Changes in photochemical and antioxidant enzyme activities in maize (Zea mays L.) leaves exposed to excess copper

Changes in photosynthetic and antioxidant activities in maize (Zea mays L.) leaves of cultivars 3223 and 31G98 exposed to excess copper (Cu) were investigated. Cu treatment reduced the shoot and root length of both cultivars. No significant difference of Cu accumulation in the roots of both cultivars was observed while the cultivar 3223 accumulated significantly higher Cu in leaves than 31G98. The observed decreases in effective quantum efficiency of PSII, ETR and qP indicate an over excitation of photochemical system in 3223 compared to 31G98. The leaf chlorophyll and carotenoid contents of both cultivars decreased with increasing Cu concentration. A far higher production of anthocyanins in 31G98 has been observed than that of 3223. At 1.5 mM Cu concentration, all antioxidant enzyme activities increased in leaves of the cultivar 31G98 while there were no significant changes in SOD and GR activities in 3223 compared to the control except increased APX and POD activities. The lower Cu accumulation in leaves and higher antioxidant enzyme activities in 31G98 suggested an enhanced tolerance capacity of this cultivar to protect the plant from oxidative damage.     

Indication of airborne pollution by birch and spruce in the vicinity of copper smelter

The aim of the study was determination of air pollution impact of the copper smelter in Bor and its surroundings (Serbia) by assessing the suitability of birch (Betula pendula Roth.) and spruce (Picea abies L.) for the purposes of biomonitoring and comparing it with previously published data from the same study area. The concentrations of Cu, Zn, Pb and Mn in leaves/needles, branches, roots and soil were determined. Sampling was performed during 2009 in two zones with high load of air pollution due to copper mining and smelting activities, and one background zone. Metal accumulation and translocation was evaluated in terms of biological factors. In addition, plant enrichment factor was calculated. According to the results, plant foliage was not enriched through soil, which indicates absorption from the air, with both species acting as excluders of Cu, Pb, Zn and Mn. Leaves were more enriched with all the metals than needles, indicating a better response of birch to airborne pollution than spruce. Cluster analysis showed different level of pollution at the sites, while correlations between Cu and Pb obtained by Principal Component Analysis indicated their anthropogenic origin. Regarding previously published results, beside birch leaves, pine needles (which showed higher level of response to pollution compared to linden leaves) could be applied in air biomonitoring surveys near copper smelters.     

Accumulation, speciation and cellular localization of copper in Sesbania drummondii

Growth, accumulation and intracellular speciation and distribution of copper (Cu) in Sesbania drummondii was studied using scanning-electron microscopy (SEM), X-ray absorption near edge structure (XANES) and extended X-ray absorption fine structure (EXAFS). The growth of seedlings was assessed in terms of biomass accumulation. The growth of the seedling was enhanced by 73.5% at a low Cu concentration (50 mg l⁻¹) compared to the control treatment. Additionally, seedling growth was inhibited by 18% at 300 mg l⁻¹ Cu with respect to the control. Copper concentration in roots and shoots was increased with increasing Cu concentration in the growth solution. The accumulation of Cu was found to be higher in roots than in the shoots. At a concentration of 300 mg l⁻¹ Cu, the roots accumulated 27,440 mg Cu kg⁻¹ dry weight (dw) while shoots accumulated 1282 mg Cu kg⁻¹ dw. Seedlings were assessed for photosynthetic activity by measuring chlorophyll a fluorescence parameters: F_v/F_m and F_v/F₀ values. Photosynthetic integrity was not affected by any of the Cu treatments. The X-ray absorption spectroscopic (XAS) studies showed that Cu was predominantly present as Cu(II) in Sesbania tissue. In addition, from the XAS studies it was shown that the Cu exists in a mixture of different coordination states consisting of Cu bound to sugars and small organic acids with some possible precipitated copper oxide. From the EXAFS studies, the coordination of Cu was determined to have four equatorial oxygen(nitrogen) ligands at 1.96 Å and two axial oxygen ligands at 2.31 Å. Scanning-electron microscopy studies revealed the distribution of Cu within the seedlings tissues, predominantly accumulated in the cortical and vascular (xylem) regions of root tissues. In the stem, most of the Cu was found within the xylem tissue. However, the deposition of Cu within the leaf tissues was in the parenchyma. The present study demonstrates the mechanisms employed by S. drummondii for Cu uptake and its biotransformation.     

Effects of silicon and copper on bamboo grown hydroponically

Due to its high growth rate and biomass production, bamboo has recently been proven to be useful in wastewater treatment. Bamboo accumulates high silicon (Si) levels in its tissues, which may improve its development and tolerance to metal toxicity. This study investigates the effect of Si supplementation on bamboo growth and copper (Cu) sensitivity. An 8-month hydroponic culture of bamboo Gigantocloa sp. “Malay Dwarf ” was performed. The bamboo plants were first submitted to a range of Si supplementation (0–1.5 mM). After 6 months, a potentially toxic Cu concentration of 1.5 μM Cu²⁺ was added. Contrary to many studies on other plants, bamboo growth did not depend on Si levels even though it absorbed Si up to 218 mg g^?1 in leaves. The absorption of Cu by bamboo plants was not altered by the Si supplementation; Cu accumulated mainly in roots (131 mg kg^?1), but was also found in leaves (16.6 mg kg^?1) and stems (9.8 mg kg^?1). Copper addition did not induce any toxicity symptoms. The different Cu and Si absorption mechanisms may partially explain why Si did not influence Cu repartition and concentration in bamboo. Given the high biomass and its absorption capacity, bamboo could potentially tolerate and accumulate high Cu concentrations making this plant useful for wastewater treatment.     

Toxicity of copper on rice growth and accumulation of copper in rice grain in copper contaminated soil

Pot soil experiments showed that copper (Cu) is highly toxic to rice. Rice grain yields decreased exponentially and significantly with the increase of soil Cu levels. Rice grain yield was reduced about 10% by soil Cu level of 100 mg kg(-1), about 50% by soil Cu level of 300-500 mg kg(-1) and about 90% by soil Cu concentration of 1,000 mg kg(-1). Root was more sensitive to soil Cu toxicity than other parts of rice plant at relatively lower soil Cu levels (less than 300-500 mg kg(-1)), but the growth of whole rice plant was severely inhibited at high soil Cu levels (300-500 mg kg(-1) or above). Cu concentrations in rice grain increased with soil Cu levels below 150-200 mg kg(-1), but decreased with soil Cu levels above 150-200 mg kg(-1), with peak Cu concentration at soil Cu level of 150-20 mg kg(-1). Cu was not distributed evenly in different parts of rice grain. Cu concentration in cortex (embryo) was more than 2-fold that in chaff and polished rice. More than 60% of the Cu in grain was accumulated in polished rice, about 24% in cortex (embryo), and about 12% in chaff. So, about 1/3 of the Cu in rice grain was eliminated after grain processing (chaff, cortex and embryo was removed).     

Chromium (VI) accumulation reduces chlorophyll biosynthesis, nitrate reductase activity and protein content in Nymphaea alba L

Plants of Nymphaea alba L. grown at various levels of chromium (VI) ranging from 1 to 200 microM accumulated chromium in concentration and duration-dependent manner. At all Cr levels, chromium accumulation by various plant tissues followed the order roots > leaves > rhizomes. Approximately 93% of total chromium present in the medium was accumulated by plants at lowest conentration (1 microM) used in the experiment. Chromium-induced toxicity appears at 1 microM chromium resulting in the build-up of delta-aminolevulinic acid (ALA) and reduced activities of delta-aminolevulinic acid dehydratase (ALAD) and nitrate reductase (NR), total chlorophyll (Chl) and protein contents. Ch1a was more sensitive than Ch1b to chromium toxicity. It could be inferred that chromium toxicity is not located at the level of ALA synthesis, but, probably at the ALAD activity which was more severely affected during chlorophyll biosynthesis. Finally, impaired chlorophyll biosynthesis resulted in reduced total chlorophyll content.     

The toxicity of copper,cadmium and zinc to four different Hydra (Cnidaria: Hydrozoa)

An acute toxicity study of three metals to Hydra species carried out using two different assessment methods, (i) determination of the LC50 and (ii) measurement of progressive morphological changes, demonstrated that relative toxicity decreased from copper to cadmium with zinc the least toxic for all species. The latter method revealed more details of the effect on Hydra in terms of physical damage to the polyp but both methods indicated that H. viridissima was more sensitive to copper and cadmium than H. vulgaris¹ (Zurich strain, male clone), H. vulgaris² (a dioecious strain reproducing sexually and asexually) and H. oligactis (dioecious, reproducing sexually and asexually). The responses to zinc were similar for all Hydra. The possible role of metabolic interactions between H. viridissima and its symbiotic green algae in contributing to the greater sensitivity of this polyp is discussed.     

Seasonal variations of monoterpene emissions from coniferous trees of different ages in Korea

Seasonal variations of emission rates and compositions from coniferous species were measured under controlled conditions using a vegetation enclosure method. Total emission rates and compositions of monoterpene compounds from young and adult trees in different seasons were compared.

It was found that the total emission rates and the components of monoterpene varied significantly with tree species, age, and season. Total emissions from C. japonica and P. koraiensis were higher for older trees than for younger trees; however, significantly higher emissions were found from younger trees for C. obtusa. Higher monoterpene emission rates from each plant were found in spring and summer compared with autumn and winter emissions.     

Effects of copper on Sambucus nigra L. seedlings studied by electron paramagnetic resonance and atomic absorption spectroscopies

We present in this work preliminary results on the translocation of copper, manganese and iron from soil to leaves of Sambucus nigra L. seedlings in the presence of increasing copper concentrations in the growth medium. The use of electron spin resonance spectroscopy and the comparison with atomic absorption spectroscopy is not presently common in the study of metal absorption in plants. This approach gave promising results both on the detection of some metal ions, as well as of radical species related to the presence of Cu2+ in plant tissues. Copper was found to act synergistically in manganese uptake.     

Copper ultrastructural localization, subcellular distribution, and phytotoxicity in Hydrilla verticillata (L.f.) Royle

Laboratory experiments were conducted to investigate copper (Cu) subcellular distribution and toxicity in Hydrilla verticillata. Fronds were subjected to different concentrations (15, 75, and 150 μM) of Cu for 7 days. Cu grains were found in cell walls, plasmodesmata, and within the nuclei and chloroplasts using the autometallographic technique. Subcellular fractionation of Cu-containing tissues indicated that in leaves subjected to high Cu concentrations, 59–65 % of the element was located in the cell wall fraction, followed by cell organelles (21–30 %) and the soluble fraction (10–14 %). The levels of K, P, Zn, and Mg declined under all Cu concentrations, but Ca, Mn, and Fe contents reached their peak at 15 μM Cu and decreased thereafter. F _v/F _m, F ₀, and F _m fell significantly in line with the decrease in pigment content. Cu exposure also caused significant damage to the chloroplasts, mitochondria, and nuclei, including disintegration of the chloroplasts and vacuolization of the mitochondria and nuclei, all of which suggested that Cu hastened plant senescence. The Cu maximum permissible concentration for H. verticillata was 10 μM, which was less than the existing general water quality standard. This suggested that H. verticillata could be used to assess Cu phytotoxicity.     

Assessment of comparative toxicities of lead and copper using plant assay

The acute toxicities of lead (Pb) and copper (Cu) to important crop plants Sorghum bicolor, Cucumis sativus, Triticum aestivum, and Zea mays were compared. The EC50 values (the concentration of metals in the soil that reduces the growth of shoots and roots by 50%) were derived using the Trimmed Spearman-Karber method. The EC50s-shoot (root) in mg Pb kg-1 dry soil and mg Cu kg-1 dry soil were in the range of 519 to >1280 (285-445), and 48-232 (<40-110), respectively. Those concentrations are likely to occur in some abandoned mine areas in Korea. The figures indicate that Cu is more toxic than Pb to the plants in this study, and that root growth is more sensitive to the toxicity endpoint than shoot growth in Cu- or Pb-amended soils. On the other hand, seed germination is insensitive to both Pb and Cu toxicities. The Pb- and Cu-sensitive plants were also identified. Among the plants tested, T. aestivum and S. bicolor were most sensitive to Pb and Cu, respectively. Z. mays was most resistant to both Pb and Cu. The combined effects of Pb and Cu depend on the plant species, and no general phenomenon was observed. Bioaccumulations of Pb and Cu were observed in all test species, and they are concentration-dependent. These differences in the toxicities of Pb and Cu in plant species should be taken into account in biomonitoring and ecological risk assessment.     

Runoff rates,chemical speciation and bioavailability of copper released from naturally patinated copper

The release of copper, induced by atmospheric corrosion, from naturally patinated copper of varying age (0 and 30 years) has been investigated together with its potential ecotoxic effect. Results were generated in an interdisciplinary research effort in which corrosion science and ecotoxicology aspects were combined. The aim of the investigation was to elucidate the situation when copper-containing rainwater leaves a roof in terms of runoff rate, chemical speciation, bioavailability and ecotoxicity effects. Data have been collected during a three-year field exposure conducted in the urban environment of Stockholm, Sweden. The potential environmental effects have been evaluated using a combination of a copper specific biosensor test with the bacterium Alcaligenes eutrophus and the conventional 72-h growth inhibition test with the green alga Raphidocelis subcapitata. The results show annual runoff rates between 1.0 and 1.5 g/m2 year for naturally patinated copper of varying age. The runoff rate increased slightly with patina age, which mainly is attributed to the enhanced first flush effect observed on thicker patina layers. The total copper concentration in investigated runoff samplings ranged from 0.9 to 9.7 mg/l. Both computer modeling and experimental studies revealed that the majority (60-100%) of released copper was present as the free hydrated cupric ion, Cu(H2O)6(2+), the most bioavailable copper species. However, other copper species in the runoff water, such as, e.g. Cu(OH)+ and Cu2(OH)2(2+), were also bioavailable. The copper-containing runoff water, sampled directly after release from the roof, caused significant reduction in growth rate of the green alga. It should be emphasized that the results describe the runoff situation immediately after release from the copper roof and not the real environmental ecotoxicity. Therefore the data should only be used as an initial assessment of the potential environmental effect of copper runoff from building applications. Future risk assessments should also consider dilution effects of copper, changes in its chemical speciation and bioavailability during environmental entry, and type and sensitivity of the receiving ecosystem.     

Analysis of serpentinophytes from north-east of Portugal for trace metal accumulation--relevance to the management of mine environment

In north–east of Portugal, the serpentinized area is about 8000 ha with a characteristic geology and flora. The serpentine plant community and respective soils were analyzed to examine the trace metal budget in different tissues of the plants exhibiting resistance to trace metals. One hundred and thirty five plant species belonging to 39 families and respective soils have been analyzed for total Co, Cr, Cu, Fe, Mn, Ni, Pb and Zn. Substantial amounts of Ni, Cr, Co and Mn were detected in plant tissues which are listed below: Ni: Alyssum serpyllifolium (38 105); Bromus hordeaceus (1467); Linaria spartea (492); Plantago radicata (140); Lavandula stoechas (118) and Cistus salvifolius (114); Cr: L. spartea (706.7); Ulmus procera (173.4); A. serpyllifolium (129.3); Cistus ladanifer (40.8); L. stoechas (29.5); P. radicata (27.81); Setariopsis verticillata (25.7); Plantago lanceolata (24); Digitalis purpurea (23.4); Logfia minima (23.1); Arenaria querioides (23); Hieracium peleteranum (22.7); Arenaria montana (14.5); Co: A. serpyllifolium (145.1); L. spartea (63.2); P. radicata (10.4); H. peleteranum (7.3); Lepidium heterophyllum (6.9); A. querioides (6.6); C. salvifolius (6.5); C. ladanifer (6.3); L. stoechas (6.1); Anthyllis lotoides (6.1); L. minima (6.1); Euphorbia falcata (5.7) and B. hordeaceus (5.6); Mn: A. serpyllifolium (830); L. spartea (339); L. stoechas (187.1); L. minima (182.7); Castanea sativa (125); Spergula pentandra (124); P. radicata (119); Cytisus striatus (115.4); Quercus pyrenaica (110); Teucrium scorodonia (109.4); Fraxinus vulgaris (109); Anthyllis sampaiana (108); Quercus ilex (108). The significance of serpentine flora, need for conservation of these fragile and environmentally invaluable plant resources for possible use for in situ remediation of metalliferous substrates are presented in this paper.     

Accumulation of lead, zinc, copper and cadmium by 12 wetland plant species thriving in metal-contaminated sites in China

The concentrations of lead, zinc, copper and cadmium accumulated by 12 emergent-rooted wetland plant species including different populations of Leersia hexandra, Juncus effusus and Equisetum ramosisti were investigated in field conditions of China. The results showed that metal accumulation by wetland plants differed among species, populations and tissues. Populations grown in substrata with elevated metals contained significantly higher metals in plants. Metals accumulated by wetland plants were mostly distributed in root tissues, suggesting that an exclusion strategy for metal tolerance widely exists in them. That some species/populations could accumulate relatively high metal concentrations (far above the toxic concentration to plants) in their shoots indicates that internal detoxification metal tolerance mechanism(s) are also included. The factors affecting metal accumulation by wetland plants include metal concentrations, pH, and nutrient status in substrata. Mostly concentrations of Pb and Cu in both aboveground and underground tissues of the plants were significantly positively related to their total and/or DTPA-extractable fractions in substrata while negatively to soil N and P, respectively. The potential use of these wetland plants in phytoremediation is also discussed.     

Assessment of zerovalent iron for stabilization of chromium, copper, and arsenic in soil

Stabilization of soil contaminated with trace elements is a remediation practice that does not reduce the total content of contaminants, but lowers the amounts of mobile and bioavailable fractions. This study evaluated the efficiency of Fe(0) to reduce the mobility and bioavailability of Cr, Cu, As and Zn in a chromated copper arsenate (CCA)-contaminated soil using chemical, biochemical and biotoxicity tests. Contaminated soil was stabilized with 1% iron grit. This treatment decreased As and Cr concentrations in leachates (by 98% and 45%, respectively), in soil pore water (by 99% and 94%, respectively) and in plant shoots (by 84% and 95%, respectively). The stabilization technique also restored most of analyzed soil enzyme activities and reduced microbial toxicity, as evaluated by the BioTox test. After stabilization, exchangeable and bioaccessible fractions of Cu remained high, causing some residual toxicity in the treated soil.     

In situ assays with tropical cladocerans to evaluate edge-of-field pesticide runoff toxicity

Tropical regions’ economy is usually based on agriculture, which involves an intensive use, and even frequent overuse, of pesticides. Nevertheless, not much research has been done on the impact of pesticides on tropical aquatic ecosystems, which are often contaminated by runoff-related pesticide inputs due to unpredictable and torrential rainfalls. This study aimed to: (i) adapt and evaluate a short-term sublethal in situ assay using post-exposure feeding as an endpoint, to the tropical cladoceran species Diaphanosoma brachyurum (collected at the Pedra do Cavalo dam in the Paraguaçu River basin, Bahia, Brazil), and, (ii) assess the role of the standard species Daphnia magna as an adequate laboratory surrogate. Lethal and sublethal (post-exposure feeding) responses were assessed for the two species. To evaluate these responses under environmentally realistic exposure conditions, a runoff event was simulated in an agricultural area previously contaminated with different deltamethrin concentrations. The resultant runoff water was used to set up microcosms with different dilutions, simulating the entrance of runoff water in an adjacent lentic system. An in situ assay with D. brachyurum was performed inside the microcosms, allowing to discriminate the effects due to deltamethrin from those due to other potential stressors associated with the experimental design (e.g. organism handling, load of suspended particles, microcosm design). Water samples were collected from microcosms to conduct a laboratory assay with D. magna. The in situ methodologies revealed to be suitable to conduct assays with D. brachyurum under tropical conditions, since all exposed organisms were successfully retrieved from the chambers. Furthermore, none of the potential stressors associated with the experimental design influenced the daphnids’ performance. The tropical cladoceran species, exposed under more realistic conditions, revealed to be more sensitive than the laboratory standard species: lethal effects were only observed for D. brachyurum and sublethal effects were noticed at a lower deltamethrin concentration for this species than for D. magna.     

Cyst-based toxicity tests X: comparison of the sensitivity of the acute Daphnia magna test and two crustacean microbiotests for chemicals and wastes

This paper statistically evaluates the sensitivity of the acute Daphnia magna bioassay in comparison to that of two crustacean microbiotests: the Streptoxkit F and Thamnotoxkit F, which make use of larvae of the anostracans Streptocephalus proboscideus and Thamnocephalus platyurus respectively, hatched from cysts. Regression equations were calculated for 146 data pairs, taken from 5 different studies dealing with the acute toxicity of pure chemicals, effluents, river sediments, solid wastes and monitoring wells, and sludges. All comparisons show that there is a significant relationship (p< 0.05) between the acute effects found with the D.magna and the two crustacean microbiotests; correlation coefficients ranged from 0.84 to 0.92.

In the majority of cases, effect ratios between the conventional D.magna and the microbiotests were within a factor 2 for both pure chemicals and environmental samples.

Variation coefficients for repeated tests on the reference chemical potassium dichromate indicate a good precision and hence a good degree of standardization of the microbiotest procedures. The evidence provided in this paper demonstrates that the new crustacean microbiotests can be used as low cost alternatives to the conventional D.magna acute assay.     

Mercury bioaccumulation and phytotoxicity in two wild plant species of Almadén area

Mercury is a widely distributed environmental pollutant, able to induce toxicity in living organisms, including higher plants. Some plant species are able to grow in mine sites, like the Almadén zone in Spain. Our study focus on two of these plant species, Rumex induratus and Marrubium vulgare and their responses to natural Hg exposure. Total Hg concentration in the soil below the plants could be classified as toxic, although the available fraction was low. Hg availability was higher for the M. vulgare than for the R. induratus plot. Hg concentrations in field plants of R. induratus and M. vulgare grown on these soils can be considered as phytotoxic, although no symptoms of Hg toxicity were observed in any of them. According to the BAF ([Hg]_tissue/[Hg]_avail), R. induratus showed a higher ability in Hg uptake and translocation to shoots, as well as higher concentrations of MDA and –SH:Hg ratios, so that this plant is more sensitive to Hg than M. vulgare. The resistance to Hg and the capability to extract Hg from the soil make both M. vulgare and R. induratus good candidates for Hg phytoremediation of contaminated soils.     

Accumulation of chromium (VI) from aqueous solutions using water lilies (Nymphaea spontanea)

This study describes an investigation using tropical water lilies (Nymphaea spontanea) to remove hexavalent chromium from aqueous solutions and electroplating waste. The results show that water lilies are capable of accumulating substantial amount of Cr(VI), up to 2.119 mg g(-1) from a 10 mg l(-1) solution. The roots of the plant accumulated the highest amount of Cr(VI) followed by leaves and petioles, indicating that roots play an important role in the bioremediation process. The maturity of the plant exerts a great effect on the removal and accumulation of Cr(VI). Plants of 9 weeks old accumulated the most Cr(VI) followed by those of 6 and 3 weeks old. The results also show that removal of Cr(VI) by water lilies is more efficient when the metal is present singly than in the presence of Cu(II) or in waste solution. This may be largely associated with more pronounced phytotoxicity effect on the biochemical changes in the plants and saturation of binding sites. Significant toxicity effect on the plant was evident as shown in the reduction of chlorophyll, protein and sugar contents in plants exposed to Cr(VI) in this investigation.     

Effects of elevated soil copper on phenology,growth and reproduction of five ruderal plant species

The repeated use of copper (Cu) fungicides to control vine downy mildew, caused by Plasmopara viticola, has been responsible for the heavy increase of Cu concentration in the upper layers of vineyard soils. To determine the effects of elevated soil Cu on plant development, we created an artificial soil gradient with Cu enrichments ranging from 0 to 400 mg kg-1. On this gradient, and for five ruderal plant species commonly found in vineyards in southern France (Poa annua L., Dactylis glomerata L., Senecio vulgaris L., Hypochoeris radicata L., and Andryala integriflolia L.), we quantified survival, growth, and reproduction throughout one flowering season. High concentrations of Cu in the soil resulted in low survival, low total plant biomass, delay in flowering and fruiting, and low seed set. However, the effects differed among species. Furthermore, high soil Cu concentrations had contrasting effects on patterns of resource allocation depending on the plant species.