Complexation of Hg with phytochelatins is important for plant Hg tolerance

Three-week-old alfalfa (Medicago sativa), barley (Hordeum vulgare) and maize (Zea mays) were exposed for 7 d to 30 μm of mercury (HgCl(2) ) to characterize the Hg speciation in root, with no symptoms of being poisoned. The largest pool (99%) was associated with the particulate fraction, whereas the soluble fraction (SF) accounted for a minor proportion (<1%). Liquid chromatography coupled with electro-spray/time of flight mass spectrometry showed that Hg was bound to an array of phytochelatins (PCs) in root SF, which was particularly varied in alfalfa (eight ligands and five stoichiometries), a species that also accumulated homophytochelatins. Spatial localization of Hg in alfalfa roots by microprobe synchrotron X-ray fluorescence spectroscopy showed that most of the Hg co-localized with sulphur in the vascular cylinder. Extended X-ray Absorption Fine Structure (EXAFS) fingerprint fitting revealed that Hg was bound in vivo to organic-S compounds, i.e. biomolecules containing cysteine. Albeit a minor proportion of total Hg, Hg-PCs complexes in the SF might be important for tolerance to Hg, as was found with Arabidopsis thaliana mutants cad2-1 (with low glutathione content) and cad1-3 (unable to synthesize PCs) in comparison with wild type plants. Interestingly, high-performance liquid chromatography-electrospray ionization-time of flight analysis showed that none of these mutants accumulated Hg-biothiol complexes.     

南极生物粪对汞的生物放大作用

首次报道了南极地区金图企鹅、阿德利企鹅、黑背鸥和巨海燕 4种海鸟 ,以及象海豹、威德尔海豹、毛皮海狮等 3种海豹新鲜粪便中的全 Hg含量 ,并对比了该区风化土壤的 Hg含量 ,其中 Hg含量水平依次为 :土壤背景值 <黑背鸥粪 <金图企鹅粪 <阿德利企鹅粪 <威德尔海豹粪 <象海豹粪 <巨海燕粪 <毛皮海狮粪 ,可见 ,相对于背景土壤生物粪明显富集 Hg元素。此外 ,结合上述粪便样品的 δ15 N测定结果 ,讨论了南极生物粪对 Hg的生物放大作用。研究表明 ,随着 δ15 N升高 ,即生物营养级的升高 ,其粪便的全 Hg含量有逐步增高的趋势 ,这证实了生物粪和生物肌体一样存在对 Hg的生物放大作用。文中还对南极阿德雷岛一个经过企鹅粪污染的 Y2湖泥芯剖面上 15个样品的 2 6种元素进行了聚类分析 ,结果表明 ,沉积物中的汞和企鹅粪的标型元素具有同源性 ,即沉积物中的汞主要是由企鹅粪带入的 ,由于生物粪对汞的生物富集作用 ,生物粪的混入会造成湖泊沉积物中汞含量的增高     

G-quadruplex DNAzyme-based Hg2+ and cysteine sensors utilizing Hg2+-mediated oligonucleotide switching

A simple and sensitive colorimetric Hg(2+) detection method is reported, based on the Hg(2+)-mediated structural switch of an unlabeled oligonucleotide strand. In the absence of Hg(2+), the oligonucleotide strand forms a stem-loop. A G-rich sequence in the strand is partially caged in the stem-loop structure and cannot fold into a G-quadruplex. In the presence of Hg(2+), T-Hg(2+)-T coordination chemistry leads to the formation of another stem-loop structure and the release of the G-rich sequence. The released sequence folds into a G-quadruplex, which binds hemin to form catalytically active G-quadruplex DNAzymes. This is detected as an absorbance increase in a H(2)O(2)-2,2'-azinobis(3-ethylbenzothiozoline)-6-sulfonic acid (ABTS) reaction system using UV-vis absorption spectroscopy. This simple colorimetric sensor can detect aqueous Hg(2+) at concentrations as low as 9.2 nM with high selectivity. Based on the strong binding interaction between Hg(2+) and the sulfur-containing amino acid cysteine (Cys), and the competition between Cys and a oligonucleotide for Hg(2+), the proposed Hg(2+)-sensing system can be further exploited as a Cys-sensing method. The method has a detection limit for Cys of 19 nM.     

汞对土壤酶活性的影响

利用室内模拟方法,研究了重金属Hg对不同土样脲酶、转化酶和中性磷酸酶活性的影响.结果表明,Hg可显著地抑制土壤脲酶和转化酶的活性,但不同土样Hg对两种酶活性的抑制程度有很大差别.HgCl₂浓度与两种酶活性之间的关系均可用对数方程很好地描述(P<0.05).4个土样的脲酶ED₅₀(生态剂量)分别为87.99、5.47、24.05和19.88 mg·kg^-1;转化酶的ED₅₀分别为76.68、727.49、236.52和316.59 mg·kg^-1.脲酶对Hg污染比转化酶敏感;有机质对土壤酶活性有一定的保护作用.除连续2年施用大量有机肥的草甸棕壤土样中Hg对中性磷酸酶有显著的激活作用外(P<0.05),其它土样无显著变化,表明中性磷酸酶活性对Hg污染反应不敏感.     

不同Hg浓度下水稻中Hg的分布累积特征

利用土壤盆栽实验方法,研究了土壤中Hg的形态分布,以及不同Hg浓度下,水稻不同生长时期各组织中Hg的分布规律和累积特征.土壤Hg存在的形态为:有机结合态＞残渣态＞＞氧化态＞＞溶解与可交换态≈特殊吸附态,在水稻的生长过程中,土壤中的Hg呈现从残渣态向有机结合态转化的趋势,有机结合态Hg平均占比为61.7％,是土壤Hg最重要的存在形态.Hg在水稻不同部位的浓度分布呈现W根＞W叶＞W茎＞W穗＞W籽粒,分析表明,水稻根、茎和籽粒中的Hg与土壤各形态Hg浓度呈显著或极显著正相关,但在水稻生长后期叶片中Hg与土壤Hg浓度的相关性不显著,叶片Hg与大气Hg进行交换起主要作用,改变了累积状况.随着土壤Hg浓度的增加和生长期的延长,根对Hg的束缚能力逐渐增加,根部Hg累积量增大,所占比例上升,而茎和叶的累积量相对稳定.     

Reduction of Hg(II) to Hg(0) by Biogenic Magnetite from two Magnetotactic Bacteria

Understanding the biogeochemical cycle of the highly toxic element mercury (Hg) is necessary to predict its fate and transport. In this study, we determined that biogenic magnetite isolated from Magnetospirillum gryphiswaldense MSR-1 and Magnetospirillum magnetotacticum MS-1 was capable of reducing inorganic mercury [Hg(II)] to elemental mercury [Hg(0)]. These two magnetotactic bacteria (MTB) lacked mercuric resistance operons in the genomes. However, they revealed high resistance to Hg(II) under atmospheric conditions and an even higher resistance under microaerobic conditions (1% O₂ and 99% N₂). Neither strain reduced Hg(II) to Hg(0) under atmospheric conditions. However, a slow rate (0.05–0.21 µM·d^?1) of Hg(II) loss occurred from late log phase to stationary phase in two MTBs' culture media under microaerobic conditions. Increased Hg(II) entered both cells under microaerobic conditions relative to atmospheric conditions. The majority of Hg(II) was still blocked by the cell membrane. Hg(II) reduction was more effective when biogenic magnetite was extracted out, with or without the magnetosome membrane envelope. When magnetosome membrane was present, 8.55–13.53% of 250 nM Hg(II) was reduced to Hg(0) by 250 mg/L biogenic magnetite suspension within 2 hours. This ratio increased to 55.07–64.70% while magnetosome membrane was removed. We concluded that two MTBs contributed to the reduction of Hg(II) to Hg(0) at a slow rate in vivo. Such reduction was more favorable to occur when biogenic magnetite is released from dead cells. It proposed a new biotic pathway for the formation of Hg(0) in aquatic systems.     

Acclimation of aquatic microbial communities to Hg(II) and CH3Hg+ in polluted freshwater ponds

The relationship of mercury resistance to the concentration and chemical speciation of mercurial compounds was evaluated for microbial communities of mercury-polluted and control waters. Methodologies based on the direct viable counting (DVC) method were adapted to enumerate mercury-resistant communities. Elevated tolerance to Hg(II) was observed for the microbial community of one mercury-polluted pond as compared to the community of control waters. These results suggest an in situ acclimation to Hg(II). The results of the methylmercury resistance-DVC assay suggested that minimal acclimation to CH₃Hg⁺ occurred since similar concentrations of CH₃HgCl inhibited growth of 50% of organisms in both the control and polluted communities. Analyses of different mercury species in pond waters suggested that total mercury, but not CH₃Hg⁺ concentrations, approached toxic levels in the polluted ponds. Thus, microbial acclimation was specific to the chemical species of mercury present in the water at concentrations high enough to cause toxic effects to nonacclimated bacterial communities.     

Biotransformation of Hg(II) by Cyanobacteria

The biotransformation of Hg(II) by cyanobacteria was investigated under aerobic and pH-controlled culture conditions. Mercury was supplied as HgCl₂ in amounts emulating those found under heavily impacted environmental conditions where bioremediation would be appropriate. The analytical procedures used to measure mercury within the culture solution, including that in the cyanobacterial cells, used reduction under both acid and alkaline conditions in the presence of SnCl₂. Acid reduction detected free Hg(II) ions and its complexes, whereas alkaline reduction revealed that meta-cinnabar (β-HgS) constituted the major biotransformed and cellularly associated mercury pool. This was true for all investigated species of cyanobacteria: Limnothrix planctonica (Lemm.), Synechococcus leopoldiensis (Racib.) Komarek, and Phormidium limnetica (Lemm.). From the outset of mercury exposure, there was rapid synthesis of β-HgS and Hg(0); however, the production rate for the latter decreased quickly. Inhibitory studies using dimethylfumarate and iodoacetamide to modify intra- and extracellular thiols, respectively, revealed that the former thiol pool was required for the conversion of Hg(II) into β-HgS. In addition, increasing the temperature enhanced the amount of β-HgS produced, with a concomitant decrease in Hg(0) volatilization. These findings suggest that in the environment, cyanobacteria at the air-water interface could act to convert substantial amounts of Hg(II) into β-HgS. Furthermore, the efficiency of conversion into β-HgS by cyanobacteria may lead to the development of applications in the bioremediation of mercury.     

Biotransformation of Hg(II) by cyanobacteria

The biotransformation of Hg(II) by cyanobacteria was investigated under aerobic and pH-controlled culture conditions. Mercury was supplied as HgCl(2) in amounts emulating those found under heavily impacted environmental conditions where bioremediation would be appropriate. The analytical procedures used to measure mercury within the culture solution, including that in the cyanobacterial cells, used reduction under both acid and alkaline conditions in the presence of SnCl(2). Acid reduction detected free Hg(II) ions and its complexes, whereas alkaline reduction revealed that meta-cinnabar (beta-HgS) constituted the major biotransformed and cellularly associated mercury pool. This was true for all investigated species of cyanobacteria: Limnothrix planctonica (Lemm.), Synechococcus leopoldiensis (Racib.) Komarek, and Phormidium limnetica (Lemm.). From the outset of mercury exposure, there was rapid synthesis of beta-HgS and Hg(0); however, the production rate for the latter decreased quickly. Inhibitory studies using dimethylfumarate and iodoacetamide to modify intra- and extracellular thiols, respectively, revealed that the former thiol pool was required for the conversion of Hg(II) into beta-HgS. In addition, increasing the temperature enhanced the amount of beta-HgS produced, with a concomitant decrease in Hg(0) volatilization. These findings suggest that in the environment, cyanobacteria at the air-water interface could act to convert substantial amounts of Hg(II) into beta-HgS. Furthermore, the efficiency of conversion into beta-HgS by cyanobacteria may lead to the development of applications in the bioremediation of mercury.     

Spectroscopic characterization of rat kidney Hg,Cu-metallothionein

Absorption, circular dichroism (CD), magnetic circular dichroism (MCD) and emission spectra are reported for rat kidney Hg,Cu-metallothionein isoform 3 isolated following induction of the metallothionein with HgCl2. While the absorption spectrum is featureless, both the CD and MCD spectra show resolved bands that arise from the Cu-thiolate and Hg-thiolate groups. The emission spectrum at 77 K is much more complicated than would be expected for a copper (I)-containing metallothionein. It is suggested the emission only arises from the copper-thiolate groups but that the presence of the mercury results in copper ions in several different environments depending on the nature of the nearest neighbour.     

Hg2+ uptake in a cyanobacterium

The uptake of Hg²⁺ and its regulation in the cyanobacteriumNostoc calcicola Bréb. was studied. Hg²⁺ uptake pattern consisted of two distinct phases: (a) rapid binding of the cation to the negatively charged cell surface (first 10 min) and (b) its subsequent metabolism-dependent intracellular import, at least up to 40 min (saturating concentration 1.5 M Hg²⁺, K_m=1.0M Hg²⁺ and V_max 0.21 nmol Hg²⁺ mg^–1 protein min^–1). Hg²⁺ influx, to a major extent, depended on photosynthetically generated energy, and the supply of exogenous ATP (10 M) or DCMU (5 M) suggested the vital role of PS II-mediated energy to support the process. The significant lowering in Hg²⁺ uptake rate as well as total cellular Hg²⁺ in the presence ofp-chloromercuribenzoate (pCMB), azide (NaN₃), N,N-dicyclohexycarbodiimide (DCCD), and thiol (mercaptoethanol) indicated the role of membrane potential,-SH groups, and ATP hydrolysis in regulating Hg²⁺ transport. While Cu²⁺ antagonized Hg²⁺ intake, Ni²⁺ showed synergism.     

Reactivity of Hg(II) with superoxide: evidence for the catalytic dismutation of superoxide by Hg(II).

Mercuric ion, a well-known nephrotoxin, promotes oxidative tissue damage to kidney cells. One principal toxic action of Hg(II) is the disruption of mitochondrial functions, although the exact significance of this effect with regard to Hg(II) toxicity is poorly understood. In studies of the effects of Hg(II) on superoxide (O2-) and hydrogen peroxide (H2O2) production by rat kidney mitochondria, Hg(II) (1-6 microM), in the presence of antimycin A, caused a concentration-dependent increase (up to fivefold) in mitochondrial H2O2 production but an apparent decrease in mitochondrial O2- production. Hg(II) also inhibited O(2-)-dependent cytochrome c reduction (IC50 approximately 2-3 microM) when O2- was produced from xanthine oxidase. In contrast, Hg(I) did not react with O2- in either system, suggesting little involvement of Hg(I) in the apparent dismutation of O2- by Hg(II). Hg(II) also inhibited the reactions of KO2 (i.e., O2-) with hemin or horseradish peroxidase dissolved in dimethyl sulfoxide (DMSO). Finally, a combination of Hg(II) and KO2 in DMSO resulted in a stable UV absorbance spectrum [currently assigned Hg(II)-peroxide] distinct from either Hg(II) or KO2. These results suggest that Hg(II), despite possessing little redox activity, enhances the rate of O2- dismutation, leading to increased production of H2O2 by renal mitochondria. This property of Hg(II) may contribute to the oxidative tissue-damaging properties of mercury compounds.     

Influence of sodium selenite on203Hg absorption,distribution, and elimination in male mice exposed to methyl203Hg

To study the effects of long-term selenium supplementation on absorption, distribution, and elimination of methylmercury (MeHg) in mice, three groups of male mice (Balb/c CA) were exposed for 7 wk to 0, 0.6, and 3 ppm sodium selenite in tap water. They were then given a single oral dose of Me²⁰³Hg (2 μmol/kg) by gastric intubation, and elimination of²⁰³Hg was followed by whole-body counting for 49 d at the same Se exposure as previously. Twenty-four hours and 49 d after dosage, 6–7 animals/group were sampled for analysis of²⁰³Hg distribution in the body. Glutathione peroxidase (GSH-PX) activity in blood and selenium levels in the liver were used as measures of selenium status. Gastrointestinal absorption of Me²⁰³Hg was not influenced by the Se status of the animals. Selenium supplementation of MeHg-exposed mice caused an enhanced whole-body elimination of Hg, but selenium-supplemented animals did not have lower Hg levels in the brain and kidney than nonsupplemented animals. The effect of selenium on the accumulation, of Hg in the brain was dose-dependent, a high dose (3 ppm Se) causing a higher initial accumulation of Hg. The intracellular distribution of²⁰³Hg in the liver and kidney was not affected by Se. The results indicate that selenium treatment of MeHg-exposed mice may have a positive effection the health of the animals by decreasing the total body burden of MeHg.     

Electrochemical characterization of redox centers organized at Hg surfaces

The electrochemical characterization of a series of redox sites absorbed at Hg surface by different interactions is reported. The redox centers, based on Fe(II) and Ru(II), are incorporated, respectively, in the molecules Fe(C₅H₅)(C₅H₄)(CH₂)₄SH and [Ru(NH₃)₅NC₅H₄CH₂NHCO(CH₂)₁₀SH](PF₆), and are anchored on the Hg surface in one component self-assembled monolayers. The electrochemical behaviour of these systems indicates that redox centers are located onto a uniform, homogeneous environment at the external surface of the monolayer. We also report the electrochemical behaviour of the positively charged redox species [Ru(NH₃)₆]³⁺ when the Hg electrode surface is functionalized with a negatively charged SAM. The SAM is formed by 11-mercaptoundecanoic acid that exposes carboxylic acid groups to solutions of different pH values. At a pH lower than 4, the cyclic voltammograms show negligible current, and pH from 5 to 9, the voltammograms are essentially identical and show a well-defined redox wave. From a study of the voltammetric responses of the couple as a function of the electrolyte composition and concentration at pH 9, we suggest that the redox reaction takes place at the defects of the SAMs created by the repulsion of the -COO⁻ head groups and that the current is determined by a diffusion-controlled mechanism.