碲化镉量子点(CdTe QDs)对肝细胞的毒性效应及线粒体介导的毒性机制研究

本文探讨了碲化镉量子点(CdTe QDs)对肝细胞的毒性效应及其影响因素,为探索量子点的肝毒性机制提供一定依据。采用人肝癌细胞(Hep G2)和人正常肝细胞(L02)为细胞模型,设置0、25、50和100μmol·L-14个浓度组,采用CCK-8法检测细胞生存率,石墨炉法检测细胞内镉元素含量,采用流式细胞术,装载荧光探针DCFH-DA检测细胞内活性氧水平,采用FITC/PI检测细胞凋亡以及JC-1检测细胞ATP水平。研究结果显示:CdTe QDs诱导2种肝细胞生存率降低,细胞凋亡率升高,细胞对QDs的摄入水平具有时间依赖性,细胞内活性氧水平显著升高,线粒体膜电位降低和ATP含量显著减少,且2种肝细胞比较发现L02细胞损伤程度更为严重。CdTe QDs对2种肝细胞造成损伤,对L02细胞损伤更明显,其原因是L02细胞对CdTe QDs摄取更多,导致进入细胞的QDs引发更为严重的损伤效应。     

硫化镉量子点对人胚肝细胞L-02的毒性研究(Cadmium Sulfide Quantum Dots Induce Cytotoxicity in Human Embryo Liver Cells)

采用乳化液膜法自组合成硫化镉量子点(CdS quantum dots,CdS QDs),探讨CdS QDs的体外毒性作用及可能的作用机制.选用人胚肝细胞(L-02)作为细胞模型,采用不同浓度的CdS QDs(0.00、1.25、2.50、5.00、10.00、20.00、40.00μg·mL-1)对L-02细胞进行染毒.24h后,检测细胞内乳酸脱氢酶(LDH)释放量、谷胱甘肽(GSH)含量和超氧化物歧化酶(SOD)活力,并比较加入抗氧化剂N-乙酞半胱氨酸(NAC)后细胞存活率的变化,同时测定了细胞内外的镉离子浓度.结果表明,与空白对照组相比,CdS QDs单独染毒组细胞存活率显著降低(p<0.05或p<0.01);加入抗氧化剂NAC后,10.00、20.00、40.00μg·mL-1染毒组细胞存活率与单独染毒组相比显著上升(p<0.01).CdS QDs浓度为5.00μg·mL-1时,细胞内Cd2+的浓度略高于细胞外Cd2+的浓度,在其他浓度下,细胞外Cd2+的浓度均显著高于细胞内Cd2+的浓度.当作用浓度上升至10.00μg·mL-1时,人胚肝细胞内LDH含量显著增加,且随着作用剂量的升高,LDH含量逐渐增加.与空白对照组相比,40.00μg·mL-1CdS QDs染毒组SOD活力和20.00μg·mL-1CdS QDs染毒组GSH含量均显著降低(p<0.05).Cd2+易透过L-02细胞的细胞膜而进入细胞内,从而造成细胞损伤.氧化损伤可能是CdSQDs对L-02细胞毒性作用的机制之一.     

氧化石墨烯存在下铜离子对大型溞的毒性研究

氧化石墨烯(graphene oxide,GO)作为一种具有独特物理化学性质的新型纳米材料被广泛应用,其进入环境后可能对传统污染物的毒性造成影响。选取大型溞为受试生物,研究了GO的存在对Cu在大型溞体内的富集、毒性和抗氧化系统的影响。结果表明,GO对Cu~(2+)具有良好的吸附效果,大幅降低了试验液中Cu~(2+)浓度。1 mg·L~(-1)和2 mg·L~(-1)GO存在下,大型溞暴露于19.2μg·L~(-1)Cu~(2+)溶液72 h后,体内的金属Cu富集量由360μg·g-1干重分别降低为308μg·g-1和215μg·g-1干重。GO的存在降低了Cu~(2+)对大型溞的毒性,Cu~(2+)对大型溞的72 h-LC50值由19.2μg·L~(-1)升高至56μg·L~(-1)。Cu~(2+)单独作用时,大型溞体内SOD活性和GSH含量表现为先诱导后抑制,而MDA含量逐渐升高;当GO存在时,大型溞体内酶活性的变化趋势与上述现象类似,但含量总体低于Cu~(2+)单独暴露时的活性和含量。研究表明GO的加入减少了大型溞体内Cu的富集量,降低了Cu~(2+)对大型溞的氧化损害,对Cu~(2+)的毒性存在一定的减轻效果。     

Cu~(2+)、Zn~(2+)、Cd~(2+)、Cr~(6+)和Mn~(2+)对萼花臂尾轮虫联合急性毒性研究

为探究重金属复合污染对轮虫的毒性影响,以萼花臂尾轮虫为受试动物,选择Cu~(2+)、Zn~(2+)、Cd~(2+)、Cr6+和Mn~(2+)等5种重金属,采用水生毒理联合效应相加指数法开展了其24 h联合急性毒性作用的评价研究。结果显示,Cu~(2+)、Zn~(2+)、Cd~(2+)、Cr6+和Mn~(2+)等5种重金属对萼花臂尾轮虫24 h半数致死浓度分别为:0.00616 mg·L~(-1),12.62 mg·L-1,2.89 mg·L-1,17.29 mg·L-1和67.32 mg·L-1。联合急性毒性实验结果显示,等毒性配比的Cu~(2+)-Cr6+(0.00385-10.806 mg·L-1)和等浓度配比的Cu~(2+)-Zn~(2+)(0.0199-0.0199 mg·L-1)、Cu~(2+)-Cd~(2+)(0.0181-0.0181 mg·L-1)、Cu~(2+)-Cr6+(0.0118-0.0118 mg·L~(-1))、Zn~(2+)-Cd~(2+)(3.475-3.475 mg·L-1)二元联合测试液的作用结果显示为拮抗效应,其余二元联合测试液的作用结果则均显示是协同效应。等毒性配比的Cu~(2+)-Cr~(6+)-Mn~(2+)(0.00210-5.902-22.981 mg·L-1)和等浓度配比的Cu~(2+)-Cd~(2+)-Mn~(2+)(0.00727-0.00727-0.00727 mg·L-1)三元联合测试液的作用结果显示为拮抗效应,其余三元联合测试液的作用结果则均显示是协同效应。等浓度配比的Cu~(2+)-Zn~(2+)-Cd~(2+)-Cr6+(0.00907-0.00907-0.00907-0.00907 mg·L-1)、Cu~(2+)-Zn~(2+)-Cd~(2+)-Mn~(2+)(0.00898-0.00898-0.00898-0.00898 mg·L~(-1))、Cu~(2+)-Zn~(2+)-Cr6+-Mn~(2+)(0.00819-0.00819-0.00819-0.00819 mg·L~(-1))四元联合测试液的作用结果显示为拮抗效应,其余四元联合测试液的作用结果的则均显示是协同效应。Cu~(2+)-Zn~(2+)-Cd~(2+)-Cr~(6+)-Mn~(2+)等毒性(0.00074-1.520-0.348-2.082-8.107 mg·L~(-1))和等浓度(0.00582-0.00582-0.00582-0.00582-0.00582 mg·L-1)配比的五元联合测试液作用结果均显示是协同效应。     

硒蛋白R抑制铜离子介导的Aβ_(42)聚集和细胞毒性

阿尔茨海默病(Alzheimer's disease,AD)是一种神经退行性疾病,其主要病理特征包括脑内出现老年斑和神经纤维缠结以及神经元缺失。过渡金属离子如Fe~(2+)、Zn~(2+)、Cu~(2+)等会在AD病人脑中大量蓄集,从而促进,-淀粉样多肽(Aβ)聚集并产生大量活性氧自由基,引起细胞功能改变。硒蛋白R(Sel R)是一种蛋氨酸亚砜还原酶,可将蛋白中的蛋氨酸亚砜还原为蛋氨酸。然而Sel R与金属离子的关系尚无报道。文章对Sel R的突变体Sel R'(Sel R中硒代半胱氨酸Sec95突变为半胱氨酸Cys95)结合Cu~+的性质进行了表征,同时也对Sel R'调节Cu~+/Cu~(2+)诱导的Aβ42聚集和细胞毒性的能力进行了研究。结果表明,Sel R'可与Cu~+特异性螯合剂BCA竞争结合Cu~+,说明Sel R'具有较强的Cu~+结合能力。采用硫磺素荧光法(Th T),发现Cu~+/Cu~(2+)显著性抑制Aβ42的纤维化、诱导其形成非纤维化聚集体,而Sel R'则能够显著抑制Cu~+/Cu~(2+)诱导的Aβ42聚集形态的改变。细胞实验表明:Sel R'能显著降低Cu~+/Cu~(2+)诱导的Aβ42的细胞毒性。上述结果提示:Sel R在脑内除了还原蛋氨酸亚砜,还可通过与铜离子的结合抑制其细胞毒性作用,从而干预AD的发展。     

CASS工艺微生物抵抗含Cu~(2+)、Ni~(2+)锻造添加剂废水毒性的效果

工(产)业园区内企业在紧急情况下排放的污水中,有毒物质会影响污水处理厂活性污泥微生物的活性,甚至导致微生物的死亡.基于此,利用CASS工艺的特点检验微生物系统抗冲击毒性的能力,通过不同的配比浓度调整试验水样中Cu~(2+)、Ni~(2+)的质量浓度,监测CASS系统进出水的Cu~(2+)、Ni~(2+)、COD_(cr)指标,用以判断微生物系统是否受到影响,进而确定CASS微生物系统抗毒性冲击的临界点.结果表明:Cu~(2+)质量浓度小于3.0 mg·L~(-1),Ni~(2+)质量浓度小于5.0 mg·L~(-1),时,CASS 工艺微生物抵抗含Cu~(2+)、Ni~(2+)锻造添加剂废水的毒性能力较强,对Cu~(2+)、Ni~(2+)、COD_(cr)的去除率均可以达到80%;当Cu~(2+)质量浓度大于3.0 mg·L~(-1),Ni~(2+)质量浓度大于5.0mg·L~(-1)时,CASS工艺微生物对毒性抵抗能力显著降低,活性受到严重破坏,对CU~(2+)、Ni~(2+)、COD_(cr)去除率下降至60%以下.所以可以确定CASS工艺微生物抵抗含Cu~(2+)、Ni~(2+)锻造添加剂废水毒性的最高质量浓度限值,Cu~(2+)为3.0 mg·L~(-1),Ni~(2+)为5.0mg·L~(-1).为污水处理厂实际运行工程中避免活性污泥微生物系统受毒性冲击而导致运行瘫痪提供参考依据.     

基于Mn∶ZnS量子点-Cu~(2+)体系室温磷光猝灭-恢复方法测定水体中焦磷酸根离子

以谷胱甘肽(GSH)为稳定剂,成功合成了高性能的水溶性Mn∶ZnS量子点(Quantum dots,QDs).Mn∶ZnS QDs表面的羧基能与Cu~(2+)结合从而有效引发QDs电子转移,致使Mn∶ZnS QDs室温磷光显著猝灭.当体系中加入焦磷酸(PPi)时,由于Cu~(2+)与PPi的结合能力强于Mn∶ZnS QDs表面的羧基,Mn∶ZnS QDs磷光强度又逐渐恢复.据此建立一种基于Mn∶ZnS QDs-Cu~(2+)体系的室温磷光探针测定焦磷酸根离子的新方法.该方法灵敏、简单,线性范围为5.0×10~(-8)—1.0×10~(-4)mol·L~(-1),检测限为1.0×10~(-8)mol·L~(-1).经过干扰实验及添加回收率实验证实,该方法具有良好的选择性,满足于实际样品水体中焦磷酸的检测分析.     

生物炭修饰材料对嘉陵江(川渝段)沿岸土吸附Cu2+的影响

为了探索生物炭修饰材料对嘉陵江流域沿岸土吸附Cu~(2+)的影响,采用生物炭(B)、磁化生物炭(MB)以及50%和100%CEC十二烷基二甲基甜菜碱(BS-12)修饰MB(分别以50%BS-MB和100%BS-MB表示)作为炭修饰材料,分别将其以1%(质量比)加入嘉陵江流域(川渝段)内苍溪(CX)、南部(NB)、嘉陵(JL)和合川(HC)沿岸土中,共计形成20个混合土样(以原土作为对照),批处理法研究各样品对Cu~(2+)的等温吸附和热力学特征,并对比不同温度、pH值和离子强度下的吸附差异.结果表明,不同混合土样对Cu~(2+)吸附等温线均呈"L"型且符合Langmuir模型,最大吸附量q_m保持在62.20—308.88 mmol·kg~(-1)之间.相同生物炭修饰材料添加下Cu~(2+)吸附量表现为JLNBCXHC的趋势.20—40℃范围内,各混合土样对Cu~(2+)的吸附量均随温度的升高而增加,表现为增温正效应.离子强度从0.01 mol·L~(-1)增加到0.1 mol·L~(-1),各混合土样(除HC外)对Cu~(2+)的吸附量均呈现先增后降的趋势.pH值升高有利于混合土样对Cu~(2+)的吸附.各混合土样对Cu~(2+)的吸附是一个自发、吸热和熵增的反应过程,且CEC和比表面积是决定混合土样对Cu~(2+)吸附效果的关键.     

PCB77和Cd共同暴露诱导的线粒体损伤及其介导的联合毒性效应

本文以人肝癌细胞系SMMC-7721为生物模型,研究并比较了PCB77和Cd~(2+)对SMMC-7721细胞的单一与联合毒性.通过细胞活性检测发现,当Cd~(2+)暴露浓度为20μmol·L-1,PCB77暴露浓度为25μmol·L~(-1),暴露时间为48 h时,两种污染物产生明显的联合毒性.二者共同暴露虽然不影响细胞膜的完整性,但引起了线粒体膜电位下降、膜通透性显著增加,进而诱导细胞凋亡,同时胞内ROS水平升高.结果表明,PCB77和Cd~(2+)的共同暴露可引发联合肝细胞毒性,其毒性机理是线粒体膜电位下降和膜通透性增加,进而引发细胞凋亡.     

四溴双酚A对人体正常肝细胞毒性效应及作用机制

四溴双酚A(TBBPA)作为目前用量最大的一种溴系阻燃剂,在含TBBPA用品的生产、使用和废弃处置过程中,能够通过多种途径进入环境介质,造成持久性污染,危害生态系统和人体健康.为探明TBBPA对人体健康的潜在毒性效应及作用机制,选取人体正常肝细胞L02作为模型,通过分析暴露后细胞形态、存活率、胞内活性氧(ROS)含量、DNA损伤及细胞凋亡等变化.结果表明,TBBPA暴露导致L02细胞形态发生明显改变、存活率显著降低,细胞彗星实验拖尾现象明显增强;随着暴露浓度的升高,L02细胞胞内ROS含量、丙二醛(MDA)含量和氧化型谷胱甘肽/还原型谷胱甘肽(GSSG/GSH)比值均呈现剂量依赖性增加.40μmol·L-1暴露条件下胞内ROS含量升高3.1倍;20 μmol·L-1和40 μmol·L-1暴露条件下,细胞凋亡率分别增加了32倍和4.8倍.推测TBBPA暴露对L02细胞的毒性效应作用机制为,暴露引起细胞氧化应激水平升高,ROS升高再引起DNA损伤程度增强,最终导致细胞凋亡率增加.上述研究结果将为评估TBBPA的毒性效应和健康风险提供科学依据.     

Nano-titanium dioxide (TiO2)-induced changes affecting Cu2+-mediated alterations in bacterium Bacillus subtilis and α-amylase

Titanium dioxide (TiO₂) nanoparticles possess the potential to coexist with Copper (Cu²⁺) in soil. The individual and combined toxicity of these two chemicals was evaluated using the bacterium Bacillus subtilis, a known soil model bacterium. Cu²⁺ (6.25–50?µg?mL^?1) alone produced toxicity to bacteria as evidenced by the decreased cell viability and deceased α-amylase production. The addition of TiO₂ (50?mg?mL^?1) enhanced the Cu²⁺-induced decrease in cell viability but elevated amylase activity. TiO₂ did not markedly affect the growth rate and lag period. A primary cause of TiO₂ increasing Cu²⁺ toxicity is presumed to be associated with hydroxyl radical formation, while increased amylase activity is considered to arise from Cu²⁺ facilitating TiO₂ degradation ability.     

水体酸化条件下Cu(II)对斑马鱼胚胎的毒性效应

为评价由酸雨、酸性矿山排水等环境污染导致的水体酸化及水体重金属联合作用对水生生物的生态毒性效应,研究了不同pH值(pH=3、4、5、6、7和7.8)条件下Cu2+对斑马鱼胚胎发育的影响。结果表明,酸性水体及Cu2+单一存在时,酸对斑马鱼胚胎24h半数效应浓度值EC50为pH=3.65,Cu2+(pH=7.8)对斑马鱼胚胎24h-EC50为0.267mg·L-1;当水体酸化及水体中的Cu2+共存时,较低的pH对Cu2+的生物毒性起协同作用,表现为随溶液pH的降低,各浓度Cu2+对斑马鱼胚胎的24h凝结率显著增高(P24h致死率=0.001),而斑马鱼胚胎96h孵化率显著降低(P96h孵化率=0.002),且不同浓度的Cu2+之间的生物毒性效应存在显著性差异(P24h致死率=0.0321;P96h孵化率=0.0028)。这说明酸性水体和Cu2+都显著影响斑马鱼胚胎的发育,且Cu2+在酸性水体中对斑马鱼胚胎的毒性显著增强。因此,在受重金属Cu2+污染的地区,如同时受到酸雨或酸性矿山排水等较低pH值和Cu2+的双重胁迫,较低浓度的Cu2+就能够对水生生物的生殖发育及水生生态系统产生严重的影响和危害。     

Low-cost and efficient adsorbent derived from pyrolysis of Jatropha curcas seeds for the removal of Cu2+ from aqueous solutions

Biochar, is a low-cost material that can be used as an alternative adsorbent for the removal of heavy metals. In this study, a low-cost and efficient adsorbent synthesised from Jatropha curcas seeds was used for the uptake of Cu²⁺ from aqueous solutions. The as-prepared adsorbent was characterised by scanning electron microscopy and Brunauer–Emmett–Teller analysis post calcination at 500 °C, its BET surface area and total pore volume were 39.62?m²?g^?1 and 0.049?m³?g^?1, respectively. Subsequently, the effects of initial pH of the solution, contact time, and adsorbent material dosage on the adsorption of Cu²⁺ by the prepared adsorbent were investigated. The as-prepared adsorbent exhibited a high performance, with a maximum adsorption amount of 32.895?mg?g^?1 for Cu²⁺ at pH 5.0 and 25 °C, owing to the presence of ?OH, C=O, C–O, Si-O-Si, and O-Si-O on its surface. The predominant Cu²⁺ adsorption mechanism was assumed to be ion exchange. Notably, the Cu²⁺ adsorption could attain equilibrium within 90?min. In addition, the fact that the Langmuir model was a better fit than the Freundlich model for the isotherm data of Cu²⁺ adsorption by the as-prepared adsorbent suggested that the adsorption of Cu²⁺ was a monolayer adsorption process.     

Single and combined effects of heavy metals and hormones on lysosomes of haemolymph cells from the mussel Mytilus galloprovincialis

Effects of heavy metals on lysosomes were studied in living cells from the mussel (Mytilus galloprovincialis Lam.). Haemolymph cells were obtained from the mussel adductor muscle, stained with neutral red (NR), and analysed by digital imaging to evaluate NR retention times within lysosomes. Exposure to Hg²⁺, Cd²⁺ and Cu²⁺ induced a reduction of NR retention time, indicating lysosomal membrane destabilisation. The intensity of these effects was correlated with the metal affinity for sulfhydryls. In contrast, Zn²⁺ showed no effect on lysosomes. Moreover, 200 μM Zn²⁺ protected lysosomes against the effects of Cd²⁺ and Cu²⁺, but not against Hg²⁺. Cell loading with the fluorescent pH probe Lyso Sensor followed by digital imaging showed a rise of lysosomal pH induced by Cd²⁺ and Hg²⁺, while Zn²⁺ prevented the effect of Cd²⁺ and also partially that of Hg²⁺. The different protective effect of Zn²⁺ against Hg²⁺ suggests a dual action of Hg²⁺ on lysosomes, possibly involving both membrane destabilisation and proton pump inhibition. Cell exposure to 17 β-estradiol also caused a reduction of NR retention time, which was synergistic to that of Hg²⁺. This suggests a common pathway between metals and hormone, possibly involving Ca²⁺ signaling. Received: 17 November 1999 / Accepted: 29 June 2000     

Interaction between peat, humic acid and aqueous metal ions

Analysis of peat samples from four regions of the British Isles indicates that the concentrations of Al, Pb and common transition metals tend, as expected, to be higher in regions subject to industrial pollution, but that the concentrations of the nontransition metals Na, K, Mg, Ca and Zn tend to be higher in regions remote from industrial pollution. Humic acids were extracted from the most polluted and least polluted of the peat samples and some characteristics of these acids were compared with those of two commercial acids. Values for stability constants of complexes formed between humic acids extracted from peat and Cu²⁺, Zn²⁺ or Pb²⁺ have been obtained by an ionexchange equilibrium method. Of the three metal ions examined, Pb²⁺ was found to form the most stable humic acid complexes, followed by Cu²⁺: this order agrees with the findings of Irving and Williams and of Bunzl. Implications to the binding of actinide and other heavy metal ions in aqueous humic acid are discussed.     

铜离子对中国花鲈幼鱼的毒性研究

为给中国花鲈(Lateolabrax maculatus)养殖提供污染生物学的理论数据,研究了中国花鲈幼鱼的铜离子中毒症状和半致死浓度(LC50);检测了幼鱼肝脏的谷胱甘肽(GSH)和丙二醛(MDA)含量及超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷丙转氨酶(GPT)和谷草转氨酶(GOT)活力;分析了铜离子胁迫对...     

纳米硫化镉量子点细胞毒性作用机制

为初步探讨硫化镉量子点(CdS QDs)的细胞毒性作用机制,采用MTT毒性实验比较了CdS QDs和常规CdS对仓鼠肺细胞(CHL)的毒性效应以及细胞内外活性氧水平.结果表明,1)在较低暴露浓度(≤20μg·mL-1)时,CdS QDs细胞毒性显著高于常规CdS,而在较高暴露浓度(>20μg·mL-1)时,两者相差不大.2)在较低暴露浓度(≤40μg·mL-1)时,添加N-乙酰半胱氨酸(NAC)可显著降低CdS QDs的细胞毒性,而在较高暴露浓度(>40μg·mL-1)时,添加NAC对CdS QDs的细胞毒性没有明显影响.添加NAC对常规CdS细胞毒性没有显著影响.综合实验结果推测CdS QDs的细胞毒性与暴露剂量有关:在低浓度(<20μg·mL-1)时,主要是活性氧的氧化损伤作用;在中等浓度(20~40μg·mL-1)时,活性氧和Cd2+的释放共同作用;在高浓度(>40μg·mL-1)时,则是Cd2+的释放占主导地位.     

Effects of cadmium on intracellular cation homoeostasis in the yeast Saccharomyces cerevisiae

Previous studies have demonstrated that cadmium can induce biochemical and physiological changes in yeast Saccharomyces cerevisiae. However, studies on the influence of cadmium on the ion balance in the cell and the interaction between cadmium and other ions are still relatively few in number. By using inductively coupled plasma-atomic emission spectrometry, the contents of some cations, including Zn²⁺, Ca²⁺, Fe³⁺, Cu²⁺, Mg²⁺, K⁺, and Na⁺ were measured. The data showed that the levels of Zn²⁺ and Fe³⁺ were increased, while those of Cu²⁺, K⁺, and Na⁺ were decreased after cadmium treatment. Afterwards, using the drop test assay, the interactions between cadmium and the selected ions were investigated. The results suggested that the cytotoxicity of cadmium could be attributable to the interference of cadmium with the intracellular cation homoeostasis. Calcium channel transporter Cch1 participates in the intracellular uptake of cadmium. Additionally, Zn²⁺, Ca²⁺, Fe³⁺, Mg²⁺, and K⁺ can rescue the toxic effect of cadmium in yeast.     

Iron (Fe2+)-induced toxicity produces morphological and physiological changes in roots in Panax ginseng grown in hydroponics

ABSTRACT

Rusty roots markedly influence on ginseng cultivation, and this phenomenon often attributed to iron (Fe) induced toxicity. To examine the physiological mechanisms underlying Fe-initiated toxicity as evidenced by rusty roots in Panax ginseng, morphological and physiological changes in roots were investigated in hydroponics using Fe²⁺ concentrations of 50 (control), 100, 200, 400 or 600 µM. Compared with control, reddish-brown deposition at the root surface increasingly appeared as Fe²⁺ concentration increased (≥200 µM). The pH also rose as Fe levels were elevated. Higher external Fe²⁺ concentrations produced changes in root organelles and cell structures. Structural alterations in mitochondria due to excess Fe storage, protoplast shrinkage and cell vacuolation as well as formation of central vacuole with deposits in roots were observed. In addition, apparent cell wall thickening, cell wall folding and shrinkage, damage of cell membranes and a large amount of cell debris occurred at higher external Fe²⁺ concentrations (≥400 µM). The Fe²⁺ mediated damage resulting in morphological and physiological changes in ginseng roots was concentration and pH dependent.