Copper Toxicity in Acidic Phytoplankton: Impacts of Labile Cu Trafficking and Causes of Mitochondria Dysfunction.

Most studies on Cu toxicity relied on indirect physicochemical parameters to predict Cu toxicity resulting from adverse impacts. This study presents a systematic and intuitive picture of Cu toxicity induced by exogenous acidification in phytoplankton Chlamydomonas reinhardtii. We first showed that acidification reduced the algal resistance to environmental Cu stress with a decreased growth rate and increased Cu bioaccumulation. To further investigate this phenomenon, we employed specific fluorescent probes to visualize the intracellular labile Cu pools in different algal cells. Our findings indicated that acidification disrupted the intracellular labile Cu trafficking, leading to a significant increase in labile Cu(I) pools. At the molecular level, Cu toxicity resulted in the inhibition of the Cu(I) import system and activation of the Cu(I) export system in acidic algal cells, likely a response to the imbalance in intracellular labile Cu trafficking. Subcellular analysis revealed that Cu toxicity induced extensive mitochondrial dysfunction and impacted the biogenesis and assembly of the respiratory chain complex in acidic algal cells. Concurrently, we proposed that the activation of polyP synthesis could potentially regulate disrupted intracellular labile Cu trafficking. Our study offers an intuitive, multilevel perspective on the origins and impacts of Cu toxicity in living organisms, providing valuable insights on metal toxicity.

Copper stress in rice: Perception, signaling, bioremediation and future prospects.

Copper (Cu) is an indispensable micronutrient for plants, animals, and microorganisms and plays a vital role in different physiological processes. However, excessive Cu accumulation in agricultural soil, often through anthropogenic action, poses a potential risk to plant health and crop productivity. This review article provided a comprehensive overview of the available information regarding Cu dynamics in agricultural soils, major sources of Cu contamination, factors influencing its mobility and bioavailability, and mechanisms of Cu uptake and translocation in rice plants. This review examined the impact of Cu toxicity on the germination, growth, and photosynthesis of rice plants. It also highlighted molecular mechanisms underlying Cu stress signaling and the plant defense strategy, involving chelation, compartmentalization, and antioxidant responses. This review also identified significant areas that need further research, such as Cu uptake mechanism in rice, Cu signaling process, and the assessment of Cu-polluted paddy soil and rice toxicity under diverse environmental conditions. The development of rice varieties with reduced Cu accumulation through comprehensive breeding programs is also necessary. Regulatory measures, fungicide management, plant selection, soil and environmental investigation are recommended to prevent Cu buildup in agricultural lands to achieve sustainable agricultural goals.

Physiological, Transcriptome, and Metabolome Analyses Reveal the Tolerance to Cu Toxicity in Red Macroalgae Gracilariopsis lemaneiformis.

Heavy metal copper (Cu) will inevitably impact the marine macroalgae Gracilariopsis lemaneiformis (G. lemaneiformis), which is a culture of economic importance along China's coastline. In this study, the detoxification mechanism of Cu stress on G. lemaneiformis was revealed by assessing physiological indicators in conjunction with transcriptome and metabolome analyses at 1 d after Cu stress. Our findings revealed that 25 µM Cu stimulated ROS synthesis and led to the enzymatic oxidation of arachidonic acid residues. This process subsequently impeded G. lemaneiformis growth by suppressing photosynthesis, nitrogen metabolism, protein synthesis, etc. The entry of Cu ions into the algae was facilitated by ZIPs and IRT transporters, presenting as Cu2+. Furthermore, there was an up-regulation of Cu efflux transporters HMA5 and ABC family transporters to achieve compartmentation to mitigate the toxicity. The results revealed that G. lemaneiformis elevated the antioxidant enzyme superoxide dismutase and ascorbate-glutathione cycle to maintain ROS homeostasis. Additionally, metabolites such as flavonoids, 3-O-methylgallic acid, 3-hydroxy-4-keto-gama-carotene, and eicosapentaenoic acid were up-regulated compared with the control, indicating that they might play roles in response to Cu stress. In summary, this study offers a comprehensive insight into the detoxification mechanisms driving the responses of G. lemaneiformis to Cu exposure.

Comparative effectiveness of carbon nanoparticles and biochar in alleviating copper stress in corn (Zea mays L.).

The application of carbon nanoparticles (CNPs) and biochar in agriculture for improving plant health and soil quality and alleviating metal stress offers alternative approaches to meet the ever-increasing demand for food. However, poor understanding of their roles in improving crop production under Cu stress represents a significant obstacle to their wide application in agriculture. To clarify how CNPs and biochar affect corn (Zea mays L.) seed germination, seedling growth, plant health, and nutrient uptake under different Cu stress levels, soil-less Petri-dish and greenhouse soil-based bioassays were conducted. The results revealed that CNPs and biochar stimulated corn seed germination and seedling growth. Besides, they were effective in immobilizing Cu2+ sorption in sandy soil and alleviating Cu stress for plant growth, as shown by the increased plant height and dry biomass. The plant nutrient uptake efficiency (NUE) was significantly increased by CNPs, with a maximum increase of 63.1% for N and 63.3% for K at the highest Cu2+ stress level (400 mg Cu2+ L-1). In contrast, non-significant effects on NUE were observed with biochar treatments regardless of Cu stress levels. Interestingly, CNPs significantly increased plant uptake of Cu in the Petri dish test, while biochar inhibited plant uptake of Cu under both experimental conditions. Principle component analysis (PCA) and Pearson correlation analysis indicated that CNPs mitigated Cu stress mainly by elevating antioxidant enzyme activities, enhancing plant photochemical efficiency, and increasing plant uptake of N and K, while biochar was more likely to reduce bioavailability and uptake of Cu in the plant. These findings have great implications for the application of CNPs and biochar as plant growth stimulators and de-toxicity agents in agriculture.

Bioimaging tools reveal copper processing in fish cells by mitophagy.

As an essential trace metal, copper (Cu) regulation, distribution and detoxification among different cellular organelles remain much unknown. In the current study, bioimaging tool was used in visualizing the locations of Cu among different organelles in fish fin cells isolated from rabbitfish Siganus fuscescens. Exposure concentration of Cu directly affected the Cu bioaccumulation and toxicity. When the exposure dosage of Cu reached 100 µM, it began to damage the cells and affect the cell viability after 10 min of exposure. Remarkably, while various Cu concentrations (50∼150 µM) initially reduced the cell viability, they did not lead to a further loss in viability over extended exposure period. Upon entry to the cells, Cu was mainly targeted to the mitochondria whose number, size and network responded immediately to the incoming Cu. However, Cu toxicity did not increase time-dependently, strongly indicating that these mitochondria damaged by Cu could be removed and its cytotoxicity could be relieved. Bioimaging results showed that lysosomes interacted with the mitochondria, which were subsequently digested within a few minutes. Meanwhile the lysosomal number increased, and the size and pH of lysosomes decreased. These reactions were in line with the observed mitophagy, suggesting that mitochondrial Cu could be detoxified, and the damaged mitochondria were removed by lysosome via mitophagy. By further purifying the cellular organelles, the mitochondrial and lysosomal Cu amounts were quantified and found to be in line with the imaging results. The present study suggested that excessive mitochondrial Cu could be removed via mitophagy to relieve the Cu toxicity.

Molecular mechanism of Cu metal and drought stress resistance triggered by Porostereum spadiceum AGH786 in Solanum lycopersicum L.

Rapid industrialization and global warming have threatened the plants with multiple abiotic stresses, such as heavy metals and drought stress. For crop cultivation, the conventional approach of cleaning the soils by excavation is very costly and not feasible for large scale. Establishing toxin-free and drought-resistant crops is a major challenge in the environment under natural and anthropogenic pressure. In the past decades, copper contamination of agricultural land has become an emerging concern. For dry land reclamation, several new strategies, including bioremediation (phytoremediation and microbial remediation), have been used. Owing to the potential of Cu hyperaccumulators, the current project aims to enhance the drought tolerance and the phytoremediation potential of Solanum lycopersicum L. with the inoculation of copper and 12% polyethylene glycol (PEG)-induced drought stress-tolerant endophytic fungus Porostereum spadiceum AGH786 under the combined stress of copper heavy metal and PEG-induced drought stress. When S. lycopersicum L. was watered with individual stress of copper (Cu) concentration (400 ppm) in the form of copper sulfate (CuSO4.5H2O), 12% PEG-induced drought stress and the combined stress of both negatively affected the growth attributes, hormonal, metabolic, and antioxidant potential, compared with control. However, the multistress-resistant AGH786 endophytic fungus ameliorated the multistress tolerance response in S. lycopersicum L. by positively affecting the growth attributes, hormonal, metabolic, and antioxidant potential, and by restricting the root-to-shoot translocation of Cu and inducing its sequestration in the root tissues of affected plants. AGH786-associated plants exhibited a reduction in the severity of copper (Cu) and drought stress, with higher levels of SlCOPT (Cu transporters) and SlMT (metallothionine) gene expressions in root and shoot tissues, indicating that AGH786 contributed to resistance to copper metal toxicity and drought stress in the host S. lycopersicum L.

Photosynthetic Characteristics and Chloroplast Ultrastructure Responses of Citrus Leaves to Copper Toxicity Induced by Bordeaux Mixture in Greenhouse.

Bordeaux mixture (Bm) is a copper (Cu)-based pesticide that has been widely used for controlling citrus scab and citrus canker. However, frequent spraying of Bm is toxic to citrus. To our knowledge, few studies are available that discuss how the photosynthetic characteristics and chloroplast ultrastructure of citrus leaves are affected by Cu toxicity induced by excessive Bm. In the study, two-year-old seedlings of Citrus grandis (C. grandis) and Citrus sinensis (C. sinensis), which were precultured in pots, were foliar-sprayed with deionized water (as control) or Bm diluted 500-fold at intervals of 7 days for 6 times (4 times as recommended by the manufacturer) to investigate the leaf Cu absorption, photosynthesis, chloroplast ultrastructure and antioxidant enzymatic activities. Bm foliar-sprayed 6 times on citrus seedlings increased the leaf Cu content, decreased the photosynthetic pigments content and destroyed the chloroplast ultrastructure, which induced leaf chlorosis and photosynthetic inhibition. A lower Cu absorption, a higher light photon-electron transfer efficiency, a relative integrity of chloroplast ultrastructure and a promoted antioxidant protection contributed to a higher photosynthetic activity of C. grandis than C. sinensis under excessive spraying of Bm. The present study provides crucial references for screening and selecting citrus species with a higher tolerance to Cu toxicity induced by excessive Bm.

Superior antioxidant and anti-ischemic neuroprotective effects of cerebrolysin in heat stroke following intoxication of engineered metal Ag and Cu nanoparticles: A comparative biochemical and physiological study with other stroke therapies.

Military personnel are often exposed to high environmental heat associated with industrial or ambient abundance of nanoparticles (NPs) affecting brain function. We have shown that engineered metal NPs Ag and Cu exacerbate hyperthermia induced brain pathology. Thus, exploration of novel drug therapy is needed for effective neuroprotection in heat stroke intoxicated with NPs. In this investigation neuroprotective effects of cerebrolysin, a balanced composition of several neurotrophic factors and active peptides fragments exhibiting powerful antioxidant and anti-ischemic effects was examined in heat stroke after NPs intoxication. In addition, its efficacy is compared to currently used drugs in post-stroke therapies in clinics. Thus, levertiracetam, pregabalin, topiramat and valproate were compared in standard doses with cerebrolysin in heat stroke intoxicated with Cu or Ag NPs (50-60nm, 50mg/kg, i.p./day for 7 days). Rats were subjected to 4h heat stress (HS) in a biological oxygen demand incubator at 38°C (Relative Humidity 45-47%; Wind velocity 22.4-25.6cm/s) that resulted in profound increase in oxidants Luminol, Lucigenin, Malondialdehyde and Myeloperoxidase, and a marked decrease in antioxidant Glutathione. At this time severe reductions in the cerebral blood flow (CBF) was seen together with increased blood-brain barrier (BBB) breakdown and brain edema formation. These pathophysiological responses were exacerbated in NPs treated heat-stressed animals. Pretreatment with cerebrolysin (2.5mL/kg, i.v.) once daily for 3 days significantly attenuated the oxidative stress, BBB breakdown and brain edema and improved CBF in the heat stressed group. The other drugs were least effective on brain pathology following heat stroke. However, in NPs treated heat stressed animals 5mL/kg conventional cerebrolysin and 2.5mL/kg nanowired cerebrolysin is needed to attenuate oxidative stress, BBB breakdown, brain edema and to improve CBF. Interestingly, the other drugs even in higher doses used are unable to alter brain pathologies in NPs and heat stress. These observations are the first to demonstrate that cerebrolysin is the most superior antioxidant and anti-ischemic drug in NPs exposed heat stroke, not reported earlier.

Modulation of Intracellular Copper Levels as the Mechanism of Action of Anticancer Copper Complexes: Clinical Relevance.

Copper (Cu) is a vital element required for cellular growth and development; however, even slight changes in its homeostasis might lead to severe toxicity and deleterious medical conditions. Cancer patients are typically associated with higher Cu content in serum and tumor tissues, indicating increased demand of cancer cells for this micronutrient. Cu is known to readily cycle between the +1 and +2 oxidation state in biological systems. The mechanism of action of Cu complexes is typically based on their redox activity and induction of reactive oxygen species (ROS), leading to deadly oxidative stress. However, there are a number of other biomolecular mechanisms beyond ROS generation that contribute to the activity of anticancer Cu drug candidates. In this review, we discuss how interfering with intracellular Cu balance via either diet modification or addition of inorganic Cu supplements or Cu-modulating compounds affects tumor development, progression, and sensitivity to treatment modalities. We aim to provide the rationale for the use of Cu-depleting and Cu-overloading conditions to generate the best possible patient outcome with minimal toxicity. We also discuss the advantages of the use of pre-formed Cu complexes, such as Cu-(bis)thiosemicarbazones or Cu-N-heterocyclic thiosemicarbazones, in comparison with the in situ formed Cu complexes with metal-binding ligands. In this review, we summarize available clinical and mechanistic data on clinically relevant anticancer drug candidates, including Cu supplements, Cu chelators, Cu ionophores, and Cu complexes.

Proteome Changes Reveal the Protective Roles of Exogenous Citric Acid in Alleviating Cu Toxicity in Brassica napus L.

Citric acid (CA), as an organic chelator, plays a vital role in alleviating copper (Cu) stress-mediated oxidative damage, wherein a number of molecular mechanisms alter in plants. However, it remains largely unknown how CA regulates differentially abundant proteins (DAPs) in response to Cu stress in Brassica napus L. In the present study, we aimed to investigate the proteome changes in the leaves of B. L. seedlings in response to CA-mediated alleviation of Cu stress. Exposure of 21-day-old seedlings to Cu (25 and 50 µM) and CA (1.0 mM) for 7 days exhibited a dramatic inhibition of overall growth and considerable increase in the enzymatic activities (POD, SOD, CAT). Using a label-free proteome approach, a total of 6345 proteins were identified in differentially treated leaves, from which 426 proteins were differentially expressed among the treatment groups. Gene ontology (GO) and KEGG pathways analysis revealed that most of the differential abundance proteins were found to be involved in energy and carbohydrate metabolism, photosynthesis, protein metabolism, stress and defense, metal detoxification, and cell wall reorganization. Our results suggest that the downregulation of chlorophyll biosynthetic proteins involved in photosynthesis were consistent with reduced chlorophyll content. The increased abundance of proteins involved in stress and defense indicates that these DAPs might provide significant insights into the adaptation of Brassica seedlings to Cu stress. The abundances of key proteins were further verified by monitoring the mRNA expression level of the respective transcripts. Taken together, these findings provide a potential molecular mechanism towards Cu stress tolerance and open a new route in accelerating the phytoextraction of Cu through exogenous application of CA in B. napus.

Main Molecular Pathways Associated with Copper Tolerance Response in Imperata cylindrica by de novo Transcriptome Assembly.

The metallophyte Imperata cylindrica inhabits copper (Cu) polluted soils in large areas from Central Chile. Here, we subjected clonal vegetative plantlets to 300 mg Cu kg-1 of substrate for 21 days to identify the main molecular pathways involved in the response to Cu stress. Transcriptomic analyses were performed for shoots and roots, with and without Cu supply. RNA-Seq and de novo transcriptome assembly were performed to identify the gene response associated with molecular mechanisms of Cu tolerance in I. cylindrica. De novo transcriptome revealed a total of 200,521 transcripts (1777 bp) comprising ~91% complete ultra-conserved genes in the eukaryote and Plantae database. The differentially expressed genes (DEGs) in roots were 7386, with 3558 of them being up-regulated and the other 3828 down-regulated. The transcriptome response in shoots was significantly less, showing only 13 up-regulated and 23 down-regulated genes. Interestingly, DEGs mainly related with actin and cytoskeleton formation, and to a minor degree, some DEGs associated with metal transporters and superoxide dismutase activity in root tissues were found. These transcriptomic results suggest that cytoskeleton could be acting as a mechanism of Cu-binding in the root, resulting in a high Cu tolerance response in this metallophyte, which deserve to be analyzed ultra-structurally. Our study contributes to reinforcing the potential of I. cylindrica as a candidate plant species to be used as a phytoremediation agent in Cu-contaminated environments.

Modulation of copper-induced antioxidant defense, Cu transport, and mitophagy by hypoxia in the large yellow croaker (Larimichthys crocea).

This study aimed to investigate the effects of hypoxia on Cu-induced antioxidant defense, Cu transport, and mitophagy in the liver of the large yellow croaker. Fish were exposed to hypoxia (3.0 mg L-1), Cu (120 µg L-1), and hypoxia (3.0 mg L-1) plus Cu (120 µg L-1) for 48 h. Hypoxia exposure increased antioxidant abilities to maintain cellular redox balance. Although Cu exposure alone improved antioxidant defense, Cu transport, and mitophagy, these stress responses could not completely neutralize Cu toxicity, as reflected by the elevated reactive oxygen species (ROS) and lipid peroxidation (LPO) and hepatic vacuoles. When compared with Cu stress alone, hypoxia increased Cu toxicity by inhibiting antioxidant defense, Cu transport, and mitophagy, leading to the increment of mortality, ROS, and LPO, and the deterioration of histological structure. The adverse effects of hypoxia on Cu-induced metal transport and mitophagy might be involved in metal-responsive element-binding transcription factor-1 (MTF-1) and Forkhead box O-3 (FoxO3) signaling pathways, respectively. Overall, hypoxia reduced antioxidant response, Cu transport, and mitophagy in fish exposed to Cu, which contributes to understanding the molecular mechanisms underlying negative effects of hypoxia on Cu toxicity in fish.

Role of Azospirillum brasilense in triggering different Fe chelate reductase enzymes in cucumber plants subjected to both nutrient deficiency and toxicity.

Azospirillum brasilense was reported to up-regulate iron (Fe) uptake mechanisms, such as Fe reduction and rhizosphere acidification, in both Fe sufficient and deficient cucumber plants (Cucumis sativus L.). Strategy I plants take up both Fe and copper (Cu) after their reduction mediated by the ferric-chelate reductase oxidase (FRO) enzyme. Interestingly, in cucumber genome only one FRO gene is reported. Thus, in the present study we applied a bioinformatics approach to identify the member of cucumber FRO gene family and allowed the identification of at least three CsFRO genes, one of which was the already identified, i.e. CsFRO1. The expression patterns of the newly identified transcripts were investigated in hydroponically grown cucumber plants treated with different Fe and Cu nutritional regimes. Gene expression was then correlated with morphological (i.e. root architecture) and physiological (Fe(III) reducing activity) parameters to shed light on: i) the CsFRO homologue responsible of the increased reduction activity in Fe-sufficient plants inoculated with A. brasilense cucumber plants, and ii) the possible effect of A. brasilense in ameliorating the symptoms of Cu toxicity in cucumber plants. The data obtained showed that all the CsFRO genes were expressed in the root tissues of cucumber plants and responded to Cu starvation, combined Cu/Fe deficiency and Cu toxicity. Only CsFRO3 was modulated by the A. brasilense in Fe-sufficient plants suggesting for the first time a different specificity of action of the three isoenzymes depending not only on the nutritional regime (either deficiency or toxicity) but also on the presence of the PGPR. Furthermore, results suggest that the PGPR could even ameliorate the stress symptoms caused by both the double (i.e. Cu and Fe) and Cu deficiency as well as Cu toxicity modulating, on one hand, the growth of the root system and, on the other hand, the root nutrient uptake.

Analysis of copper tolerant rhizobacteria from the industrial belt of Gujarat, western India for plant growth promotion in metal polluted agriculture soils.

Agricultural sites irrigated for long term with water polluted by industrial effluents containing heavy metals might adversely affect the soil microbial communities and crop yield. Hence it is important to study rhizobacterial communities and their metal tolerance in such affected agricultural fields to restore soil fertility and ecosystem. Present work deals with the study of rhizobacterial communities from plants grown in copper (Cu) contaminated agricultural fields along the industrial zone of Gujarat, India and are compared with communities from a Cu mine site. Microbial communities from rhizosphere soil samples varied in the magnitude of their Cu tolerance index indicating differences in long term pollution effects. Culture dependent denaturing gradient gel electrophoresis (CD-DGGE) of bacterial communities revealed the diverse composition at the sampling sites and a reduced total diversity due to Cu toxicity. Analysis of 16S rRNA gene diversity of Cu tolerant rhizobacteria revealed the predominance of Enterobacter spp. and Pseudomonas spp. under Cu stress conditions. Cu tolerant bacterial isolates that were able to promote growth of mung bean plants in vitro under Cu stress were obtained from these samples. Cu tolerant rhizobacterium P36 identified as Enterobacter sp. exhibited multiple plant growth promoting traits and significantly alleviated Cu toxicity to mung bean plants by reducing the accumulation of Cu in plant roots and promoted the plant growth in CuSO4 amended soils.

Copper accumulation in vineyard soils: Rhizosphere processes and agronomic practices to limit its toxicity.

Viticulture represents an important agricultural practice in many countries worldwide. Yet, the continuous use of fungicides has caused copper (Cu) accumulation in soils, which represent a major environmental and toxicological concern. Despite being an important micronutrient, Cu can be a potential toxicant at high concentrations since it may cause morphological, anatomical and physiological changes in plants, decreasing both food productivity and quality. Rhizosphere processes can, however, actively control the uptake and translocation of Cu in plants. In particular, root exudates affecting the chemical, physical and biological characteristics of the rhizosphere, might reduce the availability of Cu in the soil and hence its absorption. In addition, this review will aim at discussing the advantages and disadvantages of agronomic practices, such as liming, the use of pesticides, the application of organic matter, biochar and coal fly ashes, the inoculation with bacteria and/or mycorrhizal fungi and the intercropping, in alleviating Cu toxicity symptoms.

Dynamics of the diversity and structure of the overall and nitrifying microbial community in activated sludge along gradient copper exposures.

Diversity and composition of the microbial community, especially the nitrifiers, are essential to the treatment efficiency of wastewater in activated sludge systems. Heavy metals commonly present in the wastewater influent such as Cu can alter the community structure of nitrifiers and lower their activity. However, the dynamics of microbial community along a gradient of metal exposure have largely been unexplored, partially due to the limitations in traditional molecular methods. This study explored the dynamics regarding the diversity and community structures of overall and nitrifying microbial communities in activated sludge under intermittent Cu gradient loadings using Illumina sequencing. We created a new local nitrifying bacterial database for sequence BLAST searches. High Cu loadings (>10.9 mg/L) impoverished microbial diversity and altered the microbial community. Overall, Proteobacteria was the predominant phylum in the activated sludge system, in which Zoogloea, Thauera, and Dechloromonas (genera within the Rhodocyclaceae family of the Beta-proteobacteria class) were the dominant genera in the presence of Cu. The abundance of unclassified bacteria at the phylum level increased substantially with increasing Cu loadings. Nitrosomonas and Nitrospira were the predominant nitrifiers. The nitrifying bacterial community changed through increasing abundance and shifting to Cu-tolerant species to reduce the toxic effects of Cu. Our local nitrifying bacterial database helped to improve the resolution of bacterial identification. Our results provide insights into the dynamics of microbial community in response to various metal concentrations in activated sludge systems and improve our understanding regarding the effect of metals on wastewater treatment efficiency.

Long-term passive restoration following fluvial deposition of sulphidic copper tailings: nature filters out the solutions.

Despite the growing popularity of ecological restoration approach, data on primary succession on toxic post-mining substrates, under site environmental conditions which considerably differ from the surrounding environment, are still scarce. Here, we studied the spontaneous vegetation development on an unusual locality created by long-term and large-scale fluvial deposition of sulphidic tailings from a copper mine in a pronouncedly xerothermic, calcareous surrounding. We performed multivariate analyses of soil samples (20 physical and chemical parameters) and vegetation samples (floristic and structural parameters in three types of occurring forests), collected along the pollution gradients throughout the affected floodplain. The nature can cope with two types of imposed constraints: (a) excessive Cu concentrations and (b) very low pH, combined with nutrient deficiency. The former will still allow convergence to the original vegetation, while the latter will result in novel, depauperate assemblages of species typical for cooler and moister climate. Our results for the first time demonstrate that with the increasing severity of environmental filtering, the relative importance of the surrounding vegetation for primary succession strongly decreases.

The influence of liming on soil chemical properties and on the alleviation of manganese and copper toxicity in Juglans regia, Robinia pseudoacacia, Eucalyptus sp. and Populus sp. plantations.

Juglans regia, Robinia pseudoacacia, Eucalyptus sp. and Populus sp. plantations, suffering from Mn and Cu toxicity, were limed in order to reduce Cu and Mn solubility in soil. The purposes of the present work were: i) to study the changes in soil chemical properties after the addition of CaCO3, ii) to investigate the influence of liming on the reduction of Mn and Cu toxicity. After the addition of CaCO3 (three applications, during three successive years), pH and CaCO3 content were significantly increased, while organic C and N were significantly reduced. Exchangeable Ca concentrations have been slightly, or significantly, increased, while those of Mg have been decreased; in addition, ratios Ca/Mg and C/N have been significantly increased after liming. Impressive reductions of DTPA extractable Cu and Mn concentrations (more than 10 times in most cases) were recorded. It was also found that trees without Mn and Cu toxicity symptoms (healthy tress) before liming did not have, in many cases, significantly greater leaf Mn, Cu and Fe concentrations, than trees after soil liming (all the trees were healthy). This probably happened because excess Mn and Cu quantities had been accumulated into their root system. Finally, leaf Mn, Cu and Zn concentrations of trees suffering from toxicity were significantly decreased after soil liming, while leaf Fe concentrations, in all the plant species studied, were increased.

Copper regulates primary root elongation through PIN1-mediated auxin redistribution.

The heavy metal copper (Cu) is an essential microelement required for normal plant growth and development, but it inhibits primary root growth when in excess. The mechanism underlying how excess Cu functions in this process remains to be further elucidated. Here, we report that a higher concentration of CuSO4 inhibited primary root elongation of Arabidopsis seedlings by affecting both the elongation and meristem zones. In the meristem zone, meristematic cell division potential was reduced by excess Cu. Further experiments showed that Cu can modulate auxin distribution, resulting in higher auxin activities in both the elongation and meristem zones of Cu-treated roots based on DR5::GUS expression patterns. This Cu-mediated auxin redistribution was shown to be responsible for Cu-mediated inhibition of primary root elongation. Additional genetic and physiological data demonstrated that it was PINFORMED1 (PIN1), but not PIN2 or AUXIN1 (AUX1), that regulated this process. However, Cu-induced hydrogen peroxide accumulation did not contribute to Cu-induced auxin redistribution for inhibition of root elongation. When the possible role of ethylene in this process was analyzed, Cu had a similar impact on the root elongation of both the wild type and the ein2-1 mutant, implying that Cu-mediated inhibition of primary root elongation was not due to the ethylene signaling pathway.