Cd2+、Al3+作用下蚕豆UDS与微核相关性分析及高等植物UDS技术初探

为探讨金属离子对高等植物非按期DNA合成(UnscheduledDNASythesis,简称UDS)和微核(MCN)的诱导作用、二者之间的关联性以及利用高等植物UDS技术检测环境诱变物的可行性,利用3HTdR前体掺入法研究了Cd²⁺、Al³⁺作用下蚕豆的UDS效应。结果表明,Cd²⁺、Al³⁺均能不同程度地诱导蚕豆UDS和MCN的发生;UDS量与微核率(MCNF)之间呈负相关(r<0),但相关不显着(|r|0.05),且二者间的相关程度在Cd²⁺和Al³⁺两种金属离子作用下没有显着差别(P>0.05);利用高等植物UDS技术检测环境诱变物质,在一定受检物剂量范围内是可靠的,但超过这个剂量范围,UDS技术无法检出.     

Cd^2＋,Al^3＋作用下蚕豆UDS与微核相关性分析及高等植物UDS技？…

为探讨金属离子对高等植物非按期ＤＮＡ合成（Ｕｎｓｃｈｅｄｕｌｅｄ ＤＮＡ Ｓｙｔｈｅｓｉｓ,简称ＵＤＳ）和微核（ＭＣＮ）的诱导作用,二者之间的关联性以及利用高等培植物ＵＤＳ技术检测环境诱变物的可行性,利用＾３Ｈ－ＴｄＲ前体掺入法研究了Ｃｄ＾２＋,Ａｌ＾３＋作用下蚕豆的ＵＤＳ效应。     

表面活性剂、酸雨和Cd2+复合污染对蚕豆胚根细胞核的毒性

用蚕豆根尖微核技术研究了Cd²⁺单因子以及与表面活性剂、模拟酸雨复合污染时对植物细胞的毒性作用.结果表明,Cd²⁺浓度在0～10.0mg·L^-1范围内,对蚕豆胚根细胞微核的形成有强烈的诱导作用,Cd²⁺浓度6.0mg·L^-1时的细胞微核率为13.85‰,对照组的微核率为4.53‰,此时污染影响指数(PI)为3.06;当环境中存在表面活性剂LAS1.0mg·L^-1或pH值降到4.5和3.5时,同一Cd²⁺浓度下,蚕豆根尖细胞微核率、PI降低,同时伴有核变形,细胞中颗粒物增多,胚根组织不容易分散等症状,根的生长受到抑制,说明表面活性剂、酸雨对Cd²⁺的毒性有协同作用.pH3.5的酸雨环境中Cd²⁺对蚕豆细胞的损伤程度比pH4.5酸雨环境高.在检测高浓度、强毒性污染物的致突变效应时,应作至少3个稀释倍数,找出蚕豆根尖细胞最高微核率及PI.     

重金属离子对凡纳滨对虾肝胰脏、鳃丝和血液SOD活力的影响

研究了3种重金属离子(Cu²⁺、Zn²⁺、Cd²⁺)在96 h内对凡纳滨对虾(Litopenaeus vannamei)对肝胰脏、鳃丝和血液超氧化物歧化酶(SOD)活力的影响.结果表明,凡纳滨对虾SOD活力在3种重金属离子作用下随取样时间变化显著(P＜0.0),Cu²⁺在实验浓度范围内(0.1～1 mg·L^-1),肝胰脏、鳃丝和血液的SOD活力随时间延长呈一峰值变化,Zn²⁺在10 mg·L^-1时对肝胰脏表现为显著抑制作用,Cd²⁺在0. mg·L^-1时对肝胰脏和鳃丝起显著抑制作用,0.2 mg·L^-1对鳃丝SOD活力无显著变化(P＞0.0),其他浓度Zn²⁺(＜10 mg·L^-1)、Cd²⁺(＜0.2 mg·L^-1)对各组织器官SOD活力的影响随时间延长均呈现先升高后下降的趋势.3种重金属离子对凡纳滨对虾肝胰脏、鳃丝、血液SOD活力的影响呈现明显的剂量-时间效应关系.其SOD活力大小顺序为肝胰脏＞鳃丝＞血液,3种重金属离子对凡纳滨对虾伤害大小顺序为Cd²⁺＞Cu²⁺＞Zn²⁺.     

镉胁迫下三个萝卜栽培种蛋白质变化的双向电泳比较研究

在0.5mmol·L^-1 Cd²⁺处理时,小五缨(XWY)的发芽率下降至92%,并且随着Cd²⁺浓度的增加,发芽率逐渐降低,在1和5mmol·L^-1 Cd²⁺浓度处理时,发芽率分别降至83%和67%。象牙白(XYB)则在5mmol·L^-1 Cd²⁺浓度时,发芽率下降。卫青(WQ)在0.05mmol·L^-1 Cd²⁺浓度时,发芽率已降至83%, 1和5mmol·L^-1时则下降至58%。幼苗生长也明显受Cd²⁺处理影响,在0.05mmol·L^-1 Cd²⁺处理时, 3种栽培萝卜的幼苗生长均受到明显影响,并且随Cd²⁺浓度的增加,生长受抑加重。从发芽率和幼苗生长两种实验结果看,卫青对Cd²⁺最为敏感。双向电泳结果表明, Cd²⁺处理后3种栽培萝卜幼苗中蛋白质组分有明显变化。小五缨中, 0.1 mmol·L^-1Cd²⁺处理后,有5个蛋白质点消失, 15个新的蛋白质点被诱导产生。象牙白中, 2个蛋白质点消失, 1个蛋白质点含量减少, 13个新的蛋白质点被诱导产生。卫青中, 12个新的蛋白质点诱导产生,但没有发现蛋白质点消失现象。Cd²⁺处理后, 3种栽培萝卜中,蛋白质合成的变化与幼苗生长受抑存在明显相关性,这一实验结果对于探讨萝卜Cd²⁺害的生化机理是有重要意义的。     

利用蚕豆根尖细胞微核技术检测Cu、As污染的诱变性

应用蚕豆根尖细胞微核技术检测Cu、As及其复合污染的诱变性能,计数了蚕豆根尖细胞微核千分率(MCN‰)和污染指数(PI)并进行了F检验.结果表明,在一定浓度范围内(Cu＜200mg·L,As＜15mg·L^-1),随着Cu、As浓度的增加,MCN‰上升;24个试验处理组PI均在2以上,各处理组MCN‰有显著差异,和对照组有极显著差异(a＜0,01),表明Cu、As及其复合处理对蚕豆根尖细胞的分裂活动有明显影响.     

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。     

重金属Cd2+对拟水狼蛛体内金属硫蛋白含量及其生长发育的影响

为了明确重金属Cd²⁺对拟水狼蛛Pirata subpiraticus体内金属硫蛋白(metallothionein, MT)含量及其生长发育的影响, 在室内条件下用5个不同浓度（0, 10, 20, 40和80 μg/g）的Cd²⁺培养的黑腹果蝇Drosophila melanogaster饲喂从5种不同生境下（S1, S2, S3, S4和S5）采集的拟水狼蛛性成熟雌蛛产卵孵化的幼蛛,待幼蛛性成熟后取所得雌蛛成蛛采用原子吸收光谱法测定了Cd²⁺ 诱导下拟水狼蛛体内金属硫蛋白含量及其存活率和生长率。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, 积累量随黑腹果蝇培养基中Cd²⁺浓度的增加而增加,存在显著的浓度-效应关系。不同浓度的Cd²⁺能够诱导拟水狼蛛体内MT不同的表达,表达量与Cd²⁺浓度显著正相关(P<0.05)。当浓度低于20 μg/g时,污染点（S1, S2, S3和S4）拟水狼蛛体内MT表达量显著高于参照组S5（P<0.05）; 当高于20 μg/g时, 所有样点拟水狼蛛体内MT表达量差异不显著（P＞0.05）。拟水狼蛛存活率和成长率随着Cd²⁺浓度的升高呈下降趋势。据此认为,金属硫蛋白可能是蜘蛛耐受重金属污染的重要机制,与重金属具有一定浓度-效应关系。     

可口革囊星虫对Cd2+和Hg2+的富集及其影响

采用原子吸收、原子荧光等分析检测技术,探讨了Cd²⁺、Hg²⁺在可口革囊星虫中的富集规律及其对生长与主要营养成分的影响.结果表明: 在试验设定的胁迫浓度内,可口革囊星虫体壁肌肉对Cd²⁺、Hg²⁺的富集均随胁迫时间的延长而增加,最终达到饱和浓度;环境中Cd²⁺、Hg²⁺浓度越高,富集速度越快,达到饱和的时间越短,饱和浓度也越高.可口革囊星虫体质量增长随着重金属胁迫浓度的升高而减慢,且联合胁迫的影响程度大于单一胁迫.体壁肌肉蛋白质含量随重金属胁迫浓度的增加而升高,Cd²⁺、Hg²⁺分别在0.05和0.02 mg·L^-1胁迫浓度下达到最高,然后开始降低.联合胁迫也呈同样的规律,且影响程度更大.体壁肌肉脂肪含量随重金属胁迫浓度的增加而降低,联合胁迫下降低程度更大.     

食品添加剂和金属离子对高粱泡红色素稳定性的影响

探讨了5种常用食品添加剂和7种金属离子对高粱泡红色素稳定性的影响。结果表明:葡萄糖、蔗糖和苯甲酸钠对高粱泡红色素稳定性无不良影响,其中葡萄糖、蔗糖有不同程度的增(护)色效果;柠檬酸能显著提高色素的稳定性;而VitC能促进色素的氧化降解,有明显的破坏作用。金属离子中,Na⁺、Mg²⁺、Al³⁺、Zn²⁺等对高粱泡红色素稳定性无影响,且有一定增色作用;较高浓度(≥0.0025mol/L)Mn²⁺有一定不良影响,而Cu²⁺、Fe³⁺则有明显的破坏作用。     

Cd2+、Al3+对蚕豆(Vicia faba)DNA合成及修复的影响

利用＾３Ｈ－ＴｄＲ掺入方法,研究了不同浓度单金属离子Ｃｄ＾２＋、Ａｌ＾３＋对蚕豆ＤＮＡ合成、ＤＮＡ修复（以ＵＤＳ为指标）的影响。结果表明：在低浓度Ｃｄ＾２＾＋、Ａｌ＾３＋（Ｃｄ＾２＋浓度〈２００ｍｇ／ｌ,Ａｌ＾３＋浓度１００ｍｇ／ｌ）处理后,蚕豆ＤＮＡ合成加快,并且不同程度地诱导了ＵＤＳ的发生;但在高于此浓度的Ｃｄ＾２＋、Ａｌ＾３＋作用下,蚕豆ＤＮＡ合成受抑制,浓度越高,抑制作用超强;并且几乎不表     

Changes of reliability and efficiency of micronucleus bioassay in Vicia faba after exposure to metal contamination for several generations

Mitotic root micronucleus (MCN) frequency in Vicia faba as a bioassay, is primarily based on the extent of the sentinel Vicia response in terms of cytogenetic damage quantitatively or qualitatively to indicate the presence of mutagenic contaminants. This paper describes an investigation designed to assess changes in MCN frequencies of Vicia faba from three generation plants obtained, respectively from a reference site (RS) and a metal-contaminated experimental field (EF) in the bioassay of mutagenic Cd(2+) and NaN(3). The background value, dose-response to Cd(2+) and to NaN(3) in three generation (F(1), F(2) and F(3)) plants of the EF and the initial (F(0)) plants were determined in terms of MCN frequencies. With more generations of growing Vicia plants in the EF, a higher background value of MCN frequency, a lower slope value in the regression equation, a smaller ratio of MCN frequency between the control and treatment in the same generation and larger perturbation values were observed. This denotes that the decreased reliability and efficiency are represented in Vicia plants from the EF if the plants are used as sentinels in the bioassay of mutagenic Cd(2+) and NaN(3). It was concluded that the Vicia MCN bioassay should be used as an endpoint biomarker acceptable in biomonitoring environmental mutagens when the sentinel plants were collected from clean areas. Because no place is absolutely without pollutants, it is suggested that several seed stock centers should be established for providing sentinel Vicia if Vicia MCN bioassay is used as a biomarker to identify the global environmental status.     

A comparison of SCE test in human lymphocytes and Vicia faba: a hopeful technique using plants to detect mutagens and carcinogens

A comparison of the Vicia faba SCE with the human lymphocyte SCE test was made with regard to their capacity to detect mutagens. Twelve chemical agents that can induce sister-chromatid exchanges (SCEs) in both Vicia faba root tip cells and human lymphocytes were used. The results indicate that the plant SCE technique is a sensitive test for screening various chemical agents including food preservatives, pesticides and essences which may have mutagenic activity. A new simplified procedure was introduced for differential staining of sister chromatids in Vicia faba.     

表面活性剂、酸雨和Cd^2＋复合污染对蚕豆胚根细胞核的毒性

用蚕豆根尖微核技术研究了Cd^2+单因子以及与表面活性剂、模拟酸雨复合污染时对植物细胞的毒性作用。结果表明,Cd^2+浓度在0～10．0mg·L^-1范围内,对蚕豆胚根细胞微核的形成有强烈的诱导作用,Cd^2+浓度6．0mg·L^-1时的细胞微核率为13．85‰,对照组的微核率为4．53‰,此时污染影响指数(PI)为3．06;当环境中存在表面活性剂LAS1．0mg·L^-1或pH值降到4．5和3．5时,同-Cd^2+浓度下,蚕豆根尖细胞微核率、PI降低,同时伴有核变形,细胞中颗粒物增多,胚根组织不容易分散等症状,根的生长受到抑制,说明表面活性剂、酸雨对Cd^2+的毒性有协同作用。pH3．5的酸雨环境中Cd^2+对蚕豆细胞的损伤程度比pH4．5酸雨环境高,在检测高浓度、强毒性污染物的致突变效应时,应作至少3个稀释倍数,找出蚕豆根尖细胞最高微核率及PI。     

Genotoxicity of hydrated sulfur dioxide on root tips of Allium sativum and Vicia faba

Genotoxicity of sulfur dioxide (SO(2)) and its hydrates (bisulfite and sulfite) in human lymphocytes and other mammalian cells have been found earlier in our laboratory. In the present studies, we used Allium stavium and Vicia faba cytogenetic tests, which are the highly sensitive and simple plant bioassays. A mixture of sodium bisulfite and sodium sulfite (1:3), at various concentrations from 1 x 10(-4) to 2 x 10(-3)M was used for the treatment. Genotoxicity was expressed in terms of anaphase aberration (AA) frequencies in the Vicia-AA test and in terms of micronuclei (MCN) frequencies in both Vicia-MCN test and Alllium-MCN test. On average, the results showed a 1.7-3.9-fold increase of AA frequencies and a 3.5-4.5-fold increase of MCN frequencies in Vicia root tips as compared with the negative control. Similarly, results of Allium-MCN test also showed a significant increase in MCN frequencies in the treated samples. In addition, pycnotic cells (PNC) appeared in Allium root tips of treated groups. The frequencies of MCN, AA and PNC increased dose-dependently and the cell cycle delayed at the same time in bisulfite treated samples. Results of the present study suggest that the Vicia and Allium cytogenetic bioassays are efficient, simple and reproducible in genotoxicity studies of bisulfite.     

太湖蓝藻滤液的遗传毒性研究

蓝藻爆发是环境污染引发的重要事件之一,随之产生的蓝藻毒素又直接危及区域水安全.该论文采用蚕豆和大蒜根尖微核试验研究了太湖蓝藻暴发期间蓝藻滤液的遗传毒性.结果表明,同阴性对照相比,所有试验处理对蚕豆根尖细胞微核发生率的影响显著增加;对大蒜根尖细胞微核发生率而言除蓝藻滤液8倍稀释液的影响不显著外,其它水平效应显著高于阴性对照,而且表现出一定的剂量效应.暴发期蓝藻滤液原液对蚕豆根尖细胞微核发生率影响显著高于阳性对照(0.8mg·mL^-1环磷酰胺)的效应,从而说明蓝藻暴发时期蓝藻滤液具有较强的遗传毒性.通过微核试验效果分析,蚕豆作为植物监测系统的敏感性和稳定性都优于大蒜材料.     

Genotoxic effects of heavy metals in rat hepatocytes

The genotoxic interaction of metals, which are common environmental contaminants, was studied in cultured hepatocytes. Freshly isolated rat hepatocytes were exposed to concentrations of cadmium, copper, silver and lead salts ranging from non-cytotoxic to moderately cytotoxic (as determined by LDH release), and the incorporation of [³H]thymidine into the DNA, as a measure of repair synthesis, was followed. In addition, the uptake of metals by the nuclear fraction was determined using Inductively Coupled Plasma/Mass Spectrometry or atomic absorption spectrophotometry. The evaluation of binding of ¹⁰⁹Cd to the DNA in situ was also attempted. It was observed that after a 20 h exposure period, all the metals investigated were found in the nuclear fraction of hepatocytes, with Ag apparently being accumulated less efficiently. In parallel, Cd (0.18 to 1.8 µM) and Cu (7.9 to 78.5 µM) consistently produced a statistically significant stimulation of [³H]thymidine incorporation into the DNA, in the presence or absence of hydroxyurea while Ag was active only at the highest concentration tested (18.5 µM). In contrast, Pb failed to induce a UDS response at the levels used. Moreover, exposure of hepatocytes to 1.8 µM ¹⁰⁹CdCl₂ for 20 h led to a DNA binding ratio of 0.98 ± 0.23 ng Cd/ µg DNA. The present results support the view that the nucleus may be an important target organelle for metal toxicity.Abbreviations 2-AAF 2-acetylaminofluorene - Cd cadmium - HU hydroxyurea - lCP/MS inductively coupled plasma/mass spectrometry - Hg mercury - Ni nickel - UDS unscheduled DNA synthesis     

螺旋藻多糖对核酸内切酶活性和DNA修复合成的增强作用

本文用核酸内切酶实验和放射自显影术研究了螺旋藻水溶性多糖对DNA切除修复的效应。结果表明,该多糖能显著增强辐射引起DNA损伤的切除修复活性和程序外DNA合成(UDS)。考察切除修复的时程,发现螺旋藻多糖的存在不但能加快损伤DNA切除反应和UDS的初时速度,而且能延缓以上两个重要修复反应的饱和。     

应用蚕豆根尖细胞微核技术评价广西平乐锰矿废弃地土壤的污染状况

分别选取废弃地上的花生地、柿树地、柑桔地和桃树地等4个采样点进行调查和监测,测定了各采样点土壤浸提物的蚕豆根尖细胞微核千分率(MCN‰)和污染指数(PI),应用蚕豆根尖细胞微核技术对广西平乐锰矿废弃地土壤的污染状况进行了评价,并对造成土壤污染的原因进行了分析讨论。结果表明,用浸提法处理的4个采样点土壤的根尖细胞微核千分率(MCN‰)均明显高于对照组(P<0.05),其中柑桔地的污染程度最严重,污染指数达到了2.93;污染程度最轻的是花生地,其污染指数为1.97。这表明广西平乐锰矿废弃地土壤均存在不同程度的污染,在锰矿废弃地土壤上早期不宜种植食用的农作物。