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We evaluated the mechanisms that control Cd accumulation and distribution, and the mechanisms that protect the photosynthetic apparatus of Brachiaria decumbens Stapf. cv. Basilisk and Panicum maximum Jacq. cv. Massai from Cd‐induced oxidative stress, as well as the effects of simulated summer or winter conditions on these mechanisms. Both grasses were grown in unpolluted and Cd‐polluted Oxisol (0.63 and 3.6 mg Cd kg−1 soil, respectively) at summer and winter conditions. Grasses grown in the Cd‐polluted Oxisol presented higher Cd concentration in their tissues in the winter conditions, but the shoot biomass production of both grasses was not affected by the experimental conditions. Cadmium was more accumulated in the root apoplast than the root symplast, contributing to increase the diameter and cell layers of the cambial region of both grasses. Roots of B. decumbens were more susceptible to disturbed nutrients uptake and nitrogen metabolism than roots of P. maximum. Both grasses translocated high amounts of Cd to their shoots resulting in oxidative stress. Oxidative stress in the leaves of both grasses was higher in summer than winter, but only in P. maximum superoxide dismutase (SOD) and ascorbate peroxidase (APX) activities were increased. However, CO2 assimilation was not affected due to the protection provided by reduced glutathione (GSH) and phytochelatins (PCs) that were more synthesized in shoots than roots. In summary, the root apoplast was not sufficiently effective to prevent Cd translocation from roots to shoot, but GSH and PCs provided good protection for the photosynthetic apparatus of both grasses. [ABSTRACT FROM AUTHOR]

Cable bacteria are an emerging class of electroactive organisms that sustain unprecedented long‐range electron transport across centimeter‐scale distances. The local pathways of the electrical currents in these filamentous microorganisms remain unresolved. Here, the electrical circuitry in a single cable bacterium is visualized with nanoscopic resolution using conductive atomic force microscopy. Combined with perturbation experiments, it is demonstrated that electrical currents are conveyed through a parallel network of conductive fibers embedded in the cell envelope, which are electrically interconnected between adjacent cells. This structural organization provides a fail‐safe electrical network for long‐distance electron transport in these filamentous microorganisms. The observed electrical circuit architecture is unique in biology and can inspire future technological applications in bioelectronics. [ABSTRACT FROM AUTHOR]

The basidiomycete Suillus luteus is an important member of the ectomycorrhizal community that thrives in heavy metal polluted soils covered with pioneer pine forests. This study aimed to identify potential heavy metal chelators in S. luteus. Two metallothionein (MT) coding genes, SlMTa and SlMTb, were identified. When heterologously expressed in yeast, both SlMTa and SlMTb can rescue the Cu sensitive mutant from Cu toxicity. In S. luteus, transcription of both SlMTa and SlMTb is induced by Cu but not Cd or Zn. Several putative Cu-sensing and metal-response elements are present in the promoter sequences. These results indicate that SlMTa and SlMTb function as Cu-thioneins. Homologs of the S. luteus MTs are present in 49 species belonging to 10 different orders of the subphylum Agaricomycotina and are remarkably conserved. The length of the proteins, number and distribution of cysteine residues indicate a novel family of fungal MTs. The ubiquitous and highly conserved features of these MTs suggest that they are important for basic cellular functions in species in the subphylum Agaricomycotina. [ABSTRACT FROM AUTHOR]

Cadmium (Cd) exposure can disturb the homeostasis of essential elements. In Arabidopsis thaliana, Cd induces a squamosa promoter binding protein-like 7 (SPL7)-dependent Cu deficiency response. We investigated how Cd induces a Cu deficiency response. The Cu deficiency response consists of the active SPL7 transcription factor binding to GTAC motifs in promoters of among others several Cu transporters, a Cu chaperone, and cupro-miRNAs to regulate Cu homeostasis. We demonstrated that the addition of supplemental Cu to Cd-exposed A. thaliana plants diminished the Cu deficiency response in roots, while it even disappeared in leaves. Exposure of plants to Cd in combination with extra Cu reduced Cd levels in both roots and leaves resulting in an improved cellular oxidative state. Furthermore, we demonstrated a role for phytochelatins (PCs) in the Cd-induced Cu deficiency response, because it was reduced in roots of cad1-3 mutant plants exposed to Cd. In conclusion, a working mechanism is provided in which it is suggested that Cd increases PC levels that can complex both Cd and Cu. This results in cellular Cu deficiency and subsequently the activation of SPL7 and hence the induction of the Cu deficiency response. [ABSTRACT FROM AUTHOR]

BACKGROUND The disposal of metal-enriched plant biomass is an important and still unresolved issue associated with phytoextraction. Therefore, the aim of this study was to valorise these contaminated biomasses into valuable products, i.e. bio-chars and activated carbons ( ACs), through slow pyrolysis followed by steam activation. The effects of pyrolysis/steam activation on metal availability in the chars/ ACs were also evaluated. In addition, Cr( VI) adsorption by these ACs in aqueous solutions was studied. RESULTS Relevant chars/ ACs yields were obtained due to applied thermochemical treatments. Leachability of heavy metals from those products was limited indicating that they may be safely applied as adsorption media. Prepared ACs demonstrated good Cr( VI) adsorption capacities occurring through a 'coupled adsorption-reduction' mechanism. It was found that Cr( VI) removal strongly depended on pH of Cr( VI) solutions and was higher for ACs activated at 800 °C. Among applied isotherm adsorption models, i.e. Langmuir and Freundlich, a better fit was obtained with the Langmuir model corresponding to monolayer coverage sorption of Cr( VI) on ACs surface. CONCLUSIONS Heavy metals enriched biomass can be successfully valorised through slow pyrolysis/steam activation into value added products, i.e. ACs that can be used as efficient and safe adsorbents for Cr( VI) removal. © 2016 Society of Chemical Industry [ABSTRACT FROM AUTHOR]

The association of plants and rhizospheric bacteria provides a successful strategy to clean up contaminated soils. The purpose of this work was to enhance diesel degradation in Lupinus luteus rhizosphere by inoculation with selected bacterial strains: a diesel degrader (D), plant growth-promoting (PGP) strains, or a combination (D+PGP). Plants were set up in pots with the A or B horizon of an umbric Cambisol (ACamb and BCamb) spiked with diesel (1.25%, w/w). After 1 mo, the dissipation of diesel range organics (DRO) with respect to t = 0 (i.e., 1 wk after preparing the pots with the seedlings) concentration was significantly higher in inoculated than in noninoculated (NI) pots: The highest DRO losses were found in ACamb D+PGP pots (close to 15-20% higher than NI) and in BCamb D pots (close to 10% higher). The water-extractable DRO fraction was significantly higher at t = 30 d (15-25%) compared with t = 0 (<5%), probably due to the effects of plant root exudates and biosurfactants produced by the degrader strain. The results of this experiment reflect the importance of the partnerships between plants and bacterial inoculants and demonstrate the relevance of the effect of bacterial biosurfactants and plant root exudates on contaminant bioavailability, a key factor for enhancing diesel rhizodegradation. The association of lupine with D and PGP strains resulted in a promising combination for application in the rhizoremediation of soils with moderate diesel contamination. [ABSTRACT FROM AUTHOR]

Plant-associated bacteria are intensively investigated concerning their characteristics for plant growth promotion, biocontrol mechanisms and enhanced phytoremediation efficiency. To obtain endophytes, different sampling and isolation protocols are used although their representativeness is not always clearly demonstrated. The objective of this study was to acquire representative pictures of the cultivable bacterial root, stem and leaf communities for all B rassica napus L. individuals growing on the same field. For each plant organ, genotypic identifications of the endophytic communities were performed using three replicates. Root replicates were composed of three total root systems, whereas stem and leaf replicates needed to consist of six independent plant parts in order to be representative. Greater variations between replicates were found when considering phenotypic characteristics. Correspondence analysis revealed reliable phenotypic results for roots and even shoots, but less reliable ones for leaves. Additionally, realistic Shannon- Wiener biodiversity indices were calculated for all three organs and showed similar Evenness factors. Furthermore, it was striking that all replicates and thus the whole plant contained P seudomonas and B acillus strains although aboveground and belowground plant tissues differed in most dominant bacterial genera and characteristics. [ABSTRACT FROM AUTHOR]

Gentle remediation options (GRO) are risk management strategies/technologies that result in a net gain (or at least no gross reduction) in soil function as well as risk management. They encompass a number of technologies, including the use of plant (phyto-), fungi (myco-), and/or bacteria-based methods, with or without chemical soil additives or amendments, for reducing contaminant transfer to local receptors by in situ stabilization, or extraction, transformation, or degradation of contaminants. Despite offering strong benefits in terms of risk management, deployment costs, and sustainability for a range of site problems, the application of GRO as practical on-site remedial solutions is still in its relative infancy, particularly for metal(loid)-contaminated sites. A key barrier to wider adoption of GRO relates to general uncertainties and lack of stakeholder confidence in (and indeed knowledge of) the feasibility or reliability of GRO as practical risk management solutions. The GREENLAND project has therefore developed a simple and transparent decision support framework for promoting the appropriate use of gentle remediation options and encouraging participation of stakeholders, supplemented by a set of specific design aids for use when GRO appear to be a viable option. The framework is presented as a three phased model or Decision Support Tool (DST), in the form of a Microsoft Excel-based workbook, designed to inform decision-making and options appraisal during the selection of remedial approaches for contaminated sites. The DST acts as a simple decision support and stakeholder engagement tool for the application of GRO, providing a context for GRO application (particularly where soft end-use of remediated land is envisaged), quick reference tables (including an economic cost calculator), and supporting information and technical guidance drawing on practical examples of effective GRO application at trace metal(loid) contaminated sites across Europe. This article introduces the decision support framework. ©2015 Wiley Periodicals, Inc. [ABSTRACT FROM AUTHOR]

The impact of metal pollution on plant communities has been studied extensively in the past, but little is known about the effects of metal pollution on fungal communities that occur in metal-polluted soils. Metal-tolerant ecotypes of the ectomycorrhizal fungus S uillus luteus are frequently found in pioneer pine forests in the Campine region in Belgium on metal-polluted soils. We hypothesized that metal pollution would play an important role in shaping below-ground fungal communities that occur in these soils and that S uillus luteus would be a dominant player. To test these hypotheses, the fungal communities in a young pine plantation in soil polluted with zinc, and cadmium were studied using 454 amplicon pyrosequencing. Results show that zinc, cadmium and soil organic matter content were strongly correlated with the fungal community composition, but no effects on fungal diversity were observed. As hypothesized, S . luteus was found to be a dominant member of the studied fungal communities. However, other dominant fungal species, such as Sistotrema sp., W ilcoxina mikolae and C adophora finlandica were found as well. Their presence in metal-polluted sites is discussed. [ABSTRACT FROM AUTHOR]

Although the importance of plant-associated microorganisms for plant growth and health was getting more recognition recently, the role of seed-associated microorganisms, and especially seed endophytic bacteria, still is underestimated. Nevertheless, these associations could be beneficial for germination and seedling establishment as seed endophytic bacteria are already present in these very early plant growth stages. Moreover, bacteria with beneficial characteristics can be selected by the plants and could be transferred via the seed to benefit the next generation. In this paper, the current literature concerning bacterial endophytes that have been isolated from seeds of different plant species is reviewed. Their colonization routes, localization inside seeds and mode of transmission as well as their role and fate during germination and seedling development are discussed. At the end, some examples of bacterial seed endophytes with applications as a plant growth-promoting or biocontrol agent are given. [ABSTRACT FROM AUTHOR]

The presence of explosives in soils and the interaction with drought stress and nutrient limitation are among the environmental factors that severely affect plant growth on military soils. In this study, we seek to isolate and identify the cultivable bacteria of a 2,4-dinitrotoluene ( DNT) contaminated soil ( DS) and an adjacent grassland soil ( GS) of a military training area aiming to isolate new plant growth-promoting ( PGP) and 2,4- DNT-degrading strains. Metabolic profiling revealed disturbances in Ecocarbon use in the bare DS; isolation of cultivable strains revealed a lower colony-forming-unit count and a less diverse community associated with DS in comparison with GS. New 2,4- DNT-tolerant strains were identified by selective enrichments, which were further characterized by auxanography for 2,4- DNT use, resistance to drought stress, cold, nutrient starvation and PGP features. By selecting multiple beneficial PGP and abiotic stress-resistant strains, efficient 2,4- DNT-degrading consortia were composed. After inoculation, consortium UHasselt Sofie 3 with seven members belonging to B urkholderia, V ariovorax, B acillus, P seudomonas and R alstonia species was capable to successfully enhance root length of A rabidopsis under 2,4- DNT stress. After 9 days, doubling of main root length was observed . Our results indicate that beneficial bacteria inhabiting a disturbed environment have the potential to improve plant growth and alleviate 2,4- DNT stress. [ABSTRACT FROM AUTHOR]

To date, almost no information is available in roots and shoots of the model plant Arabidopsis thaliana about the hierarchic relationship between metal accumulation, phytohormone levels, and glutathione/phytochelatin content, and how this relation affects root development. For this purpose, specific concentrations of cadmium, copper and zinc, alone or in triple combination, were supplied for 12 days to in vitro growing seedlings. The accumulation of these metals was measured in roots and shoots, and a significant competition in metal uptake was observed. Microscopic analyses revealed that root morphology was affected by metal exposure, and that the levels of trans‐zeatin riboside, dihydrozeatin riboside, indole‐3‐acetic acid and the auxin/cytokinin ratio varied accordingly. By contrast, under metal treatments, minor modifications in gibberellic acid and abscisic acid levels occurred. Real‐time polymerase chain reaction analysis of some genes involved in auxin and cytokinin synthesis (e.g. AtNIT in roots and AtIPT in shoots) showed on average a metal up‐regulated transcription. The production of thiol‐peptides was induced by all the metals, alone or in combination, and the expression of the genes involved in thiol‐peptide synthesis (AtGSH1, AtGSH2, AtPCS1 and AtPCS2) was not stimulated by the metals, suggesting a full post‐transcriptional control. Results show that the Cd/Cu/Zn‐induced changes in root morphology are caused by a hormonal unbalance, mainly governed by the auxin/cytokinin ratio. [ABSTRACT FROM AUTHOR]

Uranium (U) causes oxidative stress in Arabidopsis thaliana plants grown at pH 5.5. However, U speciation and its toxicity strongly depend on environmental parameters, for example pH. It is unknown how different U species determine U uptake and translocation within plants and how they might affect the oxidative defense mechanisms of these plants. The present study analyzed U uptake and oxidative stress-related responses in A. thaliana (Columbia ecotype) under contrasted U chemical speciation conditions. The 18-d-old seedlings were exposed for 3 d to 25 µM U in a nutrient solution of which the pH was adjusted to 4.5, 5.5, 6.5, or 7.5. Results indicate that there is a different rate of U uptake and translocation at the different pHs, with high uptake and low translocation at low pH and lower uptake but higher translocation at high pH. After U exposure, an increased glutathione reductase activity and total glutathione concentration were observed in U-exposed roots, pointing toward an important role for glutathione in the root defense system against U either by chelation or by antioxidative defense mechanisms. In leaves, antioxidative defense mechanisms were activated on U exposure, indicated by increased superoxide dismutase and catalase activity. As it seems that U toxicity is influenced by pH, it is important to consider site-specific characteristics when making U risk assessments. Environ Toxicol Chem 2013;32:2125-2133. © 2013 SETAC [ABSTRACT FROM AUTHOR]

Plants suffering from abiotic stress are commonly facing an enhanced accumulation of reactive oxygen species ( ROS) with damaging as well as signalling effects at organellar and cellular levels. The outcome of an environmental challenge highly depends on the delicate balance between ROS production and scavenging by both enzymatic and metabolic antioxidants. However, this traditional classification is in need of renewal and reform, as it is becoming increasingly clear that soluble sugars such as disaccharides, raffinose family oligosaccharides and fructans - next to their associated metabolic enzymes - are strongly related to stress-induced ROS accumulation in plants. Therefore, this review aims at extending the current concept of antioxidants functioning during abiotic stress, with special focus on the emanate role of sugars as true ROS scavengers. Examples are given based on their cellular location, as different organelles seem to exploit distinct mechanisms. Moreover, the vacuole comes into the picture as important player in the ROS signalling network of plants. Elucidating the interplay between the mechanisms controlling ROS signalling during abiotic stress will facilitate the development of strategies to enhance crop tolerance to stressful environmental conditions. [ABSTRACT FROM AUTHOR]

The hypothesis that mitogen-activated protein kinase ( MAPK) signalling is important in plant defences against metal stress has become accepted in recent years. To test the role of oxidative signal-inducible kinase ( OXI1) in metal-induced oxidative signalling, the responses of oxi1 knockout lines to environmentally realistic cadmium ( Cd) and copper ( Cu) concentrations were compared with those of wild-type plants. A relationship between OXI1 and the activation of lipoxygenases and other initiators of oxylipin production was observed under these stress conditions, suggesting that lipoxygenase-1 may be a downstream component of OXI1 signalling. Metal-specific differences in OXI1 action were observed. For example, OXI1 was required for the up-regulation of antioxidative defences such as catalase in leaves and Fe-superoxide dismutase in roots, following exposure to Cu, processes that may involve the MEKK1- MKK2- WRKY25 cascade. Moreover, the induction of Cu/Zn superoxide dismutases in Cu-exposed leaves was regulated by OXI1 in a manner that involves fluctuations in the expression of miRNA398. These observations contrast markedly with the responses to Cd exposure, which also involves OXI1-independent pathways but rather involves changes in components mediating intracellular communication. [ABSTRACT FROM AUTHOR]

Phytoextraction has been reported as an economically and ecologically sound alternative for the remediation of metal-contaminated soils. Willow is a metal phytoextractor of interest because it allows to combine a gradual contaminant removal with production of biomass that can be valorized in different ways. In this work two willow clones growing on a metal-contaminated site were selected: ' Belgisch Rood' ( BR) with a moderate metal extraction capacity and ' Tora' ( TO) with a twice as high metal accumulation. All cultivable bacteria associated with both willow clones were isolated and identified using 16SrDNA ARDRA analysis followed by 16SrDNA sequencing. Further all isolated bacteria were investigated for characteristics that might promote plant growth (production of siderophores, organic acids and indol acetic acid) and for their metal resistance. The genotypic and phenotypic characterization of the isolated bacteria showed that the TO endophytic bacterial population is more diverse and contains a higher percentage of metal-resistant plant growth promoting bacteria than the endophytic population associated with BR. We hypothesize that the difference in the metal accumulation capacity between BR and TO clones might be at least partly related to differences in characteristics of their associated bacterial population. [ABSTRACT FROM AUTHOR]

Adaptive Zn and Cd tolerance have evolved in populations of the ectomycorrhizal fungus Suillus luteus. When exposed to high concentrations of both metals in vitro, a one-sided antagonism was apparent in the Zn- and Cd-tolerant isolates. Addition of high Zn concentrations restored growth of Cd-stressed isolates, but not vice versa. The antagonistic effect was not detected in a S. luteus isolate from non-contaminated land and in Paxillus involutus. The fungi were inoculated on pine seedlings and subsequently exposed to ecologically relevant Zn and Cd concentrations in single and mixed treatments. The applied doses severely reduced nutrient acquisition of non-mycorrhizal pines and pines inoculated with metal-sensitive S. luteus. Highest translocation of Zn and Cd to shoots occurred in the same plants. Seedlings inoculated with fungi collected from the polluted site reduced metal transfer to their host and maintained nutrient acquisition under high metal exposure. The isolate showing highest tolerance in vitro also offered best protection in symbiosis. The antagonistic effect of high Zn on Cd toxicity was confirmed in the plant experiment. The results indicate that a Zn- and Cd-polluted soil has selected ectomycorrhizal fungi that are able to survive and protect their phytobiont from nutrient starvation and excessive metal uptake. [ABSTRACT FROM AUTHOR]

The genetic structure of populations of the ectomycorrhizal basidiomycete Suillus luteus in heavy metal polluted and nonpolluted areas was studied. Sporocarps were collected at nine different locations and genotyped at four microsatellite loci. Six of the sampling sites were severely contaminated with heavy metals and were dominated by heavy metal-tolerant individuals. Considerable genetic diversity was found within the geographical subpopulations, but no reduction of the genetic diversity, current or historic, was observed in subpopulations inhabiting polluted soils. The genetic differentiation between the geographical subpopulations was low, and no evidence for clustering of subpopulations from polluted soils vs. subpopulations from nonpolluted soils was found. These results indicate that heavy metal pollution has a limited effect on the genetic structure of S. luteus populations, and this may be due to the high frequency of sexual reproduction and extensive gene flow in S. luteus, which allows rapid evolution of the tolerance trait while maintaining high levels of genetic diversity. [ABSTRACT FROM AUTHOR]

We investigated the effect of cadmium (Cd) on leaves of 3-week-old Arabidopsis thaliana with a particular interest on glutathione (GSH) production and consumption, and antioxidative defence. Plants were exposed to either 1 or 10 μM Cd for 1 week. Several genes and correspondent proteins known to participate in the regulation of redox states in plants were analysed in Cd-stressed and unstressed plants. Cd induced a significant increase in the messenger RNA level of genes involved in GSH synthesis (gsh1 and gsh2) and phytochelatin synthase (pcs1). We observed a significant decrease of reduced GSH in Cd-treated plants. In parallel, an increase of phytochelatin (PC), predominantly PC2 was observed. Cd treatment increased the accumulation of glutathione disulphide (GSSG), keeping the GSH/GSSG ratio lower than in control plants. The accumulation of GSSG was accompanied by a decrease of the glutathione reductase (gr) transcript level, while the activities of GR and nicotinamide nucleotide phosphate-reducing enzymes were significantly enhanced. The antioxidative defence mechanism related to the ascorbate (AsA)–GSH cycle was studied in parallel. A general increase of reactive oxygen species scavenging enzymes such as ascorbate peroxidase, catalase or superoxide dismutase was observed. Our data suggest that the plants respond to Cd stress by the AsA–GSH defence network at both transcriptional and enzymatic level. Taken together, it appears that Arabidopsis plants exposed to 1 μM Cd were able to adopt a new metabolic equilibrium, allowing them to cope with this metal. However, when exposed to 10 μM Cd, loss of cellular redox homeostasis resulted in oxidative stress and toxicity. [ABSTRACT FROM AUTHOR]

• We examined the subcellular cadmium (Cd) localization in roots and leaves of wild-type Arabidopsis thaliana (ecotype Columbia) exposed to environmentally relevant Cd concentrations. • Energy-dispersive X-ray microanalysis (EDXMA) was performed on high-pressure frozen and freeze-substituted tissues. • In the root cortex, Cd was associated with phosphorus (Cd/P) in the apoplast and sulfur (Cd/S) in the symplast, suggesting phosphate and phytochelatin sequestration, respectively. In the endodermis, sequestration of Cd/S was present as fine granular deposits in the vacuole and as large granular deposits in the cytoplasm. In the central cylinder, symplastic accumulation followed a distinct pattern illustrating the importance of passage cells for the uptake of Cd. In the apoplast, a shift of Cd/S granular deposits from the middle lamella towards the plasmalemma was observed. Large amounts of precipitated Cd in the phloem suggest retranslocation from the shoot. In leaves, Cd was detected in tracheids but not in the mesophyll tissue. • Extensive symplastic and apoplastic sequestration in the root parenchyma combined with retranslocation via the phloem confirms the excluder strategy of Arabidopsis thaliana. [ABSTRACT FROM AUTHOR]

Investigates the zinc tolerance of four ectomycorrhizal fungi, Suillus luteus, Suillus bovinus, Rhizopogon luteolus and Paxillus involutus. Correlation between zinc concentration in the needles of pines and zinc smelters distance; Distribution of zinc tolerance in the Suillus luteus populations; Impact of heavy metal pollution on plant communities and populations.

Here we investigated zinc tolerance of ectomycorrhizal Scots pine (Pinus sylvestris) seedlings. An ectomycorrhizal genotype of Suillus bovinus, collected from a Zn-contaminated site and showing adaptive Zn tolerance in vitro, was compared with a nonadapted isolate from a nonpolluted area. A dose-response experiment was performed. Dynamics of plant and fungal development, and phosphate and ammonium uptake capacity, were assessed under increasing Zn stress. Effects of Zn on transpiration, nutrient content and Zn accumulation were analysed. Significant Zn-inoculation interaction effects were observed for several responses measured, including uptake rates of phosphate and ammonium; phosphorus, iron and Zn content in shoots; transpiration; biomass of external mycelia; and fungal biomass in roots. The Zn-tolerant S. bovinus genotype was particularly efficient in protecting pines from Zn stress. The growth of a Zn-sensitive genotype from a normal wild-type population was inhibited at high Zn concentrations, and this isolate could not sustain the pines' acquisition of nutrients. This study shows that well adapted microbial root symbionts are a major component of the survival strategy of trees that colonize contaminated soils. [ABSTRACT FROM AUTHOR]

Sequestration of Pb by synthetic chelates has been reported to increase bioavailability, uptake, and translocation of this metal in plants. In this work the potential phytotoxic effects of Pb-EDTA were investigated in Phaseolus vulgaris L. cv. Limburgse vroege plants grown on hydroponics. Addition of 50 µM Pb-EDTA to the nutrient solution caused a significant induction of syringaldazine peroxidase (SPOD; EC 1.11.1.7) in roots and primary leaves and guaiacol peroxidase (GPOD; EC 1.11.1.7) in leaves. Addition of 100 µM Pb-EDTA further exacerbated ascorbate peroxidase (APOD; EC 1.11.1.11), GPOD, dehydroascorbate reductase (DHAR; EC 1.8.5.1), glutathione reductase (GR; EC 1.6.4.2) and malic enzyme (ME; EC 1.1.1.40) in roots and APOD and ME in primary leaves. Addition of 200 µM Pb-EDTA also induced DHAR in leaves. This induction of peroxidases (SPOD, GPOD, APOD), enzymes of the ascorbate-glutathione cycle (DHAR, GR in roots) and of an NADP+ reducing enzyme in roots and primary leaves indicates that oxidative stress has been initiated. At 200 µM Pb-EDTA, chlorophyll a and b content in leaves was significantly reduced while visible effects on root morphology and shoot length were observed, while no significant morphological effects were found in the leaves, confirming the sensitive character of the measured enzymes as plant stress indicators. Elevation of the Pb-EDTA concentration in the growth medium significantly reduced the content of Ca, Fe, Mn and Zn taken up by plants, probably due to ion leakage as a result of observed toxicity. Addition of up to 200 µM EDTA increased chelation of divalent cations in nutrient solution resulting in reduced plant uptake of Zn, Cu, Fe and Mn. This did not result in phytotoxicity. [ABSTRACT FROM AUTHOR]

Summary • Change in the copper (Cu) sensitivity of Pinus sylvestris is presented, in response to the ectomycorrhizal fungi, Suillus bovinus and Thelephora terrestris , common mycobionts on metal contaminated sites. • Seedlings grown under phosphorus (P) limitation were exposed to a range of Cu concentrations. Plant and fungal development, P nutrition, sorption of Cu on roots and external mycelia as well as transfer of Cu to shoots were assessed. • Root growth and P nutrition were severely inhibited in nonmycorrhizal pines at elevated Cu compared with mycorrhizal plants. Excess Cu had little effect on the development of mycorrhizal roots and mycelia. Thelephora terrestris was less sensitive to Cu stress than S. bovinus . The extraradical mycelium of S. bovinus retained large amounts of Cu. However, binding of Cu in fungal tissue was not a prerequisite for low Cu sensitivity since T. terrestris absorbed considerably less Cu that S. bovinus . • Both ectomycorrhizal fungi protect P. sylvestris against Cu toxicity; a benefit that was not due to a metal dilution effect. The mechanisms of mycorrhizal amelioration of Cu toxicity are probably diverse and species-dependent. [ABSTRACT FROM AUTHOR]

Copper-imposed oxidative stress and antioxidative defence responses were investigated in the primary leaves of Phaseolus vulgaris L. plants grown on hydroponics containing 50 μM CuSO[sub 4]. Copper mainly accumulates in roots; therefore, an increase of the copper content in the leaves was only observed 48 h after the start of the copper supply. Nevertheless, an increase of the thiobarbituric acid reactive metabolites (TBArm) content, an indication of stress, occurred immediately following copper application. Because the ascorbate-glutathione pathway is considered as a major antioxidative defence mechanism, the evolution of the enzymes and the related metabolites involved in this pathway were studied in the primary leaves as a function of plant copper assimilation. The capacities of monodehydroascorbate reductase (EC 1.6.5.4), dehydroascorbate reductase (EC 1.8.5.1), and glutathione reductase (EC 1.6.4.2) were increased before elevated amounts of copper could be detected in the leaves. The early enhancement of glutathione reductase was only temporary. After copper accumulation in the leaves, a second increase of the glutathione reductase capacity and also an increase of the ascorbate peroxidase capacity (EC 1.11.1.11) were observed. These changes in enzymatic capacity modified the level of the metabolites involved. Increase of the ascorbate pool and maintenance in its reduced form was observed immediately after the start of the treatment. In the beginning of the experiment, the glutathione disulphide/reduced glutathione ratio was higher in the treated plants as compared to the controls. However, towards the end of the experiment, the total glutathione pool, as well as the reduced glutathione content, increased, resulting in a lower ratio value for the treated plants. In conlusion, copper-imposed oxidative stress, as well as the antioxidative defence response in the leaves, appears to be biphasic. An indirect preventive effect on the antioxidative defence system was observed during the first phase before the leaf copper content increased. A root‐to‐shoot signalling system appears to be involved. Direct oxidation by copper of reduced cell metabolites occurred during the second phase when the leaf copper content was enhanced. [ABSTRACT FROM AUTHOR]

Twenty-one isolates of the ectomycorrhizal fungus Suillus luteus were screened for their tolerance to the heavy metals Zn, Cd, Cu and Ni, measured as inhibition of radial growth and biomass production. Two populations from even-aged pine stands were investigated: 10 isolates were obtained from an area polluted with high levels of Zn, Cd and Cu, and 11 isolates were obtained from a control population located in a nearby unpolluted area. RFLP patterns of the internal transcribed spacer region of the isolates confirmed the morphological identification of the carpophores. All isolates were maintained on basic medium without elevated metals to avoid phenotypically acquired metal tolerance. The in vitro Zn and Cd tolerance of the S. luteus isolates from the polluted habitat were significantly higher than the tolerances measured in the isolates from the nonpolluted site. This observation suggests that the elevated soil metal concentrations might be responsible for the evolution of adaptive Zn and Cd tolerance. Tolerance was maintained in an isolate not exposed to elevated metals for 3 yr. The two S. luteus populations did not differ in tolerance to Cu and Ni. The mechanisms for the adaptive Zn and Cd tolerance are not identical as there was no correlation between response to the two metals; the most Zn-tolerant isolate was the most sensitive for Cd in the metal-tolerant population. Zinc did not accumulate in basidiocarp tissue, whereas Cd levels in basidiocarps were significantly higher in the population on the polluted site. Inter-simple sequence-repeat fingerprints showed that 90% of the isolates were from different individuals. The genetic variation in the population from the unpolluted site was considerably larger than that observed at the polluted site. [ABSTRACT FROM AUTHOR]

The involvement of the ascorbate-glutathione cycle in the defence against Cu-induced oxidative stress was studied in the roots of Phaseolus vulgaris L. cv. Limburgse vroege. All the enzymes of this cycle [ascorbate peroxidase (APOD), 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] were increased, and the total ascorbate and glutathione pools rose after a 15 μM root Cu treatment. In the first hours after the start of the experiment, the accumulation of dehydroascorbate (DHA), formed as a result of a Cu-mediated direct oxidation of ascorbate (AA), was limited by a non-enzymatic reduction using glutathione (GSH) as the reductant. At 24 h, the enzyme capacities of both DHAR and GR were increased to maintain the redox status of the AA and GSH pools. After 72 h of Cu application, the DHAR capacity was inhibited and MDHAR was responsible for maintaining the AA pool in its reduced form. Although the GR capacity was enhanced after 72 h in the treated plants, the GSSG/GSH ratio was increased. This could be due to direct participation of GSH in the detoxification of Cu through reduction and complexation. [ABSTRACT FROM AUTHOR]

Eight microsatellite loci were isolated from the ectomycorrhizal basidiomycete Suillus luteus using a dual-suppression-polymerase chain reaction (PCR) method. In a test sample of 40 isolates, the total number of alleles per locus and the expected heterozygosities ranged from five to 16 and from 0.532 to 0.811, respectively. These highly polymorphic markers allow an accurate description of the genetic diversity and structure of S. luteus populations. [ABSTRACT FROM AUTHOR]

Total metal content by itself is insufficient as a measure to indicate actual environmental risk. Understanding the mobility of heavy metals in the soil and their speciation in the soil solution is of great importance for accurately assessing environmental risks posed by these metals. In a first explorative study, the effects of general soil characteristics on Cd mobility were evaluated and expressed in the form of empirical formulations. The most important factors influencing mobility of Cd proved to be pH and total soil content. This may indicate that current legislation expressing the requirement for soil sanitation in Flanders (Belgium) as a function of total soil content, organic matter, and clay does not successfully reflect actual risks. Current legal frameworks focusing on total content, therefore, should be amended with criteria that are indicative of metal mobility and availability and are based on physicochemical soil properties. In addition, soil-solution speciation was performed using two independent software packages (Visual Minteq 2.23 and Windermere Humic Aqueous model VI [WHAM VI]). Both programs largely were in agreement in concern to Cd speciation in all 29 soils under study. Depending on soil type, free ion and the organically complexed forms were the most abundant species. Additional inorganic soluble species were sulfates and chlorides. Minor species in solution were in the form of nitrates, hydroxides, and carbonates, the relative importance of which was deemed insignificant in comparison to the four major species. [ABSTRACT FROM AUTHOR]

The present study investigated the acute toxic effects of a metal-loaded Bauxsol™ reagent (MLBR), containing more than 6,450 mg kg-1 of bound metal, on the earthworm Eisenia fetida. Bauxsol is manufactured by Virotec Global Solutions of Gold Coast, Queensland, Australia. The E. fetida were exposed to 0, 10, 20, 40, 60, and 80% treatments of MLBR plus cow manure for 28 d. Good motility and no mortality of E. fetida were found in all treatments after 28 d of exposure. The greatest bioaccumulation of metals by E. fetida occurred in the 20 and 40% MLBR treatments. The bioaccumulation of metals in the worms and bioaccumulation factors (BAFs) were less than the reported toxicity thresholds to cause mortality and less than the reported BAF for moderately contaminated soils, indicating that metals bound to Bauxsol reagents are mostly nonbioavailable. Analyses of the 20% MLBR treatment at 28 d using a sequential extraction procedure showed that more than 95% of the metals are bound within the Fe/Mn oxide fractions. However, changes occurred in metal fractionation after exposure to E. fetida for Cd/Cr, and Fe/Mn. The data also showed that the exchangeable (1 M MgCl2) and the toxicity characteristic leaching procedure extractant are useful as indicators of metal bioavailability from MLBR to E. fetida. [ABSTRACT FROM AUTHOR]