Heat shock proteins in the mechanisms of stress adaptation in Baikal amphipods and palearctic <Emphasis Type="Italic">Gammarus lacustris</Emphasis> Sars. I. HSP70 family

The participation of HSP70 family proteins in the mechanisms of stress adaptation under thermal and toxic stress in Baikal endemic and palearctic organisms was investigated. The four Lake Baikal endemic species Gmelinoides fasciatus (Stebb.), Eulimnogammarus cyaneus (Dyb.), E. vittatus (Dyb.), Ommatogammarus fl avus (Dyb.) were compared with the palearctic fauna representative Gammarus lacustris Sars. The type of synthesis of HSP70 family heat shock proteins under thermal (exposure at a temperature of 20, 25, and 30°C) and toxic (exposure in CdCl₂ solutions with the concentrations 50, 10, 5, 0.5, and 0.05 mg/l) stress was determined. All species under investigation were shown to have a common trend towards HSP70 content increase; therewith some species-specific characteristics of the type of synthesis of the protein under investigation were observed. The conclusion has been made on HSP70 participation in the mechanisms of thermal and toxic resistance in the studied amphipod species.     

Effects of short-term heat stress on oxidative damage and responses of antioxidant system in Lilium longiflorum

This paper aims to determine the changes in reactive oxygen species (ROS) and the responses of the lily (Lilium longiflorum L.) antioxidant system to short-term high temperatures. Plants were exposed to three levels of heat stress (37°C, 42°C, 47°C) for 10 h when hydrogen peroxide (H₂O₂) and superoxide (O₂⁻) production rate along with membrane injury indexes, and changes in antioxidants were measured. Compared with the control (20°C), electrolyte leakage and MDA concentration varied slightly after 10 h at 37°C and 42°C, while increased significantly at 47°C. During 10 h at 37°C and 42°C, antioxidant enzyme activities, such as SOD, POD, CAT, APX and GR, were stimulated and antioxidants (AsA and GSH concentrations) maintained high levels, which resulted in low levels of O₂⁻ and H₂O₂ concentration. However, after 10 h at 47°C, SOD, APX, GR activities and GSH concentration were similar to the controls, while POD, CAT activities and AsA concentration decreased significantly as compared with the control, concomitant with significant increase in O₂⁻ and H₂O₂ concentrations. In addition, such heat-induced effects on antioxidant enzymes were also confirmed by SOD and POD isoform, as Cu/ZnSOD maintained high stability under heat stress and the intensity of POD isoforms reduced with the duration of heat stress, especially at 47°C. It is concluded that in lily plants, the oxidative damage induced by heat stress was related to the changes in antioxidant enzyme activities and antioxidants.     

Localization of antioxidant enzymes in the cellular compartments of sorghum leaves

The localization of antioxidant enzymes between the mesophyll and bundle sheath cells were determined in sorghum (Sorghum vulgare L.) leaves. The activity of antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT), guaiacol peroxidase (POD), ascorbate peroxidase (APX) and glutathione reductase (GR) were assayed in whole leaf, mesophyll and bundle sheath fractions of sorghum leaves subjected to water-limited conditions. Drought was imposed by withholding water and the plants were maintained at different water potentials ranging from 0.5–2.0 MPa. The purity of the isolates was tested using the marker enzymes like RuBPcase and PEPcase. GR was mostly localized in mesophyll fraction, while SOD, APX and peroxidase were located in bundle sheath cells. Catalase was found to be equally distributed between the two cell types. Under water stress conditions, most of the SOD activity was found in the bundle sheath tissues. Little or no activity of the enzymes CAT, APX or POD was found in the mesophyll extracts when exposed to water stress. GR activity increased when exposed to low water regimes. From this study, it is clear that antioxidants are differentially distributed between the mesophyll and bundle sheath cells in sorghum leaves. Under water stress conditions, the mesophyll cells showed less damage from oxidative stress when compared to the bundle sheath cells. This is critical for determining the sensitivity of sorghum to extreme climatic conditions.     

光质对白芨组培苗生理特性及酶基因表达的影响

为探讨光质对白芨(Bletilla striata)组培苗生长发育的影响,对不同光质下白芨组培苗的生长特征、抗氧化酶活性以及酶基因表达进行了研究。结果表明,蓝光和红光对白芨生长有显著促进作用,绿光的作用不明显。除了CAT外,不同光质处理白芨的APX、POD、SOD活性均呈上升趋势,且黄光处理的白芨SOD和APX活性最高,红光处理的POD活性最高,绿光处理的抗氧化酶活性比其他光质的低,蓝光处理35~45 d对抗氧化酶基因表达具有促进作用。因此,红光和黄光促进白芨生根组培苗的长高和生根;不同光质处理总体上提高了白芨氧化酶活性;白芨抗氧化酶基因的表达在蓝光处理下最大。     

Influence of sodium orthovanadate on the production of astaxanthin from green algae Haematococcus lacustris

Astaxanthin production is commonly induced under stress conditions such as nutrient deficiency (N or P), high light stress, and variations of temperature, high NaCl concentrations, and other factors. The objective of the present study is the analysis of the effect of oxidative stress by sodium orthovanadate (SOV), a nonspecific inhibitor of protein tyrosine phosphatases, on the cells growth and astaxanthin production of H. lacustris. In the presence of SOV (lower than 5.0 mM), maximum growth of H. lacustris obtained was 2.4 × 10⁵ cells/mL in MBBM medium at 24°C under continuous illumination (40 μE/m²/s) of white fluorescent light, with continuous aeration of CO₂ (0.2 vvm). Total carotenoids accumulated per cell biomass unit treated with 2.5 mM SOV has approximately shown 2.5 folds higher than the control after short period of SOV induction time as 2 days, despite that cells were grown under normal light. Meanwhile, maximal astaxanthin production from H. lacustris was 10.7 mg/g biomass in MBBM with 5 days of continuous illumination at 40 μE/m²/s, which has been established as optimal light intensity for the control culture of H. lacustris. Treating algae H. lacustris with sodium orthovanadate showed promoting the accumulation of astaxanthin by advancing either the inhibition of dephosphorylation or synthesis of ATP. Its potential role of PTPases in microalgae H. lacustris is discussed. The first two authors are equally contributed to this work.     

Antioxidant responses of Propylaea japonica (Coleoptera: Coccinellidae) exposed to high temperature stress

Temperature is one of the most important environmental factors, and is responsible for a variety of physiological stress responses in organisms. Induced thermal stress is associated with elevated reactive oxygen species (ROS) generation leading to oxidative damage. The ladybeetle, Propylaea japonica (Thunberg) (Coleoptera: Coccinellidae), is considered a successful natural enemy because of its tolerance to high temperatures in arid and semi-arid areas in China. In this study, we investigated the effect of high temperatures (35, 37, 39, 41 and 43 °C) on the survival and activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT), peroxidases (POD), glutathione-S-transferases (GST), and total antioxidant capacity (TAC) as well as malondialdehyde (MDA) concentrations in P. japonica adults. The results indicated that P. japonica adults could not survive at 43 °C. CAT, GST and TAC were significantly increased when compared to the control (25 °C), and this played an important role in the process of antioxidant response to thermal stress. SOD and POD activity, as well as MDA, did not differ significantly at 35 and 37 °C compared to the control; however, there were increased levels of SOD, POD and MDA when the temperature was above 37 °C. These results suggest that thermal stress leads to oxidative stress and antioxidant enzymes play important roles in reducing oxidative damage in P. japonica adults. This study represents the first comprehensive report on the antioxidant defense system in predaceous coccinellids (the third trophic level). The findings provide useful information for predicting population dynamics and understanding the potential for P. japonica as a natural enemy to control pest insects under varied environmental conditions.     

Heat shock proteins in the mechanisms of stress adaptation in Baikal amphipods and Palaearctic <Emphasis Type="Italic">Gammarus lacustris</Emphasis> Sars II. Small HSP family

The role of low-molecular-weight (small) heat shock proteins (sHSPs) in the mechanisms of thermal and toxic resistance in fresh-water organisms was investigated. The four endemic species of Lake Baikal: Gmelinoides fasciatus (Stebb.), Eulimnogammarus cyaneus (Dyb.), E. vittatus (Dyb.), and Ommatogammarus flavus (Dyb.), and Gammarus lacustris Sars, a representative of the Palaearctic fauna, were studied. The effect of the temperature factor was evaluated by exposure of amphipods to temperatures of 20, 25, and 30°C, and the effect of the toxic factor, by exposure to cadmium chloride solutions with concentrations of 50, 10, 5, 0.5, and 0.05 mg/l. All the species showed a common trend of increasing content of sHSPs; species-specific features were observed in the synthesis of the proteins. It was concluded that sHSPs participate in the mechanisms of thermal and toxic resistance in the investigated amphipod species.     

Fungal pathogen-induced changes in the antioxidant systems of leaf peroxisomes from infected tomato plants

Peroxisomes, being one of the main organelles where reactive oxygen species (ROS) are both generated and detoxified, have been suggested to be instrumental in redox-mediated plant cell defence against oxidative stress. We studied the involvement of tomato (Lycopersicon esculentum Mill.) leaf peroxisomes in defence response to oxidative stress generated upon Botrytis cinerea Pers. infection. The peroxisomal antioxidant potential expressed as superoxide dismutase (SOD, EC 1.15.1.1), catalase (CAT, EC 1.11.1.6) and glutathione peroxidase (GSH-Px, EC 1.11.1.19) as well as the ascorbate-glutathione (AA-GSH) cycle activities was monitored. The initial infection-induced increase in SOD, CAT and GSH-Px indicating antioxidant defence activation was followed by a progressive inhibition concomitant with disease symptom development. Likewise, the activities of AA-GSH cycle enzymes: ascorbate peroxidase (APX, EC 1.11.1.11), monodehydroascorbate reductase (MDHAR, EC 1.6.5.4), dehydroascorbate reductase (DHAR, EC 1.8.5.1) and glutathione reductase (GR, EC 1.6.4.2) as well as ascorbate and glutathione concentrations and redox ratios were significantly decreased. However, the rate and timing of these events differed. Our results indicate that B. cinerea triggers significant changes in the peroxisomal antioxidant system leading to a collapse of the protective mechanism at advanced stage of infection. These changes appear to be partly the effect of pathogen-promoted leaf senescence.     

根边缘细胞缓解土荆芥淋溶途径化感作用对苦荞麦代谢扰动的机制

土荆芥(Chenopodium ambrosioides)可通过淋溶途径的化感作用抑制周围植物的种子萌发和幼苗生长,而根边缘细胞(Root border cells, RBCs)对此具有缓解效应。为探讨这一效应的分子机制,以苦荞麦(Fagopyrum tataricum)为研究对象,测定了在土荆芥水浸提液处理前及处理后,保留RBC组和移除RBC组根尖活性氧(ROS)、超氧阴离子(O^-₂)和丙二醛(MDA)含量以及抗氧化酶[超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)]活性的差异。再利用转录组测序分析上述处理对其代谢通路的影响,并进行qRT-PCR验证。结果表明：土荆芥水浸提液处理后,苦荞麦根尖ROS、O^-₂和MDA的含量以及抗氧化酶(POD和CAT)活性明显提高,且RBC移除组升高更多。通路分析表明,土荆芥水浸提液处理后,移除和保留RBCs组的苯丙素合成、α-亚麻酸代谢、类黄酮合成和谷胱甘肽代谢通路都显著发生改变。其中,苯丙素合成通路以抑制为主,且移除RBCs后受到抑制的程度...     

Regulation of enzymes involved in C4 photosynthesis and the antioxidant metabolism by UV-B radiation in Egeria densa,a submersed aquatic species

Egeria densa, a submersed aquatic species, was exposed to different treatments under UV-B radiation, and the response of phosphoenolpyruvate carboxylase (PEPC) and NADP-malic enzyme (NADP-ME) was determined. Exposure to UV-B radiation for 4 h per day over 7–16 days caused an increase in both enzymes, together with an increase in the activity of some isoforms of several enzymes involved in the antioxidant metabolism, such as superoxide dismutase (SOD), ascorbate peroxidase (APX), catalase (CAT) and peroxidase (POD). The content of chlorophylls and carotenoids was considerably decreased, suggesting that degradation or repression of the synthesis of these molecules may be occurring after UV-B exposure. Reactive oxygen species (ROS) were also required for UV-B induction of PEPC and NADP-ME, as the addition of ascorbic acid before UV-B treatment prevented the induction of these enzymes, while salicylic acid was not effective in inducing NADP-ME but increased the expression of the lower molecular mass isoform of PEPC. On the other hand, damage to the photosynthetic machinery may be occurring after exposure to UV-B radiation for 8 per day over 1–2 days, as indicated by a decrease in the levels of Rubisco, PEPC and NADP-ME. Some of the enzymes involved in the antioxidant metabolism, such as CAT and APX, were also sensitive to continuous exposure, evidenced by a decrease in their activity. In this way, in E. densa, several enzymes involved in different metabolic pathways showed a distinct response, depending on the UV-B treatment. This revised version was published online in June 2006 with corrections to the Cover Date.     

Proline and betaine provide protection to antioxidant and methylglyoxal detoxification systems during cold stress in <Emphasis Type="Italic">Camellia sinensis</Emphasis> (L.) O. Kuntze

The aim of this study was to monitor the influence of proline and betaine exposure on antioxidant and methylglyoxal (MG) detoxification system during cold stress in Camellia sinensis (L.) O. Kuntze. Cold stress enhanced MG and lipid peroxidation levels in tea bud (youngest topmost leaf). This increase was resisted upon the exposure of tea bud to proline and betaine. Exposure of tea bud with proline and betaine also help in maintaining thiol/disulfide ratio during cold stress. Proline exposure enhanced glutathione-S-transferase and glutathione reductase (GR) activity, while betaine exposure increased only GR activity during cold stress. Furthermore, effect of proline/betaine was studied on glyoxalase pathway enzymes that are involved in MG detoxification and comprise of two enzymes glyoxalase I and glyoxalase II. Both proline and betaine showed protective effect on glyoxalase I and activating effect on glyoxalase II during cold stress in tea bud. This investigation, therefore, suggest that proline and betaine might provide protection to cold stress in tea by regulating MG and lipid peroxidation formation as well as by activating or protecting some of antioxidant and glyoxalase pathway enzymes.     

Physiological responses to gradual drought stress in the diploid hybrid Pinus densata and its two parental species

Pinus densata is a homoploid hybrid species, originating from P. tabuliformis × P. yunnanensis. The physiological fitness of this natural hybrid compared to its two parental species remains unknown. In this study, we investigated physiological responses of the three species by exposing artificially breed seedlings of each to drought stress lasting 28 days. Our results suggest that, in all three species, drought affected the contents of the plants’ chlorophyll, stomatal conductance, TBARS, hydrogen peroxide, and free proline and increased the activities of antioxidant enzymes, including superoxide dismutase (SOD; EC 1.15.1.1), catalase (CAT; EC 1.11.1.6), and peroxidase (POD; EC 1.11.1.7). The drought stress also induced significant changes in the activity of ascorbate peroxidase (APX; EC 1.11.1.11), monodehydroascorbate reductase (MDHAR; EC 1.6.5.4), dehydroascorbate reductase (DHAR; EC 1.8.5.1), glutathione reductase (GR; EC 1.6.4.2), and levels of ascorbate and glutathione in the ascorbate–glutathione cycle. The hybrid species P. densata appeared to achieve greater drought tolerance and exhibit hybrid superiority in antioxidant processes and other related physiological traits compared to the two parental species, although a few of the hybrid’s measured variables were similar to those of P. tabuliformis. However, P. yunnanensis was more sensitive to drought and appeared to have the lowest resistance to such stress. These physiological differences are largely consistent with the species’ habitat preferences, which may reflect their early genetic divergences and niche differentiation. These findings provide important information for management and forest restoration efforts of these species in the future.     

盐旱交叉胁迫对灰胡杨(Populus pruinosa)幼苗生长和生理生化特性的影响

以一年生灰胡杨幼苗为试验材料,利用田间控盐控水的方法,进行干旱和盐胁迫试验,通过测定生长和生理生化指标探讨幼苗在盐旱交叉胁迫下的生长发育及适应规律,旨在阐明干旱及盐交叉胁迫下植物抗旱抗盐机理。研究结果表明:在盐、旱及交叉胁迫下,灰胡杨幼苗抗氧化酶活性、MDA和脯氨酸含量与对照存在显著差异(P0.05)。(1)在8、11 g/L和15 g/L盐处理下,灰胡杨幼苗相对高生长、相对枝长和冠幅增量均受到抑制,且差异显著(P0.05),而干旱胁迫和盐旱交互胁迫下差异不显著。(2)在盐胁迫、盐旱交叉胁迫下,随着胁迫程度的加重,抗氧化酶SOD、POD、CAT活性表现出先增加后降低的趋势,三者协调一致;仅干旱胁迫时,抗氧化酶SOD、POD、CAT活性显著增加;(3)在盐、旱及其盐旱交叉胁迫下,脯氨酸含量呈上升趋势,MDA含量则表现出先降低后升高趋势,这与抗氧化酶活性先升高后降低的趋势相对应。因此,抗氧化酶活性对缓解脂膜过氧化的伤害具有一定限度,MDA含量与抗氧化酶活性呈负相关,灰叶胡杨幼苗在盐旱交叉胁迫下表现出一定的耐性。     

Antioxidant responses of citrus red mite, Panonychus citri (McGregor) (Acari: Tetranychidae), exposed to thermal stress

Relatively low or high temperatures are responsible for a variety of physiological stress responses in insects and mites. Induced thermal stress was recently associated with increased reactive oxygen species (ROS) generation, which caused oxidative damage. In this study, we examined the time-related effect of the relatively low (0, 5, 10, and 15 °C) or high (32, 35, 38, and 41 °C) temperatures on the activities of antioxidant enzymes including superoxide dismutase (SOD), catalase (CAT), peroxidases (POX), and glutathione-S-transferase (GST), and the total antioxidant capacity (TEAC) of the citrus red mite, Panonychus citri (McGregor). The malondialdehyde (MDA) concentration, as a marker of lipid peroxidation in organisms, was also measured in the citrus red mite under thermal stress conditions. Results showed that SOD and GST activities were significantly increased and play an important role in the process of antioxidant response to thermal stress. Lipid peroxidation levels increased significantly (P < 0.001) and changed in a time-dependent manner. CAT and POX activity, as well as TEAC, did not vary significantly and play a minor role to remove the ROS generation. These results suggest that thermal stress leads to oxidative stress and antioxidant enzymes play an important role in reducing oxidative damage in the citrus red mite.     

Antioxidant activity in Spalax ehrenbergi: a possible adaptation to underground stress

The blind subterranean mole rat Spalax ehrenbergi superspecies has evolved adaptive strategies to cope with underground stress. Hypoxia is known to stimulate reactive oxygen species generation; however, mechanisms by which Spalax counteracts oxidative damage have not been investigated before. We studied in Spalax the oxidative status of the Harderian gland (HG), an organ which is particularly vulnerable to oxidative stress in many rodents. With regard to the sexual dimorphism found in this gland, differences between males and females were determined and compared to the surface-dwelling Syrian hamster. Our results show, for the first time, that Spalax exhibits remarkably low biomolecular damage, which implies the existence of physiological strategies to avoid oxidative damage under fluctuating O₂ and CO₂ levels existing in the mole rat’s subterranean niche. Correspondingly, main antioxidant enzymes, such as superoxide dismutase (SOD), catalase, and glutathione reductase (GR), exhibited high activities in both genders; in particular, remarkably high levels were measured in SOD. SOD and GR activities showed statistically significant differences between sexes. Melatonin, an important circadian agent is also a very important antioxidant molecule and is synthesized in the Harderian glands (HGs) of Spalax. Therefore, the possible interaction between antioxidant enzymes and melatonin is suggested.Joint senior authorship: Aaron Avivi and Ana Coto-Montes     

Osmoregulation and osmoprotection in the leaf cells of two olive cultivars subjected to severe water deficit

In this study, we compared the efficacy of defense mechanisms against severe water deficit in the leaves of two olive (Olea europaea L.) cultivars, ‘Chemlali’ and ‘Meski’, reputed drought resistant and drought sensitive, respectively. Two-year old plants growing in sand filled 10-dm³ pots were not watered for 2 months. Changes in chlorophyll fluorescence parameters and malondialdehyde content as leaf relative water content (RWC) decreased showed that ‘Chemlali’ was able to maintain functional and structural cell integrity longer than ‘Meski’. Mannitol started to accumulate later in the leaves of ‘Chemlali’ but reached higher levels than in the leaves of ‘Meski’. The latter accumulated several soluble sugars at lower dehydration. ‘Chemlali’ leaves also accumulated larger quantities of phenolic compounds which can improve its antioxidant response. Furthermore, the activity of three antioxidant enzymes catalase (CAT), peroxidase (POD) and ascorbate peroxidase (APX) increased as leaf RWC decreased. However, differences were observed between the two cultivars for CAT and POD but not for APX. The activity of the first two enzymes increased earlier in ‘Meski’, but reached higher levels in ‘Chemlali’. At low leaf hydration levels, ‘Chemlali’ leaves accumulated mannitol and phenolic compounds and had increased CAT and POD activities. These observations suggest that ‘Chemlali’ was more capable of maintaining its leaf cell integrity under severe water stress because of more efficient osmoprotection and antioxidation mechanisms.     

Hymenolepis diminuta: experimental studies on the antioxidant system with short and long term infection periods in the rats

Many helminths cause long-lasting infections, living for several years in mammalian hosts reflecting a well balanced coexistence between host and parasite. There are many possible explanations as to how they can survive for lengthy periods. One possibility is their antioxidant systems, which can serve as defence mechanisms against host-generated oxygen radicals. Therefore, the aim of this experimental study was to examine the antioxidant system in Hymenolepisdiminuta during short (1.5 months young tapeworms) and long (1.5 years old tapeworms) term infection in the rat small intestine.The strobilae of H. diminuta tapeworms (14 young and three old) were divided into three pieces: the anterior part, containing the genital primordiae in the immature segments; the medial part, containing the early uterus in the mature, hermaphroditic proglottids and the terminal part with the mature gravid uterus in the gravid segments. Supernatants of these fragments were used for determination of markers of oxidative stress: concentration of thiobarbiturate reactive substances (TBARS) and of reduced glutathione (GSH), and the activity of antioxidant enzymes: superoxide dismutase (SOD1 and SOD2), catalase (CAT), glutathione peroxidases (GSHPxs), glutathione transferase (GST) and glutathione reductase (GSHR).The results indicated changes in levels of oxidative stress markers and antioxidant enzyme activity in both the young and old forms of H. diminuta. Relatively high activity of SOD (particularly in the anterior part of young tapeworms) was observed, as was increased activity of total GSHPx and a relatively high concentration of GSH in all parts of the tapeworms. These are caused by exposure to increased amount of ROS, which are produced during the inflammatory state. Due to the high activity of antioxidant enzymes, the anterior section of young and old tapeworms is equipped with a very effective antioxidant system. Old organisms also effectively resist oxidative stress due to reduced levels of lipid peroxidation and the high activity of GST, all of which suggest good adaptation to the hostile environment in the host’s intestine.     

The effect of heated effluent on the occurrence and the reproduction of the freshwater limpetsAncylus fluviatilis Müller, 1774,Ferrissia wautieri (Mirolli, 1960) andAcroloxus lacustris (L., 1758) in two Dutch water bodies

The discharge of cooling water favours the occurrence of freshwater limpets. Reproduction ofAncylus fluviatilis is stimulated by the higher temperature of the cooling water; the life cycle ofFerrissia wautieri (nearly exclusively found in the outlet areas) does not, however, differ from that under natural temperature regimes. ForAcroloxus lacustris this is not clear as detailed studies on the life cycle under natural temperature circumstances are lacking. Acroloxus lacustris andFerrissia wautieri could be found in large numbers on hard substrates (stones), as well as on plant material in sheltered as well as in exposed situations. In this respect they differ fromAncylus fluviatilis, which is restricted in its occurrence to hard substrates and exposed situations.It is not very clear whether the occurrence ofAncylus fluviatilis is also influenced by some form of competition for food byBithynia tentaculata; both feed on diatoms. Further experimental studies of this question should yield interesting results.The changes in numbers of the three freshwater limpet species are mainly due to reproduction and migration, both largely governed by temperature and day length.It was possible to reconstruct the influence of cooling water in the Bergumermeer area by means of the ratio between the numbers ofAcroloxus lacustris andFerrissia wautieri. The share ofFerrissia was highest near the outiet and decreased with the distance from this station.     

Relationship between copper- and zinc-induced oxidative stress and proline accumulation in<Emphasis Type="Italic"> Scenedesmus</Emphasis> sp.

A 4-h exposure of Scenedesmus sp. to Cu or Zn enhanced intracellular levels of both test metals and proline. The level of intracellular proline increased markedly up to 10 µM Cu, but higher concentrations were inhibitory. However, intracellular proline consistently increased with increasing concentration of Zn in the medium. Cu and Zn induced oxidative stress in the test alga by increasing lipid peroxidation and membrane permeability, and by reducing SH content. Pretreatment of the test alga with 1 mM proline for 30 min completely alleviated Cu-induced lipid peroxidation, minimized K⁺ efflux and also reduced depletion of the SH pool. But proline pretreatment could only slightly reduce Zn-induced oxidative stress. Interestingly, proline pretreatment increased the level of Cu (25–54%) and Zn (19–49%) inside the cells. It did not affect the activities of superoxide dismutase, ascorbate peroxidase or catalase, but improved glutathione reductase activity under Cu and Zn stress. A comparison of the effects of proline pretreatment on lipid peroxidation by Cu, Zn, methyl viologen and ultraviolet-B radiation suggests that proline protects cells from metal-induced oxidative stress by scavenging reactive oxygen species rather than by chelating metal ions. Pretreatment of cells with a known antioxidant (ascorbate) and a hydroxyl radical scavenger (sodium benzoate) considerably reduced metal-induced lipid peroxidation and proline accumulation. However, sodium benzoate had a very mild effect on Zn-induced lipid peroxidation and proline accumulation. The present study demonstrates that proline possibly acts by detoxifying reactive oxygen species, mainly hydroxyl radicals, rather than by improving the antioxidant defense system under metal stress.Abbreviations APOX Ascorbate peroxidase - CAT Catalase - GR Glutathione reductase - MDA Malondialdehyde - MV Methyl viologen - ROS Reactive oxygen species - SH Sulphydryl - SOD Superoxide dismutase - UV-B Ultraviolet-B radiation