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.     

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.     

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。     

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.     

不同炭化温度制备的蚕丝被废弃物生物炭对重金属Cd2+的吸附性能

以蚕丝被废弃物为原料,在300、500和700 ℃高温缺氧条件下热解炭化制备成3种生物炭(BC300、BC500和BC700).利用扫描电镜(SEM)、傅里叶红外光谱仪(FT-IR)、X-射线衍射仪(XRD)、比表面积分析仪等对其理化性质进行表征,并研究了不同温度下制备的生物炭对溶液中Cd²⁺的吸附特性.结果表明: 随着炭化温度上升,BET比表面积、pH、灰分均增大,生物炭表面形态结构越来越不规则.XRD结果显示:不同温度下获得的生物炭中均含有一定量的方解石,FT-IR光谱图上的峰主要为-OH和方解石典型的吸收峰;pH对生物炭吸附Cd²⁺的影响不大;Langmuir方程能更好地拟合3种生物炭对Cd²⁺的吸附等温过程,其最大吸附量分别为25.61、52.41和91.07 mg·g^-1.3种生物炭对Cd²⁺吸附过程均更符合准二级动力学方程,且BC700对Cd²⁺的吸附效果最佳.进一步研究离子浓度及阳离子共存对BC700吸附Cd²⁺的影响,结果显示: NaCl浓度越高,对Cd²⁺的吸附抑制越明显;共存阳离子中,Ca²⁺和Mg²⁺对Cd²⁺的吸附抑制更明显,而K⁺几乎无影响.因此,以蚕丝被废弃物制备的生物炭作为去除水体中Cd²⁺的吸附剂具有较强的应用潜力.     

NaHCO3胁迫下3种灌木Na+、K+、Ca2+的吸收及转运

通过盆栽试验,采用原子吸收分光光度法和非损伤微测技术,研究了NaHCO₃胁迫(300 mmol·L^-1)对大洋洲滨藜、四翅滨藜和宁夏枸杞3种灌木离子吸收及运转的影响.结果表明: 随着NaHCO₃浓度升高,两种滨藜和宁夏枸杞叶片中Na⁺含量升高,300 mmol·L^-1NaHCO₃胁迫下,宁夏枸杞叶肉细胞Na⁺的外排增加,两种滨藜净Na⁺外排降低;随着胁迫时间的延长,大洋洲滨藜和宁夏枸杞叶片的K⁺含量下降,Na⁺/K⁺升高,四翅滨藜叶片K⁺含量升高,Na⁺/K⁺降低;随着浓度的升高,宁夏枸杞叶片积累Ca²⁺减少,Na⁺/Ca²⁺高于对照,叶肉细胞Ca²⁺外排;两种滨藜叶Ca²⁺含量总体呈升高趋势,叶肉细胞Ca²⁺表现为内流.在NaHCO₃胁迫下,3种灌木通过不同的策略来消除Na⁺毒害.宁夏枸杞叶片Na⁺的积累抑制了对Ca²⁺的吸收;两种滨藜Ca²⁺的内流促使细胞质中游离Ca²⁺增加,增加的细胞质\[Ca²⁺\]cyt防治质膜H⁺ ATPase去极化,限制K⁺的外排,从而维持细胞内Na⁺/K⁺的平衡,其中四翅滨藜调控Na⁺/K⁺平衡的能力较强.     

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.     

油桃花芽破眠过程中H2O2代谢与Ca2+转运的关系

利用化学测定法分析高温、单氰胺和TDZ 3种破眠处理对“曙光”油桃休眠花芽H₂O₂代谢的主要影响,利用非损伤微测技术检测H₂O₂对休眠芽Ca²⁺转运的影响,研究H₂O₂在芽休眠解除过程中的调控作用.结果表明： 在深休眠时期,高温和单氰胺处理均能诱导芽内H₂O₂含量升高和过氧化氢酶(CAT)活性降低,并具有显著的破眠作用;TDZ对H₂O₂含量及CAT、过氧化物酶(POD)活性影响不大,破眠效果较差.休眠花芽原基组织钙通道活跃,对外源Ca²⁺呈吸收状态.外源H₂O₂可诱导休眠花芽原基组织Ca²⁺转运发生变化,低浓度H₂O₂降低Ca²⁺吸收速率,高浓度H₂O₂使组织对Ca²⁺的转运由吸收转变为释放.这表明休眠芽内H₂O₂信号和Ca²⁺信号相关联,通过诱导H₂O₂积累调控Ca²⁺信号可能在高温和单氰胺打破休眠的信号转导过程中起重要作用.     

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⁺.     

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     

Zn、Cd及其复合对小麦幼苗吸收Ca、Fe、Mn的影响

研究了溶液培养条件下Cd、Zn及其复合对小麦幼苗吸收Ca、Fe、Mn的影响.结果表明,小麦幼苗对Zn、Cd的吸收随溶液中Cd²⁺、Zn²⁺浓度的升高而增加,Cd、Zn同时存在时与其单独作用时幼苗对它们的吸收不同,Zn影响幼苗对Cd的吸收,Cd对Zn的吸收起抑制作用.Ca、Mn的吸收随溶液中Cd²⁺、Zn²⁺浓度升高而呈下降趋势,在Cd单独处理组和Zn单独处理组中Fe的吸收随Cd²⁺、Zn²⁺浓度升高而增加,但在Zn+Cd处理组中,Fe的吸收则呈下降趋势,其效应方式还与作物具体部位有关.     

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.     

Apoplastic and membrane‐associated Ca2+ in leaves and roots as affected by boron deficiency

A combination of fluorescein‐isothiocyanate (FITC), coumarin‐benzothiazol (BTC), and chlorotetracycline (CTC) fluorescence was used to simultaneously monitor apoplastic pH, apoplastic free Ca²⁺, and plasma membrane‐bound Ca²⁺. As early boron deficiency reactions supposedly include alterations of plasma membrane‐bound transport processes besides rapid effects on cell wall physical properties, the corresponding changes were followed in leaves and roots of Vicia faba L. cv. Troy.
Boron deficiency did not alter the apoplastic pH, but it reduced plasma membrane‐bound Ca²⁺ in roots at 4 h and leaves at 3 days after starting the deficiency treatment. The decrease in plasma membrane‐bound Ca²⁺ coincided with an increase in apoplastic free Ca²⁺ and K⁺, and occurred before the first visible symptoms were noticed.
It is proposed that less Ca²⁺ is bound to the plasma membrane due to a reduction of specific Ca²⁺‐binding sites (borate esters with vic ‐diols or polyhydroxy‐carboxylates) before plasma membrane integrity deteriorates.     

Involvement of intracellular Ca2+ in blue light-dependent proton pumping in guard cell protoplasts from Vicia faba

Recent studies have suggested that Ca²⁺/calmodulin (CaM) or CaM-like proteins may be involved in blue light (BL)-dependent proton pumping in guard cells. As the increase in cytosolic concentration of Ca²⁺ is required for the activation of CaM and CaM-like proteins, the origin of the Ca²⁺ was investigated by measuring BL-dependent proton pumping with various treatments using guard cell protoplasts (GCPs) from Vicia faba . BL-dependent proton pumping was affected neither by Ca²⁺ channel blockers nor by changes of Ca²⁺ concentration in the medium used for the GCPs. Addition of Ca²⁺ ionophores and an agonist to GCPs did not induce proton pumping. However, BL-dependent proton pumping was inhibited by 10 m M caffeine, which releases Ca²⁺ from the intracellular stores, and by 10 μ M 2,5-di-( tert -butyl)-1,4-benzohydroquinone (BHQ) and 10 μ M cyclopiazonic acid (CPA), inhibitors of Ca²⁺-ATPase in the sarcoplasmic and endoplasmic reticulum (ER). By contrast, the inhibitions were not observed by 10 μ M thapsigargin, an inhibitor of animal ER-type Ca²⁺-ATPase. The inhibitions by caffeine and BHQ were reversible. Light-dependent stomatal opening in the epidermis of Vicia was inhibited by caffeine, BHQ, and CPA. From these results, we conclude that the Ca²⁺ thought to be required for BL-dependent proton pumping may originate from intracellular Ca²⁺ stores, most likely from ER in guard cells, and that this origin of Ca²⁺ may generate a stimulus-specific Ca²⁺ signal for stomatal opening.     

Effect of ultraviolet radiation on accumulation and leakage of 86Rb+ in guard cells of Vicia faba

The effects of UV-C (254 nm), UV-A (365 nm) and broad-band UV (280–380 nm) on guard cells of Vicia faba L. cv. Long Pod were investigated in the presence of white light (450 μmol m⁻² s⁻¹). UV-C (7 μmol m⁻² s⁻¹) was found to cause leakage of ⁸⁶Rb⁺ from guard cells, while UV-A (0.3 μmol m⁻² s⁻¹) stimulated increased uptake in these cells. A relatively small stimulatory effect was observed by broad-band UV (3 μmol m⁻² s⁻¹) during the first 30 min of irradiation with an apparent equilibration of influx and efflux thereafter. Leakage of ⁸⁶Rb⁺ from guard cells continued despite the removal of UV-C and an increase in the amount of white light from 450 to 1500 μmol m⁻² s⁻¹, suggesting that membranes were irreversibly damaged. Irradiation of guard cells with UV-C for 30, 45 and 90 min indicated that these cells began to be affected already by 30 min UV-C irradiation.     

Barium Evokes Glutamate Release from Rat Brain Synaptosomes by Membrane Depolarization: Involvement of K+, Na+, and Ca2+ Channels

Abstract: During K⁺ -induced depolarization of isolated rat brain nerve terminals (synaptosomes), 1 m M Ba²⁺ could substitute for 1 m M Ca²⁺ in evoking the release of endogenous glutamate. In addition, Ba²⁺ was found to evoke glutamate release in the absence of K⁺-induced depolarization. Ba²⁺ (1–10 m M ) depolarized synaptosomes, as measured by voltage-sensitive dye fluorescence and [³H]-tetraphenylphosphonium cation distribution. Ba²⁺ partially inhibited the increase in synaptosomal K⁺ efflux produced by depolarization, as reflected by the redistribution of radiolabeled ⁸⁶Rb⁺. The release evoked by Ba²⁺ was inhibited by tetrodotoxin (TTX). Using the divalent cation indicator fura-2, cytosolic [Ca²⁺] increased during stimulation by approximately 200 n M , but cytosolic [Ba²⁺] increased by more than 1 μ M . Taken together, our results indicate that Ba²⁺ initially depolarizes synaptosomes most likely by blocking a K⁺ channel, which then activates TTX-sensitive Na⁺ channels, causing further depolarization, and finally enters synaptosomes through voltage-sensitive Ca²⁺channels to evoke neurotransmitter release directly. Though Ba²⁺-evoked glutamate release was comparable in level to that obtained with K⁺-induced depolarization in the presence of Ca²⁺, the apparent intrasynaptosomal level of Ba²⁺ required for a given amount of glutamate release was found to be several-fold higher than that required of Ca²⁺.     

Puccinellia tenuiflora maintains a low Na+ level under salinity by limiting unidirectional Na+ influx resulting in a high selectivity for K+ over Na+

Puccinellia tenuiflora is a useful monocotyledonous halophyte that might be used for improving salt tolerance of cereals. This current work has shown that P. tenuiflora has stronger selectivity for K⁺ over Na⁺ allowing it to maintain significantly lower tissue Na⁺ and higher K⁺ concentration than that of wheat under short- or long-term NaCl treatments. To assess the relative contribution of Na⁺ efflux and influx to net Na⁺ accumulation, unidirectional ²²Na⁺ fluxes in roots were carried out. It was firstly found that unidirectional ²²Na⁺ influx into root of P. tenuiflora was significantly lower (by 31–37%) than in wheat under 100 and 150 m m NaCl. P. tenuiflora had lower unidirectional Na⁺ efflux than wheat; the ratio of efflux to influx was similar between the two species. Leaf secretion of P. tenuiflora was also estimated, and found the loss of Na⁺ content from leaves to account for only 0.0006% of the whole plant Na⁺ content over 33 d of NaCl treatments. Therefore, it is proposed that neither unidirectional Na⁺ efflux of roots nor salt secretion by leaves, but restricting unidirectional Na⁺ influx into roots with a strong selectivity for K⁺ over Na⁺ seems likely to contribute to the salt tolerance of P. tenuiflora .