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Background and aims: Metallophytes are plants that can tolerate extreme metal concentrations in the soil in which they grow. The Dugald River zinc (Zn)-lead (Pb) gossan in Queensland (Australia) is one of the largest metal deposits in the world with a surface gossan formed after weathering over millions of years. It hosts a range of metallophytes which may have potential to be used in mine site rehabilitation. This study aimed to investigate the soil-plant relationships of metallophytes on the Dugald River gossan. Methods: Plant samples and associated rooting soil samples were collected across the gossan and then analysed for metal concentrations. Soil-plant metal relationships were subsequently explored to characterise the species in relation to metal uptake behaviour. Results: The metallophyte grass, Eriachne mucronata, dominated the gossan, yet there appeared to be no direct relationship between the occurrence of metallophytes and prevailing soil metal concentrations. Using transformation-based redundancy analysis (tb-RDA), two groups of metals, copper (Cu) and Zn-Cadmium (Cd), have been identified to be the primary metals driving species distribution. Crotalaria novae-hollandiae, was able to accumulate high concentrations of each of these metals in its leaves, with up to 16,200 mg Zn kg−1, 545 mg Cu kg−1 and 170 mg Cd kg−1. Conclusions: Soil metal concentrations alone are not suitable indications for metallophyte distribution or composition in a polymetallic environment. Crotalaria novae-hollandiae can tolerate high concentrations of metals and accumulate Zn-Cu-Cd above the respective hyperaccumulation thresholds; the species can be described for the first time as a strong polymetallic indicator-type metallophyte.

Background and aims: Phosphorus is one of the major nutrients that directly or indirectly affects all aspects of plant growth. Tropical nickel hyperaccumulators, including Phyllanthus rufuschaneyi and Rinorea cf. bengalensis from Borneo Island (in the Malaysian state of Sabah), have evolved to grow in extremely P-impoverished ultramafic soils. This study aimed to establish the response of the root and shoot ionome of these two agromining ‘metal crops’ to soil P availability. Methods: We undertook a soil P dosing trial on P. rufuschaneyi and R. cf. bengalensis in Sabah (Malaysia) over a period of 12 months. We measured the elemental concentrations in the soil solution and roots, as well as in the developed and developing stems and leaves. Results: The results show that root and shoot P accumulation increased markedly as soil P availability increased by 80-fold in P. rufuschaneyi, whereas R. cf. bengalensis did not increase as strongly to P supply, despite a 135-fold increase in soil solution P. The contrasting observations on the root and shoot P accumulation patterns in these species suggests distinct P acquisition strategies in these species. The non-responsiveness of R. cf. bengalensis to increasing soil P availability may be related to its possible association with mycorrhizal fungi. Conclusion: The findings of this study reveal the complexities of phosphate uptake, transport and accumulation in these ‘metal crops’. This information is essential to develop appropriate nutrient management in nickel agromining operations.

Nickel hyperaccumulation in Blepharidium guatemalense Standl. (Rubiaceae) was found in the tropical forests of south-eastern Mexico. This study aimed to document the geographic extent of nickel hyperaccumulation in this species, to understand its process of hyperaccumulation and to explore nickel distribution within the tissues of this plant. To accomplish these objectives, a complete non-destructive elemental screening of herbarium specimens was performed with a hand-held X-ray fluorescence spectrometer. Besides, rhizosphere soils and plant tissues were collected in Mexico and analyzed for physical–chemical parameters. Finally, elemental distribution maps of nickel and other elements in plant tissues were obtained by X-ray fluorescence spectroscopy and microscopy. This study revealed that Blepharidium guatemalense is distributed throughout Chiapas, Tabasco and Campeche, reaching the maximum nickel concentration in leaves (4.3 wt%) followed by roots and seeds (2.0 wt%) and bark (1.8 wt%). Simultaneous hyperaccumulation of cobalt and nickel was found in 15% of the herbarium specimens. Blepharidium guatemalense has uncommon re-distribution mechanisms via phloem since this tissue is the highest nickel-enriched from all parts of the plant (from roots to leaves). A high total nickel (mean of 610 µg g−1) was found in rhizosphere soils even though no evidence of ophiolite emplacement in that area has been reported. Blepharidium guatemalense represents the first hypernickelophore (> 1 wt% Ni) to be reported as growing in soils that are neither ultramafic nor enriched by anthropogenic pollutants.

Background The fern Dicranopteris linearis is a hyperaccumulator of rare earth elements (REEs), aluminium (Al) and silicon (Si). However, the physiological mechanisms of tissue-level tolerance of high concentrations of REE and Al, and possible interactions with Si, are currently incompletely known. Methods A particle-induced X-ray emission (μPIXE) microprobe with the Maia detector, scanning electron microscopy with energy-dispersive spectroscopy and chemical speciation modelling were used to decipher the localization and biochemistry of REEs, Al and Si in D. linearis during uptake, translocation and sequestration processes. Results In the roots >80 % of REEs and Al were in apoplastic fractions, among which the REEs were most significantly co-localized with Si and phosphorus (P) in the epidermis. In the xylem sap, REEs were nearly 100 % present as REEH3SiO42+, without significant differences between the REEs, while 24–45 % of Al was present as Al-citrate and only 1.7–16 % Al was present as AlH3SiO42+. In the pinnules, REEs were mainly concentrated in necrotic lesions and in the epidermis, and REEs and Al were possibly co-deposited within phytoliths (SiO2). Different REEs had similar spatial localizations in the epidermis and exodermis of roots, the necrosis, veins and epidermis of pinnae of D. linearis. Conclusions We posit that Si plays a critical role in REE and Al tolerance within the root apoplast, transport within the vascular bundle and sequestration within the blade of D. linearis.

Background and Aims The aim of this study was to test the frequency distributions of foliar elements from a large dataset from Kinabalu Park (Sabah, Malaysia) for departure from unimodality, indicative of a distinct ecophysiological response associated with hyperaccumulation. Methods We collected foliar samples (n = 1533) comprising 90 families, 198 genera and 495 plant species from ultramafic soils, further foliar samples (n = 177) comprising 45 families, 80 genera and 120 species from non-ultramafic soils and corresponding soil samples (n = 393 from ultramafic soils and n = 66 from non-ultramafic soils) from Kinabalu Park (Sabah, Malaysia). The data were geographically (Kinabalu Park) and edaphically (ultramafic soils) constrained. The inclusion of a relatively high proportion (approx. 14 %) of samples from hyperaccumulator species [with foliar concentrations of aluminium and nickel (Ni) >1000 μg g–1, cobalt, copper, chromium and zinc >300 μg g–1 or manganese (Mn) >10 mg g–1] allowed for hypothesis testing. Key Results Frequency distribution graphs for most elements [calcium (Ca), magnesium (Mg) and phosphorus (P)] were unimodal, although some were skewed left (Mg and Mn). The Ni frequency distribution was bimodal and the separation point for the two modes was between 250 and 850 μg g–1. Conclusions Accounting for statistical probability, the established empirical threshold value (>1000 μg g–1) remains appropriate. The two discrete modes for Ni indicate ecophysiologically distinct behaviour in plants growing in similar soils. This response is in contrast to Mn, which forms the tail of a continuous (approximately log-normal) distribution, suggestive of an extension of normal physiological processes.

The extend of biogeochemical cycling of nickel (Ni) by tropical hyperaccumulator plants in their native habitat is largely unknown, although these unusual plants are suspected to play a major role in the recycling of this element in ultramafic ecosystems. In this study, we have assessed the biogeochemical cycling of Ni (and other elements, including mineral nutrients) by a tropical Ni hyperaccumulator plant, i.e., Phyllanthus rufuschaneyi, which is one of the most promising species for tropical Ni agromining. The study site was a young secondary forest in Sabah (Malaysia) where Phyllanthus rufuschaneyi occurs as the dominant species on an ultramafic Cambisol. For 2 years, we monitored a 100-m2 plot and collected information on weather, biomass increase, soil fertility, water fluxes to the soil and litter fluxes for a wide range of elements, including Ni. The Ni cycle is mainly driven by internal fluxes, notably the degradation and recycling of Ni-rich litter. Over the period of investigation, the Ni litter flux corresponded to the total Ni stock of the litter (5.2 g m−2 year−1). The results further show that Ni turnover varies significantly with the accumulation properties of the plant cover. This points to the major influence of Ni hyperaccumulator plants in building up Ni available stocks in the topsoils, as has also been shown in temperate ultramafic systems. Litterfall and throughfall contribute substantially to the cycling of phosphorus, sulphur and potassium in this ecosystem, with throughfall contributing 2-, 220- and 20-fold higher to the respective nutrient fluxes relative to litterfall. The magnesium:calcium ratio far exceeded 1 in the soil, but was < 1 in the leaves of Phyllanthus rufuschaneyi. The insights from this study should be taken into account when designing tropical agromining operations; as Ni stocks could be more labile than initially thought. The removal of Ni- and nutrients-rich biomass will likely affect available Ni (and major nutrients) for the next cropping seasons, and requires sustainable fertilisation, to be utilized to replenish depleted major nutrients. These findings also have major ecological implications.

International audience; Aims: The agronomy of tropical ‘metal crops’ had not yet been tested until a recent pot trial was undertaken to test the growth and nickel (Ni) yield of two promising Ni ‘metal crops’ (Phyllanthus rufuschaneyi and Rinorea cf. bengalensis) from Sabah, Malaysia. The present study investigates the growth and Ni yield in P. rufuschaneyi in response to inorganic and organic fertilisation under field conditions.Methods: A two-year field experiment was undertaken on an ultramafic Cambisol in Sabah. Two experiments were set up with different types of fertilisers, either organic or inorganic. Ten 9-m2 plots were amended with either NPK (370 kg ha−1) or N (120 kg ha−1) fertilisation and were monitored during the first year. Another experiment with 25-m2 plots was set up to test three treatments of organic amendment (0, 1 or 2 kg m−2 of shredded P. rufuschaneyi biomass).Results: The results show that despite considerable variability, Ni concentration was generally lower in plant shoots amended with N only (i.e. urea). However, treatments showed no statistical effect on plant biomass and Ni yield in the short-term, even though organic treatments had a strong positive effect on the availability of Ni.Conclusion: The short-term effects of inorganic and organic treatments on Ni yield of the tropical ‘metal crop’ Phyllanthus rufuschaneyi were not apparent (75 kg Ni ha−1 yr−1). The field trials are ongoing to assess long-term effects. The preliminary results suggest that with appropriate agronomic systems and plant breeding, the Ni yield could be substantially increased.

Aims: Nickel hyperaccumulator plants require highly evolved mechanisms to avoid cellular-level toxicity to cope with the high prevailing concentrations of nickel in their seeds and seedlings. This study aimed to investigate tissue-level distribution and redistribution of Ni and other physiologically relevant elements during the germination of hyperaccumulator plants. Methods : Berkheya coddii Roessler (Asteraceae) and Odontarrhena muralis sensu latu (Waldst. & Kit.) Endl. (O. chalcidica), formerly Alyssum murale (Waldst. & Kit.) (Brassicaceae), were germinated for several days at room temperature before using synchrotron micro X-ray Fluorescence Spectroscopy (µXRF) to obtain high-sensitivity and high-resolution elemental images of live/hydrated plants at various stages of seed germination and seedling growth. Results: The results show that fruits and seeds of both species were highly enriched in nickel. In Berkheya coddii, nickel was located in the cotyledons, the micropylar area, the seed coat, and the point of attachment of the pappus with the cypsela body. In Odontarrhena muralis seeds, nickel occurred in the cotyledons and hypocotyl. The emergence of true leaves initiates nickel and calcium redistribution within seedlings in both species. Conclusions: The diversity in physiological responses to nickel in Berkheya coddii and Odontarrhena muralis does not only occur at the mature stage, but is inherent to both species as seed elemental storage and tolerance mechanisms during seedling development differ.

International audience; Background and aims: Pioneer plants may improve the ecological restoration of degraded ultramafic areas by plant-soil interaction processes. In this study, we assess the effect of the pioneer actinorhizal tree C. terminale (Casuarinaceae) on the recovery of plant communities and soil functions on degraded tropical ultramafic sites.Methods: Soil and plant samples were collected along a tree-age gradient in two degraded ultramafic sites in Sabah (Northern Borneo, Malaysia): a Technosol and a Leptosol. Chemical composition of plants and soils, and biological activity of soils were assessed at both sites. Plant colonisation was assessed by plot vegetation surveys.Results: An improvement in soil fertility parameters (pH reduction from 8.5 to 6.8, an increase in the concentrations of several nutrients and enhanced soil enzyme activities) was observed along the C. terminale age gradient. However, plant cover and diversity was only improved around mature trees at the site that was not impacted by mining.Conclusion: C. terminale promotes the recovery of several soil functions, mainly related to the storage and recycling of N, P, K, S. Besides plant-soil feedback, other environmental factors (i.e. exposition to sunlight, drought) may play an important role on revegetation of ultramafic soils.

Ultramafic (i.e. serpentine) soils are widespread in the Balkans and particularly in Albania. They account for a large part of plant endemism in that region and host several hyperaccumulator species, which are characterized by leaf nickel concentrations frequently above 1%. This rich nickel hyperaccumulating flora could serve as candidate to be used in phytoextraction and agromining. Despite recent interest in metal hyperaccumulating plants and agromining, very few studies have investigated the bacterial diversity and the influence of environmental factors on microbial gene profiles in the rhizosphere of hyperaccumulator plants growing on ultramafic soils. Because rhizospheric bacteria could be crucial to the success of phytoremediation, we studied a total of 48 nickel-hyperaccumulating plants which were sampled from four species that are widespread in Albania: Noccaea ochroleuca, Odontarrhena smolikana, O. rigida and O. chalcidica. All samples were taken from the ultramafic regions of Librazhd and Pogradec in eastern Albania in October 2015. Our study shows that Proteobacteria, Actinobacteria and Acidobacteria dominated the soil bacterial communities. Of these three phyla, only Proteobacteria was relatively abundant. This study underlines the influence of soil Cation Exchange Capacity on the bacterial community's diversity and structure. Based on the predicted metagenomes, the genes belonging to amino acid, lipid and carbohydrate metabolisms were identified as major gene families. Our study sheds some light on our understanding of how bacterial communities are structured within and affect the rhizosphere of hyperaccumulator plants from ultramafic soils in Albania.

Purposes To analyze the main edaphic factors that differentiate ultramafic from typical Cerrado environments and act as agents of the development of biochemical and morphological mechanisms of species adaptation to these harsh environments; and to determine the genetic diversity of three Cerrado’s shrubland native species (Justicia lanstyakii, Euploca salicoides, and Oxalis hirsutissima). Methods We conducted chemical analysis of metal contents in soils, as well as on elemental composition, and analysis of DNA extracted from leaf tissues of the three species found in both environments. In leaves of E. salicoides grown in both environments we evaluated the changes in the levels of non-structural carbohydrates (NSC), and total proteins. Results The accessions obtained in ultramafic soils were closer with each other, indicating genetic similarity and major differences in relation to the accessions collected in the Cerrado area. These differentiations probably are related with higher adaptation to soils rich in metals, mainly Ni in ultramafic, and Al in the Cerrado environments. The highest levels of NSC were observed in plants grown in ultramafic soils, including raffinose, which is related to responses to metal detoxification and drought. The allocation of Ni in the trichomes, which does not affect important processes of plant metabolism, is another mechanism developed by E. salicoides to overcome the hyperaccumulation of Ni in their tissues. Conclusions These findings can help select seed collection sites representative of the genetic diversity of native plant species for restoring degraded areas or for phytoremediation of metals.

This article has been accepted for publication in Metalomics Published by Oxford University Press. DOI: 10.1093/mtomcs/mfab045; International audience; Plants have developed a diversity of strategies to take up and store essential metals in order to colonize various types of soils including mineralized soils. Yet, our knowledge of the capacity of plant species to accumulate metals is still fragmentary across the plant kingdom. In this study, we have used the X-Ray Fluorescence technology to analyze metal concentration in a wide diversity of species of the Neotropical flora that was not extensively investigated so far. In total, we screened more than 11 000 specimens representing about 5000 species from herbaria in Paris and Cuba. Our study provides a large overview of the accumulation of metals such as manganese, zinc and nickel in the Neotropical flora. We report 30 new nickel hyperaccumulating species from Cuba, including the first records in the families Connaraceae, Melastomataceae, Polygonaceae, Santalaceae and Urticaceae. We also identified the first species from this region of the world that can be considered as manganese hyperaccumulators in the genera Lomatia (Proteaceae), Calycogonium (Melastomataceae), Ilex (Aquifoliaceae), Morella (Myricaceae) and Pimenta (Myrtaceae). Finally, we report the first zinc hyperaccumulator, Rinorea multivenosa (Violaceae), from the Amazonas region. The identification of species able to accumulate high amounts of metals will become instrumental to support the development of phytotechnologies in order to limit the impact of soil metal pollution in this region of the world.

Aims Agromining aims to improve the fertility of naturally metal-rich soils by extracting metals, such as nickel (Ni), using hyperaccumulator plants. Ultramafic soils are characterized by low fertility levels, limiting hyperaccumulator yields. Here, we characterize the potential benefits for phytoextraction efficiency of cocropping a Ni-hyperaccumulator (Odontarrhena chalcidica) and a legume (Vicia sativa), following a two-year field experiment. Methods A two-year field experiment was set up in an ultramafic zone in North-West Spain. Three treatments were tested: co-cropping, fertilized control with ammonium nitrate and non-fertilized control. Results Over the 2 years, co-cropping increased O. chalcidica’s biomass by 24% and 403% compared to fertilized and non-fertilized controls, respectively. Moreover, co-cropping had higher Ni-yields for both years, while fertilization had a negative effect on soil parameters. A non-metric multidimensional scaling analysis of the operational taxonomic units showed that the soil bacterial diversity changed over time. Soil exchangeable Ni and organic carbon influenced the phyla’s relative abundance. Metabolic genes were dominant and their relative abundances increased over time with co-cropping. Conclus i o n Pluriannua l co-cropping of a hyperaccumulator with a legume improved both hyperaccumulator and Ni yields. In contrast, mineral fertilization was shown to be detrimental to some soil microbial parameters. Thus, ameliorating agromining by replacing mineral fertilizers would combine an eco-efficient strategy with sustainable metal recovery., This work was supported by French National Research Agency through the national “Investissements d’avenir” program, reference ANR-10-LABX- 21 - LABEX RESSOURCES21, through the “Agromine” project, reference ANR-14-CE04-0005, through the European ERA-net FACCE_SURPLUS project “AGRONICKEL: Developing Ni agromining on ultramafic land in Europe”, reference ANR-15- SUSF-0003-05, through the “Application of Nickel hyperaccumulating plants in phytomining processes” program, reference I-LINK0900 and the LIFE Environment and Resource Efficiency Programme (Life Agromine; LIFE15 ENV/FR/ 000512).

The rare phenomenon of plant manganese (Mn) hyperaccumulation within the Australian flora has previously been detected in the field, which suggested that the tree genus Gossia (Myrtaceae) might contain new Mn hyperaccumulators. We conducted the first growth experiment on Gossia using a multi-factorial dosing trial to assess Mn, cobalt (Co), and zinc (Zn) (hyper)accumulation patterns in selected Gossia species (G. fragrantissima and G. punctata) after a systematic assessment of elemental profiles on all holdings of the genus Gossia at the Queensland Herbarium using handheld X-ray fluorescence spectroscopy. We then conducted detailed in situ analyses of the elemental distribution of Mn, Co, Zn and other elements at the macro (organ) and micro (cellular) levels with laboratory- and synchrotron-based X-ray fluorescence microscopy (XFM). Gossia pubiflora and Gossia hillii were newly discovered to be Mn hyperaccumulator plants. In the dosing trial, G. fragrantissima accumulated 17 400 µg g−1 Mn, 545 µg g−1 Co, and 13 000 µg g−1 Zn, without signs of toxicity. The laboratory-based XFM revealed distinct patterns of accumulation of Co, Mn, and Zn in G. fragrantissima, while the synchrotron XFM showed their localization in foliar epidermal cells, and in the cortex and phloem cells of roots. This study combined novel analytical approaches with controlled experimentation to examine metal hyperaccumulation in slow-growing tropical woody species, thereby enabling insight into the phenomenon not possible through field studies.

Aims: The tropical nickel (Ni) hyperaccumulator plants, Rinorea cf. bengalensis and Phyllanthus rufuschaneyi, are locally common on ultramafic soils in the Malaysian state of Sabah on the island of Borneo. The aim of this study was to determine whether the structure and diversity of R. cf. bengalensis rhizosphere microbial communities were dependant on the Ni concentrations in the different rhizosphere soils studied. Methods: Rinorea cf. bengalensis and Phyllanthus rufuschaneyi were subjected to a randomized block design experiment in which four Ni dose levels were applied (0, 60, 240, 600 mg Ni kg−1) × 5 replicates per level for 12 months cultivation. At the end of the trial foliar elemental concentrations were measured. In addition, rhizosphere soils were collected for elemental and microbial analyses. The microbial analysis consisted of measuring microbial enzymatic activities and a diversity analysis based on Illumina sequencing. Results: The Illumina sequencing analysis yielded 2,649,138 sequences grouped under 2430 different Operational Taxonomic Units (OTUs) belonging to 25 phyla. No impact of the Ni amendments was shown for the four microbial enzymatic activities tested, even at the highest Ni dose level (600 mg Ni kg−1). Higher microbial enzymatic activities were found in the rhizosphere of R. cf. bengalensis relative to P. rufuschaneyi. Nickel concentrations appeared to have a high impact on the P. rufuschaneyi rhizosphere soils, whereas no effect was found for the rhizosphere bacterial community diversity of R. cf. bengalensis. Conclusions: There may be a significant role of rhizodeposits associated with R. cf. bengalensis that protect and preserve the microbial communities in the rhizosphere, regardless of the prevailing nickel concentrations.

The aim of this study was to assess the potential of the woody nickel hyperaccumulator species Blepharidium guatemalense (Standl.) Standl. for agromining in southeastern Mexico. Pot trials consisting of nickel dosing (0, 20, 50, 100, and 250 mg Ni kg−1), and synthetic and organic fertilization were conducted. Field trials were also undertaken with different harvesting regimes of B. guatemalense. Foliar nickel concentrations increased significantly with rising nickel additions, with a 300-fold increase at 250 mg Ni kg−1 treatment relative to the control. Synthetic fertilization strongly increased nickel uptake without any change in plant growth or biomass, whereas organic fertilization enhanced plant shoot biomass with a negligible effect on foliar nickel concentrations. A 5-year-old stand which was subsequently harvested twice per year produced the maximum nickel yield tree−1yr−1, with an estimated total nickel yield of 142 kg ha−1yr−1. Blepharidium guatemalense is a prime candidate for nickel agromining on account of its high foliar Ni concentrations, high bioconcentration (180) and translocation factors (3.3), fast growth rate and high shoot biomass production. Future studies are needed to test the outcomes of the pot trials in the field. Extensive geochemical studies are needed to identify potential viable agromining locations. Novelty Statement Our research team is a pioneer in the discovery of metal hyperaccumulator plants in Mesoamerica with at least 13 species discovered in the last 2 years. This study is the first to assess the potential of nickel agromining (phytomining) in Mexico (and in all the American continent), using one of the strongest nickel hyperaccumulators reported so far. The promising results of this study are the basis for optimal agricultural management of Blepharidium guatemalense.

Substantial unrealized opportunities exist for economic Ni agromining in the tropics. However, until recently this technology has remained relatively unexploited in this part of the world. In this chapter, we discuss the progress of tropical Ni agromining in two regions, namely Southeast Asia and the neotropical region. Significant advances have been made in Ni agromining operations in Southeast Asia, particularly Sabah (Malaysia), in the past few years: (i) exploring for suitable locations, (ii) screening for hyperaccumulator plants in native flora, (iii) selecting candidate hyperaccumulator species with high biomass production and shoot Ni concentrations (‘metal crops’), (iv) testing the agronomy of ‘metal crops’ to be used in viable agromining operations, and (v) demonstration of real-life agromining operations at field scale. The two most promising ‘metal crops’ in Sabah are Phyllanthus rufuschaneyi and Rinorea cf. bengalensis. However, Ni agromining developments in neotropical regions are still in their infant stages. Preliminary investigations have led to the discovery of a potential ‘metal crop’, Blepharidium guatemalense, having highly desirable attributes for possible Ni agromining. It is envisaged that tropical Ni agromining could be a productive alternative land use in these regions, and as such will significantly improve the livelihood of potential local ‘metal farmers’.

International audience; The hyperaccumulator Pycnandra acuminata is a New Caledonian rainforest tree known to have the highest concentration of nickel in any living organism with 25 wt% in its latex. All trees (with diameter >10 cm) and soil profiles in a 0.25-hectare permanent plot were sampled to assess the biogeochemical compartmentalisation of nickel in a dense stand of Pycnandra acuminata trees. Nickel stable isotope analysis permitted insights into the cycling of nickel in this ecosystem. The total tree biomass of the plot was ~281 tonnes ha-1, approximately 0.44 kg of cobalt, 49.1 kg of manganese, 257 kg of nickel and 6.76 kg of zinc. Nickel stable isotope analysis identified the biotic origin of the nickel in the soil upper layers with P. acuminata shoots enriched in lighter nickel isotopes. The δ60Ni latex signature suggests that long-distance transport, radial xylem and phloem loading are at play in Pycnandra acuminata.

International audience; Background and aims: The genus Denhamia(Celastraceae) includes fifteen Australian species, many of which have a propensity for manganese (Mn) (hyper)accumulation. Among the key aims of this study were to: i) elucidate Mn accumulation in D. bilocularis, D. celastroides, D. pittosporoides and D. silvestris under controlled conditions; ii) examine the in situ distributions of Mn and other elements in tissues of i) above, and also in two other species growing in their natural habitat, i.e. D. silvestris and D. cunninghamii; iii) test numerous Denhamia herbarium specimens for Mn accumulation using portable X-ray fluorescence spectroscopy (XRF).Methods: Portable XRF and laboratory micro-XRF were used to examine Mn accumulation and foliar distribution patterns in several Australian Denhamia species. These techniques were variously applied to fresh field material and experimentally-raised plants treated with Mn (250 μg g−1, 500 μg g−1 and 1000 μg g−1) and dry herbarium material.Results: The findings revealed D. bilocularis as a new Mn hyperaccumulator, with foliar Mn concentrations of up to 15,300 μg g−1 in herbarium samples, and 13,700 μg g−1 in experimentally grown plants. Laboratory μXRF maps consistently showed that foliar Mn accumulation was localized at the extremities of wild D. cunninghamii and D. silvestris leaves; whereas it was found in the vasculature of experimentally grown D. bilocularis and D. pittosporoides leaves.Conclusions: This study presents evidence that Mn accumulation across the Australian Denhamia species is taxonomically variable, however further work is required.

International audience; 17 Aims. Rinorea cf. bengalensis is a Ni hyperaccumulator which occurs in Sabah (Malaysia), on Borneo Island, 18 that is able to accumulate considerable amounts of Ni and influences the Ni cycle in surface soil layers, both in 19 terms of Ni concentration and Ni isotopic composition. In this study, the biogeochemical processes underpinning 20 Ni isotopic fractionation in the soil-plant system and the mechanisms regulating Ni homeostasis in R. cf. 21 bengalensis plants were elucidated. 22 Methods. Two specimens of R. cf. bengalensis of different ages and associated surface soils were collected from 23 ultramafic soils in Sabah. Soil mineralogy, Ni concentrations, speciation and isotopic signatures were 24 subsequently determined in plant and soil samples. 25 Results. Nickel in R. cf. bengalensis leaves is mainly complexed with citrate. Soil Ni available fractions have 26 different δ 60 Ni values depending on the Ni bearing phases. Rinorea cf. bengalensis specimens take up lighter Ni 27 isotopes and a pronounced isotopic fractionation within the plant is observed, especially in the young specimen. 28 Conclusions. The results suggest that the observed fractionation in the young plant can be attributable to kinetic 29 effects (lighter isotopes move faster), which become less evident in the older specimen, as Ni is redistributed and 30 homogenized through phloem loading and unloading processes. 31 2

Selenium (Se), a trace element essential for human and animal biological processes, is deficient in many agricultural soils. Some extremely rare plants can naturally accumulate extraordinarily high concentrations of Se. The native legume Neptunia amplexicaulis, endemic to a small area near Richmond and Hughenden in Central Queensland, Australia, is one of the strongest Se hyperaccumulators known on Earth, with foliar concentrations in excess of 4000 μg Se g−1 previously recorded. Here, we report on the Se distribution at a whole plant level using laboratory micro X-ray Fluorescence Microscopy (μXRF) and scanning electron microscopy (SEM-EDS), as well as on chemical forms of Se in various tissues using liquid chromatography-mass spectrometry (LC-MS) and synchrotron X-ray absorption spectroscopy (XAS). The results show that Se occurs in the forms of methyl-selenocysteine and seleno-methionine in the foliar tissues, with up to 13 600 μg Se g−1 total in young leaves. Selenium was found to accumulate primarily in the young leaves, flowers, pods and taproot, with lower concentrations present in the fine-roots and stem and the lowest present in the oldest leaves. Trichomes were not found to accumulate Se. We postulate that Se is (re)distributed in this plant via the phloem from older leaves to newer leaves, using the taproot as the main storage organ. High concentrations of Se in the nodes (pulvini) indicate this structure may play an important a role in Se (re)distribution. The overall pattern of Se distribution was similar in a non-Se tolerant closely related species (Neptunia gracilis), although the prevailing Se concentrations were substantially lower than in N. amplexicaulis.

Background and Aims Hybanthus austrocaledonicus (Violaceae) is a nickel (Ni) hyperaccumulator endemic to New Caledonia. One of the specimens stored at the local herbarium had a strip of bark with a remarkably green phloem tissue attached to the sheet containing over 4 wt% Ni. This study aimed to collect field samples from the original H. austrocaledonicus locality to confirm the nature of the green ‘nickel-rich phloem’ in this taxon and to systematically assess the occurrence of Ni hyperaccumulation in H. austrocaledonicus and Hybanthus caledonicus populations. Methods X-ray fluorescence spectroscopy scanning of all collections of the genus Hybanthus (236 specimens) was undertaken at the Herbarium of New Caledonia to reveal incidences of Ni accumulation in populations of H. austrocaledonicus and H. caledonicus. In parallel, micro-analytical investigations were performed via synchrotron X-ray fluorescence microscopy (XFM) and scanning electron microscopy with X-ray microanalysis (SEM-EDS). Key Results The extensive scanning demonstrated that Ni hyperaccumulation is not a characteristic common to all populations in the endemic Hybanthus species. Synchrotron XFM revealed that Ni was exclusively concentrated in the epidermal cells of the leaf blade and petiole, conforming with the majority of (tropical) Ni hyperaccumulator plants studied to date. SEM-EDS of freeze-dried and frozen-hydrated samples revealed the presence of dense solid deposits in the phloem bundles that contained >8 wt% nickel. Conclusions The occurrence of extremely Ni-rich green phloem tissues appears to be a characteristic feature of tropical Ni hyperaccumulator plants.

New Caledonia is a global biodiversity hotspot known for its metal hyperaccumulator plants. X-ray fluorescence technology (XRF) has enabled non-destructive and quantitative determination of elemental concentrations in herbarium specimens from the ultramafic flora of the island. Specimens belonging to six major hyperaccumulator families (Cunoniaceae, Phyllanthaceae, Salicaceae, Sapotaceae, Oncothecaceae and Violaceae) and one to four specimens per species of the remaining ultramafic taxa in the herbarium were measured. XRF scanning included a total of c. 11 200 specimens from 35 orders, 96 families, 281 genera and 1484 species (1620 taxa) and covered 88.5% of the ultramafic flora. The study revealed the existence of 99 nickel hyperaccumulator taxa (65 known previously), 74 manganese hyperaccumulator taxa (11 known previously), eight cobalt hyperaccumulator taxa (two known previously) and four zinc hyperaccumulator taxa (none known previously). These results offer new insights into the phylogenetic diversity of hyperaccumulators in New Caledonia. The greatest diversity of nickel hyperaccumulators occur in a few major clades (Malphigiales and Oxalidales) and families (Phyllanthaceae, Salicaceae, Cunoniaceae). In contrast, manganese hyperaccumulation is phylogenetically scattered in the New Caledonian flora.

Albanian taxa and populations of the genus Odontarrhena are most promising candidates for research on metal tolerance and Ni-agromining, but their genetic structure remains unknown. We investigated phylogenetic relationships and genetic differentiation in relation to distribution and ploidy of the taxa, anthropic site disturbance, elevation, soil type, and trace metals at each population site. After performing DNA sequencing of selected accessions, we applied DNA-fingerprinting to analyze the genetic structure of 32 populations from ultramafic and non-ultramafic outcrops across Albania. Low sequence divergence resulted in poorly resolved phylograms, but supported affinity between the two diploid serpentine endemics O. moravensis and O. rigida. Analysis of molecular variance (AMOVA) revealed significant population differentiation, but no isolation by distance. Among-population variation was higher in polyploids than in diploids, in which genetic distances were lower. Genetic admixing at population and individual level occurred especially in the polyploids O. chalcidica, O. decipiens, and O. smolikana. Admixing increased with site disturbance. Outlier loci were higher in serpentine populations but decreased along altitude with lower drought and heat stress. Genetic variability gained by gene flow and hybridization at contact zones with &ldquo, resident&rdquo, species of primary ultramafic habitats promoted expansion of the tetraploid O. chalcidica across anthropogenic sites.

Plant and soil 457(1-2), 293 - 320 (2020). doi:10.1007/s11104-020-04714-x, AimsThis study aimed to establish elemental profiles and to spatially resolve the elemental distribution in five New Caledonian woody Ni hyperaccumulator plant species (Geissois pruinosa var. pruinosa, Homalium francii, Hybanthus austrocaledonicus, Psychotria gabriellae, and Pycnandra acuminata) originating from the Cunoniaceae, Salicaceae, Violaceae, Rubiaceae, and Sapotaceae families respectively.MethodsUsing synchrotron-based micro-X-ray Fluorescence (��XRF) imaging of different plant tissues, from the roots to the shoots and reproductive organs, this study aimed to clarify how distribution patterns of nickel, and other physiologically relevant elements, differ between these species.ResultsThe results show that the tissue-level and cellular-level distribution of nickel in P. gabriellae, H. austrocaledonicus, G. pruinosa var. pruinosa, and H. francii conform with the majority of studied Ni hyperaccumulator plant species globally, including (temperate) herbaceous species, with localization mainly in epidermal cells and phloem bundles. However, P. acuminata has nickel-rich laticifers, which constitute an independent network of cells that is parallel to the vascular bundles and are the main sink for nickel.ConclusionsSynchrotron-based micro-X-ray Fluorescence (��XRF) is a powerful method for investigating how metal hyperaccumulation influences acquisition and spatial distribution of a wide range of elements. This non-invasive method enables investigation into the in vivo distribution of multiple elements and the structure and organisation of cells (e.g. laticifers)., Published by Springer Science + Business Media B.V, Dordrecht [u.a.]

The Island of Borneo is a major biodiversity hotspot, and in the Malaysian state of Sabah, ultramafic soils are extensive and home to more than 31 endemic nickel hyperaccumulator plants. The aim of this study was to characterize the structure and the diversity of the rhizosphere bacterial communities of several of these nickel hyperaccumulator plants and factors that affect these bacterial communities in Sabah. The most abundant phyla were Proteobacteria, Acidobacteria and Actinobacteria. At family level, Burkholderiaceae and Xanthobacteraceae (Proteobacteria phylum) were the most abundant families in the hyperaccumulator rhizospheres. Redundancy analysis based on soil chemical analyses and relative abundances of the major bacterial phyla showed that abiotic factors of the studied sites drove the bacterial diversity. For all R. aff. bengalensis rhizosphere soil samples, irrespective of studied site, the bacterial diversity was similar. Moreover, the Saprospiraceae family showed a high representativeness in the R. aff. bengalensis rhizosphere soils and was linked with the nickel availability in soils. The ability of R. aff. bengalensis to concentrate nickel in its rhizosphere appears to be the major factor driving the rhizobacterial community diversity unlike for other hyperaccumulator species.

Plant and soil 455, 289 ��� 303 (2020). doi:10.1007/s11104-020-04629-7, $Aims$Knowledge on the ecophysiology of cobalt hyperaccumulator species is limited. The nickel hyperaccumulator Rinorea cf. bengalensis from Borneo can accumulate high concentrations of cobalt in nature. This study investigates the cobalt accumulation potential of Rinorea cf. bengalensis in relation to nickel concentrations in soils and the subsequent tissue and cellular-level distributions of cobalt, nickel and major cations.$Methods$Seedlings of Rinorea cf. bengalensis were grown in mixed treatments on ultramafic soil containing a high concentration of available nickel. Cobalt and nickel salts were then added to the soil to study their interactions. The tissue and cellular-level distributions of cobalt, calcium, nickel, and potassium were investigated using synchrotron-based X-ray fluorescence microscopy.$Results$The maximum foliar cobalt concentration reached 1200 ��g g$^{���1}$. Accumulation of cobalt competed with nickel accumulation although nickel seems to stimulate cobalt phloem translocation. Plants suffered toxicity in the treatment with the highest soil cobalt concentration. Cobalt and nickel have contrasting distribution patterns in the leaves of Rinorea cf. bengalensis, with cobalt mainly excreted on the surface of the leaves, whereas nickel is localised in foliar epidermal cells.$Conclusions$Rinorea cf. bengalensis can accumulate high concentrations of cobalt, but is intrinsically more tolerant to nickel. It does not rely on a similar sequestration mechanism for both metals, which could explain the lesser tolerance for cobalt. Nickel appears to be essential for the plant to tolerate high cobalt concentrations. Further studies intending to develop agronomic practices are needed to determine the viability of Rinorea cf. bengalensis for nickel-cobalt agromining., Published by Springer Science + Business Media B.V, Dordrecht [u.a.]

The fern Dicranopteris linearis (Gleicheniaceae) from China is a hyperaccumulator of rare earth element (REE), but little is known about the ecophysiology of REE in this species. This study aimed to clarify tissue-level and organ-level distribution of REEs via synchrotron-based X-ray fluorescence microscopy (XFM). The results show that REEs (La + Ce) are mainly colocalized with Mn in the pinnae and pinnules, with the highest concentrations in necrotic lesions and lower concentrations in veins. In the cross sections of the pinnules, midveins, rachis, and stolons, La + Ce and Mn are enriched in the epidermis, vascular bundles, and pericycle (midvein). In these tissues, Mn is localized mainly in the cortex and mesophyll. We hypothesize that the movement of REEs in the transpiration flow in the veins is initially restricted in the veins by the pericycle between vascular bundle and cortex, while excess REEs are transported by evaporation and cocompartmentalized with Mn in the necrotic lesions and epidermis in an immobile form, possibly a Si-coprecipitate. The results presented here provide insights on how D. linearis regulates high concentrations of REEs in vivo, and this knowledge is useful for developing phytotechnological applications (such as REE agromining) using this fern in REE-contaminated sites in China.

International audience; Gold mining generates trace elements in the environment that are often associated with adverse effects on human health. This study determines trace elements levels in water, soil and plants (Datura inoxia and Calotropis procera) in Komabangou area, the main artisanal gold mining site in Niger. The collected samples were pre-treated before trace element analysis by inductively coupled plasma mass spectrometry (ICP-MS) for soils samples and by inductively coupled plasma optical emission spectroscopy (ICP-OES) for waters and plants samples. For soils, concentrations reached up to 555 mg kg(-1) and 468 mg kg(-1) for As and Zn, respectively. In waters, highest levels were observed for Zn (540 mg L-1), followed in decreasing order by Cu (7.4 mg L-1) and Al (4.27 mg L-1). As and Zn concentrations were 134 mg kg(-1) and 388 mg kg(-1); 49 mg kg(-1) and 46 mg kg(-1) respectively in leaves of Datura inoxia and Calotropis procera from gold site, whereas As and Zn, represented only 0.56 mg kg(-1) and 0.48 mg kg(-1); 6.40 mg kg(-1) and 46 mg kg(-1) respectively in leaves of control species. In fact, results obtained revealed both contaminations by trace elements of soil and water for the majority of samples taken in gold zone with an exceeding of limit values recommended by WHO in particular for As, Cr, Ni, Pb and Zn. Therefore, the establishment of a monitoring and treatment program for polluted soils and waters in this gold zone would be necessary to preserve human health.

International audience; Agromining is an emerging technology that utilizes selected ‘metal crops' (= hyperaccumulator plants) to extract valuable target metals from unconventional resources for profit from mineralised soils. Growth characteristics, shoot metal concentrations, and agrominable locations are important considerations in economic agromining. Globally, the greatest potential for nickel (Ni) agromining exists in the tropics. However, the agronomic systems of tropical ‘metal crops' have not been previously tested. Furthermore, it is currently unknown whether nutrient dosing of prospective tropical agromining Ni ‘metal crops’ could possibly cause a shoot Ni‐dilution effect which may ultimately limit economically viable Ni yields. We undertook a pioneering study on Ni uptake and growth responses to nutrient dosing in two promising tropical ‘metal crops' (Phyllanthus rufuschaneyi and Rinorea cf. bengalensis). The experiment consisted of a large randomised block growth trial in large pots over 12 months in Sabah (Malaysia). At 3‐month intervals, the plants were exposed to soluble treatments that altered available concentrations of nitrogen (N), phosphorus (P), and potassium (K). We found strong positive growth responses to N and P additions in P. rufuschaneyi, whereas K additions had negative growth effects. In R. cf. bengalensis, all treatments had positive growth effects. The increases in biomass in response to nutrient dosing did not significantly reduce shoot Ni concentrations in both species, with the exception of N addition in P. rufuschaneyi. This study reveals that Ni uptake and growth responses to nutrient dosing are species‐dependent, primarily influenced by the ecophysiology of the species. Inorganic fertilization could possibly be an important component of the management of local ‘metal crops' to be used in viable commercial agromining in the tropics, but this needs to be tested in the field with different formulations of N, P, and K.

International audience; PremiseHyperaccumulation of heavy metals in plants has never been documented from Central America or Mexico. Psychotria grandis, P. costivenia, and P. glomerata (Rubiaceae) have been reported to hyperaccumulate nickel in the Greater Antilles, but they also occur widely across the neotropics. The goals of this research were to investigate the geographic distribution of hyperaccumulation in these species and explore the phylogenetic distribution of hyperaccumulation in this clade by testing related species.MethodsPortable x‐ray fluorescence (XRF) spectroscopy was used to analyze 565 specimens representing eight species of Psychotria from the Missouri Botanical Garden herbarium.ResultsNickel hyperaccumulation was found in specimens of Psychotria costivenia ranging from Mexico to Costa Rica and in specimens of P. grandis from Guatemala to Ecuador and Venezuela. Among related species, nickel hyperaccumulation is reported for the first time in P. lorenciana and P. papantlensis, but no evidence of hyperaccumulation was found in P. clivorum, P. flava, or P. pleuropoda. Previous reports of hyperaccumulation in P. glomerata appear to be erroneous, resulting from taxonomic synonymy and specimen misidentification.ConclusionsHyperaccumulation of nickel by Psychotria is now known to occur widely from southern Mexico through Central America to northwestern South America, including some areas not known to have ultramafic soils. Novel aspects of this research include the successful prediction of new hyperaccumulator species based on molecular phylogeny, use of XRF technology to nondestructively obtain elemental data from herbarium specimens, and documentation of previously unknown areas of ultramafic or nickel‐rich soil based on such data.

Sabah (Malaysia) on the Island of Borneo has a high plant diversity (>8000 species) occurring on a wide range of soils, including ultramafic soils which are known to host hyperaccumulator plants. In this study a new approach (“Herbarium X-ray Fluorescence Ionomics”) was used to obtain elemental data from herbarium specimens using non-destructive X-ray Fluorescence spectroscopy analysis. In total ~7300 specimens were thus analysed for nickel, cobalt, and manganese concentrations at the Herbarium of the Forest Research Centre (FRC) in Sepilok, Sabah. The measurements led to recording a total of 759 specimens (originating from 17 families in 30 genera and 74 species) as trace element hyperaccumulators, including 28 nickel hyperaccumulator species (in 10 families, 17 genera), 12 cobalt hyperaccumulator species (in 3 families, 7 genera), and 51 manganese hyperaccumulator species (in 12 families, 24 genera). The outcomes of this research demonstrate that X-ray Fluorescence scanning is highly useful approach for hyperaccumulator plant discovery in herbarium collections that has the potential to add vast numbers of hyperaccumulating taxa to the global inventory.

On the basis of our previous field survey, we postulate that the pattern and degree of zinc (Zn) isotope fractionation in the Zn hyperaccumulator Noccaea caerulescens (J. & C. Presl) F. K. Mey may reflect a relationship between Zn bioavailability and plant uptake strategies. Here, we investigated Zn isotope discrimination during Zn uptake and translocation in N. caerulescens and in a nonaccumulator Thlaspi arvense L. with a contrasting Zn accumulation ability in response to low (Zn-L) and high (Zn-H) Zn supplies. The average isotope fractionations of the N. caerulescens plant as a whole, relative to solution (Δ(66)Znplant-solution), were -0.06 and -0.12‰ at Zn-L-C and Zn-H-C, respectively, indicative of the predominance of a high-affinity (e.g., ZIP transporter proteins) transport across the root cell membrane. For T. arvense, plants were more enriched in light isotopes under Zn-H-A (Δ(66)Znplant-solution = -0.26‰) than under Zn-L-A and N. caerulescens plants, implying that a low-affinity (e.g., ion channel) transport might begin to function in the nonaccumulating plants when external Zn supply increases. Within the root tissues of both species, the apoplast fractions retained up to 30% of Zn mass under Zn-H. Moreover, the highest δ(66)Zn (0.75‰-0.86‰) was found in tightly bound apoplastic Zn, pointing to the strong sequestration in roots (e.g., binding to high-affinity ligands/precipitation with phosphate) when plants suffer from high Zn stress. During translocation, the magnitude of isotope fractionation was significantly greater at Zn-H (Δ(66)Znroot-shoot = 0.79‰) than at Zn-L, indicating that fractionation mechanisms associated with root-shoot translocation might be identical to the two plant species. Hence, we clearly demonstrated that Zn isotope fractionation could provide insight into the internal sequestration mechanisms of roots when plants respond to low and high Zn supplies.

The fern Pteris vittata has been the subject of numerous studies because of its extreme arsenic hyperaccumulation characteristics. However, information on the arsenic chemical speciation and distribution across cell types within intact frozen-hydrated Pteris vittata fronds is necessary to better understand the arsenic biotransformation pathways in this unusual fern. While 2D X-ray absorption spectroscopy imaging studies show that different chemical forms of arsenic, As(III) and As(V), occur across the plant organs, depth-resolved information on arsenic distribution and chemical speciation in different cell types within tissues of Pteris vittata have not been reported. By using a combination of planar and confocal μ-X-ray fluorescence imaging and fluorescence computed μ-tomography, we reveal, in this study, the localization of arsenic in the endodermis and pericycle surrounding the vascular bundles in the rachis and the pinnules of the fern. Arsenic is also accumulated in the vascular bundles connecting into each sporangium, and in some mature sori. The use of 2D X-ray absorption near edge structure imaging allows for deciphering arsenic speciation across the tissues, revealing arsenate in the vascular bundles and arsenite in the endodermis and pericycle. This study demonstrates how different advanced synchrotron X-ray microscopy techniques can be complementary in revealing, at tissue and cellular levels, elemental distribution and chemical speciation in hyperaccumulator plants.

International audience; Ultramafic derived soils are characterized by low nutrient soils, a low Ca:Mg ratio, and high metal contents such as Ni, Co and Cr. The vegetation growing on these soils is highly adapted and includes both Ni hyperaccumulator and accumulator species. Today, approximately 530 Ni hyperaccumulator species are listed worldwide and the Ni concentration can be extremely high, e.g. up to 25% in latex from Pycnandra acuminata (Sapotaceae), a tree found in New Caledonia. The aim of this study is to identify the potential role of Ni hyperaccumulator plants in the Ni biogeochemical cycle at the soil surface by using Ni isotopes. A set of Ni hyperaccumulator and Ni accumulator plants as well as topsoils were sampled on the Barro Alto and Niquelândia ultramafic complexes (Goiás State, Brazil). Three Ni hyperaccumulator plants were collected: Justicia lanstyakii, Heliotropium aff. salicoides, Cnidoscolus aff. urens, as well as one Ni accumulator plant, Manihot sp. The isotopic compositions of the whole plants were determined and compared to those of the bulk topsoils and DTPA-extractable Ni. The topsoils exhibited δ60Ni values ranging from −0.30 ± 0.06‰ to 0.16 ± 0.05‰. The DTPA-extractable Ni in the topsoils ranged from 94 to 623 mg kg−1, i.e. 0.9–4.9% of the total soil Ni and was found to be isotopically heavier than the corresponding topsoil (from −0.30 ± 0.05‰ to 0.34 ± 0.08‰). The δ60Ni values for the Ni accumulator plants showed an enrichment in heavy Ni isotopes in the aerial parts of the plant compared to the roots, whereas similar δ60Ni values for the roots, stems and aerial parts suggested that no significant fractionation results from Ni uptake and translocation in Ni hyperaccumulator plants. Moreover, the aerial parts (i.e. leaves and flowers) from all of the plants analyzed showed the highest Ni concentrations and the heaviest δ60Ni values up to 1.21 ± 0.05‰. The enrichment in heavy Ni isotopes in the leaves (0.09 ± 0.06‰

International audience; Ultramafic soils are typically enriched in nickel (Ni), chromium (Cr), and cobalt (Co) and deficient in essential nutrients, making them unattractive for traditional agriculture. Implementing agromining systems in ultramafic agricultural soils represent an ecological option for the sustainable management and re-valorisation of these low-productivity landscapes. These novel agroecosystems cultivate Ni-hyperaccumulating plants which are able to bioaccumulate this metal in their aerial plant parts; harvested biomass can be incinerated to produce Ni-enriched ash or “bio-ore” from which Ni metal, Ni ecocatalysts or pure Ni salts can be recovered. Nickel hyperaccumulation has been documented in ∼450 species, and in temperate latitudes these mainly belong to the family Brassicaceae and particularly to the genus Odontarrhena (syn. Alyssum pro parte). Agromining allows for sustainable metal recovery without causing the environmental impacts associated with conventional mining activities, and at the same time, can improve soil fertility and quality and provide essential ecosystem services. Parallel reductions in Ni phytotoxicity over time would also permit cultivation of conventional agricultural crops. Field studies in Europe have been restricted to Mediterranean areas and these only evaluated the Ni-hyperaccumulator Odontarrhena muralis s.l. Two recent EU projects (Agronickel and LIFE-Agromine) have established a network of agromining field sites in ultramafic regions with different edapho-climatic characteristics across Albania, Austria, Greece and Spain. Soil and crop management practices are being developed so as to Kidd et al. Sustainable Agromining Systems for Nickel Recovery optimize the Ni agromining process; field studies are evaluating the potential benefits of fertilization regimes, crop selection and cropping patterns, and bioaugmentation with plant-associated microorganisms. Hydrometallurgical processes are being up-scaled to produce nickel compounds and energy from hyperaccumulator biomass. Exploratory techno-economic assessment of Ni metal recovery by pyrometallurgical conversion of O. muralis s.l. shows promising results under the condition that heat released during incineration can be valorized in the vicinity of the processing facility.

Background Nickel hyperaccumulator plants are of much interest for their evolution and unique ecophysiology, and also for potential applications in agromining—a novel technology that uses plants to extract valuable metals from soil. The majority of nickel hyperaccumulators are known from ultramafic soils in tropical regions (Cuba, New Caledonia and Southeast Asia), and one genus, Phyllanthus (Phyllanthaceae), is globally the most represented taxonomic entity. A number of tropical Phyllanthus-species have the potential to be used as ‘metal crops’ in agromining operations mainly because of their ease in cultivation and their ability to attain high nickel concentrations and biomass yields. Results One of the most promising species globally for agromining, is the here newly described species Phyllanthus rufuschaneyi. This species can be classified in subgenus Gomphidium on account of its staminate nectar disc and pistillate entire style and represents the most western species of this diverse group. The flower structure indicates that this species is probably pollinated by Epicephala moths. Conclusions Phyllanthus rufuschaneyi is an extremely rare taxon in the wild, restricted to Lompoyou Hill near Kinabalu Park in Sabah, Malaysia. Its utilization in agromining will be a mechanism for conservation of the taxon, and highlights the importance of habitat and germplasm preservation if rare species are to be used in novel green technologies.

International audience; Aims This study evaluated the effect of phloem translocation on Ni accumulation in the hyperaccumulator Noccaea caerulescens. Methods The first experiment assessed the metal and organic compound concentrations in phloem sap. Leaves were cut from plants grown in nutrient solutions added with 100 mu M Ni or Zn, and then used for phloem sap extraction by means of the EDTA-stimulated exudation method. In the second experiment, Ni-61(2+) was applied to old leaves as a foliar spray to assess bidirectional movement of Ni in phloem. Results In the first experiment, enrichment of Ni or Zn was found in phloem exudates, indicating high Ni and Zn phloem loading and translocation capacity in N. caerulescens. Amino acids, e.g. nicotianamine and histidine, were present at low concentrations in exudates, which are insufficient for Ni and Zn chelation. On the contrary, concentrations of organic acids, especially malate, were high in all treatments, which may be involved in Ni and Zn chelation in phloem. The second experiment showed that 89 % of Ni-61 exported from old leaves was translocated upward to young leaves, whereas only 11% moved downward to roots. Conclusions Phloem sap of N. caerulescens is enriched in Ni and malate, the majority of which moves upward to young tissues. Phloem translocation may play an important role for Ni accumulation in young leaves of N. caerulescens.

International audience; Nickel (Ni) agromining of ultramafic soils has been proposed as an eco-friendly option for metal recovery, which can also improve the fertility and quality of these low productive soils. The selection of adequate plant species and the analysis of their performance under the different climatic conditions are of interest for optimising the process and evaluating its full viability. A one-year field experiment was carried out to evaluate the viability of the two Ni-hyperaccumulating Mediterranean species, Alyssum murale and Leptoplax emarginata, for agromining purposes in ultramafic soils under a humid-temperate climate. Field plots of 50 m(2) were established and the soil was fertilised with gypsum and inorganic NPK fertilisers prior to cropping. Alyssum murale produced a slightly higher Ni yield than L. emarginata, but Ni bioaccumulation was dependent on the plant phenological stage for both species, being maximal at mid-flowering (4.2 and 3.0 kg Ni ha(-1), respectively). In both species, Ni was mainly stored in the leaves, especially in leaves of vegetative stems, but also in flowers and fruits in the case of L. emarginata. The main contributors to Ni yield of A. murale were flowering stems and their leaves, while for L. emarginata they were flowering stems and fruits. Implementing the agromining system increased soil nutrient availability, and modified microbial community structure and metabolic activity (due to fertilisation and plant root activity). The soil bacterial communities were dominated by Proteobacteria, Actinobacteria, Acidobacteria and Chloroflexi, and the agromining crops modified the relative abundance of some phyla (increasing Proteobacteria, Bacteroidetes and Nitrospirae and reducing Acidobacteria and Planctomycetes). Cultivating A. murale increased the densities of total culturable bacteria, while L. emarginata selected Ni-tolerant bacteria in its rhizosphere. In summary, both species showed great potential for their use in Ni agromining systems, although optimising soil and crop management practices could improve the phytoextraction efficiency. (C) 2018 Elsevier B.V. All rights reserved.

International audience; Nickel (Ni) agromining aims to phytoextract heavy metals using hyperaccumulators whilst at the same time rehabilitating ultramafic soils. After removing the bioavailable metal, ultramafic soils are improved in terms of their agronomic properties with the aim of future agricultural uses. The low fertility of ultramafic soils can be compensated by integrating legumes already used in traditional agro-systems because of their importance in soil nitrogen enrichment. However, few studies have evaluated the potential profits of legumes on Ni agromining and their potential benefits on soil biological fertility. Here, we characterized the effect of a crop rotation with two plants, a legume (Vicia sativa) and a hyperaccumulator (Alyssum murale), on the phytoextraction efficiency and on soil structure and biofunctioning. A pot experiment was set up in controlled conditions to grow A. murale and four treatments were tested: rotation with V. sativa (Ro), fertilized mono-culture (FMo), non-fertilized mono-culture (NFMo) and bare soil without plants (BS). No significant difference was found between the Ro and NFMo treatments for the dry biomass yield. However, the Ro treatment showed the highest Ni concentrations ([Ni]) in A. murale shoots compared to FMo and NFMo treatments. The Ro treatment plants had more than twice as many leaves [Ni] compared to FMo. Soil physico-chemical analyses showed that the Ro treatment was better structured and showed the highest presence of bacterial micro-aggregates, as well as less non-aggregated particles. Legumes integration in Ni-agromining systems could be a pioneering strategy to reduce chemical inputs and to improve soil biofunctioning and thus fertility.

Most of the research dedicated to agromining has focused on cultivating a single hyperaccumulator plant, although plant diversity has been shown to positively modify soil characteristics. Hence, we compared the effect of cropping a nickel-hyperaccumulator Alyssum murale with a legume (Vicia sativa) to A. murale's mono-culture, on the bacterial diversity and physico-chemical characteristics of an ultramafic soil. A pot experiment with 5 replicates was conducted in controlled conditions for 11 months. The treatments studied were: co-cropping and rotation vs. mineral fertilization controls and bare soil. The introduction of legumes induced a clearly positive effect on the soil's microbial biomass carbon and nitrogen. Arylsulfatase and urease activities tended to be enhanced in the co-cropping and rotation treatments and to be lessened in the mineral fertilization treatments. However, β-glucosidase and phosphatase activities were seen to decrease when legumes were used. Our results showed that the rotation treatment induced a higher organic matter content than the fertilized control did. Actinobacteria was the most-represented bacterial phyla and had lower relative abundance in treatments associating legumes. Conversely, the relative abundance of Acidobacteria and Gemmatimonadetes phyla increased but not significantly in treatments with legumes. The relative abundance of Chloroflexi phylum was shown to be significantly higher for the fertilized rotation control. The relative abundance of β-Proteobacteria subphylum increased but not significantly in treatments with legumes. NMDS analysis showed a clear separation between planted treatments and bare soil and between co-cropping and rotation and fertilized controls. Shannon index showed reduction in microbial diversity that was mainly due to chemical inputs in the soil. This study showed that these new cropping systems influenced both the bacterial diversity and the physico-chemical characteristics of an ultramafic soil. In addition, this study provides evidence that mineral fertilization can negatively impact bacterial communities and some of their functions linked to biogeochemical cycles.

Metal hyperaccumulator plants represent a unique biological resource for scientific research and practical applications. Though essential, however, an adequate knowledge of the systematics of these plants is often missing. This is the case of Odontarrhena, a large but taxonomically critical group of nickel hyperaccumulators from Eurasia. We present a study on this genus in Albania, to fill a gap in our knowledge of this group from a major centre of diversity of metallicolous flora, and to contribute updated information to the Global Hyperaccumulator Database. Morphological and karyological analyses of material from field collections across all major serpentine outcrops in the country, in different years and seasons, allowed to delimit seven taxa: O. albiflora, O. chalcidica, O. moravensis, O. sibirica, O. decipiens, O. smolikana subsp. glabra and O. rigida. The three latter taxa have been long neglected and were resurrected in view of their clear distinctness, while commonly accepted taxa such as O. bertolonii subsp. scutarina and O. markgrafii were reduced to synonymy of O. chalcidica due to the lack of consistent differentiation. Polyploidy was prevalent, while diploid complements were typical of the two vicariant endemics O. rigida and O. moravensis. Types are indicated or newly designated for each entity, and nomenclatural issues are addressed based on in-depth studies of literature and herbarium material. Revised descriptions, phenology, habitat and distribution data are given for each taxon, as well as original iconographies and chromosome counts. A revised identification key is provided. Shoot nickel concentrations were determined to assess accumulation levels of taxa and populations in natural conditions and their potential for phytoextraction of this metal from the soil. With ca. 23000 and 17000 µg of Ni g-1 of shoot dry weight, respectively, the tetraploids O. chalcidica and O. decipiens were the most promising candidates, especially the latter for its robust habit.

International audience; The majority of nickel hyperaccumulator plant species have been discovered by screening using a field spot test based on dimethylglyoxime. Recently, the use of a portable X-ray fluorescence spectroscopy instruments has enabled non-destructive analyses of existing herbarium collections. Given that the family Phyllanthaceae globally has the greatest numbers of hyperaccumulators, all available specimens from this family, including the speciose genus Antidesma, at the Forest Research Centre Herbarium in Sabah, Malaysia, were analysed. The results reveal 9 new manganese hyperaccumulators in the genus Antidesma, including Antidesma puncticulatum, with manganese concentrations reaching up to 46,400 A mu g g(-1). Prior to this study, only one manganese hyperaccumulator had been recorded in Sabah. The present study is the first to discover nickel hyperaccumulator plant species (four species) in the genus Antidesma, including Antidesma montis-silam with concentrations reaching up to 32,700 A mu g g(-1). Further collection and analyses of plant material of Antidesma montis-silam in the native habitat confirmed the high foliar nickel concentrations. The high nickel hyperaccumulating characteristic and ostensibly fast growth rate of Antidesma montis-silam infer potential for use in agromining technology. Some species in the genus Antidesma exhibit co-accumulation of manganese and nickel. This case-study shows how herbarium XRF screening can lead to discovery of taxa with unique properties.

Agromining aims to rehabilitate contaminated or natural metal-rich soils (ultramafic soils) by extracting metals of high economic importance, such as nickel (Ni), using hyperaccumulator plants and then to recover these metals for industrial purposes. Ultramafic soils are characterized by low fertility levels and this can limit yields of hyperaccumulators and metal phytoextraction. Here, we characterized the potential benefits for phytoextraction efficiency of co-cropping two plants: a Ni-hyperaccumulator (Alyssum murale; Brassicaceae) and a legume (Vicia sativa; Fabaceae). A field experiment with 3 replicates was set up in an ultramafic zone in North West Spain. Four treatments were tested: co-cropping (“Co”), fertilized mono-culture (“FMo”), non-fertilized mono-culture (“NFMo”) and bulk soil (“BS”). “FMo” and “Co” treatments increased the biomass yields of A. murale by 453% and 417% respectively, compared to “NFMo”. “Co” treatment generated 35% and 493% higher Ni-yields than “FMo” and “NFMo”, respectively. Most of the microbial analyses showed that introducing V. sativa (“Co” treatment) into the cropping system had beneficial effects. “Co” treatment significantly modified the phenotypical structure of bacterial communities and raise the relative abundance of the phylum Bacteroidetes and reduced that of Actinobacteria. In addition, non-metric multidimensional scaling analysis of the operational taxonomic units (OTUs) showed that “Co” was clearly separate from all other treatments. Thus, this study showed that co-cropping a hyperaccumulator with a legume in Ni-agromining systems not only improves plant biomass and Ni-yields, but also enhanced some soil microbial enzymatic activities. Ameliorating agromining by replacing fertilizers would combine eco-efficient or sustainable metal recovery with soil fertility/quality improvement.

Hyperaccumulator plants have the unique ability to concentrate specific elements in their shoot in concentrations that can be thousands of times greater than in normal plants. Whereas all known zinc hyperaccumulator plants are facultative hyperaccumulators with only populations on metalliferous soils hyperaccumulating zinc (except for Arabidopsis halleri and Noccaea species that hyperaccumulate zinc irrespective of the substrate), the present study discovered that Dichapetalum gelonioides is the only (zinc) hyperaccumulator known to occur exclusively on ‘normal’ soils, while hyperaccumulating zinc. We recorded remarkable foliar zinc concentrations (10 730 µg g−1, dry weight) in Dichapetalum gelonioides subsp. sumatranum growing on ‘normal’ soils with total soil zinc concentrations of only 20 µg g−1. The discovery of zinc hyperaccumulation in this tropical woody plant, especially the extreme zinc concentrations in phloem and phloem-fed tissues (reaching up to 8465 µg g−1), has possible implications for advancing zinc biofortification in Southeast Asia. Furthermore, we report exceptionally high foliar nickel concentrations in D. subsp. tuberculatum (30 260 µg g−1) and >10 wt% nickel in the ash, which can be exploited for agromining. The unusual nickel and zinc accumulation behaviour suggest that Dichapetalum-species may be an attractive model to study hyperaccumulation and hypertolerance of these elements in tropical hyperaccumulator plants.

Ultramafic soils display high concentrations of nickel and a number of nutrient deficiencies. Nickel-hyperaccumulator plants, such as Alyssum murale , have evolved in these environments and developed specific metal homeostasis, showing concentrations of nickel (Ni) sometimes exceeding 1% in their aerial biomass. Rhizosphere bacterial communities associated with Ni-hyperaccumulator plants can differ from those of non-accumulating plants growing on the same site. Among the edaphic factors that could influence the phylogenetic structure of the bacterial communities, altitude and metal-bioavailability such as Ni in particular, could be significant. Our objectives were to understand the specific changes in the structure of the A. murale rhizosphere bacterial community that occurred across two gradients: elevation and Ni geochemistry, using a high-throughput sequencing technique (454-pyrosequencing). In this study, Chloroflexi was the major phylum present, with 53–77% of relative abundance. Moreover, we found that the higher the soil's chemically-available Ni contents, the higher was the relative abundance of Proteobacteria (particularly Alphaproteobacteria ) and Actinobacteria . In contrast, the abundance of Chloroflexi decreased with increasing levels of available Ni. Our results demonstrate that the chemical-availability of Ni in the studied soil drives the bacterial community diversity in the rhizosphere of A. murale, regardless of elevation gradient and other soil physicochemical parameters.

International audience; Globally the discovery of hyperaccumulator plants has been hindered by systematic screening of plant species, and is highly biased towards Ni hyperaccumulators. This is mainly due to the existence of a reagent paper test that is only specific to nickel (based on dimethylglyoxime) such that more than 400 of the approximately 500 known hyperaccumulators species are for Ni. New technical advances now permit massive screening of herbarium specimens using non-destructive, portable X-Ray Fluorescence Spectroscopy (XRF), an approach that has already led to the discovery of numerous hyperaccumulator species new to science. The elemental distribution in selected hyperaccumulator plant tissues can then be further studied using techniques such as desktop or synchrotron micro-XRF, nuclear microprobe (PIXE), scanning/transmission electron microscopy with energy-dispersive spectroscopy (SEM/TEM-EDS), secondary ion mass spectrometry (SIMS) or laser ablation inductively-coupled plasma mass spectrometry (LA-ICP-MS). The use of histochemical dyes combined with light microscopy further aids in the identification of anatomical and structural features of the studied plant tissues.

International audience; In Mexico, ultramafic complexes are present in different regions from the northwest (Baja California Norte) to the southeast (Chiapas). In this paper, we present the results of the exploration of three ultramafic (serpentine) habitats in central and southern Mexico: Cuicatlan-Concepcin Papalo (Oaxaca), Tehuitzingo-Tecomatlan (Puebla), and San Juan de Otates (Guanajuato). Previous geology studies showed that these complexes are mainly made up of serpentinized peridotites. Soil analyses demonstrated typical ultramafic characteristics such as high content of Mg in relation to Ca, and high concentrations of Fe, Cr, Co, and Ni. Soil samples from Oaxaca and Puebla had similar Ni contents around 2300 mg kg(-1), while samples of Guanajuato showed the lowest Ni levels with an average of 200 mg kg(-1) as well as for other metals such as Co, Cr, Mn, and Zn. During this study, 83 plant specimens were collected, of which 52 were identified at genus level and 40 at species level. The collected plants belong to 19 different families such as Anacardiaceae, Fabaceae, Acanthaceae, Asteraceae, Sterculiaceae, and Verbenaceae which are also widely present in other ultramafic areas in Iran, Brazil, Sri Lanka, and Costa Rica. Only two Mexican endemic species are included in the collection. Ni hyperaccumulators were not detected at any of the studied sites. Therefore, hyperaccumulation, as a tolerance mechanism of the flora in response to ultramafic geochemical stress, does not seem to be developed in Central Mexico, as observed in the close Costa Rican site of Santa Elena.

This review synthesizes contemporary understanding of copper–cobalt (Cu–Co) tolerance and accumulation in plants. Accumulation of foliar Cu and Co to > 300 μg g−1 is exceptionally rare globally, and known principally from the Copperbelt of Central Africa. Cobalt accumulation is also observed in a limited number of nickel (Ni) hyperaccumulator plants occurring on ultramafic soils around the world. None of the putative Cu or Co hyperaccumulator plants appears to comply with the fundamental principle of hyperaccumulation, as foliar Cu–Co accumulation is strongly dose-dependent. Abnormally high plant tissue Cu concentrations occur only when plants are exposed to high soil Cu with a low root to shoot translocation factor. Most Cu-tolerant plants are Excluders sensu Baker and therefore setting nominal threshold values for Cu hyperaccumulation is not informative. Abnormal accumulation of Co occurs under similar circumstances in the Copperbelt of Central Africa as well as sporadically in Ni hyperaccumulator plants on ultramafic soils; however, Co-tolerant plants behave physiologically as Indicators sensu Baker. Practical application of Cu–Co accumulator plants in phytomining is limited due to their dose-dependent accumulation characteristics, although for Co field trials may be warranted on highly Co-contaminated mineral wastes because of its relatively high metal value.

International audience; Phytomining is the conception of agro-metallurgical production chains based on cropping hyperaccumulator plants on contaminated or naturally rich (ultramafic) soils to produce high value metal compounds. This study aimed to evaluate the effect of enhancing multispecies hyperaccumulator covers with Plant Growth Promoting Rhizobacteria (PGPR) on Ni phytoextraction. Plant growth promoting rhizobacteria were isolated from the rhizosphere of two different plant associations (Bornmuellera tymphaea - Noccaea tymphaea (NB) and Bornmuellera tymphaea - Alyssum murale (AB)) collected from natural conditions. The screening of isolates from both plant associations for their PGPR traits revealed two PGPR strains (AB30 and NB24) affiliated to Variovorax paradoxus. Inoculation of mesocosms containing the same plant associations with these selected PGPR was performed after 5 months of culture. The biomass of the different plant parts and respective concentrations of nickel in the plants were recorded 1 month after inoculation. Root biomass of the inoculated plant associations was significantly higher than the respective non-inoculated ones. The total biomass of the inoculated plant association Bornmuellera tymphaea - Noccaea tymphaea was significantly higher than the total biomass of all others. Similarly, shoot biomass of the inoculated plant association B. tymphaea - N. tymphaea was significantly higher than that of all other covers. Results showed an increase in Ni uptake when plants were inoculated with PGPR strains, when compared with uninoculated plant associations. According to the plant cover, the inoculation increased Ni amounts in roots of 105.8 and 66.4 % respectively in ABi and NBi covers, and 39.9 and 79.6 % in the shoots. The combination of the hyperaccumulator plants N. tymphaea-B.tymphaea inoculated by one of the two PGPR strains (strain NB24), isolated from the rhizosphere of this mixed cover, seemed to be an interesting option for an efficient Ni phytoextraction to be tested in the field.