Root-exuded specialized metabolites reduce arsenic toxicity in maize.

By releasing specialized metabolites, plants modify their environment. Whether and how specialized metabolites protect plants against toxic levels of trace elements is not well understood. We evaluated whether benzoxazinoids, which are released into the soil by major cereals, can confer protection against arsenic toxicity. Benzoxazinoid-producing maize plants performed better in arsenic-contaminated soils than benzoxazinoid-deficient mutants in the greenhouse and the field. Adding benzoxazinoids to the soil restored the protective effect, and the effect persisted to the next crop generation via positive plant-soil feedback. Arsenate levels in the soil and total arsenic levels in the roots were lower in the presence of benzoxazinoids. Thus, the protective effect of benzoxazinoids is likely soil-mediated and includes changes in soil arsenic speciation and root accumulation. We conclude that exuded specialized metabolites can enhance protection against toxic trace elements via soil-mediated processes and may thereby stabilize crop productivity in polluted agroecosystems.

Training the next generation of plastics pollution researchers: tools, skills and career perspectives in an interdisciplinary and transdisciplinary field.

Plastics pollution research attracts scientists from diverse disciplines. Many Early Career Researchers (ECRs) are drawn to this field to investigate and subsequently mitigate the negative impacts of plastics. Solving the multi-faceted plastic problem will always require breakthroughs across all levels of science disciplinarity, which supports interdisciplinary discoveries and underpins transdisciplinary solutions. In this context, ECRs have the opportunity to work across scientific discipline boundaries and connect with different stakeholders, including industry, policymakers and the public. To fully realize their potential, ECRs need to develop strong communication and project management skills to be able to effectively interface with academic peers and non-academic stakeholders. At the end of their formal education, many ECRs will choose to leave academia and pursue a career in private industry, government, research institutes or non-governmental organizations (NGOs). Here we give perspectives on how ECRs can develop the skills to tackle the challenges and opportunities of this transdisciplinary research field and how these skills can be transferred to different working sectors. We also explore how advisors can support an ECRs' growth through inclusive leadership and coaching. We further consider the roles each party may play in developing ECRs into mature scientists by helping them build a strong foundation, while also critically assessing problems in an interdisciplinary and transdisciplinary context. We hope these concepts can be useful in fostering the development of the next generation of plastics pollution researchers so they can address this global challenge more effectively.

After the sun: a nanoscale comparison of the surface chemical composition of UV and soil weathered plastics.

Once emitted into the environment, macro- (MaP), micro- (MP) and nanoplastics (NP) are exposed to environmental weathering. Yet, the effects of biogeochemical weathering factors occurring in the soil environment are unknown. As the transport, fate, and toxicity of MP and NP depend directly on their surface properties, it is crucial to characterize their transformation in soils to better predict their impact and interactions in this environment. Here, we used scanning transmission x-ray micro spectroscopy to characterize depth profiles of the surface alteration of environmental plastic debris retrieved from soil samples. Controlled weathering experiments in soil and with UV radiation were also performed to investigate the individual effect of these weathering factors on polymer surface alteration. The results revealed a weathered surface on a depth varying between 1 µm and 100 nm in PS, PET and PP environmental plastic fragments naturally weathered in soil. Moreover, the initial step of surface fragmentation was observed on a PS fragment, providing an insight on the factors and processes leading to the release of MP and NP in soils. The comparison of environmental, soil incubated (for 1 year) and UV weathered samples showed that the treatments led to different surface chemical modifications. While the environmental samples showed evidence of alteration involving oxidation processes, the UV weathered samples did not reveal oxidation signs at the surface but only decrease in peak intensities (indicating decrease of the number of chemical C bonds). After a one-year incubation of samples in soil no clear aging effects were observed, indicating that the aging of polymers can be slow in soils. Supplementary Information: The online version contains supplementary material available at 10.1186/s43591-023-00066-2.

Soil (microbial) disturbance affect the zinc isotope biogeochemistry but has little effect on plant zinc uptake.

Zinc (Zn) is an important micronutrient but can be toxic at elevated concentrations. We conducted an experiment to test the effect of plant growth and soil microbial disturbance on Zn in soil and plants. Pots were prepared with and without maize and in an undisturbed soil, a soil that was disturbed by X-ray sterilization and a soil that was sterilized but reconditioned with the original microbiome. The Zn concentration and isotope fractionation between the soil and the soil pore water increased with time, which is probably due to physical disturbance and fertilization. The presence of maize increased the Zn concentration and isotope fractionation in pore water. This was likely related to the uptake of light isotopes by plants and root exudates that solubilized heavy Zn from the soil. The sterilization disturbance increased the concentration of Zn in the pore water, because of abiotic and biotic changes. Despite a threefold increase in Zn concentration and changes in the Zn isotope composition in the pore water, the Zn content and isotope fractionation in the plant did not change. These results have implications for Zn mobility and uptake in crop plants and are relevant in terms of Zn nutrition.

Geochemistry of groundwater and metal(loid) behavior in the costal aquifers of the Maharlu Lake, Iran.

Groundwater in coastal aquifers of the Maharlu Lake, southern Iran, has suffered from quality degradation and salinity increases in recent decades. These aquifers are important sources for drinking and agricultural uses in area around the lake. Hydrogeochemical analyses were carried out to identify the geochemical processes controlling groundwater chemistry and heavy metal concentrations in the coastal aquifer of the Maharlu Salt Lake. A total number of 54 water samples (45 samples from three coastal aquifers and 9 water samples from lake water) were collected for laboratory analysis of major ions and heavy metals three times from November-2014 to July-2015. In addition, physiochemical parameters of temperature, electrical conductivity, pH, and redox potential were measured in the field. The findings indicate three major controls of groundwater chemistry in the coastal aquifers of the Maharlu Lake including (1) lake water intrusions, (2) evaporites and carbonate dissolution-precipitation, and (3) input of organic matter probably linked to the seepage of domestic sewage with resulting sulfate reduction. In contrast, dissolution of minerals in surface sediments and direct evaporation of groundwater in the coastal aquifers seem to be insignificant based on geochemical ratios and water table depth. Concentrations of B, Li, Co, Cr, Cu, Ni, and U increase, but Zn concentrations decrease in groundwater due to the lake water intrusions into the costal aquifers. Gypsum dissolution and consequent carbonate reactions cause B, Co, Cr, Cu, Ni, Zn, and U enrichment in the groundwater samples. These findings can provide water managers and local authorities with a comprehensive framework of the coastal groundwater geochemistry, allowing a better understanding of the effects of current management practices and the implementation of mitigation approaches such as reduction of groundwater extraction to limit further lake saltwater intrusion and water resources deterioration.

Identification and characterisation of individual nanoplastics by scanning transmission X-ray microscopy (STXM).

Nanoplastics (NP) are of environmental and human health concern. We tested a novel NP extraction method and scanning transmission X-ray spectro-microscopy (STXM) in combination with near-edge X-ray absorption fine-structure spectroscopy (NEXAFS) to image and identify individual NP in environmental and food matrices. We (1) discussed the potential of STXM compared to other methods potentially suitable for NP analysis, (2) applied the method on NP suspensions of eight of the most common polymers, (3) analyzed environmental water and soil samples spiked with NP and (4) characterized NP in tea water infused in plastic teabags and unspiked soil samples. Here we show that STXM has methodological advantages and that polymers give characteristic spectra, which allows NP identification in environmental and food matrices. For soils we deliver a visual and spectroscopic characterization of NP, proving their presence and highlighting their diversity. Thus, STXM, can be used for the detection and characterisation of NP in different types of matrices.

Microplastics in agricultural drainage water: A link between terrestrial and aquatic microplastic pollution.

Microplastic (MP) contamination has been reported to be higher in terrestrial compared to aquatic environments. This is probably due to the fact that plastic items are mostly produced and used in terrestrial environments and have a longer residence time. However, there are several links between the terrestrial and aquatic environments. We analyzed drainage water samples from agricultural soils in the Seeland, a heavily drained agriculturally intensive area in Switzerland for its MP (>100 µm) concentration and composition. We found MP in relevant numbers (mean 10.5 ± 9.5 N L-1). The polymers were mainly PA and PE, and the size distribution showed an exponential increase with decreasing particle size. The results show considerable MP concentrations in drainage water and imply a transport of MP in soils down to the drainage pipes. Given the large areas drained both in Switzerland and globally, it is proposed that MP leaching from soil can be a significant source of MP to aquatic ecosystems. Such a contribution should be considered when dealing with MP cycling on a local to global scale.

A Systematic Analysis of Metal and Metalloid Concentrations in Eight Zebrafish Recirculating Water Systems.

Metals and metalloids are integral to biological processes and play key roles in physiology and metabolism. Nonetheless, overexposure to some metals or lack of others can lead to serious health consequences. In this study, eight zebrafish facilities collaborated to generate a multielement analysis of their centralized recirculating water systems. We report a first set of average concentrations for 46 elements detected in zebrafish facilities. Our results help to establish an initial baseline for trouble-shooting purposes, and in general for safe ranges of metal concentrations in recirculating water systems, supporting reproducible scientific research outcomes with zebrafish.

Tracing the fate of phosphorus fertilizer derived cadmium in soil-fertilizer-wheat systems using enriched stable isotope labeling.

Applying mineral phosphorus (P) fertilizers introduces a considerable input of the toxic heavy metal cadmium (Cd) into arable soils. This study investigates the fate of P fertilizer derived Cd (Cddff) in soil-wheat systems using a novel combination of enriched stable Cd isotope mass balances, sequential extractions, and Bayesian isotope mixing models. We applied an enriched 111Cd labeled mineral P fertilizer to arable soils from two long-term field trials with distinct soil properties (a strongly acidic pH and a neutral pH) and distinct past mineral P fertilizer application rates. We then cultivated wheat in a pot trial on these two soils. In the neutral soil, Cd concentrations in the soil and the wheat increased with increasing past mineral P fertilizer application rates. This was not the case in the strongly acidic soil. Less than 2.3% of freshly applied Cddff was taken up by the whole wheat plant. Most of the Cddff remained in the soil and was predominantly (>95% of freshly applied Cddff) partitioned into the easily mobilizable acetic acid soluble fraction (F1) and the potentially mobile reducible fraction (F2). Soil pH was the determining factor for the partitioning of Cddff into F1, as revealed through a recovery of about 40% of freshly applied Cddff in F1 in the neutral pH soil compared with about 60% in the strongly acidic soil. Isotope mixing models showed that F1 was the predominant source of Cd for wheat on both soils and that it contributed to over 80% of the Cd that was taken up by wheat. By tracing the fate of Cddff in entire soil-plant systems using different isotope source tracing approaches, we show that the majority of Cddff remains mobilizable and is potentially plant available in the subsequent crop cycle.

Specific and conserved patterns of microbiota-structuring by maize benzoxazinoids in the field.

BACKGROUND: Plants influence their root and rhizosphere microbial communities through the secretion of root exudates. However, how specific classes of root exudate compounds impact the assembly of root-associated microbiotas is not well understood, especially not under realistic field conditions. Maize roots secrete benzoxazinoids (BXs), a class of indole-derived defense compounds, and thereby impact the assembly of their microbiota. Here, we investigated the broader impacts of BX exudation on root and rhizosphere microbiotas of adult maize plants grown under natural conditions at different field locations in Europe and the USA. We examined the microbiotas of BX-producing and multiple BX-defective lines in two genetic backgrounds across three soils with different properties. RESULTS: Our analysis showed that BX secretion affected the community composition of the rhizosphere and root microbiota, with the most pronounced effects observed for root fungi. The impact of BX exudation was at least as strong as the genetic background, suggesting that BX exudation is a key trait by which maize structures its associated microbiota. BX-producing plants were not consistently enriching microbial lineages across the three field experiments. However, BX exudation consistently depleted Flavobacteriaceae and Comamonadaceae and enriched various potential plant pathogenic fungi in the roots across the different environments. CONCLUSIONS: These findings reveal that BXs have a selective impact on root and rhizosphere microbiota composition across different conditions. Taken together, this study identifies the BX pathway as an interesting breeding target to manipulate plant-microbiome interactions. Video Abstract.

Global distribution of oxygenated polycyclic aromatic hydrocarbons in mineral topsoils.

Hazardous oxygenated polycyclic aromatic hydrocarbons (OPAHs) originate from combustion (primary sources) or postemission conversion of polycyclic aromatic hydrocarbons (PAHs) (secondary sources). We evaluated the global distribution of up to 15 OPAHs in 195 mineral topsoils from 33 study sites (covering 52° N-47° S, 71° W-118 °E) to identify indications of primary or secondary sources of OPAHs. The sums of the (frequently measured 7 and 15) OPAH concentrations correlated with those of the Σ16EPA-PAHs. The relationship of the Σ16EPA-PAH concentrations with the Σ7OPAH/Σ16EPA-PAH concentration ratios (a measure of the variable OPAH sources) could be described by a power function with a negative exponent <1, leveling off at a Σ16EPA-PAH concentration of approximately 400 ng g-1 . We suggest that below this value, secondary sources contributed more to the OPAH burden in soil than above this value, where primary sources dominated the OPAH mixture. This was supported by a negative correlation of the Σ16EPA-PAH concentrations with the contribution of the more readily biologically produced highly polar OPAHs (log octanol-water partition coefficient <3) to the Σ7OPAH concentrations. We identified mean annual precipitation (Spearman ρ = .33, p < .001, n = 143) and clay concentrations (ρ = .55, p < .001, n = 33) as important drivers of the Σ7OPAH/Σ16EPA-PAH concentration ratios. Our results indicate that at low PAH contamination levels, secondary sources contribute considerably and to a variable extent to total OPAH concentrations, whereas at Σ16EPA-PAH contamination levels >400 ng g-1 , there was a nearly constant Σ7OPAH/Σ16EPA-PAH ratio (0.08 ± 0.005 [SE], n = 80) determined by their combustion sources.

250-year records of mercury and trace element deposition in two lakes from Cajas National Park, SW Ecuadorian Andes.

Historical records of trace elements in lake sediments provide source-to-sink information about potentially toxic pollutants across space and time. We investigated two lakes located at different elevations in the Ecuadorian Andes to understand how trace element fluxes are related to (i) geology, (ii) erosion in the watersheds, and (iii) local point sources and atmospheric loads. In remote Lake Fondococha (4150 m a.s.l.), total Hg fluxes stay constant between ca. 1760 and 1950 and show an approximately 4.4-fold increase between pre-1950 and post-1950 values. The post-1950 increase in fluxes of other trace elements (V, Cr, Co, Ni, Cu, Zn, As, Cd, and Pb) is lower (2.1-3.0-fold) than for Hg. Mostly lithogenic sources and enhanced soil erosion contribute to their post-1950 increase (lithogenic contribution: > 85%, Hg: ~ 58%). Average post-1950 Hg fluxes are approximately 4.3 times higher in peri-urban Lake Llaviucu (3150 m a.s.l.) than in the remote Lake Fondococha. Post-1950 fluxes of the other trace elements showed larger differences between Lakes Fondococha and Llaviucu (5.2 < 25-29.5-fold increase; Ni < Pb-Cd). The comparison of the post-1950 average trace element fluxes that are derived from point and airborne sources revealed 5-687 (Hg-Pb) times higher values in Lake Llaviucu than in Lake Fondococha suggesting that Lake Llaviucu's proximity to the city of Cuenca strongly influences its deposition record (industrial emissions, traffic, caged fishery). Both lakes responded with temporary drops in trace element accumulations to park regulations in the 1970s and 1990s, but show again increasing trends in recent times, most likely caused by increase in vehicular traffic and openings of copper and gold mines around Cajas National Park.

Variations of sedimentary Fe and Mn fractions under changing lake mixing regimes, oxygenation and land surface processes during Late-glacial and Holocene times.

Global spread of anoxia in aquatic ecosystems has become a major issue that may potentially worsen due to global warming. The reconstruction of long-term hypolimnetic anoxia records can be challenging due to lack of valid and easily measurable proxies. The sedimentary Mn/Fe ratio measured by X-ray fluorescence (XRF) is often used as a proxy for past lake redox conditions. Yet the interpretation of this ratio can be problematic when Fe and Mn accumulation is not solely redox driven. We used the varved sediments of Lake Moossee (Switzerland) to examine the partitioning of Fe and Mn in seven fractions by sequential extraction under various oxygen conditions over the last 15,000 years. We combined this data with XRF scans and an independent diagnostic proxy for anoxia given by a hyperspectral imaging (HSI)-inferred record of bacteriopheophytin, to validate the use of the XRF-Mn/Fe ratio as redox proxy. In the 15,000-year long record, Fe was bound to humins and amorphous, crystalline, sulfide and residual forms. Mn was mainly present in carbonate and amorphous forms. Higher erosion, prolonged anoxia, diagenesis and humic matter input affected Fe and Mn accumulation. Under holomixis the XRF-Mn/Fe ratio successfully reflected lake redox conditions. Periods with higher detrital Fe input obscured the applicability of the ratio. During phases of permanent anoxia, intensified early diagenetic processes trapped Mn in the sediments in carbonate, crystalline oxide and humic forms. Our study shows that the single use of the XRF-Mn/Fe ratio is often not conclusive for inferring past lake redox conditions. The application of the XRF-Mn/Fe as a proxy for anoxia requires taking into account the individual lake characteristics and changes in lake environmental conditions, which affect the accumulation of Fe and Mn in the sediments.

A method for extracting soil microplastics through circulation of sodium bromide solutions.

Microplastics (MP) have been recently found in soil environments. These MP might have adverse effects at high concentrations and thus efficient extraction and analysis of MP from soil is needed. Here we propose a new method of NaBr solution circulation for extracting soil MP. A device for the circular extraction of soil MP was developed. This device included a separation, vacuum filtration, and solution recovery system. It was then utilized to test separation efficiency of soil MP with three economic and environmentally friendly extraction reagents: NaCl, CaCl2 and NaBr solutions. The separation was tested with ten different types of polymers, three different size classes and three different shapes of MP. Extraction with NaBr showed the highest recovery rates ranging from 85% to 100%. After extraction the samples were treated with H2O2 and analyzed by micro-Fourier transform infrared spectroscopy. The developed method was assessed for its potential influence on MP and no significant changes in the integrity of multiple MP were found. Finally, the established method was used to analyze MP in four types of soil: farmland, yellow-brown, paddy and floodplain soil from the suburb of Shanghai. Results showed that the mean abundance of MP was 136.6-256.7 item kg-1. Various MP including PP (40%), PE (35.5%), Acrylic (15.6%), PET (6.7%) and PA (2.2%) were found. With this paper, we provide an alternative method through NaBr solution circulation for the extraction of soil MP.

The Fate of Zn in Agricultural Soils: A Stable Isotope Approach to Anthropogenic Impact, Soil Formation, and Soil-Plant Cycling.

The supplementation of Zn to farm animal feed and the excretion via manure leads to an unintended Zn input to agricultural systems, which might compromise the long-term soil fertility. The Zn fluxes at three grassland sites in Switzerland were determined by a detailed analysis of relevant inputs (atmospheric deposition, manure, weathering) and outputs (seepage water, biomass harvest) during one hydrological year. The most important Zn input occurred through animal manure (1076-1857 g ha-1 yr-1) and Zn mass balances revealed net Zn accumulations (456-1478 g ha-1 yr-1). We used Zn stable isotopes to assess the importance of anthropogenic impacts and natural long-term processes on the Zn distribution in soils. Soil-plant cycling and parent material weathering were identified as the most important processes, over the entire period of soil formation (13 700 years), whereas the soil pH strongly affected the direction of Zn isotopic fractionation. Recent anthropogenic inputs of Zn only had a smaller influence compared to the natural processes of the past 13 700 years. However, this will probably change in the future, as Zn stocks in the 0-20 cm layer will increase by 22-68% in the next 100 years, if Zn inputs remain on the same level as today.

Sorption kinetics of isotopically labelled divalent mercury (196Hg2+) in soil.

Understanding the sorption kinetics of Hg2+ is the key to predicting its reactivity in soils which is indispensable for environmental risk assessment. The temporal change in the solubility of 196Hg2+ spikes (6 mg kg-1) added to a range of soils with different properties was investigated and modelled. The sorption of 196Hg2+ displayed a biphasic pattern with a rapid initial (short-term) phase followed by a slower (time-dependent) one. The overall reaction rate constants ranged from 0.003 to 4.9 h-1 and were significantly correlated (r = 0.94) to soil organic carbon (SOC). Elovich and Spherical Diffusion expressions compellingly fitted the observed 196Hg2+ sorption kinetics highlighting their flexibility to describe reactions occurring over multiple phases and wide timeframes. A parameterized Elovich model from soil variables indicated that the short-term sorption is solely controlled by SOC while the time-dependent sorption appeared independent of SOC and decreased at higher pH values and Al(OH)3 and MnO2 concentrations. This is consistent with a rapid chemical reaction of Hg2+ with soil organic matter (SOM) which is followed by a noticeably slower phase likely occurring through physical pathways e.g. pore diffusion of Hg2+ into spherical soil aggregates and progressive incorporation of soluble organic-Hg into solid phase. The model lines predicted that in soils with >4% SOC, Hg2+ is removed from soil solution over seconds to minutes; however, in soils with <2% SOC and higher pH values, Hg2+ may remain soluble for months and beyond with a considerable associated risk of re-emission or migration to the surrounding environments.

Using isotopes to trace freshly applied cadmium through mineral phosphorus fertilization in soil-fertilizer-plant systems.

Applications of mineral phosphorus (P) fertilizer can lead to cadmium (Cd) accumulation in soils and can increase Cd concentrations in edible crop parts. To determine the fate of freshly applied Cd, a Cd source tracing experiment was conducted in three soil-fertilizer-wheat systems by using a mineral P fertilizer labeled with the radio isotope 109Cd and by exploiting natural differences in Cd stable isotope compositions (δ114/110Cd). Source tracing with stable isotopes overestimated the proportion of Cd in plants derived from the P fertilizer, because the isotope ratios of the sources were not sufficiently distinct from those of the soils. Despite indistinguishable extractable Cd pools between control and treatments, the addition of P fertilizer resulted in a more negative apparent isotope fractionation between soil and wheat. Overall, the radio isotope approach provided more robust results and revealed that 6.5 to 15% of the Cd in the shoot derived from the fertilizer. From the introduced Cd, a maximum of 2.2% reached the wheat shoots, whilst 97.8% remained in the roots and soils. The low recoveries of the fertilizer derived Cd suggest that continuous P fertilizer application in the past decades can lead to a build-up of a residual Cd pool in soils.

Towards an understanding of the Cd isotope fractionation during transfer from the soil to the cereal grain.

Cd in soils might be taken up by plants, enter the food chain and endanger human health. This study investigates the isotopic fractionation of major processes during the Cd transfer from soils to cereal grains. Thereto, soil, soil solution, wheat and barley plants (roots, straw and grains) were sampled in the field at three study sites during two vegetation periods. Cd concentrations and δ114/110Cd values were determined in all samples. The composition of the soil solution was analyzed and the speciation of the dissolved Cd was modelled. Isotopic fractionation between soils and soil solutions (Δ114/110Cd20-50cm-soil solution = -0.61 to -0.68‰) was nearly constant among the three soils. Cd isotope compositions in plants were heavier than in soils (Δ114/110Cd0-20cm-plants = -0.55 to -0.31‰) but lighter than in soil solutions (Δ114/110Cdsoil solution-plants = 0.06-0.36‰) and these differences correlated with Cd plant-uptake rates. In a conceptual model, desorption from soil, soil solution speciation, adsorption on root surfaces, diffusion, and plant uptake were identified as the responsible processes for the Cd isotope fractionation between soil, soil solution and plants whereas the first two processes dominated over the last three processes. Within plants, compartments with lower Cd concentrations were enriched in light isotopes which might be a consequence of Cd retention mechanisms, following a Rayleigh fractionation, in which barley cultivars were more efficient than wheat cultivars.

Zinc isotope fractionation during grain filling of wheat and a comparison of zinc and cadmium isotope ratios in identical soil-plant systems.

Remobilization of zinc (Zn) from shoot to grain contributes significantly to Zn grain concentrations and thereby to food quality. On the other hand, strong accumulation of cadmium (Cd) in grain is detrimental for food quality. Zinc concentrations and isotope ratios were measured in wheat shoots (Triticum aestivum) at different growth stages to elucidate Zn pathways and processes in the shoot during grain filling. Zinc mass significantly decreased while heavy Zn isotopes accumulated in straw during grain filling (Δ66 Znfull maturity-flowering  = 0.21-0.31‰). Three quarters of the Zn mass in the shoot moved to the grains, which were enriched in light Zn isotopes relative to the straw (Δ66 Zngrain-straw -0.21 to -0.31‰). Light Zn isotopes accumulated in phloem sinks while heavy isotopes were retained in phloem sources likely because of apoplastic retention and compartmentalization. Unlike for Zn, an accumulation of heavy Cd isotopes in grains has previously been shown. The opposing isotope fractionation of Zn and Cd might be caused by distinct affinities of Zn and Cd to oxygen, nitrogen, and sulfur ligands. Thus, combined Zn and Cd isotope analysis provides a novel tool to study biochemical processes that separate these elements in plants.