracellular concentration of the toxic ions. In Gram-negative bacteria, the efflux pump consists of a two-component ATPase complex. ArsA is the ATPase subunit and is associated with an integral membrane subunit, ArsB. Arsenate is enzymatically reduced to arsenite (the substrate of ArsB and the activator of ArsA) by the small cytoplasmic ArsC polypeptide. In Gram-positive bacteria, comparable arsB and arsC genes (and proteins) are found, but arsA is missing. In addition to the wide spread plasmid arsenic resistance determinant, a few bacteria confer resistance to arsenite with a separate determinant for enzymatic oxidation of more-toxic arsenite to less-toxic arsenate. In contrast to the detailed information on the mechanisms of arsenic resistance in bacteria, little work has been reported on this subject in algae and fungi.     

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.