Cadmium-induced changes in antioxidant enzymes in suspension culture of soybean cells

Cadmium (Cd), similarly to other heavy metals, inhibits plant growth. We have recently showed that Cd(2+) either stimulates (1-4 microM) or inhibits (>/= 6 microM) growth of soybean (Glycine max L.) cells in suspension culture (Sobkowiak & Deckert, 2003, Plant Physiol Biochem. 41: 767-72). Here, soybean cell suspension cultures were treated with various concentrations of Cd(2+) (1-10 microM) and the following enzymes were analyzed by native electrophoresis: superoxide dismutase (SOD), catalase (CAT), peroxidase (POX) and ascorbate peroxidase (APOX). We found a significant correlation between the cadmium-induced changes of soybean cell culture growth and the isoenzyme pattern of the antioxidant enzymes. The results suggest that inhibition of growth and modification of antioxidant defense reactions appear in soybean cells when Cd(2+) concentration in culture medium increases only slightly, from 4 to 6 microM.     

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.     

Responses of maternal siblings ofPinus densiflora to simulated add rain

Because most research on acid rain does not consider the genetic composition of seeds and the inherent variations in seed mass and germination date, the potentially diverse patterns of plant response are neglected within a population. Therefore, we examined the effects of both family identity and simulated acid rain onPinus densiflora seedlings over a 34-week period, beginning at germination. We were able to evaluate the importance of family and pH treatment independent of seed mass and germination date by considering the latter two variables as covariates. Seeds were collected from six randomly chosen trees that were assumed to represent different genotypes. After germination in a controlled environment, the seedlings were moved outdoors. Each was then subjected twice a week to 40 mL of simulated acid rain, at a pH of either 3.0 or 5.6. We examined variations in germination date, seedling height, needle numbers and lengths, and root and shoot masses. The maternal family effect was significant for all characters over the entire test period. Strong effects of pH treatment were detected only at the later growth stages. Artificial rain with a pH of 3.0 enhanced growth rates, particularly aboveground, compared with seedlings treated with pH 5.6 rain. However, regardless of pH, certain groups of maternal siblings consistently performed better in almost all parameters. The exception was for mean germination date, which differed among families only at a pH of 3.0. Therefore, although we demonstrated that acid rain affected both the date of germination as well as seedling growth rates inP. densiflora, the responses differed among maternal families, and were also associated with seed mass and germination date.     

Effects of mercury and cadmium on the activities of antioxidative enzymes in the seedlings ofPhaseolus aureus

Phaseolus aureus Roxb. was exposed to HgCl₂ and Cd(NO₃)₂ either at the germination stage in concentration 0.5, 5 and 25 μM for 48 and 96 h, or at the seedling stage (5^th day of germination) in concentration 0.5, 5 and 20 μM for 6, 24 and 48 h. The germination and the growth of roots (germination stage treatment) were less in Hg than in Cd treatment. The seedlings (seedling stage treatment) were, however, more susceptible to Cd than Hg. Both root and leaf tissues of the plant treated at the germination stage showed enhanced lipid peroxidation and activities of the antioxidative enzymes (catalase, guaiacol peroxidase and ascorbate peroxidase), except the catalase in leaf in 25 μM Cd treatment. At seedling stage the content of malondialdehyde increased significantly only in the leaf tissue, during 6 h exposure. The activities of all the enzymes exhibited an increasing trend in both the tissue of the seedlings, particularly the leaf, at least after 24 and 48 h, except the catalase whose activity declined in response to Cd. Active involvement of the guaiacol and ascorbate peroxidases, rather than catalase, in scavenging cellular H₂O₂ was indicated. It was concluded that the two metals had little primary damaging effect on membranes.     

Combined Effect and Mechanism of Acidity and Lead Ion on Soybean Biomass

Heavy metal pollution and soil acidification are serious global environmental issues. The combined pollution from acidification and heavy metal has become a new environmental issue in regions where the two issues simultaneously occur. However, studies on combined pollution are still limited. In the current study, we investigated the combined effect and mechanism of acidity and heavy metal [lead ion (Pb²⁺)] on soybean biomass as well as on growth, nitrogen nutrition, and antioxidant system in soybean roots. Results showed that the combined treatment with acidity and Pb²⁺ decreased the soybean biomass. At pH 4.5, the soybean biomass in the combined treatment with acidity and 0.9 mmol L^?1 Pb²⁺ was lower than that in the combined treatment with acidity and Pb²⁺ at 0.3 or 1.5 mmol L^?1. This result was also observed at pH 3.5 and 3.0. The combined treatment with acidity and Pb²⁺ also resulted in the following consequences: root growth inhibition; decrease in nitrate, ammonium, and malondialdehyde contents; increase in nitrite reductase activity; and decrease in peroxidase activity. The extent at which the test indexes decreased/increased in the combined treatment was higher than that in the single acidity treatment. The correlation analysis results indicated that the decrease in the soybean biomass in the combined treatment with acidity and Pb²⁺ resulted from the decrease in the root growth, nitrate–nitrogen assimilation, and peroxidase activity.     

Effects of simulated acid rain on germination,seedling growth and oxidative metabolism of recalcitrant‐seeded Trichilia dregeana grown in its natural seed bank

Increased air pollution in a number of developing African countries, together with the reports of vegetation damage typically associated with acid precipitation in commercial forests in South Africa, has raised concerns over the potential impacts of acid rain on natural vegetation in these countries. Recalcitrant (i.e. desiccation sensitive) seeds of many indigenous African species, e.g. must germinate shortly after shedding and hence, may not be able to avoid exposure to acid rain in polluted areas. This study investigated the effects of simulated acid rain (rainwater with pH adjusted to pH 3.0 and 4.5 with 70:30, H₂SO₄:HNO₃) on germination, seedling growth and oxidative metabolism in a recalcitrant‐seeded African tree species Trichilia dregeana Sond., growing in its natural seed bank. The results suggest that acid rain did not compromise T. dregeana seed germination and seedling establishment significantly, relative to the control (non‐acidified rainwater). However, pH 3.0 treated seedlings exhibited signs of stress typically associated with acid rain: leaf tip necrosis, abnormal bilobed leaf tips, leaf necrotic spots and chlorosis, reduced leaf chlorophyll concentration, increased stomatal density and indications of oxidative stress. This may explain why total and root biomass of pH 3.0 treated seedlings were significantly lower than the control. Acid rain also induced changes in the species composition and relative abundance of the different life forms emerging from T. dregeana's natural seed bank and in this way could indirectly impact on T. dregeana seedling establishment success.     

Effects of simulated acid rain on the physiology of carmine spider mite, Tetranychus cinnabarinus (Boisduvals) (Acari: Tetranychidae)

Abstract:  The physiological effect of simulated acid rain sprayed on carmine spider mite Tetranychus cinnabarinus (Boisduvals) and host plant, were measured in a series of laboratory trials. We examined potential changes in three kinds of protective enzymes [peroxidase (POD), superoxide dismutase (SOD) and catalase (CAT)] and three hydrolases [acid phosphatase (ACP), alkaline phosphatase (ALP) and carboxylesterase (CarE)] in response to changes in pH values of simulated acid rain at different time of exposure. POD, SOD and CAT activities increased significantly with the increase in the acidity of the acid rain, reaching the highest levels at pH 4.0 or 3.0, and then declined. Changes in ACP activity were similar to those observed in the protective enzymes. The increasing extent of the activities of these four enzymes after 30 and 45 days treatment became smaller than that after 15 days treatment . ALP activities decreased as pH value declined. There were no significant changes in CarE activities after 15 and 45 days, but that in pH 4.0 and 3.0 decreased after 30 days. The enhanced anti-oxidation enzyme levels (POD, SOD and CAT) and ACP activities in pH 4.0 and 3.0 reduced the effects of these toxic products on mites, resulting in the strengthening of the defensive power, and increase in survival and reproductive power of the mites, thus leading to an increase in the density of mites on host plant. From these results, we inferred that POD, SOD, CAT and ACP might be relevant to population changes of mites under acid rain pressure.     

重金属对土壤中萝卜种子发芽与根伸长抑制的生态毒性

高等植物是生态系统中的基本组成部分。一个平衡、稳定的生态系统生产健康、优良的高等植物。反之 ,一个不稳定或受到外来污染的生态系统 ,对高等植物的生长可带来不利和可见的负面影响。因此 ,利用高等植物的生长状况监测土壤污染程度 ,是从生态学角度衡量土壤健康状况 ,评价土壤质量的重要方法之一[4 ,6] 1) 。土壤生态毒理学评价方法是对化学分析方法的重要补充。目前已建立的高等植物毒理试验有三种方法 ,即 1根伸长试验 ;2种子发芽试验 ;3早期植物幼苗生长试验[3 ,5,6,10 ] 。最初 ,这类试验主要用于纯化学品的毒性检验 ,但随着对土壤…     

镉胁迫对矮牵牛种子萌发、幼苗生长及抗氧化酶活性的影响

采用营养液培养的方法研究了不同浓度镉Cd（0、1、2和4 mmol·L^-1）处理对矮牵牛（Petunia hybrida）种子萌发、幼苗生长及生理特性的影响。结果发现：与对照相比,较低浓度（1 mmol·L^-1）Cd处理下矮牵牛种子萌发和幼苗生长均显著被抑制,当Cd浓度增至4 mmol·L^-1时完全抑制了矮牵牛种子的萌发。Cd胁迫在低浓度时激活矮牵牛幼苗叶片过氧化物酶（POD）、过氧化氢酶（CAT）活性,而高浓度产生抑制效应;抗坏血酸过氧化物酶（APX）和超氧化物歧化酶（SOD）活性与对照相比均显著升高,丙二醛（MDA）含量无显著变化。不同浓度的乙二胺四乙酸(EDTA)的加入使4 mmol·L^-1 Cd胁迫下矮牵牛幼苗叶片POD和APX活性增加,但不影响SOD和CAT活性;此外,6 mmol·L^-1 EDTA使Cd胁迫下矮牵牛幼苗叶片中MDA含量增加。结果表明,Cd胁迫抑制矮牵牛种子萌发和幼苗生长,却使幼苗叶片POD、CAT和APX活性增加,EDTA的加入能够进一步提高Cd胁迫幼苗POD和APX活性。因此,矮牵牛幼苗在Cd胁迫下具有较强的抗氧化损伤的能力。     

Cadmium inhibits delta-aminolevulinate dehydratase from rat lung in vitro: interaction with chelating and antioxidant agents

The effect of cadmium (Cd(2+)) on delta-aminolevulinate dehydratase (delta-ALA-D) activity from rat lung in vitro was investigated. delta-ALA-D activity, a parameter for metal intoxication, has been reported as a target of Cd(2+) in different tissues. The protective effect of monotherapies with dithiol chelating (meso-2,3-dimercaptosuccinic acid (DMSA) and 2,3-dimercaptopropane-1-sulfonic acid (DMPS)) or antioxidant agents (ascorbic acid, diphenyl diselenide (PhSe)(2), and N-acetylcysteine (NAC)) was evaluated. The effect of a combined therapy (dithiol chelatingxantioxidant agent) was also studied. Zinc chloride (ZnCl(2)) and dithiothreitol (DTT) were used to investigate the mechanisms involved in cadmium, chelating and antioxidant effects on delta-ALA-D activity. Cadmium inhibited rat lung delta-ALA-D activity at low concentrations. DTT (3mM), but not ZnCl(2) (100microM), protected the inhibition of enzyme activity caused by Cd(2+). Chelating agents were not effective in restoring the enzyme activity. DMPS and DMSA presented inhibitory effect on enzyme activity. DTT restored the inhibition caused by both chelating agents, but ZnCl(2) restored only the inhibitory effect induced by DMSA. These compounds caused a marked potentiation of delta-ALA-D inhibition induced by Cd(2+). ZnCl(2) did not restore inhibition of enzyme activity caused by Cd(2+) plus chelating agents. Conversely, DTT restored the inhibition induced by Cd(2+)/DMSA, but not by Cd(2+)/DMPS. Antioxidants were not effective in ameliorating delta-ALA-D inhibition induced by Cd(2+), whereas ascorbic acid potentiated the enzyme inhibition induced by this metal. A combined effect of Cd(2+)xDMPSx(PhSe)(2) and Cd(2+)xDMPSxNAC was observed. There was no combined effect of Cd(2+)xchelatorxantioxidants when DMSA was used. This study demonstrated that Cd(2+)inhibited delta-ALA-D activity and chelating and antioxidant agents, alone or combined, did not restore the enzyme activity. In contrast, these compounds potentiated the inhibition induced by Cd(2+) in rat lung.     

酸雨和UV-B对玉米幼苗光合速率和抗氧化酶活性的影响

以玉米( Zea mays L.)的两个品种渝糯7号和苏玉糯为研究材料, 研究酸雨与UV-B辐射单一因子和复合处理对植物光合作用及抗氧化酶活性的影响机理, 模拟酸雨[pH6.5(A0), 4.5(A1)和3.5(A2)]和增强UV-B辐射[0(B0), 2.88(B1)和5 76(B2)kJ·m-2·d-1]及其复合作用下, 玉米叶片光合速率, 膜脂过氧化程度, 抗氧化酶活性的变化.结果表明, 与对照相比, 酸雨或增强UV-B辐射单因素处理下两个品种的玉米幼苗所测生理指标有相似的变化, 基本表现为: 净光合速率和过氧化氢酶(CAT)活性降低, 质膜透性与丙二醛(MDA)含量增加, 过氧化物酶(POD)活性亦增加, 超氧化物歧化酶(SOD)活性随处理时间延长先升高后降低.复合处理下, 两个品种的玉米幼苗所测生理指标与对照相比也与单因素处理表现相似, 且随处理组合的不同, 复合胁迫下各指标的效果可能表现为协同效应, 但更多地表现为拮抗作用.以上结果说明, 模拟UV-B辐射升高和酸雨pH值降低促进了玉米叶片活性氧的代谢速率, 导致了抗氧化酶系统的紊乱, 抑制了光合作用.在复合处理下, 两个因子对不同指标在不同处理时间和复合水平下, 表现出一定的协同或拮抗效应可能与作物对环境变化产生一定的抗逆性和适应性有关.在不同时期两玉米品种对紫外辐射与酸雨敏感性不同.与苏玉糯相比, 渝糯7号对外界环境变化的抗性更大.     

The effect of some factors of polluted environment on catalase responses and resistance of microbial isolates against toxic oxidative stress

The properties of bacterial isolates from polluted environments which are characterized by increased levels of oxidative stress do not reflect only the level of contaminants, but also arise as a consequence of many permanently changed conditions. The survival rate of Comamonas terrigena N3H isolates from an environment with elevated levels of H(2)O(2) is correlated with stimulation of catalase. The response of bacterial catalase to the effect of phenol in exogenous conditions was affected by the presence of an additional contaminant, Cd(2+). An isolate of Aspergillus niger selected from river sediment containing 363?mg/kg As, 93?mg/kg Sb at pH 5.2-4.8 grew on Czapek-Dox agar ~1.6 times faster than an isolate of the same species from coal dust sediment with approximately the same level of pollution (400?mg/kg As) but somewhat lower pH (3.3-2.8). It also exhibited differences in the microscopic characteristics of its mycelial structures. Both isolates exhibited a higher tolerance to the exogenic toxic effects of metals (As(5+), Cd(2+), and Cu(2+) at 5, 25, or 50?mg/L) than a control culture, but the differences in tolerance between them were only slight. These laboratory results suggest that there are complicated relationships which may exist in the "in situ" environment.     

Hydrogen Sulfide Alleviates Aluminum Toxicity in Germinating Wheat Seedlings

Protective role of hydrogen sulfide (H2S) on seed germination and seedling growth was studied in wheat (Triticum) seeds subjected to aluminum (Al3+) stress. We show that germination and seedling growth of wheat is inhibited by high concentrations of AICI3. At 30 mmol/L AICI3 germination is reduced by about 50% and seedling growth is more dramatically inhibited by this treatment. Pre-incubation of wheat seeds in the H2S donor NaHS alleviates AICI3-induced stress in a dose-dependant manner at an optimal concentration of 0.3 mmol/L. We verified that the role of NaHS in alleviating Al3+ stress could be attributed to H2S/HS- by showing that the level of endogenous H2S increased following NaHS treatment. Furthermore, other sodium salts containing sulfur were ineffective in alleviating Al3+ stress. NaHS pretreatment significantly increased the activities of amylases and esterases and sustained much lower levels of MDA and H2O2 in germinating seeds under Al3+ stress. Moreover, NaHS pretreatment increased the activities of guaiacol peroxidase, ascorbate peroxidase, superoxide dismutase and catalase and decreased that of lipoxygenase. NaHS pretreatment also decreased the uptake of Al3+ in AICI3-treated seed. Taken together these results suggest that H2S could increase antioxidant capability in wheat seeds leading to the alleviation of Al3+ stress.     

Cadmium stress affects seed germination and seedling growth in <Emphasis Type="Italic">Sorghum bicolor</Emphasis> (L.) Moench by changing the activities of hydrolyzing enzymes

Seed germination, one of the most important phases in the life cycle of a plant, is highly responsive to existing environment. Hydrolyzing enzymes play a major role in the mobilization of food reserves by hydrolyzing carbohydrates, proteins and fats. This paper reports on the effect of Cd toxicity on seed germination and the activities of hydrolyzing enzymes, like acid phosphatases (ACPs), proteases and α-amylases in Sorghum bicolor (L.) Moench. The metal uptake by embryonic axes and seeds was quantified. We found that sorghum could tolerate up to 0.5 mM Cd. At concentrations above 3.0 mM, seed germination was adversely affected with a complete cessation of seedling growth. All investigated hydrolyzing enzymes exhibited a significant decrease in activity with increasing Cd concentrations. The isozyme profiles indicated the loss of one or two isozymes of ACP, induction of a new isozyme for total protease (at 3.0 mM Cd) and a decline in the intensity of α-amylase isozymes. SEM studies revealed that Cd affected a change in root hair density. SEM investigations also confirmed the assay results of the inhibition of starch mobilization from endosperm. This suggested an inhibition of the hydrolysis of reserve carbohydrates and translocation of hydrolyzed sugars, ultimately resulting in decreased germination and disruption of seedling growth. Because sorghum is an important dryland crop, its response to the presence of Cd in agro-ecosystems and Cd-induced phytotoxicity during seed germination and seedling growth needs critical investigation.     

汞对萝卜种子发芽及幼苗某些生理特性的影响

研究了重金属汞离子(Hg^2 )对萝卜种子发芽及幼苗某些生理特性的影响。结果表明,随着汞离子浓度的增加和处理时间的延长,萝卜种子的发芽率和发芽指数以及幼苗的生长势和生长量均明显下降。幼苗子叶叶绿素a、b的含量下降。幼苗根系、茎叶过氧化物酶的活性则明显升高。     

外源水杨酸对NaCl胁迫下大豆种子萌发和幼苗生长生理的影响

以大豆种子、幼苗为试验材料,采用砂培的方法,研究了0.2mmol·L-1外源水杨酸(SA)对100mmol·L-1 NaCl胁迫下大豆种子萌发、幼苗形态及生物量、膜脂过氧化和抗氧化酶活性的影响。结果显示:NaCl胁迫下,大豆种子萌发和幼苗生长受到显著抑制,且随着胁迫时间的延长(0~3d),大豆幼苗相对电解质渗漏率、硫代巴比妥酸活性产物(TBARS)含量显著升高,超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)活性均明显降低。外源SA促进NaCl胁迫下大豆种子萌发和根茎生长,增加幼苗生物量积累,降低幼苗叶片相对电解质渗漏率和TBARS含量,增强其叶片SOD、CAT、APX活性。研究表明,NaCl胁迫能显著抑制大豆种子萌发和幼苗生长,而一定浓度的外源SA能有效提高NaCl胁迫下大豆种子活力及幼苗抗氧化酶活性,减轻膜脂过氧化程度,缓解NaCl胁迫所造成的伤害,提高大豆幼苗抗盐胁迫的能力。     

重金属对油菜种子萌发和胚根生长的影响

分析了Hg2 、Cd2 、Ni2 、Co2 、Zn2 5种重金属离子对油菜种子萌发和胚根伸长的影响,以及金属离子K 、Mg2 和Ca2 与重金属的交互作用。结果表明:(1)重金属对油菜种子萌发的抑制作用依次为Hg2 >Cd2 和Co2 >Ni2 >Zn2 ,而对胚根生长的毒害作用依次为Hg2 >Cd2 >Co2 >Ni2 >Zn2 。(2)萌发率为40%以上时,K 和Ca2 可以提高Ni2 、Zn2 和Co2 胁迫下油菜种子的萌发率,却进一步降低了Hg2 、Cd2 胁迫下种子的萌发;Mg2 可以提高Ni2 、Zn2 、Cd2 和Co2 胁迫下种子的萌发率,但对Hg2 毒害却没有缓解。(3)胚根伸长率达到60%以上时,K 和Mg2 增强了Ni2 、Hg2 、Cd2 和Co2 对胚根生长的抑制,而Ca2 则缓解了Zn2 、Ni2 和Co2 对胚根生长的抑制作用。研究结果对于重金属复合污染土壤中植物种子的萌发和定植具有理论和实践意义。     

重金属对土壤中小麦种子发芽与根伸长抑制的生态毒性

测定了4种土壤（红壤、草甸棕壤、暗棕壤和栗钙土）条件下,Cu,Zn,Pb和Cd单一污染对小麦种子发芽与根伸长抑制主及其复合污染效应（暗棕壤条件下）。结果表明,同一浓度下,重金属对小麦根伸长抑制率均明显大于对种子发芽抑制率,植物根对金属污染的生态毒性比种子发芽敏感,土壤有机质和土壤N含量与Cu,Zn,Pb和Cd污染对小麦根伸长抑制率显著负相关（R^2OR=0.91,R^2K-N=0.92),土壤PH和阳离子交换量与Cu,Zn,Pb和Cd污染对小麦根伸长抑制率的相关性不显著（R^2PH=0.62,R^2CEC=0.60),在单一污染对小麦根伸长为刺激作用浓度（较低浓度）或为抑制作用浓度下（较高浓度）,复合污染均表现为协同作用。^     

Study on the mechanism of cerium nitrate effects on germination of aged rice seed

Attempts were made to promote germination of naturally and artificially aged rice seeds by treating them with cerium nitrate. The germination rate, germination index, and vigor index of aged rice seed were significantly increased by cerium. It was because the treatments of aged rice seed with cerium nitrate enhanced respiratory rate and activities of superoxide dismutase, catalase, and peroxidase, and decreased superoxide O₂ ⁻ and malondialdehyde contents that plasma membrane permeability was reduced. It was suggested that cerium be used for the seed treatment before sowing.