取食加Cd2+食物后拟水狼蛛发育历期、耐饥力和体内Cd2+含量的变化

为明确食物中重金属离子在蜘蛛体内的传递、生物放大及对其生长历期和耐饥力的影响,采用原子吸收光谱法检测了拟水狼蛛Pirata subpiraticus取食加Cd²⁺食物后体内Cd²⁺含量变化情况,测定了Cd²⁺对其发育历期和耐饥力的间接影响。结果表明：食物中的Cd²⁺能够通过食物链进行传递并在拟水狼蛛体内积累,积累量随拟水狼蛛龄期的增长而增加,积累量与消耗的黑腹果蝇Drosophila melanogaster数量极显著正相关(P<0.01), 与消耗食物中Cd²⁺含量显著正相关(P<0.05); 对食物中Cd²⁺的吸收率为65.4%,生物放大因子为1.8。成蛛改喂无Cd²⁺的黑腹果蝇后在观察的4周内体内Cd²⁺含量变化不明显。食物中的Cd²⁺能导致拟水狼蛛的发育历期显著延长、耐饥力显著降低。研究结果可为进一步研究环境中Cd²⁺沿土壤-昆虫-天敌传递、放大和生理耐受等提供理论依据。     

镉污染对不同生境拟水狼蛛氧化酶和金属硫蛋白应激的影响

为探讨镉(Cd)对机体抗氧化功能及金属硫蛋白(MT)的影响,在室内分别用不添加Cd2 和添加浓度为20 mgkg-1 Cd2 培养基培养的黑腹果蝇来饲喂4种不同生境下(S1,S2,S3和S4)拟水狼蛛,于饲喂5d、10d和20d后,分别测定其体内MT和丙二醛 (MDA)的含量及超抗氧化酶(GST、SOD和CAT)活性。结果表明：1,不同生境拟水狼蛛用不添加Cd2 培养基培养的黑腹果蝇饲喂后,不添加Cd2 对照组拟水狼蛛镉的积累量和MT含量无显著变化,但均显著低于添加Cd2 污染组。添加Cd2 污染组拟水狼蛛镉的积累量和MT含量都显著高于对照组,且均随着饲喂时间的延长而显著升高,具有明显的时间–效应关系(p<0.05)。2,在饲喂5d和10d后,不添加Cd饲喂的拟水狼蛛MDA含量和抗氧化酶系差异都不显著。添加Cd2 污染组(S1,S2和S3)MDA含量显著高于对照组(S4),MDA含量与饲喂时间呈显著正相关(p<0.05);GST、SOD和CAT等抗氧化酶活性污染组显著低于对照组,与饲喂时间呈显著负相关(p<0.05);饲喂20d后,污染组MDA含量和抗氧化酶（SOD和CAT）活性均与对照组无显著差异,但GST活性差异显著。     

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。     

不同生境汞对拟水狼蛛抗氧化系统的影响

以拟水狼蛛为研究对象,采用原子吸收光谱法测定了不同生境汞在土壤和蜘蛛体内的含量,研究了不同生境汞对拟水狼蛛抗氧化酶的影响.结果 表明:4个采集样点(S1、S2、S3和S4)拟水狼蛛体内重金属汞的含量差异显著(P<0.05).在重金属胁迫下,重金属汞含量高(S1、S2和S3)的拟水狼蛛体内GSH含量显著高于对照组(S4),是对照组的2.4~3.8倍.GSH含量与重金属汞含量显著正相关(r2=0.9624,P<0.01).对于GST、CAT和SOD,重金属汞含量高则酶活性低,GST、CAT和SOD酶活性与重金属汞含量显著负相关.因此检测拟水狼蛛体内的酶活性变化即可知环境中重金属汞的污染程度,提示拟水狼蛛可以作为重金属污染的重要监测指示生物.     

硫化氢通过抑制镉离子内流降低拟南芥体内的镉毒性

硫化氢（H₂S）作为一种新兴的气体信号分子,在植物体内主要由半胱氨酸脱巯基酶（CDes）降解半胱氨酸产生。已有报道表明,H₂S信号与植物激素共同作用增强植物的镉（Cd）耐受。然而,H₂S信号响应重金属Cd胁迫的作用机制尚缺乏系统研究。本文以拟南芥为实验材料,从不同水平探究H₂S分子对Cd胁迫诱导氧化应激的保护作用。结果表明,CDes基因表达量和H₂S的产率随CdCl₂浓度升高而逐渐增加。重金属Cd胁迫导致幼苗干重降低约33%、体内过氧化氢显著增加、丙二醛含量升高约110%、超氧化物歧化酶活性增加约100%、谷胱甘肽还原酶活性和过氧化氢酶活性分别下降27%和21%,还原性谷胱甘肽含量随之显著降低。生理浓度NaHS（H₂S供体）预处理显著缓解以上Cd胁迫产生的影响,使恢复到对照水平。同时,H₂S处理可显著下调质膜中Cd转运蛋白（HMA4和IRT1）的表达,同时上调液泡膜中MRP3和CAX2的表达。利用非损伤微测技术测定植物根系Cd²⁺的流动速度和流动方向。结果显示,生理浓度的H2S显著抑制Cd^{2 +}内流,最终表现为植物叶片和根中的Cd含量显著降低,分别下降了15%和38.4%。总之,在Cd胁迫条件下,H₂S信号可激活植物体内的抗氧化酶促和非酶促系统,以清除细胞内H₂O₂。H₂S对Cd²⁺转运和液泡区式化的调节,降低了体内Cd²⁺的浓度,减小Cd毒性对植物生长的影响。为理解农作物应对重金属胁迫的机制提供了新的思路。     

硫化氢通过抑制镉离子内流降低拟南芥体内的镉毒性

硫化氢（H₂S）作为一种新兴的气体信号分子,在植物体内主要由半胱氨酸脱巯基酶（CDes）降解半胱氨酸产生。已有报道表明,H₂S信号与植物激素共同作用增强植物的镉（Cd）耐受。然而,H₂S信号响应重金属Cd胁迫的作用机制尚缺乏系统研究。本文以拟南芥为实验材料,从不同水平探究H₂S分子对Cd胁迫诱导氧化应激的保护作用。结果表明,CDes基因表达量和H₂S的产率随CdCl₂浓度升高而逐渐增加。重金属Cd胁迫导致幼苗干重降低约33%、体内过氧化氢显著增加、丙二醛含量升高约110%、超氧化物歧化酶活性增加约100%、谷胱甘肽还原酶活性和过氧化氢酶活性分别下降27%和21%,还原性谷胱甘肽含量随之显著降低。生理浓度NaHS（H₂S供体）预处理显著缓解以上Cd胁迫产生的影响,使恢复到对照水平。同时,H₂S处理可显著下调质膜中Cd转运蛋白（HMA4和IRT1）的表达,同时上调液泡膜中MRP3和CAX2的表达。利用非损伤微测技术测定植物根系Cd²⁺的流动速度和流动方向。结果显示,生理浓度的H2S显著抑制Cd^{2 +}内流,最终表现为植物叶片和根中的Cd含量显著降低,分别下降了15%和38.4%。总之,在Cd胁迫条件下,H₂S信号可激活植物体内的抗氧化酶促和非酶促系统,以清除细胞内H₂O₂。H₂S对Cd²⁺转运和液泡区式化的调节,降低了体内Cd²⁺的浓度,减小Cd毒性对植物生长的影响。为理解农作物应对重金属胁迫的机制提供了新的思路。     

拟水狼蛛对食物中镉的吸收和排泄及生物学响应

为明确拟水狼蛛对食物中镉的吸收和排泄及生物学响应,采用原子吸收光谱法检测了连续3代拟水狼蛛对食物中Cd²⁺的吸收和排泄情况,并测定了Cd²⁺对其生长发育、繁殖力和耐饥力的影响。结果表明:食物中过量的Cd²⁺能够通过食物链进行传递并在拟水狼蛛体内积累,积累量随拟水狼蛛代数的增加而增加,第2代和第3代拟水狼蛛体内Cd²⁺积累量浓度显著高于第1代中Cd²⁺积累量,第2代和第3代间差异不显著,积累量与消耗的黑腹果蝇数量极显著正相关,P<0.01;与消耗食物中Cd含量显著正相关,P<0.05。连续3代拟水狼蛛分别吸收了食物中65.4%、68.5%和69.1%的Cd,生物营养级放大因子分别为1.71、2.12和2.17。连续3代拟水狼蛛Cd排泄量极低。Cd处理能显著改变拟水狼蛛背胸甲宽、幼蛛存活率、生长历期和繁殖力。随拟水狼蛛受胁迫代数的增加,幼蛛存活率显著减少,生长历期显著延长。卵袋重量显著减轻,卵数目显著减少,卵体积显著升高。Cd处理还能显著降低拟水狼耐饥力。研究结果可为进一步研究环境中Cd沿土壤-昆虫-天敌传递、放大和生理耐受提供更为充分的理论依据。     

河南南阳不同生境土壤重金属含量及其对拟水狼蛛生物学特性的影响

为了探明重金属含量对拟水狼蛛Pirata subpiraticus生物学特性的影响, 本研究于2008年7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150头拟水狼蛛样本, 采用原子吸收光谱法测定了5种不同生境下雌拟水狼蛛体内重金属含量, 并测定了雌蛛头胸甲宽、体重、卵袋重量、卵数目和卵体积等生物学指标。结果表明：不同样点拟水狼蛛重金属含量与土壤重金属含量显著正相关, 4个重金属污染区与宝天曼自然保护区（Baotianman, BTM）拟水狼蛛体内Cd、Cu和Zn的含量差异极显著(P<0.001), 桐柏铜矿（Tongbai Tongkuang, TBTK）和南阳油田(Nanyang Youtian, NYYT) 的Pb含量与BTM的差异极显著(P<0.001), 主成分分析表明Cd、Cu和Zn是反应重金属污染的主要指标。在重金属胁迫下, 4个重金属污染区与保护区的拟水狼蛛头胸甲宽、雌体重量、卵袋重量、卵数目和卵体积差异均达到极显著（P<0.001）或显著水平（P<0.05）, 主成分分析表明背胸甲宽、卵袋重量和卵体积是重要生物学指标。 拟水狼蛛的背胸甲宽、卵袋重量和卵的体积与重金属含量显著相关, 重金属含量高的地方拟水狼蛛背胸甲宽较小, 雌性体重、卵袋重量和卵数目较小, 卵的体积大, 因此说明拟水狼蛛处在逆环境中时, 动物为保持总的繁殖力不变的前提下, 后代个体的大小与数目间满足某种平衡, 即达到协同进化的目的, 拟水狼蛛可以作为重金属污染的监测指示生物。     

不同藻密度下Cd2+浓度对萼花臂尾轮虫生命表统计学参数的影响

为了比较不同食物密度下污染物浓度对受试生物的慢性毒性,筛选出以轮虫为受试生物对水环境中Cd污染进行监测的敏感指标,研究了在不同斜生栅藻密度(1.0×10⁶、3.0×10⁶和5.0 ×10⁶ cells·ml^-1)下不同浓度(2.5、5.0、10.0、20.0和40.0 μg·L^-1)Cd²⁺对萼花臂尾轮虫生命表统计学参数的影响.结果表明：(25±1) ℃下Cd²⁺对轮虫的24 h LC₅₀为37.7 μg·L^-1.与各藻密度下的对照组相比,当藻密度为1.0×10⁶ cells·ml^-1时,20.0和40.0 μg·L^-1的Cd²⁺显著延长了轮虫的世代时间,5.0 μg·L^-1的Cd²⁺显著提高了轮虫的后代混交率;当藻密度为3.0×10⁶ cells·ml^-1时,除了5.0 μg·L^-1外,其他浓度的Cd²⁺显著降低了轮虫的后代混交率;而当藻密度为5.0×10⁶ cells·ml^-1时,Cd²⁺浓度对轮虫的所有生命表统计学参数均无显著影（P>0.05）.藻密度对轮虫的世代时间、生命期望、净生殖率和后代混交率均有显著影响（P<0.05）,Cd²⁺浓度对轮虫的世代时间和后代混交率有显著影响（P<0.05）,两者交互作用对后代混交率有极显著影响（P<0.01）.轮虫的世代时间和后代混交率是在1.0×10.6和3.0×10.6 cells·ml^-1食物密度下对Cd²⁺污染比较敏感的参数,其中后代混交率最敏感.     

不同藻密度下Cd2+浓度对萼花臂尾轮虫生命表统计学参数的影响

为了比较不同食物密度下污染物浓度对受试生物的慢性毒性,筛选出以轮虫为受试生物对水环境中Cd污染进行监测的敏感指标,研究了在不同斜生栅藻密度(1.0×10⁶、3.0×10⁶和5.0 ×10⁶ cells·ml^-1)下不同浓度(2.5、5.0、10.0、20.0和40.0 μg·L^-1)Cd²⁺对萼花臂尾轮虫生命表统计学参数的影响.结果表明：(25±1) ℃下Cd²⁺对轮虫的24 h LC₅₀为37.7 μg·L^-1.与各藻密度下的对照组相比,当藻密度为1.0×10⁶ cells·ml^-1时,20.0和40.0 μg·L^-1的Cd²⁺显著延长了轮虫的世代时间,5.0 μg·L^-1的Cd²⁺显著提高了轮虫的后代混交率;当藻密度为3.0×10⁶ cells·ml^-1时,除了5.0 μg·L^-1外,其他浓度的Cd²⁺显著降低了轮虫的后代混交率;而当藻密度为5.0×10⁶ cells·ml^-1时,Cd²⁺浓度对轮虫的所有生命表统计学参数均无显著影（P>0.05）.藻密度对轮虫的世代时间、生命期望、净生殖率和后代混交率均有显著影响（P<0.05）,Cd²⁺浓度对轮虫的世代时间和后代混交率有显著影响（P<0.05）,两者交互作用对后代混交率有极显著影响（P<0.01）.轮虫的世代时间和后代混交率是在1.0×10.6和3.0×10.6 cells·ml^-1食物密度下对Cd²⁺污染比较敏感的参数,其中后代混交率最敏感.     

Phosphorus content and growth of fenugreek as affected by cadmium application

Changes in growth and phosphorus content in plants and seeds of fenugreek with increasing cadmium concentration was evaluated. Root length and shoot length ranged from 11.63 to 27.72 and from 9.70 to 54.78 cm, respectively. With the increasing Cd²⁺ concentration there was a significant decrease in root and shoot length, and fresh mass. Various phosphorus fractions of shoot decreased with increasing Cd²⁺ concentration except lipid P and nucleic acid P which increased at 65 and 95 d after sowing and protein P only increased at vegetative stage. In seeds (60 d after flowering) lipid P increased except at 2.5 μg(Cd²⁺) g^?1 (soil) while protein P decreased.     

Analysis of Metal-Regulated Metallothionein and Heat Shock Gene Expression in HeLa-Derived Cadmium-Resistant Cells

The expression of metallothionein (MT) and heat shock protein gene families was investigated in normal and in HeLa-derived cadmium-resistant cells, named H454. In the absence of amplification of MT genes H454 cells accumulated elevated concentrations of cadmium ions and synthesized higher levels of MT proteins than unselected HeLa cells. Northern blot analyses revealed higher levels of MT mRNAs in the resistant cells than in wild-type cells after Cd²⁺and Zn²⁺exposure. Evaluation of the cytotoxic potential of the different metals confirmed the high resistance to cadmium of the H454 cells. Two proteins of the heat shock family, hsp70 and GRP78, were synthesized in Cd²⁺-exposed H454 cells at levels comparable to the ones present in Cd²⁺-treated normal cells. Northern blot analyses of the mRNA levels corresponding to these proteins revealed elevated expression of both hsp70 and GRP78 mRNAs in H454 cells upon exposure to cadmium ions and no response to zinc induction. These data suggest the existence in the H454 cells of a cadmium-specific pathway of regulation of MT and heat shock genes.     

Comparative studies of zinc metabolism in cultured chinese hamster cells with differing metallothionein-induction capacities

Previous work in our laboratory led to the isolation of a cadmium (Cd)-resistant variant (Cd^r2C10) of the line CHO Chinese Hamster cell having a 10-fold greater resistance to the cytotoxic action of Cd²⁺ compared with the CHO cell. This resistance was attributed to an increased capacity of the Cd²⁺-resistant Cd^r2C10 subline to induce synthesis of the Cd²⁺- and Zn²⁺-binding protein(s), metallothionein(s) (MT). Evidence that Cd²⁺ behaves as an analog of the essential trace metal, Zn²⁺, especially as an inducer of MT synthesis, suggested that the Cd^r and CHO cell types could be employed to investigate cellular Zn²⁺ metabolism. In the present study, measurements were made to compare CHO and Cd^r cell types for (a) growth as a function of the level of ZnCl₂ added to the culture medium, (b) uptake and subcellular distribution of Zn²⁺, and (c) capacity to induce MT synthesis. The results of these measurements indicated that (a) both CHO and Cd^r cell types grew normally (T _d≊16–18 h) during exposures to Zn²⁺ at levels up to 100 μM added to the growth medium, but displayed abrupt growth inhibition at higher Zn²⁺ levels, (b) Cd^r cells incorporate fourfold more Zn²⁺ during a 24-h exposure to the maximal subtoxic level of Zn²⁺ and (c) the CHO cell lacks the capacity to induce MT synethesis while the Cd^r cell is proficient in this response during exposure to the maximal subtoxic Zn²⁺ level. These findings suggest that (a) the CHO and Cd^r cell systems will be useful in further studies of cellular Zn²⁺ metabolism, especially in comparisons of Zn²⁺ metabolism in the presence and absence of induction of the Zn²⁺-sequestering MT and (b) a relationship exists between cellular capacity to induce MT synthesis and capacity for cellular Zn²⁺ uptake.     

Cadmium-induced changes in growth and cell cycle gene expression in suspension-culture cells of soybean

The toxic effects of cadmium on growth and development of living organisms are well documented. However, the molecular mechanisms responsible for the inhibition of plant growth by cadmium are still not completely understood. We determined the effects of cadmium concentrations in the range of 1–11 μM on the growth of Glycine max L. cv. Navico suspension-culture cells, as well as on the expression of two cell cycle genes: cyclin B1 and cyclin-dependent type A kinase (CDK-A). There was no detectable decrease in cell viability at any tested Cd²⁺ concentrations. The lower concentrations of Cd²⁺ (1–4 μM) stimulated cell culture growth; however, this did not correspond with increased expression of cell cycle genes. The inhibition of cell growth was observed at concentration of Cd²⁺ higher than 6 μM. Interestingly, it correlated well with the decreased cyclin B1 mRNA levels, but had no significant effect on the levels of CDK-A mRNA.     

Effect of some heavy metal ions on copper-induced metallothionein synthesis in the yeast Saccharomyces cerevisiae

Copper-induced metallothionein (MT) synthesis in Saccharomyces cerevisiae was investigated in order to associate this exclusively with Cu²⁺ in vivo, when cultured in nutrient medium containing other heavy metal ions. Expression of the CUP1 promoter/lacZ fusion gene was inhibited by all heavy metal ions tested, especially Cd²⁺ and Mn²⁺. By adding Cd²⁺ and Mn²⁺ at 10 M concentration, the -galactosidase activity decreased by about 80% and 50% of the maximum induction observed with 1 mM CuSO₄, respectively. Furthermore, cell growth was markedly inhibited by combinations of 1 mM-Cu²⁺ and 1 M-Cd²⁺. Therefore, the yeast S. cerevisiae could not rely on MT synthesis as one of the copper-resistance mechanisms, when grown in a Cd²⁺ environment. In contrast, the presence of Mn²⁺ in the nutrient medium showed alleviation rather than growth inhibition by high concentrations of Cu²⁺. The recovery from growth inhibition by Mn²⁺ was due to decreased Cu²⁺ accumulation. Inhibitory concentrations of Co²⁺, Ni²⁺ and Zn²⁺ on expression of the CUP1p/lacZ fusion gene were at least one order of magnitude higher than that of Cd²⁺ and Mn²⁺. These results are discussed in relation to Cu²⁺ transport and Cu-induced MT synthesis in the copper-resistance mechanism of the yeast S. cerevisiae.     

Cadmium Cation Increases the Production and mRNA Levels of Insulin-Like Growth Factor-Binding Protein-1 in HepG2

The production of insulin-like growth factor-binding protein-1 (IGFBP-1) in HepG2 was increased by cadmium cation (Cd²⁺) at 3 μM, but not by other divalent cations. The mRNA level of IGFBP-1 was also increased by the administration of 3 μM of Cd²⁺. These results suggest that Cd²⁺ impacts the gene expression of IGFBP-1, which leads to production of IGFBP-1.     

The effect of cadmium on potato (Solanum tuberosum L.) shoot culture growth

The uptake of cadmium and its effect on the growth of potato shoot tips grownin vitro were followed in dependence on cadmium concentration in nutrient medium. Concentration of 10 ⁻⁶ M Cd ²⁺ did not substantially affect potato plantlet growth dynamics; but the concentration of 10⁻³ M Cd²⁺ showed a strong growth inhibitory effect accompanied with increased cadmium accumulation in both root and shoot tissues.     

Bioaccumulation and effect of cadmium in the photosynthetic apparatus of Prosopis juliflora

In the present study Prosopis juliflora plants grown in hydroponics solution were exposed to 50,100 and 1000 μM CdCl₂. The cadmium uptake, transport and toxicity on the photosynthetic activities in the plants were measured at 48 h after starting cadmium treatments. The results showed that the concentration of Cd²⁺ in P. juliflora tended to increase with addition of Cd²⁺ to hydroponics solution. However, the increase of Cd²⁺ in roots and leaves varied largely. In this sense, the accumulation of Cd²⁺ in P. juliflora roots increased significantly in proportion with the addition of this metal. In contrast a relatively low level of Cd²⁺ transportation index, and bioaccumulation factor were found in P. juliflora at 48 h after of treatments. On the other hand the maximum photochemical efficiency of photosystem II (Fv/Fm) and the activity of photosystem II (Fv/Fo) ratios in P. juliflora leaf treated with Cd²⁺ not showed significantly changes during the experiment. These results suggested that the photosynthetic apparatus of P. juliflora was not the primary target of the Cd²⁺ action. Further studies will be focused in understanding the participation of the root system in Prosopis plants with the rhizosphere activation and root adsorption to soil Cd²⁺ under natural conditions.