Mycorrhizal symbiosis and the nitrogen nutrition of forest trees.

Terrestrial plants form primarily mutualistic symbiosis with mycorrhizal fungi based on a compatible exchange of solutes between plant and fungal partners. A key attribute of this symbiosis is the acquisition of soil nutrients by the fungus for the benefit of the plant in exchange for a carbon supply to the fungus. The interaction can range from mutualistic to parasitic depending on environmental and physiological contexts. This review considers current knowledge of the functionality of ectomycorrhizal (EM) symbiosis in the mobilisation and acquisition of soil nitrogen (N) in northern hemisphere forest ecosystems, highlighting the functional diversity of the fungi and the variation of symbiotic benefits, including the dynamics of N transfer to the plant. It provides an overview of recent advances in understanding 'mycorrhizal decomposition' for N release from organic or mineral-organic forms. Additionally, it emphasises the taxon-specific traits of EM fungi in soil N uptake. While the effects of EM communities on tree N are likely consistent across different communities regardless of species composition, the sink activities of various fungal taxa for tree carbon and N resources drive the dynamic continuum of mutualistic interactions. We posit that ectomycorrhizas contribute in a species-specific but complementary manner to benefit tree N nutrition. Therefore, alterations in diversity may impact fungal-plant resource exchange and, ultimately, the role of ectomycorrhizas in tree N nutrition. Understanding the dynamics of EM functions along the mutualism-parasitism continuum in forest ecosystems is essential for the effective management of ecosystem restoration and resilience amidst climate change. KEY POINTS: • Mycorrhizal symbiosis spans a continuum from invested to appropriated benefits. • Ectomycorrhizal fungal communities exhibit a high functional diversity. • Tree nitrogen nutrition benefits from the diversity of ectomycorrhizal fungi.

Post-mining ecosystem reconstruction.

The global decade of restoration brings into sharp focus the need to rehabilitate lands damaged by mining, to provide safe, stable, and productive landscapes. For the majority of mines, the required final land use is some form of natural, semi-natural or managed ecosystem, such as agriculture, aquaculture or forestry. Mining activities lead to new highly altered landscapes that require rehabilitation. These comprise various on-land stores of waste material and mined land itself. The repair of damaged ecosystems is described by many terms including restoration, rehabilitation, revegetation, ecological restoration, and reclamation. These terms overlap in meaning, have regional biases, and all fall short of what is really required: ecosystem reconstruction. This requires a highly multidisciplinary approach drawing on many disciplines including geotechnical engineering, social science, soil science, law, hydrology, botany, geology, pollination biology, financial planning, alongside ecology. Ideally, mine rehabilitation should be progressive, start early in the life of the mine, and employ a strict regime of characterising and tracking waste materials for use in creating safe and stable post-mining landscapes. These actions will limit risks and optimise outcomes, especially when waste materials contain toxic metals or have high levels of acidity, alkalinity or salinity. Some mine sites are appropriate for the restoration of native ecosystems and biodiversity that existed pre-mining, but many, including landscape features created from waste materials, are not. Criteria for successful land rehabilitation are complex, multivariate, and highly contingent on the agreed final land use. Future advances in mine rehabilitation include the use of geomorphic landscape design and emerging thinking on cradle-to-cradle mining. This primer will discuss the complex factors that need to be considered in ecosystem reconstruction after mining and outlines approaches for optimising land rehabilitation outcomes.

Sustainable water management in rice cultivation reduces arsenic contamination, increases productivity, microbial molecular response, and profitability.

Arsenic (As) and silicon (Si) are two structurally competitive natural elements where Si minimises As accumulation in rice plants, and based on this two-year field trial, the study proposes adopting alternating wetting and drying (AWD) irrigation as a sustainable water management strategy allowing greater Si availability. This field-based project is the first report on AWD's impact on As-Si distribution in fluvio-alluvial soils of the entire Ganga valley (24 study sites, six divisions), seasonal variance (pre-monsoon and monsoon), rice plant anatomy and productivity, soil microbial diversity, microbial gene ontology profiling and associated metabolic pathways. Under AWD to flooded and pre-monsoon to monsoon cultivations, respectively, greater Si availability was achieved and As-bioavailability was reduced by 8.7 ± 0.01-9.2 ± 0.02% and 25.7 ± 0.09-26.1 ± 0.01%. In the pre-monsoon and monsoon seasons, the physiological betterment of rice plants led to the high rice grain yield under AWD improved by 8.4 ± 0.07% and 10.0 ± 0.07%, proving the economic profitability. Compared to waterlogging, AWD evidences as an optimal soil condition for supporting soil microbial communities in rice fields, allowing diverse metabolic activities, including As-resistance, and active expression of As-responsive genes and gene products. Greater expressions of gene ontological terms and complex biochemical networking related to As metabolism under AWD proved better cellular, genetic and environmental responsiveness in microbial communities. Finally, by implementing AWD, groundwater usage can be reduced, lowering the cost of pumping and field management and generating an economic profit for farmers. These combined assessments prove the acceptability of AWD for the establishment of multiple sustainable development goals (SDGs).

Species-specific effects of mycorrhizal symbiosis on Populus trichocarpa after a lethal dose of copper.

Poplars have been identified as heavy metals hyperaccumulators and can be used for phytoremediation. We have previously established that their symbiosis with arbuscular mycorrhizal fungi (AMF) may alter their uptake, tolerance and distribution to excess concentrations of heavy metals in soils. In this study we hypothesised that mycorrhizal symbiosis improves the tolerance of poplars to lethal copper (Cu) concentrations, but this influence may vary among different AMF species. We conducted an experiment in a growth chamber with three Cu application levels of control (0 mg kg-1), threshold-lethal (729 mg kg-1) and supra-lethal (6561 mg kg-1), and three mycorrhizal treatments (non-mycorrhizal, Rhizophagus irregularis, and Paraglomus laccatum) in a completely randomized design with six replications. The poplars did not grow after application of 729 mg Cu kg-1 substrate, and mycorrhizal symbiosis did not help plants to tolerate this level of Cu. This can be explained by the toxicity suffered by mycorrhizal fungi. Translocation of Cu from roots to shoots increased when plants were colonised with R. irregularis and P. laccatum under threshold-lethal and supra-lethal applications of Cu, respectively. This result shows that mycorrhizal mediation of Cu partitioning in poplars depends on the fungal species and substrate Cu concentration. Multi-model inference analysis within each mycorrhizal treatment showed that in plants colonised with R. irregularis, a higher level of mycorrhizal colonisation may prevent Cu transfer to the shoots. We did not observe this effect in P. laccatum plants probably due to the relatively low colonisation rate (14%). Nutrient concentrations in roots and shoots were impacted by applied substrate Cu levels, but not by mycorrhizas. Magnesium (Mg), potassium (K), and manganese (Mn) concentrations in roots reduced with enhancing applied substrate Cu due to their similar ionic radii with Cu and having common transport mechanism. Synergistic effect on shoot concentration between applied substrate Cu levels and Mg, K, calcium, iron (Fe), and zinc was observed. Root Cu concentration was inversely related with root K and Mn concentrations, and shoot Cu concentration had a positive correlation with shoot Fe and K concentrations. Overall, mycorrhizal symbiosis has the potential to enhance plant health and their resilience to Cu toxicity in contamination events. However, it is important to note that the effectiveness of this symbiotic relationship varies among different mycorrhizal species and is influenced by the level of contamination.

Fragment size and diversity of mulches affect their decomposition, nutrient dynamics, and mycorrhizal root colonisation.

Plant-based mulch has been proposed as a sustainable way of maintaining soil fertility. However, the role of mulch diversity, quality, and size in decomposition dynamics, and their effect on crop yield, has not been fully explored. We investigated how mulch quality, proxied by the constituent plant species diversity, and residue size drive mulch decomposition, nutrient release, crop nutrition, and yield. A rhizotron experiment was set up with barley as a model crop, with the addition of mulch of two particle sizes (1.5 and 30 cm) and four different plant residue mixes of differing biodiversity (17, 12, 6, and 1 species) in a fully factorial design. Soil nutrient dynamics were measured at advanced decomposition stages, together with residue quality, arbuscular mycorrhizal fungal (AMF) root colonisation, and crop yield. Residue mass loss was significantly affected by its chemical composition. Initial NDF content was more restricted factor in C and N mineralisation than C:N or lignin. Long residues retained significantly higher C and N content, than short residues. Crop yield was not affected by residue type or size. Residue size significantly affected barley growth rate, influencing seed protein content. Soil available K was significantly increased by residues with a higher initial C:N ratio. Short residues resulted in higher soil Zn. Residues of higher diversity resulted inhigher AMF root colonisationof the barley plants. Generally, long residue mulches maintain higher fertilisation capacity at advanced stage of decomposition than short ones, without a deleterious effect on crop yield. Further investigation should evaluate the effect of continuous application of long residue mulches on soil fertility and microbial symbiosis.

Soil-to-plant transfer of 40K, 238U and 232Th and radiological risk assessment of selected mining sites in Nigeria.

One of the major route through which humans are exposed to ionizing radiation is via food chain, which is consequent of soil-to-plant transfer of radionuclides. This work reported the activity concentrations of 40K, 238U and 232Th in samples of water, soil and guinea corn grains collected from Beryllium and Gold mining sites in Kwara, Nigeria. In-situ measurements at approximately 1 m in the air was carried out using a well-calibrated portable Gamma Spectrometer (Super Spec RS-125), while the soil, water and the guinea corn samples were analyzed using a '3 × 3' inch lead-shielded NaI (Tl) detector. The measured activity concentrations of the natural radionuclides in the soil from both mines are lower than the in-situ measurements. This was attributed to the contribution from other terrestrial materials on-site. The estimated mean transfer factors (TFs) for 40K, 238U and 232Th are 0.21, 0.17 and 0.31, and 0.46, 0.19 and 0.28 respectively for the Beryllium and Gold mining sites. While the TFs for 238U and 232Th exceeded the mean value of 0.0062 and 0.0021 for 238U and 232Th respectively, the TFs for 40K are well below the 0.74 for cereals grains provided by International Atomic Energy Agency (IAEA). The radiation impact assessment using the Monte Carlo simulations reveals values that were generally less than the global average values provided by the United Nations Scientific Committee on the Effects of Atomic Radiation (UNSCEAR). Hence, the risk of cancer inducement due to radiation exposure is within the acceptable limits for both mining sites.

Phosphorus uptake and toxicity are delimited by mycorrhizal symbiosis in P-sensitive Eucalyptus marginata but not in P-tolerant Acacia celastrifolia.

Many plant species from regions with ancient, highly weathered nutrient-depleted soils have specialized adaptations for acquiring phosphorus (P) and are sensitive to excess P supply. Mycorrhizal associations may regulate P uptake at high external P concentrations, potentially reducing P toxicity. We predicted that excess P application will negatively impact species from the nutrient-depleted Jarrah forest of Western Australia and that mycorrhizal inoculation will reduce P toxicity by regulating P uptake. For seedlings of the N2-fixing legume Acacia celastrifolia and the tree species Eucalyptus marginata, we measured growth at P concentrations of 0-90 mg kg-1 soil and in relation to inoculation with the arbuscular mycorrhizal fungus (AMF) Rhizophagus irregularis. Non-inoculated A. celastrifolia maintained leaf P concentrations at <2 mg g-1 dry mass (DM) across the range of external P concentrations. However, for non-inoculated E. marginata, as external P concentrations increased, leaf P also increased, reaching >9 mg g-1 DM at 30 mg P kg-1 soil. Acacia celastrifolia DM increased with increasing external P concentrations, while E. marginata DM was maximal at 15 mg P kg-1 soil, declining at higher external P concentrations. Neither DM nor leaf P of A. celastrifolia was affected by inoculation with AMF. For E. marginata, even at 90 mg P kg-1 soil, inoculation with AMF resulted in leaf P remaining <1 mg g-1 DM, and DM being maintained. These data strengthen the evidence base that AMF may not only facilitate P uptake at low external P concentrations, but are also important for moderating P uptake at elevated external P concentrations and maintaining plant P concentrations within a relatively narrow concentration range.

A critical review of Pongamia pinnata multiple applications: From land remediation and carbon sequestration to socioeconomic benefits.

Pongamia pinnata (L.) Pierre (Pongamia) is a tree native to Southeast Asia. Recently, interest in Pongamia focused on its potential as a biofuel source as its seeds contain around 40% oil. However, Pongamia has multiple applications beyond biofuel production. It is a legume, can form symbiotic associations with mycorrhizal fungi, has been shown to be tolerant to drought, salinity, and heavy metals in soil, and has potential to mitigate climate change. Additionally, Pongamia oil has medicinal properties, can be used as biopesticide, insect repellent, to produce soap, and as a source of edible grade vegetable oil. The seed cake can be used as a source of bioenergy, food and feed protein, and organic fertiliser, and the flowers are a good source of pollen and nectar. Pongamia can also bring socio-economic benefits as its ability to restore degraded and contaminated land provides opportunities for local communities through novel valorisation pathways. These multiple applications have potential to form part of a circular bioeconomy in line with sustainable development goals. Although research on the multiple applications of Pongamia has grown considerably, knowledge gaps remain and these need to be addressed so that the full potential of Pongamia can be achieved. Further understanding of the mechanisms underlying its resilience to abiotic stresses, phytoremediation potential and biotic interactions should be a priority, and co-ordinated breeding efforts will be key. Here, we critically review the available literature on Pongamia and highlight gaps in knowledge in which future research should focus on to ensure that the full potential of this versatile tree can be achieved. We conclude that Pongamia can potentially form part of a circular bioeconomy and that harnessing the multiple applications of Pongamia in a holistic manner, with collaboration among key stakeholders, is crucial for the successful application of its benefits far beyond biofuel production.

Applying cover crop residues as diverse mixtures increases initial microbial assimilation of crop residue-derived carbon.

Increasing the diversity of crops grown in arable soils delivers multiple ecological functions. Whether mixtures of residues from different crops grown in polyculture contribute to microbial assimilation of carbon (C) to a greater extent than would be expected from applying individual residues is currently unknown. In this study, we used 13C isotope labelled cover crop residues (buckwheat, clover, radish, and sunflower) to track microbial assimilation of plant residue-derived C using phospholipid fatty acid (PLFA) analysis. We also quantified microbial assimilation of C derived from the soil organic matter (SOM) because fresh residue inputs also prime the decomposition of SOM. To consider the initial stages of residue decomposition, and preclude microbial turnover, we compared a quaternary mixture of residues with the average effect of their four components 1 day after incorporation. Our results show that the microbial biomass carbon (MBC) in the treatment receiving the mixed residue was significantly greater, by 132% (3.61 µg C g-1), than the mean plant residue-derived MBC in treatments receiving the four individual components of the mixture. However, there was no evidence that the mixture resulted in any additional assimilation of C derived from native SOM than the average observed in individual residue treatments. We surmise that, during the initial stages of crop residue decomposition, a greater biodiversity of residues increases microbial assimilation to a greater extent than would be expected from applying individual residues either due to faster decomposition or greater carbon use efficiency (CUE). This might be facilitated by functional complementarity in the soil microbiota, permitted by a greater diversity of substrates, reducing competition for any single substrate. Therefore, growing and incorporating crop polycultures (e.g., cover crop mixtures) could be an effective method to increase microbial C assimilation in the early stages of cover crop decomposition. Highlights: The effect of mixing crop residues on assimilation of C by soil microbial biomass was investigated.The study is important due to recent interest in diverse cover crop mixtures for arable systems.Mixing crop residues enhanced the assimilation of plant residue-derived C into microbial biomass.Growing and incorporating cover crop polycultures may enhance C storage in arable soils.

Mycorrhizal type of woody plants influences understory species richness in British broadleaved woodlands.

Mature temperate woodlands are commonly dominated by ectomycorrhizal trees, whereas understory plants predominantly form arbuscular mycorrhizal associations. Due to differences in plant-fungus compatibility between canopy and ground layer vegetation the 'mycorrhizal mediation hypothesis' predicts that herbaceous plant establishment may be limited by a lack of suitable mycorrhizal fungal inoculum. We examined plant species data for 103 woodlands across Great Britain recorded in 1971 and in 2000 to test whether herbaceous plant species richness was related to the proportion of arbuscular mycorrhizal woody plants. We compared the effect of mycorrhizal type with other important drivers of woodland plant species richness. We found a positive effect of the relative abundance of arbuscular mycorrhizal woody plants on herbaceous plant species richness. The size of the observed effect was smaller than that of pH. Moreover, the effect persisted over time, despite many woodlands undergoing marked successional change and increased understorey shading. This work supports the mycorrhizal mediation hypothesis in British woodlands and suggests that increased abundance of arbuscular mycorrhizal woody plants is associated with greater understory plant species richness.

Next generation restoration metrics: Using soil eDNA bacterial community data to measure trajectories towards rehabilitation targets.

In post-mining rehabilitation, successful mine closure planning requires specific, measurable, achievable, relevant and time-bound (SMART) completion criteria, such as returning ecological communities to match a target level of similarity to reference sites. Soil microbiota are fundamentally linked to the restoration of degraded ecosystems, helping to underpin ecological functions and plant communities. High-throughput sequencing of soil eDNA to characterise these communities offers promise to help monitor and predict ecological progress towards reference states. Here we demonstrate a novel methodology for monitoring and evaluating ecological restoration using three long-term (>25 year) case study post-mining rehabilitation soil eDNA-based bacterial community datasets. Specifically, we developed rehabilitation trajectory assessments based on similarity to reference data from restoration chronosequence datasets. Recognising that numerous alternative options for microbiota data processing have potential to influence these assessments, we comprehensively examined the influence of standard versus compositional data analyses, different ecological distance measures, sequence grouping approaches, eliminating rare taxa, and the potential for excessive spatial autocorrelation to impact on results. Our approach reduces the complexity of information that often overwhelms ecologically-relevant patterns in microbiota studies, and enables prediction of recovery time, with explicit inclusion of uncertainty in assessments. We offer a step change in the development of quantitative microbiota-based SMART metrics for measuring rehabilitation success. Our approach may also have wider applications where restorative processes facilitate the shift of microbiota towards reference states.

Natural attenuation of legacy hydrocarbon spills in pristine soils is feasible despite difficult environmental conditions in the monsoon tropics.

The Kimberley region of Western Australia is a National Heritage listed region that is internationally recognised for its environmental and cultural significance. However, petroleum spills have been reported at a number of sites across the region, representing an environmental concern. The region is also characterised as having low soil nutrients, high temperatures and monsoonal rain - all of which may limit the potential for natural biodegradation of petroleum. Therefore, this work evaluated the effect of legacy petroleum hydrocarbons on the indigenous soil microbial community (across the domains Archaea, Bacteria and Fungi) at three sites in the Kimberley region. At each site, soil cores were removed from contaminated and control areas and analysed for total petroleum hydrocarbons, soil nutrients, pH and microbial community profiling (using16S rRNA and ITS sequencing on the Illumina MiSeq Platform). The presence of petroleum hydrocarbons decreased microbial diversity across all kingdoms, altered the structure of microbial communities and increased the abundance of putative hydrocarbon degraders (e.g. Mycobacterium, Acremonium, Penicillium, Bjerkandera and Candida). Microbial community shifts from contaminated soils were also associated with an increase in soil nutrients (notably Colwell P and S). Our study highlights the long-term effect of legacy hydrocarbon spills on soil microbial communities and their diversity in remote, infertile monsoonal soils, but also highlights the potential for natural attenuation to occur in these environments.

The transfer of trace metals in the soil-plant-arthropod system.

Essential and non-essential trace metals are capable of causing toxicity to organisms above a threshold concentration. Extensive research has assessed the behaviour of trace metals in biological and ecological systems, but has typically focused on single organisms within a trophic level and not on multi-trophic transfer through terrestrial food chains. This reinforces the notion of metal toxicity as a closed system, failing to consider one trophic level as a pollution source to another; therefore, obscuring the full extent of ecosystem effects. Given the relatively few studies on trophic transfer of metals, this review has taken a compartment-based approach, where transfer of metals through trophic pathways is considered as a series of linked compartments (soil-plant-arthropod herbivore-arthropod predator). In particular, we consider the mechanisms by which trace metals are taken up by organisms, the forms and transformations that can occur within the organism and the consequences for trace metal availability to the next trophic level. The review focuses on four of the most prevalent metal cations in soil which are labile in terrestrial food chains: Cd, Cu, Zn and Ni. Current knowledge of the processes and mechanisms by which these metals are transformed and moved within and between trophic levels in the soil-plant-arthropod system are evaluated. We demonstrate that the key factors controlling the transfer of trace metals through the soil-plant-arthropod system are the form and location in which the metal occurs in the lower trophic level and the physiological mechanisms of each organism in regulating uptake, transformation, detoxification and transfer. The magnitude of transfer varies considerably depending on the trace metal concerned, as does its toxicity, and we conclude that biomagnification is not a general property of plant-arthropod and arthropod-arthropod systems. To deliver a more holistic assessment of ecosystem toxicity, integrated studies across ecosystem compartments are needed to identify critical pathways that can result in secondary toxicity across terrestrial food-chains.

Cadmium stress causes differential effects on growth and the secretion of carbon-degrading enzymes in four mycorrhizal basidiomycetes.

We hypothesised that cadmium exposure would hinder growth and secretion of carbon-degrading enzymes by mycorrhizal fungi, and that this would vary according to their tolerance to cadmium stress. The enzymes measured were ß-Glucosidase, ß-Xylosidase, ß-D-cellubiosidase, N-acetyl-ß-Glucosaminidase in three strains of ectomycorrhizal fungi Hebeloma subsaponaceum, Scleroderma sp., Hebeloma sp. and a feremycorrhizal fungus Austroboletus occidentalis. Fungi were subjected to cadmium stress for 28 d (in modified Melin-Norkrans liquid medium). The results showed unanticipated differential response of enzyme activities among the fungal species, including potential hormesis effects. Austroboletus occidentalis showed an increase in enzyme activity under cadmium stress.

Bioremediation potential of Cd by transgenic yeast expressing a metallothionein gene from Populus trichocarpa.

Cadmium (Cd) is an extremely toxic environmental pollutant with high mobility in soils, which can contaminate groundwater, increasing its risk of entering the food chain. Yeast biosorption can be a low-cost and effective method for removing Cd from contaminated aqueous solutions. We transformed wild-type Saccharomyces cerevisiae (WT) with two versions of a Populus trichocarpa gene (PtMT2b) coding for a metallothionein: one with the original sequence (PtMT2b 'C') and the other with a mutated sequence, with an amino acid substitution (C3Y, named here: PtMT2b 'Y'). WT and both transformed yeasts were grown under Cd stress, in agar (0; 10; 20; 50 µM Cd) and liquid medium (0; 10; 20 µM Cd). Yeast growth was assessed visually and by spectrometry OD600. Cd removal from contaminated media and intracellular accumulation were also quantified. PtMT2b 'Y' was also inserted into mutant strains: fet3fet4, zrt1zrt2 and smf1, and grown under Fe-, Zn- and Mn-deficient media, respectively. Yeast strains had similar growth under 0 µM, but differed under 20 µM Cd, the order of tolerance was: WT < PtMT2b 'C' < PtMT2b 'Y', the latter presenting 37% higher growth than the strain with PtMT2b 'C'. It also extracted ~80% of the Cd in solution, and had higher intracellular Cd than WT. Mutant yeasts carrying PtMT2b 'Y' had slightly higher growth in Mn- and Fe-deficient media than their non-transgenic counterparts, suggesting the transgenic protein may chelate these metals. S. cerevisiae carrying the altered poplar gene offers potential for bioremediation of Cd from wastewaters or other contaminated liquids.

Identifying potential threats to soil biodiversity.

A decline in soil biodiversity is generally considered to be the reduction of forms of life living in soils, both in terms of quantity and variety. Where soil biodiversity decline occurs, it can significantly affect the soils' ability to function, respond to perturbations and recover from a disturbance. Several soil threats have been identified as having negative effects on soil biodiversity, including human intensive exploitation, land-use change and soil organic matter decline. In this review we consider what we mean by soil biodiversity, and why it is important to monitor. After a thorough review of the literature identified on a Web of Science search concerning threats to soil biodiversity (topic search: threat* "soil biodiversity"), we compiled a table of biodiversity threats considered in each paper including climate change, land use change, intensive human exploitation, decline in soil health or plastic; followed by detailed listings of threats studied. This we compared to a previously published expert assessment of threats to soil biodiversity. In addition, we identified emerging threats, particularly microplastics, in the 10 years following these knowledge based rankings. We found that many soil biodiversity studies do not focus on biodiversity sensu stricto, rather these studies examined either changes in abundance and/or diversity of individual groups of soil biota, instead of soil biodiversity as a whole, encompassing all levels of the soil food web. This highlights the complexity of soil biodiversity which is often impractical to assess in all but the largest studies. Published global scientific activity was only partially related to the threats identified by the expert panel assessment. The number of threats and the priority given to the threats (by number of publications) were quite different, indicating a disparity between research actions versus perceived threats. The lack of research effort in key areas of high priority in the threats to soil biodiversity are a concerning finding and requires some consideration and debate in the research community.

Cadmium isotope fractionation reveals genetic variation in Cd uptake and translocation by Theobroma cacao and role of natural resistance-associated macrophage protein 5 and heavy metal ATPase-family transporters.

In response to new European Union regulations, studies are underway to mitigate accumulation of toxic cadmium (Cd) in cacao (Theobroma cacao, Tc). This study advances such research with Cd isotope analyses of 19 genetically diverse cacao clones and yeast transformed to express cacao natural resistance-associated macrophage protein (NRAMP5) and heavy metal ATPases (HMAs). The plants were enriched in light Cd isotopes relative to the hydroponic solution with Δ114/110Cdtot-sol = -0.22 ± 0.08‰. Leaves show a systematic enrichment of isotopically heavy Cd relative to total plants, in accord with closed-system isotope fractionation of Δ114/110Cdseq-mob = -0.13‰, by sequestering isotopically light Cd in roots/stems and mobilisation of remaining Cd to leaves. The findings demonstrate that (i) transfer of Cd between roots and leaves is primarily unidirectional; (ii) different clones utilise similar pathways for Cd sequestration, which differ from those of other studied plants; (iii) clones differ in their efficiency of Cd sequestration. Transgenic yeast that expresses TcNRAMP5 (T. cacao natural resistance-associated macrophage gene) had isotopically lighter Cd than did cacao. This suggests that NRAMP5 transporters constitute an important pathway for uptake of Cd by cacao. Cd isotope signatures of transgenic yeast expressing HMA-family proteins suggest that they may contribute to Cd sequestration. The data are the first to record isotope fractionation induced by transporter proteins in vivo.

Helping stakeholders select and apply appraisal tools to mitigate soil threats: Researchers' experiences from across Europe.

Soil improvement measures need to be ecologically credible, socially acceptable and economically affordable if they are to enter widespread use. However, in real world decision contexts not all measures can sufficiently meet these criteria. As such, developing, selecting and using appropriate tools to support more systematic appraisal of soil improvement measures in different decision-making contexts represents an important challenge. Tools differ in their aims, ranging from those focused on appraising issues of cost-effectiveness, wider ecosystem services impacts and adoption barriers/opportunities, to those seeking to foster participatory engagement and social learning. Despite the growing complexity of the decision-support tool landscape, comprehensive guidance for selecting tools that are best suited to appraise soil improvement measures, as well as those well-adapted to enable participatory deployment, has generally been lacking. We address this gap using the experience and survey data from an EU-funded project (RECARE: Preventing and REmediating degradation of soils in Europe through land CARE). RECARE applied different socio-cultural, biophysical and monetary appraisal tools to assess the costs, benefits and adoption of soil improvement measures across Europe. We focused on these appraisal tools and evaluated their performance against three broad attributes that gauge their differences and suitability for widespread deployment to aid stakeholder decision making in soil management. Data were collected using an online questionnaire administered to RECARE researchers. Although some tools worked better than others across case studies, the information collated was used to provide guiding strategies for choosing appropriate tools, considering resources and data availability, characterisation of uncertainty, and the purpose for which a specific soil improvement measure is being developed or promoted. This paper provides insights to others working in practical soil improvement contexts as to why getting the tools right matters. It demonstrates how use of the right tools can add value to decision-making in ameliorating soil threats, supporting the sustainable management of the services that our soil ecosystems provide.

Metal-Tolerant Fungal Communities Are Delineated by High Zinc, Lead, and Copper Concentrations in Metalliferous Gobi Desert Soils.

The soil fungal ecology of the southern Gobi region of Mongolia has been little studied. We utilized the ITS1 region from soil DNA to study possible influences soil metal concentrations on soil fungal community variation. In the sample network, a distinctive fungal community was closely associated with high zinc (Zn), lead (Pb), and copper (Cu) concentrations. The pattern of occurrence suggests that high metal concentrations are natural and not a product of mining activities. The metal-associated fungal community differs little from the "normal" community in its major OTUs, and in terms of major fungal guilds and taxa, and its distinctiveness depends on a combination of many less common OTUs. The fungal community in the sites with high metal concentrations is no less diverse than that in areas with normal background levels. Overall, these findings raise interesting questions of the evolutionary origin and functional characteristics of this apparently "metal-tolerant" community, and of the associated soil biota in general. It is possible that rehabilitation of metal-contaminated mined soils from spoil heaps could benefit from the incorporation of fungi derived from these areas.

Ectomycorrhizal Fungal Communities and Their Functional Traits Mediate Plant-Soil Interactions in Trace Element Contaminated Soils.

There is an increasing consensus that microbial communities have an important role in mediating ecosystem processes. Trait-based ecology predicts that the impact of the microbial communities on ecosystem functions will be mediated by the expression of their traits at community level. The link between the response of microbial community traits to environmental conditions and its effect on plant functioning is a gap in most current microbial ecology studies. In this study, we analyzed functional traits of ectomycorrhizal fungal species in order to understand the importance of their community assembly for the soil-plant relationships in holm oak trees (Quercus ilex subsp. ballota) growing in a gradient of exposure to anthropogenic trace element (TE) contamination after a metalliferous tailings spill. Particularly, we addressed how the ectomycorrhizal composition and morphological traits at community level mediate plant response to TE contamination and its capacity for phytoremediation. Ectomycorrhizal fungal taxonomy and functional diversity explained a high proportion of variance of tree functional traits, both in roots and leaves. Trees where ectomycorrhizal fungal communities were dominated by the abundant taxa Hebeloma cavipes and Thelephora terrestris showed a conservative root economics spectrum, while trees colonized by rare taxa presented a resource acquisition strategy. Conservative roots presented ectomycorrhizal functional traits characterized by high rhizomorphs formation and low melanization which may be driven by resource limitation. Soil-to-root transfer of TEs was explained substantially by the ectomycorrhizal fungal species composition, with the highest transfer found in trees whose roots were colonized by Hebeloma cavipes. Leaf phosphorus was related to ectomycorrhizal species composition, specifically higher leaf phosphorus was related to the root colonization by Thelephora terrestris. These findings support that ectomycorrhizal fungal community composition and their functional traits mediate plant performance in metal-contaminated soils, and have a high influence on plant capacity for phytoremediation of contaminants. The study also corroborates the overall effects of ectomycorrhizal fungi on ecosystem functioning through their mediation over the plant economics spectrum.