Mechanisms of copper tolerance by Armeria maritima in Dolfrwyong Bog,north Wales--initial studies

Preliminary field studies were carried out at Dolfrwynog Bog in July 2000. Replicate samples of water, Armeria maritima plants and the soils adhering to its roots were collected and analysed for copper. Concentrations of up to 6486 mg kg^–1 of copper in the soils were recorded. Accumulation of copper by the plant as expressed by concentration factors (CF) show that it is acting mainly as a copper excluder. Of the copper that is taken up, most of it is retained within the roots with very little being transported to the shoots of the plant. Moreover, a further possible mechanism of tolerance is exhibited by the excretion of copper through its decaying leaves. Towards the use of in vitro cultures to study the copper tolerance mechanisms in A. maritima a micropropagation protocol has been developed. The ex vitro plants have been rooted and established in compost.     

Effects of copper on the latency of lysosomal hexosaminidase in the digestive cells of Mytilus edulis

Mytilus edulis collected from Tomales Bay, California, USA, during mid-winter 1979 were exposed to increased concentrations of dissolved copper under controlled laboratory conditions. A dose-dependent reduction in the latency of lysosomal hexosaminidase activity in digestive cells was induced after a 30 d exposure to copper. The half-time of the hexosaminidase staining reaction in sections of digestive gland from control mussels was 15.5 min; for mussels exposed to 25, 50, and 75 g Cu l^-1 it was 11.8, 8.5 and 5.5 min, respectively. In addition, the dyecoupled reaction product was seen earlier in sections from individuals exposed to 50 and 75 g Cu l^-1 (after 30 s) and 25 g Cu l^-1 (1 min) than in sections from control individuals (2.5 min). Copper accumulations were demonstrated histochemically to have the same distribution as the hexosaminidase reaction product, indicating that copper is sequestered in lysosomes. Copper concentrations in digestive gland tissue, were related to the concentrations of copper in the water to which the mussels were exposed.     

Effects of nanoparticles on Daphnia magna population dynamics

Spherical TiO₂ nanoparticles (npTiO₂) were prepared by controlled hydrolysis of tetraethoxy orthotitanate under a nitrogen atmosphere. ZnO nanoparticles (npZnO) were prepared using hydrothermal methods. The crystal structure, chemical, thermal and morphological properties of npZnO and npTiO₂ were characterised using Fourier Transform Infrared Spectrometer, enery-dispersive X-ray spectroscopy, X-ray diffraction, and scanning electron microscope techniques. The short- and long-term experiments were started with neonates taken from the same culture and laboratory condition. In the acute experiments, npTiO₂, npZnO, and cocktail concentrations were applied. 96h-LC₅₀ values were 1.8, 0.7, and 0.1?mg?L^?1, respectively (p?<?.05). For the chronic experiments, different npTiO₂ concentrations were performed. 21d-LC₅₀ value was 1.0?mgL^?1 (p?<?.05). Morphometry became progressively worse in concentrations of more than 1?mgL^?1 npTiO₂. Neonate and young individuals were more sensitive to death because of their low tolerance. This result was affected by population progeny and growth rates (p?<?.05). While control and 0.5?mgL^?1 npTiO₂ groups were determined as growing population, 1.5 and 2?mgL^?1 npTiO₂ groups had decreased population size as R₀ values. Consequently, the relationships between nanoparticle accumulation within Daphnia magna and its population structure and body morphometry for each concentration were important indicators. Its tolerance level to nanoparticles under laboratory conditions reflected its replacement and behaviour in the ecosystem.     

植物铜耐性机理的研究进展

主要从植物根细胞壁积累固定、细胞膜对铜的吸收控制、金属配体的螯合作用、铜在系统液泡的分隔机制及胁迫蛋白的合成5个方面,分别阐述植物对铜分子的耐性机制的研究进展,全面了解了铜在植物中的亚细胞分布、铜在植物根系到地上部分运输过程的转运机制以及植物在铜胁迫下的抗性反应等。并在此基础上提出了存在的问题以及今后研究的重点。     

狼尾草根系对阿特拉津长期胁迫的氧化应激响应

通过盆栽实验研究了抗性植物狼尾草根部丙二醛(MDA)、脯氨酸(Pro)、抗坏血酸(As A)含量及超氧化物歧化酶(SOD)、谷胱甘肽还原酶(GR)等氧化应激生理指标对不同浓度阿特拉津长期(48 d)胁迫的响应规律。结果表明:当阿特拉津胁迫浓度分别高于20 mg·kg~(-1)和50 mg·kg~(-1)时,狼尾草根系的MDA与Pro含量较对照组显著升高(P0.05);随着阿特拉津胁迫浓度的增加,狼尾草根部SOD和GR活性呈先升高后降低的趋势,其中当阿特拉津胁迫浓度为20 mg·kg~(-1)时,SOD和GR活性达到最大值;供试植物根系中As A含量与阿特拉津胁迫浓度呈正相关。综上,中低浓度(≤20 mg·kg~(-1))阿特拉津处理没有对狼尾草的根系产生明显的氧化胁迫效应,狼尾草根系的上述抗氧化应激生理指标对于发挥阿特拉津抗性起着重要的作用。     

Differential tolerance to metals among populations of the introduced bryozoan Bugula neritina

Resistance to heavy metals is a potentially important trait for introduced marine organisms, facilitating their successful invasion into disturbed natural communities. We conducted laboratory and field experiments to examine differential resistance to copper (Cu) between two source populations of the introduced bryozoan Bugula neritina, originating from a polluted (Port Kembla Harbour, NSW, Australia) and an unpolluted (Botany Bay, NSW, Australia) environment. A laboratory toxicity test was conducted to test the relative resistance of B. neritina recruits from the two sources, by measuring the attachment success, survival and growth of individuals exposed to a range of Cu concentrations (0, 25, 50 and 100 μg l⁻¹ Cu). Upon completion, reciprocal transplantation of the colonies to the original polluted and unpolluted locations was carried out to assess ongoing survival and growth of colonies in the field. B. neritina colonies originating from the polluted Port Kembla Harbour had increased resistance to Cu relative to populations from an unpolluted part of Botany Bay. There appeared to be a cost associated with increased metal tolerance. In the laboratory, Botany Bay recruits displayed significantly higher growth in control treatments and significantly poorer growth at 100 μg l⁻¹ Cu with respect to Port Kembla Harbour individuals, which showed unusually uniform and low growth irrespective of Cu concentration. No difference in attachment success or post-metamorphic survival was observed between populations. Field transplantation showed copper resistance in Port Kembla Harbour colonies constituted an advantage in polluted but not benign environments. The findings of this study provide evidence of the benefits to invasive species of pollution tolerance and suggest that human disturbance can facilitate the establishment and spread of invasive species in marine systems.     

Acclimation and tolerance of Artemia salina and Ophryotrocha labronica to cooper sulphate

The brine shrimp Artemia salina L. and the polychaete worm Ophryotrocha labronica La Greca and Bacci were acclimated in sea water with copper sulphate at concentrations of 0.1, 0.05, and 0.025 ppm Cu⁺⁺, for 3 and 2 generations, respectively. Both adults and larvae of A. salina showed a greater tolerance to 1 ppm Cu⁺⁺ after acclimation compared to controls of the same age, although this tolerance diminished in successive generations. The acclimation effect was less marked in O. labronica. In both species, tolerance to 10 ppm Cu⁺⁺ upwards was not enhanced. Growth-rate inhibition and an adverse effect on reproduction was observed, in some instances in direct relationship to the acclimation concentration. It is suggested that, in A. salina, a certain tolerance to copper may be acquired through exposure to low concentrations.     

成团泛菌(Pantoea agglomerans)对镉胁迫下蒌蒿耐受性的影响

为探讨功能微生物对镉(Cd)胁迫下蒌蒿耐受性的影响,以成团泛菌(Pantoea agglomerans)J2为对象,通过盆栽试验,研究不同Cd处理水平下J2菌对蒌蒿生长、生理、Cd富集转运与根际微生态的影响。结果表明:J2菌可显著促进蒌蒿株高、地上部干重、地下部干重、总生物量及根冠比的增加。在J2菌的作用下,蒌蒿叶绿素a、叶绿素b和总叶绿素含量以及超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性均显著增加,丙二醛(MDA)含量明显减少。J2菌可显著促进蒌蒿对Cd的富集转运,蒌蒿地上部、地下部与总Cd含量均显著增加,地上部生物富集系数(BCF)明显大于地下部,转运系数(TF)相比于对照增加了7.5%~37.4%。J2菌可显著增加蒌蒿根际土壤细菌、放线菌数量及微生物总量,明显减少真菌数量。研究表明成团泛菌J2对蒌蒿生长、生理、Cd富集转运与根际微生态可产生积极影响,在一定程度上可提高蒌蒿植株对Cd的耐受性。     

Biosorption of Cu (II) ions by indigenous copper-resistant bacteria isolated from polluted coastal environment

The concentrations of copper in water and sediment samples of Cuddalore coastal area, Tamil Nadu, India were examined. The isolation, characterization, screening, and metal sorption capabilities of copper-resistant bacteria (CURB) also were investigated. Biosorption capabilities and growth time varied among the resistant isolates. All isolates reduced more than 80% of copper from copper-treated broth. However, Bacillus cereus CURB-4 strain absorbed more copper ions (99.9%) than other resistant isolates. Changes in structural group position and synthesis of extracellular polysaccharides (EPS) of CURB-4 strain under copper stress and controlled conditions also were examined. Under copper stress condition, CURB-4 produced more EPS (71 ± 0.6%) compared to control conditions (66 ± 0.4%). Fourier transform infrared spectroscopy revealed that the changes in functional group position of EPS were observed in stressed condition. Overall, synthesis of EPS in bacteria is a protective barrier against environmental stress.     

Change in the gross biochemistry and mineral content accompanying the moult cycle in the Antarctic krill Euphausia superba

Concentrations of water, ash, protein, chitin, lipid, calcium, magnesium, sodium, potassium, strontium and copper were measured in individuals from a laboratory population of Antarctic krill, Euphausia superba Dana, over the course of a moult cycle. Significant changes in all variables were encountered. Total ash, lipid, calcium, magnesium and strontium all increased in concentration following moulting. Water, protein and copper concentrations all decreased following ecdysis and increased again towards the end of the moult cycle. The major ions sodium and potassium fluctuated around mean levels. Cast moults of E. superba were shown to be a drain on the ionic load of the krill, and the losses inherent in exuviation could account for much of the variation observed during the moult cycle.     

The common mussel Mytilus edulis as an indicator of pollution by zinc,cadmium, lead and copper. I. Effects of environmental variables on uptake of metals

The net uptake of zinc, cadmium, lead and copper by the common mussel Mytilus edulis (L.) exposed to different conditions was investigated with a view to using this species as an indicator of contamination of the marine environment by these metals. The variables studied were season, position of the mussel in the water column, water salinity, water temperature, and the simultaneous presence of all four metals. Each of these 5 variables affected the net uptake of some or all of the metals studied under some conditions. Seasonal variation in concentrations of zinc, cadmium and copper was found in samples collected at three separate locations. The relationship of seasonal variation to tissue weight and absorption route of the metals is discussed. Near to freshwater inputs of trace metals, the concentrations of zinc, cadmium and lead in mussels were found to vary according to the depth at which the mussels were collected; in summer when freshwater run-off is less, this effect was absent. Low salinities did not affect the net uptake of zinc by mussels, but increased the net uptake of cadmium and decreased that of lead. Low temperatures had no effect on the net uptake of zinc or lead; the net uptake of cadmium was unaffected by low temperatures at high salinities but was decreased by low temperatures at low salinities. The presence of the other metals had no effect on the individual net uptake of either zinc, cadmium or lead. A sampling programme was devised to eliminate the effects of these environmental variables and to allow the use of M. edulis as an indicator of zinc, cadmium and lead in marine and estuarine environments. In contrast to the other metals, the net uptake of copper by the mussel was extremely erratic, and was affected by salinity and temperature changes and by the presence of the other metals and changes in their relative concentrations. The effects of other metals on the net uptake of copper cannot be easily eliminated or allowed for; it is, therefore, suggested that the mussel should not be used as an indicator of copper in the marine environment.     

菲胁迫下湿生植物美人蕉(Canna indica)对斜生栅藻(Scenedesmus obliquus)生长的影响

多环芳烃有机污染物的环境问题日益严重,亟待需求从群落及生态系统的角度进行毒理学复合危害效应的研究。本文采用以美人蕉(Canna indica)种植根区水培养斜生栅藻(Scenedesmus obliquus)的方式,观察菲胁迫下其生长的变化。结果表明,美人蕉种植根区水对斜生栅藻生长具有化感效应,且表现为"低促高抑"的现象。而在菲胁迫下,种植根区水比例为15%和30%时随着菲浓度的增加,生长促进作用增强,且在菲浓度为1 mg·L-1出现最高值;在100%种植根区水时随着菲浓度的增加生长抑制作用增强。说明,菲胁迫使美人蕉种植根区水对藻类的化感效应增强。进行逻辑斯谛生长拟合发现,30%、1 mg·L-1处理组种群内禀增长率最高,斜生栅藻种群出现暴发增长的生态风险最大。     

Variations between winter and summer in the toxicity of copper to a population of the mysid Praunus flexuosus

 Copper toxicity was tested on a coastal population of the mysid Praunus flexuosus (Müller) from Southampton Water (Southern England) under winter and summer conditions. Ten-day toxicity tests were performed on the different life-cycle stages (female, male and juvenile) present in winter (December/February) and summer (August). The individuals were in winter or summer physiological condition and were exposed to seawater to which 0, 5, 25, 75 and 200 μg l⁻¹ copper was added. There were significantly different copper toxicity effects in winter and summer. In winter mortality was ≤ 1% at all levels of copper exposure, while in summer identical exposure levels caused mortality of up to 93%. The 96 h LC₅₀ was 30.8 μg l⁻¹ copper added in the summer. In winter, the low mortality prevented calculation of LC₅₀. There were differences in responses to copper between the life-cycle stages. Juveniles were more sensitive than adults, and were severely affected within 24 h. Females were more affected than males at lower doses and shorter exposure times. Received: 28 April 1999 / Accepted: 22 June 2000     

Influence of anoxia on adaptation of euryhaline polychaetes to hyposmotic conditions

The metabolic reactions ofArenicola marina andNereis diversicolor (both euryoxic and euryhaline polychaetes from the intertidal) to hyposmotic shock under anaerobic conditions were investigated in 1987–1988 using specimens from the East Frisian Wadden Sea. Although reductions in salinity were within the limits of tolerance under normoxic conditions, during anoxia they resulted in increased stress and a significant reduction in survival time. Both species were unable to sufficiently intensify anaerobic energy production to match the increased requirement for energy during adaptation to hyposmotic conditions. Moreover, as an adaptation to anoxic conditions, worms reduced their metabolic activity after the first 6 h. Thus, specimens remained more swollen than at the same reduction in salinity under normoxic conditions. InA. marina only the extracellular volume was reduced all be it to a limited extent.N. diversicolor probably reacted in the same or similar way. In both species, cells remained swollen. The concentrations of cellular free amino acids were not reduced; that means the main mechanism of cellular volume regulation was not activated under these conditions.     

Bioaccumulation and toxic effects of copper on growth and oxygen consumption by the postlarvae of Penaeus indicus

The aim of this study was to assess the toxic impact of copper on postlarvae (PL) of the penaeid shrimp Penaeus indicus. Tolerance, growth, oxygen consumption and metal accumulation were investigated in these PL on exposure to copper. Tolerance studies were conducted for 96 h to assess the tolerance limits of P. indicus PL exposed to different concentrations of copper using static renewal bioassay tests. Using the Probit method, the regression equation was calculated as Y=0.4899+2.3562 X, with a correlation coefficient of 0.9707. The 96 h LC₅₀ was 0.8204 ppm. The effect of sublethal (one-fifth of 96 h LC₅₀) copper on PL for short- and long-term exposures revealed a significant (p<0.05) decrease in the rate of oxygen consumption, metabolic rate, mean length, wet and dry weight of the exposed PL over their respective controls which can be attributed to a gradual and time-dependent accumulation of the metal, as noticed in the exposed PL through accumulation studies. Overall, the data suggest that on chronic exposure even sublethal concentrations of copper can reduce the metabolic rate and growth in P. indicus PL. This is perhaps the first attempt to use the wild P. indicus PL as a bioindicator of copper toxicity.     

Chemoecological studies of the exocrine glandular larval secretions of two chrysomelid species (Coleoptera):Phaedon cochleariae andChrysomela lapponica

Summary The exocrine glandular secretions of larvae of the subfamily Chrysomelinae are known to repel conspecific adults, other competitive phytophagous insects and natural enemies. InPhaedon cochleariae, the intraspecific activity of tlc fractions of the larval secretion was tested in order to examine the ecological significance of two fractions containing minor components and a fraction containing the major compound, the cyclopentanoid monoterpene (epi)chrysomelidial. InChrysomela lapponica, the defensive activity of the larval secretion against ants is known from specimens feeding upon willow or birch. The feeding preferences of larvae and adults ofC. lapponica from a Finnish and a Czech population were tested. The Finnish individuals significantly preferred feeding uponSalix borealis, whereas they hardly fed upon birch. The Czech specimens clearly preferred birch (Betula pubescens) to willow species. Application of salicin onto leaves of a willow species free of this phenolglycoside revealed that the Finnish individuals preferred feeding upon leaves with salicin. On the other hand, the Czech individuals avoided feeding upon leaves ofB. pubescens treated with salicin. The chemical composition of the glandular secretion of the Finnish larvae differed from the one of the Czech larvae. GC-MS-analyses of the secretions revealed that salicylaldehyde was the only major component of the secretion of Finnish larvae feeding upon the salicin-containing willowS. borealis. The glandular secretion of the Czech larvae feeding upon birch contained numerous esters of isobutyric acid and 2-methylbutyric acid. When Czech larvae had fed upon a salicin-containing willow (S. fragilis), the major compounds of their secretion were benzoic acid, salicylalcohol and benzoic acid esters; salicylaldehyde was only detected in traces. Thus,C. lapponica individuals from the Finland population adapted so closely to a salicincontaining willow that they clearly prefer this plant for food and that they obviously derive their main larval defensive compound (salicylaldehyde) from their host-plant.     

Field distribution and sulphide tolerance of Capitella capitata (Annelida: Polychaeta) around shallow water hydrothermal vents off Milos (Aegean Sea). A new sibling species?

The cosmopolitan polychaete Capitella capitata, known as a complex of opportunistic sibling species, usually dominates the macrobenthos of polluted or unpredictable environments. A population of C. capitata, termed Capitella sp. M, was found in a shallow water hydrothermal vent area south of Milos (Greece). Here, this population occurs close to vent outlets (termed the “transition zone”), an environment with steep gradients of temperature, salinity and pH and increased sulphide concentrations of up to 710 μM. The field distribution of C. capitata in relation to sulphide concentrations around the vent outlets was investigated and sulphide tolerance experiments were conducted on laboratory-cultured worms to elucidate possible adaptations of Capitella sp. M to these extreme environmental conditions. In order to investigate whether the population from the Milos hydrothermal vent area can be considered a distinct sibling species within the C. capitata complex, crossbreeding experiments and analysis of general protein patterns were conducted with Capitella sp. M and three other C. capitata populations of different ecological ranges. Capitella sp. M showed high resistance (median survival time: 107 ± 38 h) to anoxia plus high sulphide concentrations of 740 μM. It seems that the ability to survive high-sulphide conditions in combination with reduced interspecific competition enables the polychaete to maintain a continuous population in this rigorous habitat. From the extremely high tolerance to anoxia and sulphide, shown in both the crossbreeding experiments and the analysis of total proteins, it can be concluded that Capitella sp. M from the Milos hydrothermal vent area represents a separate sibling species within the C. capitata complex. Received: 3 March 1997 / Accepted: 12 September 1997     

Bioassay-guided isolation of allelochemicals from Avena sativa L.: allelopathic potential of flavone C-glycosides

The allelopathic potential of oat (Avena sativa L., var. Argentina, Poaceae, Cyperales) was investigated under field and laboratory conditions. In field trials, oat plants provided an effective control of weeds, showing a species-specific impact: the most abundant weed species, Picris echioides was reduced by 94% in number of individuals. Aerial parts of oat plants, harvested immediately before soil incorporation, were utilized in a bioassay-guided isolation, which was aimed at identifying the phytotoxic compounds in a methanol/water extract of the aerial parts of the plants. Further partitions of extract gave an active n-butanol portion composed of flavonoids and saponins. Phytotoxic activity was detected for the flavonoid fraction, whereas no activity was found for the saponin mixture. Germination of an indicator species, lettuce (Lactuca sativa L.), was completely inhibited at flavonoid concentrations of 6.7, 10.0, and 20.0 mg/mL, and conversely the number of abnormal seedlings was greatly increased from 2% of control to over 96% at concentrations equal to 6.7 and 10.0 mg/mL. Analysis of the flavonoid fraction by ESI-MS techniques identified two components of the mixture as isoorientin 2-O-glucoside and isovitexin 2″-O-arabinoside. Saponins were characterized as avenacoside A, avenacoside B, 26-desglucoavenacoside A, and 26-desglucoavenacoside B. Both flavone C-glycosides and saponins were isolated previously from oat.     

Mechanisms for copper tolerance in Amphora coffeaeformis-internal and external binding

Growth of the ship-fouling diatom Amphora coffeaeformis and accumulation of copper in the cells were evaluated for cultures exposed to copper. Comparisons with literature reports for other species revealed that A. coffeaeformis shows no ability to maintain normal growth rates in the presence of high cellular copper levels. This suggests that internal binding is not the principal copper tolerance mechanism for this species. In addition, the copper complexing capacity of A. coffeaeformis exudates was evaluated. Significant complexing by these exudates was demonstrated by DPASV analysis. When added to the culture medium of another species (Thalassiosira profunda), A. coffeaeformis exudates were also able to reduce copper toxicity and accumulation in the cells of that species. However, the copper tolerance of A. coffeaeformis was greater than that acquired by T. profunda grown with A. coffeaeformis exudates; thus exudate production was deemed not to be a primary tolerance mechanism. Comparison of copper accumulations inside and outside cells of A. coeffeaeformis suggests that binding at the cell surface or to mucilage may be an important factor in the tolerance of this species to copper.     

Interactions between two species of marine diatoms: Effects on their individual copper tolerance

The purpose of this study was to examine the effect of the interaction of two species of marine diatoms,Skeletonema costatum (Cleve) Greville andNitzschia thermalis (Ehrenberg) Auerswald, on their individual copper tolerances. The two species, obtained from stock cultures in 1989, were grown together at three copper concentrations (1, 4 and 5 × 10^–7 M added total copper). In the unialgal cultures that were used as controls, the two species grew as predicted from their tolerance tests. However, in mixed cultures,N. thermalis was the only species that exhibited growth, regardless of the copper concentration in the medium. Growth retardation ofS. costatum in the presence ofN. thermalis was attributed to an inhibitory exudate. The effect of the exudate appeared to have been temporary, as demonstrated by the extended lag phase and subsequent satisfactory exponential growth rate ofS. costatum. It is suggested that the exudate degraded within a period of 5 d (=lag phase) because exponential growth rate was resumed. It thus appears that the interaction between the two diatom species is more important in determining the survival ofS. costatum than its individual copper tolerance. This is not the case forN. thermalis. Such interactions would be unaccounted for in single-species toxicity tests. On the other hand, if they are known, prediction of how a community that includes these two species would respond to copper additions becomes possible.