Air quality in post-mining towns: tracking potentially toxic elements using tree leaves.

In this study, leaves of the evergreen holm oak Quercus ilex were used to assess airborne contamination of potentially toxic elements (PTEs) at five towns located on the slopes of the Mt. Amiata (central Italy), an area with a long history of mining and, more recently, an important district for the industrial exploitation of geothermal energy. PTE composition and covariance of washed and unwashed Q. ilex leaves of three different ages (6, 12 and 24 month-old) were used to identify atmospheric inputs of PTEs at residential areas, evaluate long-term adsorption and retention of PTEs by the leaves, thus providing an indication of potential human exposure. Moreover, the determination of foliar concentrations of major elements (C, N, S and P) allowed an assessment of the nutritional status of the investigated urban tree stands which excluded the existence of stress condition caused by air pollution or other disturbances. Results indicated that overall Pb, Cu, and Cd concentration were low in the investigated urban sites, if compared with similar studies conducted in larger Italian cities, denoting a low contribution of vehicular traffic to the atmospheric pathway. The five urban settlements were characterized by a specific profile of elements (Al, Ba, Hg and Sb) enriched in unwashed leaves, resulting from the distinct geochemical characteristics of the area and from diffuse (i.e., urban activity) and point sources of PTEs emission (i.e., brownfields, geothermal power plants). The latter sources primarily govern the distribution of Hg, whose contamination was found to be very localized close to a major abandoned mining area. Our data provided quantitative evidence of the spectrum of PTEs potentially impacting resident population and may prove useful in support of follow-up instrumental monitoring campaigns of air quality, as well as for human health and ecological risk assessments.

Source-specific risk assessment for cadmium in wheat and maize: Towards an enrichment model for China.

Cadmium (Cd) pollution of agricultural soil is of public concern due to its high potential toxicity and mobility. This study aimed to reveal the risk of Cd accumulation in soil and wheat/maize systems, with a specific focus on the source-specific ecological risk, human health risk and Cd enrichment model. For this we investigated more than 6100 paired soil and grain samples with 216 datasets including soil Cd contents, soil pH and grain Cd contents of 85 sites from China. The results showed that mining activities, sewage irrigation, industrial activities and agricultural practices were the critical factors causing Cd accumulation in wheat and maize cultivated sites. Thereinto, mining activities contributed to a higher Cd accumulation risk in the southwest China and Middle Yellow River regions; sewage irrigation influenced the Cd accumulation in the North China Plain. In addition, the investigated sites were classified into different categories by comparing their soil and grain Cd contents with the Chinese soil screening values and food safety values, respectively. Cd enrichment models were developed to predict the Cd levels in wheat and maize grains. The results showed that the models exhibited a good performance for predicting the grain Cd contents among safe and warning sites of wheat (R2 = 0.61 and 0.72, respectively); while the well-fitted model for maize was prone to the overestimated sites (R2 = 0.77). This study will provide national viewpoints for the risk assessments and prediction of Cd accumulation in soil and wheat/maize systems.

Cadmium concentrations in German soybeans are elevated in conurbations and in regions dominated by mining and the metal industry.

BACKGROUND: Cadmium (Cd) is a mobile heavy metal and soybeans accumulate more of this element compared to other grain crops. Because vegan and vegetarian lifestyles are increasingly popular, plant-based diets should be free from contaminants. Cd minimization has recently been identified as a breeding aim in addition to protein and oil levels. It is also important to consider the regional variability of Cd in crops. RESULTS: A large sample collection (n = 602) was subjected to Cd analyses. Soybeans were grown using a standard protocol by private gardeners in a citizen science program. Some 12.5% of samples exceeded European Union limits, although Cd concentrations and exceedances showed regional variation around the national mean (0.13 mg kg-1 ). Contamination was higher in densely populated regions, possibly as a result of the long-term emissions of heavy metal loaded dusts from industry, traffic and energy production. Cd concentrations were also elevated in regions once characterized by mining and heavy industry. Values were low overall in rural areas and agricultural regions despite potential inputs via phosphate fertilizers. CONCLUSION: Grain Cd levels are highly variable across Germany. Soybean farming may be problematic in regions with a long history of mining and heavy metal deposition. © 2018 Society of Chemical Industry.

Phytotoxicity of tin mine waste and accumulation of involved heavy metals in common buckwheat (Fagopyrum esculentum Moench).

Extraction and processing of cassiterite (SnO2) left large tailings with high concentrations of tin, tungsten, molybdenum and lithium. Information on the phytotoxicity of mine waste is important with regard to ecological hazards. Exposure studies help to identify plants useful for the stabilization of waste tips and the phytomining of metals. A greenhouse study was performed using a dilution series of mine waste and four crops, a halophytic and a metallophytic species to derive dose response curves. Based on effective doses for growth reductions, sensitivity increased in the following order: maize > common buckwheat > quinoa > garden bean. Element analyses in different species and compartments of common buckwheat grown in a mixture of standard soil and 25% of the mine waste showed that only low levels of the metals were taken up and that transfer to seed tissues was negligible. As indicated by soil metal levels prior to and after the experiment, only lithium and arsenic proved to be plant available and reached high levels in green tissues while seed levels were low. The experiment confirmed differences in the uptake of metals with regard to elements and species. Common buckwheat is a suited candidate for cultivation on metal polluted soils.

Atmospheric CO2 enrichment and drought stress modify root exudation of barley.

Rising CO2 concentrations associated with drought stress is likely to influence not only aboveground growth, but also belowground plant processes. Little is known about root exudation being influenced by elements of climate change. Therefore, this study wanted to clarify whether barley root exudation responds to drought and CO2 enrichment and whether this reaction differs between an old and a recently released malting barley cultivar. Barley plants were grown in pots filled with sand in controlled climate chambers at ambient (380 ppm) or elevated (550 ppm) atmospheric [CO2 ] and a normal or reduced water supply. Root exudation patterns were examined at the stem elongation growth stage and when the inflorescences emerged. At both dates, root exudates were analyzed for different compounds such as total free amino acids, proline, potassium, and some phytohormones. Elevated [CO2 ] decreased the concentrations in root exudates of some compounds such as total free amino acids, proline, and abscisic acid. Moreover, reduced water supply increased proline, potassium, electric conductivity, and hormone concentrations. In general, the modern cultivar showed higher concentrations of proline and abscisic acid than the old one, but the cultivars responded differentially under elevated CO2 . Plant developmental stage had also an impact on the root exudation patterns of barley. Generally, we observed significant effects of CO2 enrichment, watering levels, and, to a lesser extent, cultivar on root exudation. However, we did not find any mitigation of the adverse effects of drought by elevated CO2 . Understanding the multitude of relationships within the rhizosphere is an important aspect that has to be taken into consideration in the context of crop performance and carbon balance under conditions of climate change.

Simple and robust determination of the activity signature of key carbohydrate metabolism enzymes for physiological phenotyping in model and crop plants.

The analysis of physiological parameters is important to understand the link between plant phenotypes and their genetic bases, and therefore is needed as an important element in the analysis of model and crop plants. The activities of enzymes involved in primary carbohydrate metabolism have been shown to be strongly associated with growth performance, crop yield, and quality, as well as stress responses. A simple, fast, and cost-effective method to determine activities for 13 key enzymes involved in carbohydrate metabolism has been established, mainly based on coupled spectrophotometric kinetic assays. The comparison of extraction buffers and requirement for dialysis of crude protein extracts resulted in a universal protein extraction protocol, suitable for the preparation of protein extracts from different organs of various species. Individual published kinetic activity assays were optimized and adapted for a semi-high-throughput 96-well assay format. These assays proved to be robust and are thus suitable for physiological phenotyping, enabling the characterization and diagnosis of the physiological state. The potential of the determination of distinct enzyme activity signatures as part of a physiological fingerprint was shown for various organs and tissues from three monocot and five dicot model and crop species, including two case studies with external stimuli. Differential and specific enzyme activity signatures are apparent during inflorescence development and upon in vitro cold treatment of young inflorescences in the monocot ryegrass, related to conditions for doubled haploid formation. Likewise, treatment of dicot spring oilseed rape with elevated CO2 concentration resulted in distinct patterns of enzyme activity responses in leaves.

Effects of legacy mining on mercury concentrations in conifer needles and mushrooms in northern Palatinate, Germany.

Due to integrated pollution prevention and control measures and the reduced burning of coal, air concentrations of mercury (Hg0) are currently levelling off. In the future, however, evasion from land surfaces will probably reverse this trend. Reasons are the rising temperatures and the loss of forest cover caused by calamities, droughts, storms and wildfires. Plant leaves constitute an important matrix for the accumulation of gaseous mercury and uptake and re-volatilisation by plants depends on the species, the vitality and the age and morphology of leaf organs. It has been shown that older conifer needles show higher concentrations than young needles and Hg accumulation is increasing throughout the season. In present study, we collected branches from Norway Spruce (Picea abies) in a former cinnabar mining region in Northern Palatinate, where artisanal and small-scale mining left innumerable waste dumps. While mining, smelting and processing of the ores were terminated during WWII, high total mercury concentrations remained in the top soils locally, with presumably only small fractions being plant available. In the lab, up to seven needle age classes were analysed. 1000 needle weights increased with age and as expected, also the Hg concentrations were elevated in the older needles. Needle concentrations were higher than those reported from other national biomonitoring programs confirming the regional imprint from legacy mercury. To complement our biomonitoring study, we collected edible mushrooms in former mining areas. Hg concentrations in most samples exceeded the EU maximum residue levels (MRL), while only a few broke the existing cadmium and lead limits. Tolerable weekly intake (TWI) for inorganic mercury would be surpassed with the consumption of a small portion of mushrooms. Further studies should be performed on the outgassing of Hg from mine wastes and the incorporation of Hg in the local food web, including its methylation and biomagnification.

Tree-rings analysis to reconstruct atmospheric mercury contamination at a historical mining site.

Mercury (Hg) is a global environmental concern due to its toxicity (especially high in methylated form) and the long-range distribution of its gaseous elemental form (GEM). Hg-contaminated areas, such as abandoned mining sites, pose intrinsic difficulties for their management and heavy monitoring costs. In these environments, plant-based solutions may play a key role in the ecosystem quality assessment and support remediation strategies, combining reliability and cost-effectiveness. In this study, we adopted a biomonitoring approach by using tree rings of four different species collected in the proximity of the mining-metallurgical area of Abbadia San Salvatore, central Italy, a major former Hg mining district whose reclamation is currently in progress. Our dendrochemical analysis was aimed at identifying the historical changes of local atmospheric Hg contamination and at singling out, for the first time in the study area, other potentially toxic elements (PTEs) associated with the past mining activity. Collected cores dated back to early as 1940 and provided the temporal patterns of atmospheric Hg emission vs the produced liquid quantities, so reconstructing the historical impact of the mining site on nearby terrestrial ecosystems and resident human population. Current GEM contamination was found about twenty times lower than that of the fully operational mine periods. From a first survey on other PTEs, thallium (Tl) and lead (Pb) appeared to be potentially associated with the mining activity, thus suggesting new working assumptions for further dendrochemical analyses and for the inclusion of Pb in human biomonitoring surveys of the Mt. Amiata area, actually not present in the control list. The results prompt a more thorough assessment by tracking for a longer time span a critical site that is an ideal open-field lab to study the ecophysiology of different tree species in relation to environmental behavior of PTEs for better-assessing wildlife and human exposures.

Senescence-specific alteration of hydrogen peroxide levels in Arabidopsis thaliana and oilseed rape spring variety Brassica napus L. cv. Mozart.

In order to analyze the signaling function of hydrogen peroxide (H(2)O(2)) production in senescence in more detail, we manipulated intracellular H(2)O(2) levels in Arabidopsis thaliala (L.) Heynh by using the hydrogen-peroxide-sensitive part of the Escherichia coli transcription regulator OxyR, which was directed to the cytoplasm as well as into the peroxisomes. H(2)O(2) levels were lowered and senescence was delayed in both transgenic lines, but OxyR was found to be more effective in the cytoplasm. To transfer this knowledge to crop plants, we analyzed oilseed rape plants Brassica napus L. cv. Mozart for H(2)O(2) and its scavenging enzymes catalase (CAT) and ascorbate peroxidase (APX) during leaf and plant development. H(2)O(2) levels were found to increase during bolting and flowering time, but no increase could be observed in the very late stages of senescence. With increasing H(2)O(2) levels, CAT and APX activities declined, so it is likely that similar mechanisms are used in oilseed rape and Arabidopsis to control H(2)O(2) levels. Under elevated CO(2) conditions, oilseed rape senescence was accelerated and coincided with an earlier increase in H(2)O(2) levels, indicating that H(2)O(2) may be one of the signals to inducing senescence in a broader range of Brassicaceae.

Nitrogen Supply Drives Senescence-Related Seed Storage Protein Expression in Rapeseed Leaves.

In general, yield and fruit quality strongly rely on efficient nutrient remobilization during plant development and senescence. Transcriptome changes associated with senescence in spring oilseed rape grown under optimal nitrogen supply or mild nitrogen deficiency revealed differences in senescence and nutrient mobilization in old lower canopy leaves and younger higher canopy leaves [1]. Having a closer look at this transcriptome analyses, we identified the major classes of seed storage proteins (SSP) to be expressed in vegetative tissue, namely leaf and stem tissue. Expression of SSPs was not only dependent on the nitrogen supply but transcripts appeared to correlate with intracellular H2O2 contents, which functions as well-known signaling molecule in developmental senescence. The abundance of SSPs in leaf material transiently progressed from the oldest leaves to the youngest. Moreover, stems also exhibited short-term production of SSPs, which hints at an interim storage function. In order to decipher whether hydrogen peroxide also functions as a signaling molecule in nitrogen deficiency-induced senescence, we analyzed hydrogen peroxide contents after complete nitrogen depletion in oilseed rape and Arabidopsis plants. In both cases, hydrogen peroxide contents were lower in nitrogen deficient plants, indicating that at least parts of the developmental senescence program appear to be suppressed under nitrogen deficiency.

Divergent N Deficiency-Dependent Senescence and Transcriptome Response in Developmentally Old and Young Brassica napus Leaves.

In the spring oilseed rape (OSR) cultivar 'Mozart' grown under optimal N supply (NO) or mild N deficiency (NL) the transcriptome changes associated with progressing age until early senescence in developmentally old lower canopy leaves (leaf #4) and younger higher canopy leaves (leaf #8) were investigated. Twelve weeks old NO and NL plants appeared phenotypically and transcriptomically identical, but thereafter distinct nutrition-dependent differences in gene expression patterns in lower and upper canopy leaves emerged. In NO leaves #4 of 14-week-old compared to 13-week-old plants, ∼600 genes were up- or downregulated, whereas in NL leaves #4 ∼3000 genes were up- or downregulated. In contrast, in 15-week-old compared to 13-week-old upper canopy leaves #8 more genes were up- or downregulated in optimally N-supplied plants (∼2000 genes) than in N-depleted plants (∼750 genes). This opposing effect of N depletion on gene regulation was even more prominent among photosynthesis-related genes (PSGs). Between week 13 and 14 in leaves #4, 99 of 110 PSGs were downregulated in NL plants, but none in NO plants. In contrast, from weeks 13 to 16 in leaves #8 of NL plants only 11 PSGs were downregulated in comparison to 66 PSGs in NO plants. Different effects of N depletion in lower versus upper canopy leaves were also apparent in upregulation of autophagy genes and NAC transcription factors. More than half of the regulated NAC and WRKY transcription factor, autophagy and protease genes were specifically regulated in NL leaves #4 or NO leaves #8 and thus may contribute to differences in senescence and nutrient mobilization in these leaves. We suggest that in N-deficient plants the upper leaves retain their N resources longer than in amply fertilized plants and remobilize them only after shedding of the lower leaves.

Regional differences in plant levels and investigations on the phytotoxicity of lithium.

The growing use of lithium (Li) in industrial and energetic applications and the inability to completely recycle the alkali metal will most likely increase anthropogenic emissions and environmental concentrations in the future. Although non-essential to plants, Li(+) is an important ultra-trace element in the animal and human diet and is also used in the treatment of e.g. mental disorders. Most of the lithium is consumed with the drinking water and vegetables, but concentrations in foodstuffs vary with the geochemistry of the element. In order to identify potential risks and to avoid an overmedication due to consumption of Li rich or Li contaminated foods it is advisable to identify background levels and to derive recommended Daily Allowances (RDAs) for the element. Although Germany does not possess large amounts of primary or secondary resources of lithium, geochemical investigations (mineral and ground waters and soils) in this country confirm a wide variation of environmental concentrations with generally higher levels in the southwest. Despite the large number of soil and water data, only very few data exist on lithium concentrations in plants and its phytotoxicity. Within the scope of present study common grassland plant species were sampled in regions of SW-Germany with reportedly high geogenic levels of Li. The data are discussed with regard to literature surveys and existing reference values. Since lithium has phytotoxic effects a greenhouse experiment was performed with different Li salts (LiCl and Li2CO3) and plant species (maize, bean and buckwheat) to derive dose-response relationships for the endpoint shoot growth. While corn growth was not reduced significantly by soil concentrations of 118 ppm, EC50 values in buckwheat were 47 and 16 ppm for lithium derived from LiCl and Li2CO3, respectively.

Abundance and activity of nitrate reducers in an arable soil are more affected by temporal variation and soil depth than by elevated atmospheric [CO2].

Elevated atmospheric carbon dioxide concentrations ([CO(2) ]) might change the abundance and the function of soil microorganisms in the depth profile of agricultural soils by plant-mediated reactions. The seasonal pattern of abundance and activity of nitrate-reducing bacteria was studied in a Mini-FACE experiment planted with oilseed rape (Brassica napus). Three depths (0-10, 10-20 and 20-30 cm) were sampled. Analyses of the abundances of total (16S rRNA gene) and nitrate-reducing bacteria (narG, napA) revealed strong influences of sampling date and depth, but no [CO(2)] effects. Abundance and activity of nitrate reducers were higher in the top soil layer and decreased with depth but were not related to extractable amounts of nitrogen and carbon in soil. Dry periods reduced abundances of total and nitrate-reducing bacteria, whereas the potential activity of the nitrate reductase enzyme was not affected. Enzyme activity was only weakly correlated to the abundance of nitrate-reducing bacteria but was related to NH(4) (+) and NO(3) (-) concentrations. Our results suggest that in contrast to the observed pronounced seasonal changes, the elevation of atmospheric [CO(2) ] has only a marginal impact on nitrate reducers in the investigated arable ecosystem.

Twenty years of biological monitoring of element concentrations in permanent forest and grassland plots in Baden-Württemberg (SW Germany).

BACKGROUND, AIM AND SCOPE: Environmental monitoring of pollutants in international or local programmes has enabled authorities to evaluate the success of political measures over time. Strict environmental legislation and the introduction of cleaner technologies have already led to significant improvements of the air and water quality in many countries. Still, the discharge and deposition of anthropogenic long-range transported pollutants often remain above the critical thresholds and long-term targets defined for terrestrial and aquatic ecosystems even in EU countries. In order to determine the spatial and temporal variation of pollutant and nutrient loads in different environmental media a unique ecological response cadastre (Okologisches Wirkungskataster, OKWI) was set up in the Land of Baden-Württemberg (SW Germany) in the mid 1980s. As a part of the program a state-wide bioindicator network was established in 64 forest and 18 permanent grassland ecosystems, in which selected chemical elements were measured over time. Here, we report on the results of these analyses and discuss the general spatio-temporal trends in pollution loads. MATERIALS AND METHODS: Sixty-four forest and 18 permanent grassland plots were established in state-owned forest and nature conservation areas of SW Germany representing different landscapes and geologies of the State of Baden-Württemberg. Apart from performing vegetation relevées in marked plots of either the grassland or forest sites, plant samples were collected in intervals of 2 to 3 years following a standardised protocol. To be able to compare the different monitoring sites, four common species were chosen as indicator species in the grasslands. Later on, also bulk grassland samples were taken regardless of the species. In the forests, foliage of the dominant tree species (Fagus sylvatica, Abiea alba or Fraxinus excelsior) was sampled in the crown of marked trees and from the same species in the herb layer. The elements analysed in the plant material were the essential plant nutrients C, N, S, P, Ca, K and Mg and the metals Mn, Cd, Pb, Al, Cu, Ni and Hg. Data were analysed using descriptive and multivariate statistics and maps were produced to identify regional differences in pollutant deposition. RESULTS AND DISCUSSION: Out of the elements analysed, lead and sulphur concentrations showed the most pronounced downward trends over time in tree foliage and grassland samples with the largest decreases observed in the early 1990 s. Both the reduced lead and sulphur levels in the biomonitors reflect the successful implementation of clean air policies, i.e. the introduction of unleaded gasoline, the availability of desulphurisation technologies and the economic transition of Eastern European heavy industries. However, the decrease in sulphur concentrations was lower in beech foliage from SW Germany as compared to beech leaves from six German national parks suggesting regional differences in sulphur deposition and trends thereof. At the same time, sulphur concentrations declined more strongly in the grassland samples indicating that much of the deposited sulphur remains in the forest ecosystems while in the grassland ecosystems it is gradually removed by the frequent cutting and grazing. During the time series, the decrease in sulphur deposition coincided with a marked increase in rain pH. At the same time, the increasing nitrogen concentrations observed over time in beech leaves suggest that emissions of oxidised and reduced nitrogen are still adding to the large-scale eutrophication of SW German forests. However, N concentrations in both the tree foliage and in the bulk grassland samples were unrelated to the modelled N deposition. When also considering macronutrient concentrations and N:P and N:K ratios, the results point to serious nutrient imbalances in many beech forests, which may reduce plant vitality and tree growth in the long run. CONCLUSIONS: Biological monitoring using plants is an effective tool to address changes in the environmental quality over time and space. The success of European clean air policies and the introduction of emission reduction technologies could be mirrored by the declining sulphur and lead concentrations in the present and in other monitoring programmes. However, the changed deposition patterns, i.e. lower deposition of acids and higher deposition of reactive nitrogen, are coupled to changes in the soil chemistry and will continue to affect plant nutrition and the uptake of elements in the future. RECOMMENDATIONS AND PERSPECTIVES: Although it could be shown that deposition of sulphur and lead has declined markedly in the past 20 years, biomonitoring is still necessary to reveal changes in element concentrations and nutrient imbalances. The use of plants as bioindicators should be continued as an integral part of environmental monitoring programmes. Besides the chemical analyses, also biometric parameters, e.g. thousand needle or leaf weights, biomass production in grassland plots and stem increments in forest plots, should be included to monitor the long-term responses of European ecosystems to environmental and climatic changes.