胁迫时间与非毒性离子对重金属抑制拟南芥种子发芽及幼苗生长的影响

测定了Hg²⁺、Cd²⁺、Cu²⁺、Pb²⁺单一重金属胁迫对拟南芥种子发芽和幼苗生长的影响.结果表明,重金属对幼苗生长的毒性大于对种子发芽的毒性,以抑制种子发芽的IC₅₀为指标,4种重金属的毒性顺序为Hg²⁺＞Cd²⁺＞Pb²⁺/Cu²⁺,以幼苗生长为指标,则毒性顺序为:Cu²⁺＞Hg²⁺＞Cd²⁺/Pb²⁺,并随着胁迫时间延长,种子萌发率下降.此外,不同重金属在不同发芽时段对种子的毒性也不尽相同,Cd²⁺的毒性在种子吸水后的0～12 h大于12～24 h,而Hg²⁺毒性在12～24 h大于0～12 h,其中,种皮对减轻重金属毒性起着十分重要的作用.通过非毒性离子(Ca²⁺、Mg²⁺、K⁺、Na⁺)与重金属离子(Hg²⁺、Cd²⁺、Cu²⁺、Pb²⁺)交互作用对拟南芥种子发芽及幼苗生长效应的研究发现, mmol·L^-1的Ca²⁺、Mg²⁺、K⁺、Na⁺可以增强Hg²⁺对种子发芽的毒性,但对Cd²⁺的毒性却没有影响.对于幼苗来说,Ca²⁺、Mg²⁺、K⁺、Na⁺可以显著增强Hg²⁺的毒性,Ca²⁺可以缓解Cd²⁺的毒性,但却增加Cu²⁺的毒性,K⁺可以缓解Pb²⁺对幼苗的毒害作用.最后,本文对重金属的毒害机理进行了探讨.     

Ca2+和K+对拟南芥幼苗镉毒害的缓解作用

该文探讨了外源钙(Ca)或钾(K)处理对不同程度的镉(Cd)胁迫(0–80 μmol·L^–1)下拟南芥(Arabidopsis thaliana)幼苗的生长和生理特性的影响。综合Ca和K对不同浓度Cd胁迫下拟南芥幼苗生长、根长以及生物量的影响情况, 表明各浓度Cd胁迫下外源Ca²⁺的最适缓解浓度均为10 mmol·L^–1; 而K⁺的最适缓解浓度在低浓度(20和40 μmol·L^–1)和高浓度(60和80 μmol·L^–1)Cd胁迫下分别为10 mmol·L^–1和20 mmol·L^–1。在低浓度Cd胁迫下, 添加适宜浓度的Ca²⁺或K⁺后幼苗可溶性蛋白和丙二醛(MDA)含量以及超氧化物歧化酶(SOD)活性相比未添加Ca和K的对照组无显著变化, 而过氧化物酶(POD)活性和总酚、类黄酮、花色素苷, 酸溶性硫醇化合物、谷胱甘肽(GSH)、植物螯合肽(PCs)的含量均下降; 高浓度Cd处理下, 添加适宜浓度的Ca²⁺或K⁺后幼苗的SOD活性升高, POD活性降低, 可溶性蛋白、MDA、总酚、类黄酮、花色素苷、酸溶性硫醇化合物、GSH以及PCs的含量也均低于对照组。在各浓度Cd胁迫下, 添加外源Ca或K均使拟南芥幼苗根部细胞DNA损伤减弱, 表现为TT嘧啶二聚体的累积量显著减少(P<0.05)。以上结果表明, 在Cd胁迫(尤其是高浓度Cd胁迫)下, 外源Ca或K通过调节酚类、金属螯合物质的代谢水平以及提高拟南芥的抗氧化能力来缓解Cd对拟南芥幼苗的毒害效应, 缓解细胞DNA损伤。该研究结果不仅能够为深入探讨Ca和K对缓解重金属毒害的分子机理提供实验依据, 而且为Ca和K应用于重金属污染的防治提供参考。     

纳米银诱导拟南芥活性氧自由基的积累和抗氧化系统的改变

为了研究纳米银(AgNPs)能否诱导陆生高等植物产生氧化胁迫,本试验以陆生模式高等植物拟南芥(Arabidopsis thaliana)为材料,以本实验室合成的纳米银处理5 d后,测定拟南芥幼苗的ROS含量,MDA含量和抗氧化酶系活性。研究结果表明:在5~20 mg/L AgNPs处理下,ROS水平均显著提高,表明AgNPs引起了氧化胁迫;15 mg/L AgNPs处理使得MDA含量显著提高,表明膜受到损伤;相应的,抗氧化酶系活性发生了改变以抵抗氧化胁迫;而800μg/L Ag~+(量等同于20 mg/L AgNPs释放的银离子量)无法改变ROS水平和抗氧化酶活性。说明AgNPs可以诱导拟南芥产生氧化胁迫,造成ROS的积累,导致抗氧化系统的改变,并且这些效应无法通过纳米银释放的Ag~+作用引起。以上的结果为阐明纳米银植物毒性机理及为纳米银的生态风险评估提供了一定的依据。     

镉胁迫对家蚕脂肪体脂质过氧化物含量及抗氧化酶活性和mRNA表达的影响

【目的】探讨鳞翅目模式昆虫家蚕Bombyx mori作为重金属污染的监测指示生物在镉胁迫下的酶反应及相关的基因表达。【方法】给家蚕幼虫期全龄添食镉(Cd²⁺), 调查不同性别家蚕5龄幼虫脂肪体中脂质过氧化物丙二醛（MDA）的含量, 超氧化物歧化酶（SOD）、 过氧化氢酶（CAT）和谷胱甘肽过氧化物酶（GSH-Px）的活性及其基因表达水平的变化。【结果】Cd²⁺胁迫对雌雄家蚕MDA 含量均具有浓度效应关系, MDA含量随Cd²⁺胁迫浓度的升高而增加。Cd²⁺胁迫下, SOD和CAT活性表现为先升后降的变化趋势, Pearson相关性分析显示SOD和CAT活性变化有显著相关性(雄: R=0.770, P=0.001; 雌: R=0.854, P=0.000）。雌性家蚕脂肪体中CAT活性变化和Cat mRNA水平的表达具有正相关性（R=0.712, P=0.003）。雄性家蚕脂肪体中GSH-Px活性随Cd²⁺胁迫浓度的升高而增加, 显示浓度 效应关系, 12.5~50 mg/kg Cd²⁺胁迫组GSH-Px活性与对照相比有显著差异（P<0.05）, 其活性和GSH-Px mRNA水平的表达具有正相关性（R=0.834, P=0.000）; 雌性家蚕脂肪体中GSH-Px活性表现为先升后降的变化趋势, 12.5 mg/kg Cd²⁺胁迫组GSH-Px活性与对照相比有显著增加（P<0.01）。【结论】结果表明, 急性镉胁迫对家蚕脂肪体有明显的毒性作用, 其作用机制与脂质过氧化加剧和抗氧化酶活性变化有关。家蚕对重金属镉的解毒机制有性别相关性。     

Cu2+对铜锈微囊藻生长及叶绿素荧光特性的影响

利用浮游植物荧光仪（Photo-PAM）研究了不同Cu²⁺浓度处理不同时间（12、24、48、72、96 h）对铜锈微囊藻（Microcystis aeruginosa）生长及叶绿素荧光特性的影响。结果表明:在一定时间（96 h）内,低浓度Cu²⁺(1μmol·L^-1)处理的铜锈微囊藻的藻细胞密度以及PSII最大光合效率（Fv/Fm）、PSII实际光合效率（ΦPSII）、光能利用效率（α）与对照相比变化不显著,最大电子传递速率（Pm）显著下降;高浓度Cu²⁺(2、4、6、8μmol·L^-1)处理下,藻细胞密度及Fv/Fm、ΦPSII、α、Pm随着Cu²⁺浓度的加大和胁迫时间的延长均显著降低,浓度为6、8μmol·L^-1 Cu²⁺处理48 h,铜锈微囊藻Fv/Fm、ΦPSII、α、Pm的测得值为0。可见,高浓度Cu²⁺(≥2μmol·L^-1)处理极大地影响铜锈微囊藻的生长及叶绿素荧光特性,胁迫作用显著。     

多环芳烃荧蒽诱导拟南芥氧化胁迫

选用模式植物拟南芥为材料,以四环的多环芳烃（PAHs）荧蒽为研究对象,从植物对非生物胁迫响应紧密相关的抗氧化酶及膜保护系统的变化入手,研究了植物对多环芳烃胁迫的生理响应.结果表明：荧蒽胁迫下拟南芥经历了氧化胁迫和膜脂过氧化过程.0.75 mmol·L^-1的荧蒽使拟南芥光合作用过程受到抑制;1.00 mmol·L^-1的荧蒽使拟南芥丙二醛(MDA)含量极显著增加, 抗坏血酸过氧化物酶(APX)活性极显著下降, 膜脂过氧化作用加剧,1.25 mmol·L^-1的荧蒽使拟南芥过氧化物酶(POD)活性极显著下降,H₂O₂在细胞内累积,拟南芥明显受害.     

外源NO介导Cu胁迫下番茄GSH-PCs合成途径

一氧化氮(NO)作为生物活性分子,广泛参与各种生物和非生物胁迫.采用营养液培养,研究了Cu胁迫下外源NO介导的番茄还原型谷胱甘肽 植物螯合肽(GSH PCs)合成途径中相关酶活性及代谢产物的变化.结果表明: 与对照(CK)相比,Cu胁迫可以显著激活番茄体内γ-谷氨酰半胱氨酸合成酶(γ-ECS)、谷胱甘肽合成酶(GS)活性,根系GSH、PCs含量急剧升高,且随着处理时间的延长,γ-ECS、GS活性和GSH、PCs含量呈持续上升趋势;添加外源硝普钠(SNP, NO供体)可以进一步提高Cu胁迫下番茄根系γ-ECS、GS活性,促进GSH、PCs的合成,增强清除过氧化物的能力,并螯合过多的Cu²⁺,降低其生物毒性.叶片中的GSH-PCs代谢变化在一定程度上滞后于根系.外源丁硫氨酸-亚砜亚胺(BSO,GSH合成抑制剂)显著抑制根系γ-ECS活性,添加SNP可以显著逆转根系中BSO对GSH和PCs合成的抑制,对叶片中PCs合成的影响较小.Cu胁迫下,外源NO可能启动了某些信号机制,并通过激活或增强GSH-PCs合成途径中的酶促和非酶促系统,降低过多的Cu²⁺的生物毒性和氧化伤害.     

高浓度镉、锌及其复合作用对烟草抗氧化系统的影响

采用水培方法,研究高浓度镉(0.1 mmol·L^-1 Cd²⁺)、锌(0.15 mmol·L^-1 Zn²⁺)及其复合作用(0.1 mmol·L^-1 Cd²⁺+0.15 mmol·L^-1Zn²⁺)对烟草种子的萌发率、幼苗叶片活性氧(ROS)水平、抗氧化物浓度、抗氧化酶活性及膜脂过氧化程度的影响.结果表明: 单因子条件下,与对照相比,高浓度镉、锌处理烟草种子萌发率降低;叶片超氧自由基(O₂^-· )产生速率与过氧化氢(H₂O₂)含量升高;过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)、脱氢抗化血酸还原酶(DHAR)、单脱氢抗坏血酸还原酶(MDAR)和谷胱甘肽还原酶(GR)活性升高;谷胱甘肽(GSH)含量及其与氧化型谷胱甘肽比值(GSH/GSSG)下降;丙二醛(MDA)含量升高.与镉、锌单因子处理相比,镉、锌复合处理的烟草种子萌发率显著升高;O₂^-· 产生速率、H₂O₂和MDA含量降低;CAT、APX、MDAR活性在处理末期升高.镉、锌胁迫对烟草可造成生理水平上的损伤,且毒性效应随着处理时间的延长而增强.镉、锌复合作用可缓解镉、锌单因子胁迫对烟草幼苗的毒害.     

旱-盐复合胁迫对玉米种子萌发和生理特性的影响

分别用15% PEG、100 mmol·L^-1 NaCl及其混合溶液模拟干旱(D)、盐(S)及旱-盐复合胁迫(D+S)对玉米种子萌发及幼苗生长的影响.结果表明: 3种胁迫处理均明显抑制了种子萌发、幼芽、幼根的伸长及生物量的积累,且影响程度为D>D+S>S;幼芽及幼根中过氧化氢(H₂O₂)、超氧阴离子(O^-·₂)等活性氧含量及丙二醛(MDA)含量明显升高,质膜相对透性增大,脯氨酸、可溶性糖和可溶性蛋白等生理渗透调节物质含量显著增加,且幼芽中含量高于幼根,积累程度均为D>D+S>S.3种胁迫处理均使幼芽、幼根中的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)及抗坏血酸过氧化物酶(APX) 等抗氧化酶活性增强;其中,SOD和APX活性表现为复合胁迫介于单一胁迫之间,而POD和CAT活性表现为复合胁迫大于单一胁迫;说明旱-盐复合逆境胁迫对玉米种子萌发及幼苗生理特性的影响并不是单一胁迫的简单叠加,与单一干旱胁迫相比,旱-盐复合胁迫在一定程度上能够缓解干旱胁迫对玉米种子萌发及幼苗生长的影响.     

H2O2-NOX系统: 一种植物体内重要的发育调控与胁迫响应机制

以H₂O₂为中心的活性氧(reactive oxygen species, ROS)的产生是动植物发育与响应外界生物与非生物胁迫的普遍 特征, 其在生理和分子2个水平上调控植物的发育和对外界胁迫的响应, 并与一系列信号转导过程相关联。作为关键的ROS产生酶, 质膜NADPH氧化酶(plasma membrane NADPH oxidase, PM-NOX)在植物应对各种生物和非生物胁迫中具有重要作用, 被广泛认为是胁迫条件下植物细胞ROS产生并积累的主要来源。该文简要综述了近年来人们在植物细胞ROS产生、清除、生理功能以及PM-NOX酶的结构特征与功能等方面的研究进展, 并认为H₂O₂-NOX系统是一种植物体内普遍存在的重要发育调控与胁迫响应机制。     

Calcium Requirement for Fast Axonal Transport in Frog Motoneurons

Abstract: Calcium is required to sustain fast axonal transport in sensory neurons of frog and cat. We studied the Ca²⁺ dependence of fast axonal transport in the motoneurons of the lower spinal cord from frog. The accumulation of acetylcholinesterase at a crush on the ventral roots was used to follow axonal transport. Two types of experiments were performed: modification of the medium bathing the ventral roots, alone, and modification of the medium bathing the spinal cord and ventral roots. Incubation (17-18 h) of the ventral roots in Ca²⁺-free medium markedly inhibited acetylcholinesterase transport, a finding that demonstrates a Ca²⁺ requirement for fast axonal transport in motoneurons; when 4 m M MgCl₂ was added to the Ca²⁺-free medium, transport was also greatly reduced. During incubation of the ventral roots in normal medium supplemented with 0.18 m M CoCl₂ transport proceeded normally; but when the Co²⁺ concentration was raised to 1.8 m M , transport was diminished as drastically as in the Ca²⁺-free medium. Incubation of the spinal cord and ventral roots in medium containing 0.18 m M CoCl₂ did not reduce the accumulation of acetylcholinesterase at the crush. Similarly, accumulation of acetylcholinesterase at a crush on the dorsal root was not significantly reduced by exposure of the dorsal root ganglion and root to 0.18 m M Co²⁺. Exposure of sensory cell bodies to 0.18 m M Co₂₊ thus produces differential effects on transport of acetylcholinesterase and on transport of newly synthesized radiolabeled protein.     

盐胁迫下菊芋根系脱落酸对钠离子转运和光系统Ⅱ的影响

通过根系施加脱落酸(ABA)合成抑制剂钨酸钠,研究盐胁迫(150 mmol·L^-1 NaCl)下菊芋根系ABA信号对根系Na⁺转运、叶片Na⁺积累和光系统Ⅱ(PSⅡ)的影响。结果表明:钨酸钠抑制盐胁迫下根系ABA合成,降低根系Na⁺外排,提高根系Na⁺向叶片的转运系数。盐胁迫增加叶片Na⁺含量,没有影响叶片膜脂过氧化、PSⅡ反应中心蛋白合成和PSⅡ最大光化学效率(F_v/F_m)。根系ABA合成受抑制,显著增加盐胁迫下叶片Na⁺积累,加剧叶片膜脂过氧化,损伤PSⅡ反应中心蛋白,显著降低F_v/F_m,诱发PSⅡ光抑制。总之,盐胁迫下菊芋根系ABA信号诱导根系Na⁺外排,抑制Na⁺向地上部转运,有利于减少叶片Na⁺积累,防御PSⅡ氧化损伤。     

Transport interactions between cadmium and zinc in roots of bread and durum wheat seedlings

Field studies have shown that the addition of Zn to Cd-containing soils can help reduce accumulation of Cd in crop plants. To understand the mechanisms involved, this study used ¹⁰⁹Cd and ⁶⁵Zn to examine the transport interactions of Zn and Cd at the root cell plasma membrane of bread wheat ( Triticum aestivum L.) and durum wheat ( Triticum turgidum L. var. durum ). Results showed that Cd²⁺ uptake was inhibited by Zn²⁺ and Zn²⁺ uptake was inhibited by Cd²⁺. Concentration-dependent uptake of both Cd²⁺ and Zn²⁺ consisted of a combination of linear binding by cell walls and saturable, Michaelis-Menten influx across the plasma membrane. Saturable influx data from experiments with and without 10 µm concentrations of the corresponding inhibiting ion were converted to double reciprocal plots. The results revealed a competitive interaction between Cd²⁺ and Zn²⁺, confirming that Cd²⁺ and Zn²⁺ share a common transport system at the root cell plasma membrane in both bread and durum wheat. The study suggests that breeding or agronomic strategies that aim to decrease Cd uptake or increase Zn uptake must take into account the potential accompanying change in transport of the competing ion.     

硅对铜胁迫下小麦幼根细胞超微结构的影响

为探索硅对重金属铜胁迫下小麦的缓解作用,以矮抗58为供试材料,采用室内溶液培养法,研究不同浓度重金属铜(0、15、30 mg·L^-1)胁迫及铜胁迫下添加外源硅(50 mg·L^-1)对小麦幼苗生长、根系活力及根尖超微结构的影响.结果表明: 铜处理下,小麦幼苗根长、株高和根系活力均极显著低于对照(P＜0.01),加硅处理后小麦根长、株高和根系活力与单一铜处理相比均有所提高.铜胁迫下,小麦根尖细胞壁和细胞膜均遭到不同程度的破坏,细胞间空隙加大、细胞器消失;加硅处理后,虽然与对照相比,部分细胞和细胞器仍有变形,但细胞结构基本保持完好.综上,外源硅能在一定程度上缓解重金属铜对小麦幼苗生长及细胞组分的胁迫.
     

Efficient iron plaque formation on tea(Camellia sinensis) roots contributes to acidic stress tolerance

Tea plants grow in acidic soil, but to date, their intrinsic mechanisms of acidic stress tolerance have not been elucidated. Here, we assessed the tea plant response to growth on NHt4 nutrient media having different p H and iron levels. When grown in standard NHt4 nutrient solution(iron insufficient, 0.35 mg Là1 Fe2t), tea roots exhibited significantly lower nitrogen accumulation, plasma membrane Ht-ATPase activity, and protein levels; net Htefflux was lower at pH 4.0 and 5.0 than at pH 6.0. Addition of30 mg Là1 Fe2t(iron sufficient, mimicking normal soil Fe2tconcentrations) to the NHt4 nutrient solution led to more efficient iron plaque formation on roots and increased root plasma membrane Ht-ATPase levels and activities at p H 4.0 eland 5.0, compared to the p H 6.0 condition. Furthermore,plants grown at pH 4.0 and 5.0, with sufficient iron,exhibited significantly higher nitrogen accumulation than those grown at pH 6.0. Together, these results support the hypothesis that efficient iron plaque formation, on tea roots, is important for acidic stress tolerance. Furthermore,our findings establish that efficient iron plaque formation is linked to increased levels and activities of the tea root plasma membrane Ht-ATPase, under low pH conditions.     

铈对紫萍抗铜能力的影响

铈能减缓铜胁迫导致的紫萍(Spirodela polyrhiza L.)叶片膜透性的增加、可溶性蛋白含量和叶绿素a/b 值的下降以及超氧阴离子(O₂·)、丙二醛(MDA)的积累。与同浓度的单一铜处理相比, 加入20 mg/L 的铈后, 超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)的活性和类胡萝卜素(Car)、抗坏血酸(AsA)含量不同程度的增强或增多, 暗示铈可能通过提高紫萍活性氧清除能力以增强对铜的抗性和耐性。     

Cu ions interact with cell membranes

The influence of Cu²⁺ ions on the physical properties of resealed human erythrocyte membranes was studied by fluorescence spectroscopy. A net ordering effect was observed at the hydrophobic–hydrophilic interface both in the bulk as well as in the lipid–protein boundary. The explanation for this result was found by X-ray diffraction performed in multilayers of dimyristoylphosphatidylcholine (DMPC) and dimyristoylphosphatidylethanolamine (DMPE), representative of phospholipid classes located in the outer and inner monolayers of the human erythrocyte membrane, respectively. Cu²⁺ did not significatively affect the structure of DMPE; however, DMPC polar head and hydrocarbon chain arrangements were perturbed at low but reordered at high Cu²⁺ concentrations. These effects were respectively explained in terms of a limited and extended interaction between Cu²⁺ ions and DMPC PO₄⁻ groups. Thus, the ordering effect in the erythrocyte membrane could be based on the interaction of this cation with phosphatidylcholine phosphate groups located in its outer leaflet. This binding, besides producing a decrease of membrane fluidity, might also induce a change in its electric field. These two effects should affect the activity of membrane proteins, particularly of ion channels. In fact, it was found that increasing concentrations of Cu²⁺ ions applied to either the mucosal or serosal surface of the isolated toad skin elicited a dose-dependent decrease of the short-circuit current (SCC) and of the potential difference (PD). These results lead to the conclusion that Cu²⁺ ions inhibited Na⁺ transport across the epithelial cell membranes.     

铜处理对菠菜幼苗矿质营养吸收和细胞超微结构的影响

土壤重金属积累严重影响植物生长和生态系统平衡,探寻植物对重金属的耐性机理尤为重要.菠菜可能具有一定的耐铜性,但Cu对其矿质元素吸收、细胞超微结构等方面的耐性机理尚不明确.本研究以菠菜幼苗为研究对象,通过盆栽试验,探究不同浓度铜处理对菠菜幼苗生长、矿质元素吸收、叶片细胞超微结构等指标的影响.结果表明: 100 mg·L^-1 CuSO₄处理浓度时,菠菜幼苗根Cu²⁺积累量小于地上部,其根系生长量增加,地上部生长量稍有下降,继续增加铜处理浓度,植物体各器官生长参数均呈下降趋势.低浓度铜处理时(<400 mg·L^-1 CuSO₄),菠菜幼苗叶N、K、Ca、Mg、Fe含量增加,P含量减少;根N、P、K含量减少,Ca、Mg、Fe含量增加;叶片细胞内各细胞器清晰可见,基粒片层排列仍较为整齐,叶绿体内外膜完整.高浓度铜处理时(>600 mg·L^-1 CuSO₄),菠菜幼苗叶N含量增加,P、K、Ca、Mg、Fe含量减少;根N、P、K、Ca、Mg、Fe含量均减少;叶片细胞内叶绿体变圆,叶绿体膜变薄,基质、基粒片层变少,层堆积高度下降,细胞核解体,液泡、细胞壁中有黑色小点分布,可能是大量Cu²⁺聚集导致细胞内膨压增大所致.低浓度铜处理并未对菠菜幼苗的生长生理特性产生明显的负面影响,而高浓度铜处理并未终止菠菜幼苗的生长.说明菠菜幼苗具有一定的耐铜性.     

Differential DNA strand breaking abilities of *OH and ROS generating radiomimetic chemicals and gamma-rays: study of plasmid DNA, pMTa4, in vitro

Reactive oxygen species (ROS) interact with components of a living cell. Among them ^⋅OH is known to cause major oxidative damage to living cells and is proposed to be involved in pathogenesis including carcinogenesis. Proper understanding of consequences of such damage is, therefore, medically relevant. In this report, aqueous solution of plasmid DNA, pMTa4, has been exposed to Fenton oxidant and Haber-Weiss oxidant mediated free radical generating chemical systems, and ⁶⁰Co γ-rays in vitro either alone or in combination to study their strand breaking abilities. The exposed pMTa4 was analyzed by agarose gel electrophoresis. The results show qualitative differences in the induction of strand breaks on the plasmid pMTa4 molecule by the iron (Fe²⁺), copper (Cu²⁺) or γ-rays mediated ^⋅OH and other ROS.