镉、铜和锌胁迫下黑藻活性氧的产生及抗氧化酶活性的变化研究

研究了不同Cd、Cu、Zn处理浓度对黑藻体内活性氧()产生及对抗氧化酶(SOD、POD、CAT)活性的分子毒理学效应以探讨高等水生植物抗氧化酶对重金属胁迫的反应。结果表明,三种重金属都不同程度地加快了产生速率;Cu使SOD、POD、CAT活性下降;Cd也都减弱了SOD和POD活性,而CAT活性在0.5—5mg/L处理浓度时增加;Zn对SOD活性也为抑制作用,当浓度为0.5—5mg/L时POD和CAT活性都上升。关联度分析发现Cd、Cu和Zn胁迫下黑藻起主要保护作用的分别为SOD、POD和CAT,而SOD最易受到影响。Cd、Cu处理下的叶绿素含量也都呈下降趋势,而0.5—5mg/L的Zn浓度刺激了叶绿素合成。所有Zn处理、0.5mg/L的Cu处理和0.5—1mg/L的Cd处理的叶绿素a/b值都大于对照值。除了Cu使可溶性蛋白含量减少外,0.5—5mg/L的Zn和0.5—1mg/L的Cd都使其含量增加。综合起来,Cu的毒性最强,其次为Cd,Zn最弱。致死阈浓度分别为:Cu:0.5—1mg/L;Cd:1—2mg/L;Zn:5—6mg/L。SOD是评价重金属对沉水植物毒性效应的灵敏指标。黑藻对水环境Cu污染反应敏感。       

6种重金属对赤子爱胜蚓的急性毒性效应与风险评价

【背景】近年来,土壤重金属污染问题日益凸显,对生态环境、食品安全和人体健康构成了严重威胁,其急性毒性尚未明确。【方法】采用滤纸接触法和人工土壤法测定了铜(Cu2+)、锌(Zn2+)、镍(Ni+)、镉(Cd2+)、铅(Pb2+)和锰(Mn2+)等6种重金属对赤子爱胜蚓的急性毒性效应,并参照欧盟指令91/414/EEC标准评价了其环境风险。【结果】滤纸接触法48h测定结果表明,6种重金属对蚯蚓的LC50为3.17(2.53～3.81)～90.42(69.45～140.47)μg.cm-2,其毒性次序为Cu2+>Zn2+>Ni+>Cd2+>Pb2+>Mn2+。人工土壤法14d测定结果表明,6种重金属对蚯蚓的LC50为1347(1236～1453)～6936(6144～8930)mg.kg-1,其毒性次序为Cu2+>Cd2+>Ni+>Zn2+>Mn2+>Pb2+。风险评价结果显示,Cu2+、Zn2+、Ni+、Cd2+、Pb2+和Mn2+等6种重金属的暴露比(toxicity/exposureratio,TER)分别为3.37、4.46、8.68、1428、13.87和5.85。【结论与意义】6种重金属对土壤动物蚯蚓均具有潜在的毒性效应。Cu2+、Zn2+、Ni+、Mn2+等4种重金属对赤子爱胜蚓存在急性毒性风险,而Cd2+和Pb2+对赤子爱胜蚓的急性毒性风险水平是可接受的。该评价结果可为我国制定基于风险的土壤环境质量标准提供依据。     

Effects of heavy metals on nitrogen uptake by Paxillus involutus and mycorrhizal birch seedlings

The effects of the heavy metals Cu, Cd, Ni, Pb and Zn on [(14)C]methylamine and [(14)C]aminoisobutyric acid uptake were studied in the free-living fungus Paxillus involutus and in mycorrhizal and non-mycorrhizal birch roots. The uptake of both N sources by P. involutus was inhibited by the five metals tested. However, Cu(2+) and Pb(2+) had a greater inhibitory effect. Non-competitive inhibitions were determined between heavy metals and [(14)C]methylamine uptake. [(14)C]Methylamine uptake was reduced by one third by 2 μM Cd(2+) and Cu(2+) in non-mycorrhizal roots, whereas that of mycorrhizal roots was not affected. However, it was reduced by 30 to 80% by 200 μM Cd(2+) and Cu(2+) irrespective of the mycorrhizal status. [(14)C]Aminoisobutyric acid uptake in mycorrhizal roots was not significantly affected by Cd(2+) and Cu(2+), whereas that of non-mycorrhizal roots was decreased by 77% at 200 μM Cu(2+). [(14)C]Aminoisobutyric acid uptake was 4.5 to 6 fold higher in mycorrhizal roots, compared with non-mycorrhizal roots, even under metal exposure. The high efficiency of N acquisition by mycorrhizal birch seedlings under metal exposure might be regarded as a mechanism of stress avoidance.     

产油微藻Auxenochlorella protothecoides UTEX 2341对镉胁迫的生理响应及抗性机理

【背景】重金属镉(Cd)污染问题日益严峻。微藻是一种良好的生物吸附剂。目前研究多关注于微藻对Cd的去除率及吸附性能方面,对其抗Cd机理的研究比较少。【目的】以产油微藻Auxenochlorella protothecoides UTEX 2341为材料,研究Cd胁迫对UTEX 2341生理性状、抗性机理及产油的影响。【方法】测定微藻在0-5mmol/LCd胁迫下的生长产油情况,并进一步分析2mmol/LCd胁迫下藻体中色素、可溶性蛋白和油脂含量的变化,以及藻体亚显微结构、抗氧化酶和抗氧化剂、脂肪酸组分的改变情况。【结果】Auxenochlorella protothecoides UTEX 2341能够耐受2 mmol/L Cd胁迫,虽然其生物量和叶绿素含量在Cd胁迫下略有降低,但油脂产量显著增加,胁迫168 h时为1.60 g/L,是对照的1.77倍。此外,高浓度Cd引起了胞内活性氧的积累。抗性机理分析表明高Cd胁迫显著抑制了微藻体内的抗氧化酶活性,但非酶抗氧化剂类胡萝卜素和还原性谷胱甘肽的含量显著增加,分别是对照的1.42倍和4.5倍,从而缓解高浓度Cd对微藻细胞造成的氧化损伤,减轻Cd的毒性。脂肪酸组分分析结果表明,Cd胁迫下油酸(C18:1)含量增加,且脂肪酸成分中C16-C18的含量达96%-98%,符合生物柴油的生产标准。【结论】该研究为揭示微藻抗Cd的机理以及Cd胁迫下微藻柴油合成的调控机理提供了一定的研究基础。     

Heavy Metal Accumulation and Ecological Responses of Turfgrass to Rubbish Compost with EDTA Addition

Domestic rubbish compost is a complex-polluted system, containing multiple heavy metals,which limits its application. In the present study, Cr, Mn, Ni, Cu, Zn, Cd, and Pb accumulation and ecological responses of turfgrass to rubbish compost were investigated following the addition of EDTA. The results showed that the addition of EDTA significantly increased heavy metal accumulation in Lolium perenne L.and Festuca arundinacea L. Most heavy metal concentrations in L. perenne increased with increasing EDTA supply. The concentrations of Cr, Mn, Ni, Cu, and Cd in L. perenne were highest following the addition of 30 mmol/kg EDTA and the concentrations of Cr and Ni at this point reached concentrations of1914.17 and 521.25 μg/g, respectively. When the EDTA level was ＜ 20 mmol/kg, the accumulation of most heavy metals in F. arundinacea increased with increasing EDTA supply, but showed a tendency to decrease at EDTA concentrations ＞20 mmol/kg. The highest concentrations of Mn, Ni, Cu, and Zn in F.arundinacea reached 268.01, 110.94, 161.52 and 1 354.97 μg/g, respectively, following the addition of 20mmol/kg EDTA. The EDTA-induced increase in the accumulation of heavy metals in turfgrass was plantand metal-specific. L. perenne had a relatively high ability to accumulate Cr, Ni, and Zn. The highest Zn concentration was 2 979.58 μg/g and, following the addition of EDTA, the concentrations of the three metals were increased 26.23, 20.03, and 10.49-fold, respectively, compared with control. However, F. arundinacea showed a high ability to accumulate Cr, with the highest concentration (596.02 μg/g) seen following the addition of 30 mmol/kg EDTA; the concentration of Cr increased 15.51-fold compared with control. With EDTA addition, ecological responses of both turfgrass species showed that EDTA at concentrations ＜10mmol/kg increased seed germination and aboveground net primary production (ANP) of L. perenne and slightly inhibited those of F.arundinacea, but EDTA at concentrations ＞20 mmol/kg inhibited these parameters significantly for both species. Moreover, EDTA increased the chlorophyll and proline content at all concentrations tested. On the basis of the synthetic remediation index, the optimal EDTA concentration for turfgrass remediation of heavy metals in compost is approximately 10 mmol/kg.     

The nucleotide sequence of metallothioneins (MT) in liver of the Kafue lechwe (Kobus leche kafuensis) and their potential as biomarkers of heavy metal pollution of the Kafue River

The study determined heavy metal concentrations and MT1 nucleotide sequence [phylogeny] in liver of the Kafue lechwe. Applicability of MT1 as a biomarker of pollution was assessed. cDNA-encoding sequences for lechwe MT1 were amplified by RT-PCR to characterize the sequence of MT1 which was subjected to BLAST searching at NCBI. Phylogenetic relationships were based on pairwise matrix of sequence divergences calculated by Clustal W. Phylogenetic tree was constructed by NJ method using PHILLIP program. Metals were extracted by acid digestion and concentrations of Cr, Co, Cu, Zn, Cd, Pb, and Ni were determined using an AAS. MT1 mRNA expression levels were measured by quantitative comparative real-time RT-PCR. Lechwe MT1 has a length of 183bp, which encode for MT1 proteins of 61AA, which include 20 cysteines. Nucleotide sequence of lechwe MT1 showed identity with sheep MT (97%) and cattle MT1E (97%). Phylogenetic tree revealed that lechwe MT1 was clustered with sheep MT and cattle MT1E. Cu and Ni concentrations and MT1 mRNA expression levels of lechwe from Blue Lagoon were significantly higher than those from Lochinvar (p<0.05). Concentrations of Cd and Cu, Co and Cu, Co and Pb, Ni and Cu, and Ni and Cr were positively correlated. Spearman's rank correlations also showed positive correlations between Cu and Co concentrations and MT mRNA expression. PCA further suggested that MT mRNA expression was related to Zn and Cd concentrations. Hepatic MT1 mRNA expression in lechwe can be used as biomarker of heavy metal pollution.     

Functional expression of AtHMA4, a P1B-type ATPase of the Zn/Co/Cd/Pb subclass

Mechanisms are required by all organisms to maintain the concentration of essential heavy metals (e.g. Zn and Cu) within physiological limits and to minimise the detrimental effects of non-essential heavy metals (e.g. Cd). Heavy-metal P-type ATPases (HMAs) are a subgroup of the P-type ATPase superfamily that may contribute to metal homeostasis in plants. We cloned and characterised a member of this family, AtHMA4, from Arabidopsis thaliana that clusters with the Zn/Co/Cd/Pb subclass of HMAs on phylogenetic analysis. Sequencing of the AtHMA4 cDNA showed that it contained the conserved motifs found in all P-type ATPases and also motifs that are characteristic of heavy-metal ATPases. Escherichia coli mutants defective in the HMAs, CopA and ZntA, were used in functional complementation studies. AtHMA4 was able to restore growth at high [Zn] in the zntA mutant but not at high [Cu] in the copA mutant, suggesting a role in zinc transport. Heterologous expression of AtHMA4 in Saccharomyces cerevisiae made the yeast more resistant to Cd but did not affect sensitivity to other metals compared with vector-transformed controls. The organ specificity of AtHMA4 was analysed in Arabidopsis and showed that AtHMA4 was expressed in a range of tissues with highest expression in roots. AtHMA4 was upregulated in roots exposed to elevated levels of Zn and Mn but downregulated by Cd. Possible physiological roles of this transporter in Arabidopsis are discussed.     

泥蚶谷胱甘肽过氧化物酶基因的全长克隆与表达分析

为了探讨谷胱甘肽过氧化物酶（Glutathione peroxidase,GPx）基因在泥蚶应激反应中的作用,研究采用RACE技术克隆了泥蚶GPx基因（TgGPx）cDNA全长,其cDNA全长1195 bp,包含45 bp 5’-UTR,639 bp开放阅读框（ORF）和511 bp 3’-UTR。ORF编码212个氨基酸残基,预测蛋白分子量为24.3 kD,理论等电点为8.33,其中,第53个氨基酸U是由密码子202UGA204编码的硒代半胱氨酸（Se-Cys）。在3’-UTR上存在一段序列,形成一种独特的茎环结构,即SECIS元件。SECIS元件在密码子UGA翻译为Se-Cys的过程中起决定性作用。通过序列比对与系统进化分析,发现软体动物中也存在不同种类的GPx基因,TgGPx与GPx1和GPx2的亲缘关系较近。利用qRT-PCR技术对TgGPx在泥蚶的不同组织以及重金属刺激后的表达量进行分析,结果表明,TgGPx在泥蚶的5个组织中都有表达,但存在组织特异性,在外套膜中的表达量最高,在血细胞中的表达量最低。用重金属铅、铜、镉刺激后,TgGPx在肝胰脏中的表达量显著升高,表明TgGPx在维护机体正常功能方面及泥蚶抵御外界刺激的应激反应中发挥作用。     

Isolation and characterization of heavy-metal-mobilizing bacteria from contaminated soils and their potential in promoting Pb, Cu, and Cd accumulation by Coprinus comatus

The enhanced effect of heavy-metal-mobilizing bacteria on the uptake of Pb, Cu, and Cd by Coprinus comatus from Pb-, Cu-, and Cd-multicontaminated soil was assessed in this study. Thirteen strains, tolerating 800?mg·L(-1) Pb, 200?mg·L(-1) Cu, and 200?mg·L(-1) Cd simultaneously were selected for heavy-metal-solubilizing experiments in soil. The mobilization of heavy metals depended on the characteristics of bacteria and heavy metals. Correlation analysis demonstrated that for Pb solubilization, the acid-producing ability was the most significant factor, while for Cu and Cd, siderophores played a leading role in this process. Four strains, based on their excellent ability to solubilize heavy metal in soil, were applied in pot experiments. The results showed that all strains can promote the growth of C. comatus and meanwhile help mushroom accumulate more heavy metals (Pb, Cd, and Cu). The maximum uptake for total Pb and Cu by C. comatus was observed in inoculations with Bacillus sp. strain JSG1 (2.02- and 2.13-fold, respectively, compared with uninoculated soil), while for Cd, it was recorded in Bacillus sp. strain PB2 treated soil (2.03-fold). Therefore, this work suggests that the mushroom-bacteria interaction can be developed into a novel bioremediation strategy.     

Regulation of cellular glutathione peroxidase by different forms and concentrations of selenium in primary cultured bovine hepatocytes

The expression and activity of cellular glutathione peroxidase (GPx1) are regulated by selenium (Se). Generally speaking, organic forms of Se have less toxicity and greater bioavailability compared with inorganic forms. In this study, the effects of different forms and concentrations of Se on the regulation of mRNA level and activity of GPx1 in bovine hepatocytes were evaluated, and the optimal doses of different forms of Se that supported the full expression of GPx1 were determined. Primary cultured bovine hepatocyte monolayers derived from neonatal male Holstein calves (aged 1–2 days) were incubated for 24 h with 0 (control), 0.5, 1, 1.5, 2, 3, 4 or 5 μmol/L of Se from dl-selenomethionine (Se-Met), sodium selenite (Na₂SeO₃) or Kappa-selenocarrageenan (Se-Car). Compared with controls, a significantly lower level of release of lactic dehydrogenase (LDH) was observed at 0.5–5 μmol/L of Se-Met, 0.5–1 μmol/L of Na₂SeO₃ and 0.5 μmol/L of Se-Car, but significantly higher LDH release was observed at 2–5 μmol/L of Na₂SeO₃ and 3–5 μmol/L of Se-Car, and the response occurred in a dose-dependent manner. The intracellular content of reduced glutathione in all hepatocytes treated with Se was significantly lower than that of controls. Significant increases in GPx1 mRNA were obtained in all hepatocytes treated with Se, with maximal effects at 3 μmol/L of Se-Met, 1.5 μmol/L of Na₂SeO₃ and 2 μmol/L of Se-Car, respectively. Furthermore, 3 μmol/L of Se from Se-Met resulted in peak levels of GPx1 mRNA. After reaching a maximal level, higher Se supplementation led to a reduction of GPx1 mRNA. The activity of GPx1 showed similar patterns but of lower magnitude. We conclude that (a) the regulation of mRNA level and activity of GPx1 in primary cultured bovine hepatocytes by different forms of Se varies and (b) the optimal doses of Se to support the full expression of GPx1 in bovine hepatocytes when supplied as Se-Met, Na₂SeO₃ and Se-Car are 3, 1.5 and 2 μmol/L, respectively.     

Quantitative and Qualitative Changes in Peroxidase of Cucurbita Pepo Cultivars Stressed with Heavy Metals

Seedlings of two cultivars of zucchini (Cucurbita pepo L.) Courgette d'Italie (CI) and Courgette d'Alger (CA) were pre-treated with various concentrations of cadmium, copper and zinc for 30 d. High accumulation of heavy metals especially in the roots was showed. Peroxidase activity was affected according to the type of metal added, concentration, and the plant cultivar used. In leaves and roots of the CI control plants peroxidase activities were 50 and 17 % higher than in the CA control plants. Treatment with Cd (5 μg g⁻¹), Cu (200 μg g⁻¹), and Zn (500 μg g⁻¹) increased peroxidase activities in CA but decreased it in CI both in leaves and roots. Heavy metals tested lead also to some qualitative changes characterized by appearance of new isoforms of peroxidase. The results show the possibility to use the activities of peroxidase as biomarkers for Cd, Cu and Zn stresses. This revised version was published online in July 2006 with corrections to the Cover Date.     

重金属在拟水狼蛛体内的分布及对其体内抗氧化酶活性的影响

为了探明重金属在拟水狼蛛Pirata subpiraticus体内的分布及对其体内抗氧化酶活性的影响,本研究于2009年6月在河南南阳地区5种不同生境下,共采集50份土壤样本和300头拟水狼蛛样本,采用原子吸收光谱法测定了5种不同生境下雄雌拟水狼蛛体内重金属的分布、GSH含量以及GST,CAT和SOD的活性。结果表明：5个采集样点（S1, S2, S3, S4和S5） 拟水狼蛛体内重金属（Cd,Pb,Cu和Zn）的含量差异显著（P<0.05）。同一地点拟水狼蛛体内重金属（Cd,Pb,Cu和Zn）的含量不同身体部位差异显著（P<0.05）,头胸部>足部>腹部,同一地点雄性拟水狼蛛重金属含量显著高于雌性（P<0.05）。在重金属胁迫下,重金属含量高（S1,S2,S3和S4）的拟水狼蛛体内GSH 含量显著高于参照组（S5）,雄性GSH 含量高于雌性（P<0.05）。对于不同身体部位,GSH 含量差异显著,头胸部GSH 含量最高,其次为足部,腹部含量最低;GSH 含量与重金属含量显著正相关（r2=0.9854,P<0.05）。对于GST,CAT和SOD,重金属含量高则酶活性低,雄性酶活性显著低于雌性,不同身体部位酶活差异不显著;GST,CAT和SOD酶活性与重金属（Cd和Pb或者Pb）含量显著负相关。因此检测拟水狼蛛不同身体部位的酶活性变化就可知环境中重金属的污染程度,拟水狼蛛可以作为重金属污染的重要监测指示生物。     

Induction of several acute-phase protein genes by heavy metals: a new class of metal-responsive genes.

Acute-phase reactants, metallothioneins, and heat-shock proteins are the products of three families of genes that respond to glucocorticoids and cytokines. Metallothioneins and heat-shock proteins, however, are also stimulated by heavy metals, whereas very little is known about the effect of heavy metals on acute-phase-reactant genes. We have studied the effect of heavy metals (Hg, Cd, Pb, Cu, Ni, and Zn) and Mg on the acute-phase reactants alpha 1-acid glycoprotein, C-reactive protein, alpha 1-antitrypsin and alpha 1-antichymotrypsin. alpha 1-Acid glycoprotein and C-reactive protein mRNA levels were increased severalfold in livers of heavy-metal-treated Balb/c mice. The strongest induction was mediated by Hg, followed in order of response by Cd greater than Pb greater than Cu greater than Ni greater than Zn greater than Mg. None of the metals affected the mRNA levels of albumin, alpha 1-antitrypsin, and alpha 1-antichymotrypsin. Furthermore, failure to repress albumin, a negative acute-phase reactant, indicated that the induction of these genes was not due to a metal-mediated inflammatory response. The metals also induced alpha 1-acid glycoprotein and C-reactive protein in adrenalectomized animals, indicating that induction by the heavy metals is not mediated by the glucocorticoid induction pathway. Sequence analysis has revealed a region of homology to metal-responsive elements in the alpha 1-acid glycoprotein and C-reactive protein promoters. Additionally, an alpha 1-acid glycoprotein expression vector, pAGP(-595)CAT, responded to Hg and Cd when transfected into human HepG2 cells.(ABSTRACT TRUNCATED AT 250 WORDS)     

铜尾矿库区土壤与植物中重金属形态分析

对铜陵铜尾矿区土壤和植物中重金属形态进行了研究.结果表明,尾矿库区种植地极端贫瘠,有机质含量仅2.6～.8 g·kg^-1,而土壤Cu、Cd、Pb、Zn含量皆高于对照土壤,其中Cu含量达809.30～1 39.4 mg·kg^-1,Cd含量达3.2～6.3 mg·kg^-1,达到对照土壤30～60倍.结缕草和三叶草体内重金属含量与土壤重金属交换态及有机结合态含量成正相关,与碳酸盐结合态、铁锰氧化物结合态成显著或极显著负相关,与矿物态含量相关性不显著.在两种优势植物中,Cu、Zn、Pb均以活性较低的醋酸提取态、盐酸提取态和残渣态为主;Zn在根系和茎叶中,NaCl提取态占有较大比例,而Cd均以NaCl提取态为主.     

Pollution,sources, and risks of heavy metals in coastal waters of China

Abstract

Comprehensive information on heavy metals in coastal waters at national scale of China is limited. Therefore, this study investigated the distribution, pollution, and ecological-health risks of heavy metals in coastal waters along 18,000?km coastline of China. Total 13 target heavy metals in coastal waters along coastline of China showed drastic spatial variations with average concentrations ranging from .14 (Cd) to 136.26 (Cu) μg/L. Cu was the dominant heavy metal with the maximal concentration of 1485.92?μg/L. Three methods including heavy metal pollution index (HPI), Nemerow index (NI), and contamination degree (CD) were adopted to explore heavy metal pollution. HPI obtained the worst-case evaluation results to illustrate that heavy pollution occurred at over 50% of sampling sites. Anthropogenic sources were the main sources of heavy metals in the coastal waters. Approximately 28.13% and 9.38% of sampling sites illustrated considerable and very high ecological risks, respectively. Metals including Cu, As, and Hg were the main pollution and risk contributors. Heavy metals in coastal waters posed high cancer risks and unacceptable non-cancer risks to both adults and children. Therefore, effective control of heavy metals is necessary for regional sustainability and well-beings of residents in coastal regions of China.