Extracellular complexation of Cd in the Hartig net and cytosolic Zn sequestration in the fungal mantle of Picea abies – Hebeloma crustuliniforme ectomycorrhizas

Compartmentation of heavy metals on or within mycorrhizal fungi may serve as a protective function for the roots of forest trees growing in soils containing elevated concentrations of metals such as Cd and Zn. In this paper we present the first quantitative measurements by X‐ray microanalysis of heavy metals in high‐pressure frozen and cryosectioned ectomycorrhizal fungal hyphae. We used this technique to analyse the main sites of Cd and Zn in fungal cells of mantle and Hartig net hyphae and in cortical root cells of symbiotic Picea abies – Hebeloma crustuliniforme associations to gain new insights into the mechanisms of detoxification of these two metals in Norway spruce seedlings. The mycorrhizal seedlings were exposed in growth pouches to either 1 mM Cd or 2 mM Zn for 5 weeks. The microanalytical data revealed that two distinct Cd‐ and Zn‐binding mechanisms are involved in cellular compartmentation of Cd and Zn in the mycobiont. Whereas extracellular complexation of Cd occurred predominantly in the Hartig net hyphae, both extracellular complexation and cytosolic sequestration of Zn occurred in the fungal tissue. The vacuoles were presumed not to be a significant pool for Cd and Zn storage. Cadmium was almost exclusively localized in the cell walls of the Hartig net (up to 161 mmol kg ^{? 1} DW) compared with significantly lower concentrations in the cell walls of mantle hyphae (22 mmol kg ^{? 1} DW) and in the cell walls of cortical cells (15 mmol kg ^{? 1} DW). This suggests that the apoplast of the Hartig net is a primary accumulation site for Cd. Zinc accumulated mainly in the cell walls of the mantle hyphae (111 mmol kg ^{? 1} DW), the Hartig net hyphae (130 mmol kg ^{? 1} DW) and the cortical cells (152 mmol kg ^{? 1} DW). In addition, Zn occurred in high concentrations in the cytoplasm of the fungal mantle hyphae (up to 164 mmol kg ^{? 1} DW) suggesting that both the cell walls and the cytoplasm of fungal tissue are the main accumulation sites for Zn in P. abies resulting in decreased Zn transfer from the fungus to the root.     

Element distribution and iron speciation in mature wheat grains (Triticum aestivum L.) using synchrotron X‐ray fluorescence microscopy mapping and X‐ray absorption near‐edge structure (XANES) imaging

Several studies have suggested that the majority of iron (Fe) and zinc (Zn) in wheat grains are associated with phytate, but a nuanced approach to unravel important tissue‐level variation in element speciation within the grain is lacking. Here, we present spatially resolved Fe‐speciation data obtained directly from different grain tissues using the newly developed synchrotron‐based technique of X‐ray absorption near‐edge spectroscopy imaging, coupling this with high‐definition μ‐X‐ray fluorescence microscopy to map the co‐localization of essential elements. In the aleurone, phosphorus (P) is co‐localized with Fe and Zn, and X‐ray absorption near‐edge structure imaging confirmed that Fe is chelated by phytate in this tissue layer. In the crease tissues, Zn is also positively related to P distribution, albeit less so than in the aleurone. Speciation analysis suggests that Fe is bound to nicotianamine rather than phytate in the nucellar projection, and that more complex Fe structures may also be present. In the embryo, high Zn concentrations are present in the root and shoot primordium, co‐occurring with sulfur and presumably bound to thiol groups. Overall, Fe is mainly concentrated in the scutellum and co‐localized with P. This high resolution imaging and speciation analysis reveals the complexity of the physiological processes responsible for element accumulation and bioaccessibility.     

Bioimaging of multiple elements by high‐resolution LA‐ICP‐MS reveals altered distribution of mineral elements in the nodes of rice mutants

Inter‐vascular transfer in rice (Oryza sativa) nodes is required for delivering mineral elements to developing tissues, which is mediated by various transporters in the nodes. However, the effect of these transporters on distribution of mineral elements in the nodes at a cellular level is still unknown. Here, we established a protocol for bioimaging of multiple elements at a cellular level in rice node by laser ablation‐inductively coupled plasma‐mass spectrometry (LA‐ICP‐MS), and compared the mineral distribution profile between wild‐type (WT) rice and mutants. Both relative comparison of mineral distribution normalized by endogenous ¹³C and quantitative analysis using spiked standards combined with soft ablation gave valid results. Overall, macro‐nutrients such as K and Mg were accumulated more in the phloem region, while micro‐nutrients such as Fe and Zn were highly accumulated at the inter‐vascular tissues of the node. In mutants of nodal Zn transporter OsHMA2, Zn localization pattern in the node tissues did not differ from that of WT; however, Zn accumulation in the inter‐vascular tissues was lower in uppermost node I but higher in the third upper node III compared with the WT. In contrast, Si deposition in the mutants of three nodal Si transporters Lsi2, Lsi3 and Lsi6 showed different patterns, which are consistent with the localization of these transporters. This improved LA‐ICP‐MS analysis combined with functional characterization of transporters will provide further insight into mineral element distribution mechanisms in rice and other plant species.     

Zinc hyperaccumulation and cellular distribution in Arabidopsis halleri

Although Arabidopsis halleri ( = Cardaminopsis halleri) is known as a Zn hyperaccumulator, there have been no detailed studies on Zn accumulation, tolerance and cellular distribution in this species. In a hydroponic experiment, A. halleri grew healthily with Zn concentrations varying from 1 to 1000 μ M , without showing phytotoxicity or reduction in root or shoot dry weights. The concentration of Zn in the shoots increased from 300 μ g g ^{? 1} dry weight in the 1 μ M Zn treatment to 32 000 μ g g ^{? 1} in the 1000 μ M Zn treatment. Approximately 60% of the total Zn in the shoots were water‐soluble, and there was no evidence of Zn and P co‐precipitation. Both citric and malic acid concentrations in the shoots were not significantly affected by the Zn treatments, whereas in the roots there was a positive response in both organic acids to increasing Zn in solution. Cellular distribution of Zn, Ca and K in frozen hydrated leaf tissues was examined using energy‐dispersive X‐ray microanalysis. Zinc was sequestered in the base of trichomes, whereas the middle and upper parts of trichomes were highly enriched with Ca. Mesophyll cells appeared to have more Zn than the epidermis, probably because the latter were very small in size. Similarities and differences between A. halleri and the other well‐known Zn hyperaccumulator, Thlaspi caerulescens, are discussed.     

Bacteria associated with yellow lupine grown on a metal‐contaminated soil: in vitro screening and in vivo evaluation for their potential to enhance Cd phytoextraction

In order to stimulate selection for plant‐associated bacteria with the potential to improve Cd phytoextraction, yellow lupine plants were grown on a metal‐contaminated field soil. It was hypothesised that growing these plants on this contaminated soil, which is a source of bacteria possessing different traits to cope with Cd, could enhance colonisation of lupine with potential plant‐associated bacteria that could then be inoculated in Cd‐exposed plants to reduce Cd phytotoxicity and enhance Cd uptake. All cultivable bacteria from rhizosphere, root and stem were isolated and genotypically and phenotypically characterised. Many of the rhizobacteria and root endophytes produce siderophores, organic acids, indole‐3‐acetic acid (IAA) and aminocyclopropane‐1‐carboxylate (ACC) deaminase, as well as being resistant to Cd and Zn. Most of the stem endophytes could produce organic acids (73.8%) and IAA (74.3%), however, only a minor fraction (up to 0.7%) were Cd or Zn resistant or could produce siderophores or ACC deaminase. A siderophore‐ and ACC deaminase‐producing, highly Cd‐resistant Rhizobium sp. from the rhizosphere, a siderophore‐, organic acid‐, IAA‐ and ACC deaminase‐producing highly Cd‐resistant Pseudomonas sp. colonising the roots, a highly Cd‐ and Zn‐resistant organic acid and IAA‐producing Clavibacter sp. present in the stem, and a consortium composed of these three strains were inoculated into non‐exposed and Cd‐exposed yellow lupine plants. Although all selected strains possessed promising in vitro characteristics to improve Cd phytoextraction, inoculation of none of the strains (i) reduced Cd phytotoxicity nor (ii) strongly affected plant Cd uptake. This work highlights that in vitro characterisation of bacteria is not sufficient to predict the in vivo behaviour of bacteria in interaction with their host plants.     

Differentiation of metallicolous and non‐metallicolous Salix caprea populations based on phenotypic characteristics and nuclear microsatellite (SSR) markers

The Salicaceae family comprises a large number of high‐biomass species with remarkable genetic variability and adaptation to ecological niches. Salix caprea survives in heavy metal contaminated areas, translocates and accumulates Zn/Cd in leaves. To reveal potential selective effects of long‐term heavy metal contaminations on the genetic structure and Zn/Cd accumulation capacity, 170 S. caprea isolates of four metal‐contaminated and three non‐contaminated middle European sites were analysed with microsatellite markers using Wright's F statistics. The differentiation of populations North of the Alps are more pronounced compared to the Southern ones. By grouping the isolates based on their contamination status, a weak but significant differentiation was calculated between Northern metallicolous and non‐metallicolous populations. To quantify if the contamination and genetic status of the populations correlate with Zn/Cd tolerance and the accumulation capacity, the S. caprea isolates were exposed to elevated Cd/Zn concentrations in perlite‐based cultures. Consistent with the genetic data nested anova analyses for the physiological traits find a significant difference in the Cd accumulation capacity between the Northern and Southern populations. Our data suggest that natural populations are a profitable source to uncover genetic mechanisms of heavy metal accumulation and biomass production, traits that are essential for improving phytoextraction strategies.     

Distributions of cadmium,zinc, and polyphenols in Gamblea innovans

Gamblea innovans is a Cd- and Zn-accumulating deciduous tree widely distributed in the secondary forests of Japan. We aimed to understand the characteristics of Cd and Zn accumulation in G. innovans in order to effectively utilize the species for phytoremediation. To accomplish that, we studied the relationship between secondary metabolite concentrations and the accumulation and distributions of Cd and Zn in G. innovans leaves and basal stems using micro-X ray fluorescence (µ-XRF). Our results showed a negative correlation between Zn leaf concentrations and polyphenol/2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity. This finding might be related to stress or the manifestation of a mechanism for tolerance to Cd and Zn accumulation. In addition, we observed that Cd accumulated primarily in the apoplastic region of surface tissues such as bark and the epidermis of leaves, whereas Zn accumulated in both apoplastic and symplastic regions. Thus, it might be possible that G. innovans can distinguish between Cd and Zn and control their translocation.     

An insight into the thermodynamic characteristics of human thrombopoietin complexation with TN1 antibody

Human thrombopoietin (hTPO) primarily stimulates megakaryocytopoiesis and platelet production and is neutralized by the mouse TN1 antibody. The thermodynamic characteristics of TN1 antibody–hTPO complexation were analyzed by isothermal titration calorimetry (ITC) using an antigen‐binding fragment (Fab) derived from the TN1 antibody (TN1‐Fab). To clarify the mechanism by which hTPO is recognized by TN1‐Fab the conformation of free TN1‐Fab was determined to a resolution of 2.0 Å using X‐ray crystallography and compared with the hTPO‐bound form of TN1‐Fab determined by a previous study. This structural comparison revealed that the conformation of TN1‐Fab does not substantially change after hTPO binding and a set of 15 water molecules is released from the antigen‐binding site (paratope) of TN1‐Fab upon hTPO complexation. Interestingly, the heat capacity change (ΔCp) measured by ITC (?1.52 ± 0.05 kJ mol^?1 K^?1) differed significantly from calculations based upon the X‐ray structure data of the hTPO‐bound and unbound forms of TN1‐Fab (?1.02 ～ 0.25 kJ mol^?1 K^?1) suggesting that hTPO undergoes an induced‐fit conformational change combined with significant desolvation upon TN1‐Fab binding. The results shed light on the structural biology associated with neutralizing antibody recognition.     

Melatonin and roentgen irradiation-induced acute radiation enteritis in Albino rats: an animal model

Background: Roentgen irradiation can affect normal cells, especially the rapidly growing ones such as the mucosal epithelial cells of the small intestine. The small intestine is the most radiosensitive gastrointestinal organ and patients receiving radiotherapy directed to the abdomen or pelvis may develop radiation enteritis. Although roentgen rays are widely used for both imaging and therapeutic purposes, our knowledge about the morphological changes associated with radiation enteritis is lacking. Hypothesis: This study tries to tests the hypothesis that “the intake of melatonin can minimize the morphological features of cell damage associated with radiation enteritis”. Objectives and methods: We performed this investigation to test our hypothesis and to examine the possible radioprotective effects of melatonin in acute radiation enteritis. To achieve these goals, an animal model consisting of 60 Albino rats was established. The animals were divided into five groups: Group 1, non‐irradiated; Group 2, X‐ray irradiated (X‐ray irradiation, 8 Grays); Group 3, X‐ray irradiated‐pretreated with solvent (ethanol and phosphate buffered saline); Group 4, non‐irradiated‐group treated with melatonin, and Group 5, X‐ray irradiated‐pretreated with melatonin. The small intestines were evaluated for gross (macroscopic), histological, morphometric (light microscopy), and ultrastructural changes (transmission electron microscopy). Results: We found morphological variations among the non‐irradiated‐group, X‐ray irradiated‐group and X‐ray irradiated‐intestines of the animals pretreated with melatonin. The development of acute radiation enteritis in X‐ray irradiated‐group (Groups 2 and 3) was associated with symptoms of enteritis (diarrhea and abdominal distention) and histological features of mucosal injury (mucosal ulceration, necrosis of the epithelial cells). There was a significant reduction of the morphometric parameters (villous count, villous height, crypt height and villous/crypt height ratio). Moreover, the ultrastructural features of cell damage were evident including: apoptosis, lack of parallel arrangement of the microvilli, loss of the covering glycocalyx, desquamation of the microvilli, vacuolation of the apical parts of the cells, dilatation of the rough endoplasmic reticulum, and damage of the mitochondrial cristae. In the non‐irradiated‐group and in X‐ray irradiated‐intestines of the animals pretreated with melatonin (Group 5), these changes were absent and the intestinal mucosal structure was preserved. Conclusion: Administration of melatonin prior to irradiation can protect the intestine against X‐rays destructive effects, i.e. radiation enteritis. The clinical applications of these observations await further studies.     

Micro‐particle‐induced X‐ray emission mapping of elemental distribution in roots of a Mediterranean‐type sclerophyll,Agathosma betulina (Berg.) Pillans,colonized by Cryptococcus laurentii

The role of rhizosphere yeasts as plant nutrient‐scavenging microsymbionts in resource‐limited Mediterranean‐type heathlands is unknown. This study, therefore, focused on quantitative elemental distribution within the roots of a medicinal sclerophyll, Agathosma betulina (Berg.) Pillans, grown under nutrient‐poor conditions, and colonized by Cryptococcus laurentii. Micro‐particle‐induced X‐ray emission (PIXE) was used to assess quantitative elemental distribution within the roots of A. betulina inoculated with viable C. laurentii, as well as within roots of control plants that received autoclaved yeast. To aid in the interpretation of heterogeneous elemental distribution patterns, apoplastic barriers (Casparian bands) in root tissues were located using fluorescence microscopy. In addition, root cross‐sections were examined for endophytic C. laurentii using light and transmission electron microscopy (TEM). The average concentrations of P, Fe and Mn were significantly (P < 0.05) higher in roots of yeast‐inoculated plants, compared to control plants. Casparian bands were observed in the exodermal cells of both treatments, and the presence of these bands was correlated with elemental enrichment in the epi/exodermal‐outer cortical tissues. Light and TEM micrographs revealed that the yeast was not a root endophyte. This is the first report describing the role of a soil yeast as a plant nutrient‐scavenging microsymbiont.     

Complexation and detoxification of Zn and Cd in metal accumulating plants

Metal accumulating plants exposed to toxic levels of zinc (Zn) and cadmium (Cd) uptake metals through extracellular and intracellular complexation with inorganic and organic ligand formation. However, little is known about the nature and formation mechanism of these metal–ligand complexes. Though, Zn and Cd have many similar chemical properties, yet their complexation and compartmentalization in plants vary with plant species. In principal, the question arises what factors govern Zn and Cd partitioning in plants? What form of the metal is taken up by the root, and is further distributed and accumulated in both vegetative and reproductive tissues? Therefore, the aim of present study is to address several questions concerning the mechanisms of Zn and Cd coordination and compartmentalization in plants using X-ray absorption spectroscopy (XAS) technique. XAS allows direct determination of elemental oxidation states and coordination environments in different plant tissues. This review article briefly explains some other important techniques of XAS; EXAFS (extended X-ray absorption fine structure) and XANES (X-ray absorption near edge structure), which are employed for determining Zn and Cd complexation within the plant. Therefore, In present review, the predominant as well as the minor chemical forms of Zn and Cd present in particular plant tissue have been discussed which could give better insight towards metal accumulation and detoxification mechanisms operated in plants. This information could assist in employing suitable hyperaccumulator plants for metal phytoextraction and reclamation of metal contaminated sites.     

A Comparative Study of the Effects of Nontoxic Chloride Treatments on CdTe Solar Cell Microstructure and Stoichiometry

This work presents the first systematic comparison of the effects of a range of chlorides (CdCl₂, MgCl₂, NaCl, and NH₄Cl) on the microstructure and chemical composition of CdTe/CdS/ZnO/SnO₂ solar cells, providing valuable insight to the ubiquitous Cl‐activation process. Using X‐ray diffraction, it is shown that CdCl₂ induces the greatest extent of recrystallization (standard deviation of texture coefficients, σ, reduces from 0.93 for as‐grown CdTe to 0.43) and minimizing stress (from 178 MPa for as‐grown material to zero). MgCl₂ treatment also yields significant randomization of the CdTe texture (σ = 0.55) but NaCl treatment does not (σ = 1.10). A strong correlation between the extent of metallurgical changes induced by the chloride treatment (and consequently, device efficiency) and the dissociation energy of the cationCl bond is shown, thereby accounting for the ineffectiveness of NaCl (bond energy = 4.3 eV). From this, a mechanism for Cl activation is postulated. By X‐ray photoelectron spectroscopy it is also shown that the Te/Cd ratio at the back surface, and the Cl content at the CdTe–CdS interface, are both higher following CdCl₂‐ and MgCl₂ treatments (Te/Cd = 1.3–1.4, and 1–2 at% Cl) than following NaCl treatment (Te/Cd = 1.1, and 0 at% Cl).     

Cd speciation and localization in the hyperaccumulator Arabidopsis halleri

Arabidopsis halleri is a model plant investigated for Zn and Cd hyperaccumulation. In this work, A. halleri plants from metallicolous origin exposed to Cd and Zn were studied using an original combination of chemical and physical techniques to obtain insights on Cd hyperaccumulation mechanisms. In most cases, Zn and Cd accumulation in leaves increased with time and with their respective concentrations in hydroponics. At the rosette scale, ¹⁰⁹Cd autoradiography showed that the amount of Cd increased with time but there was no clear trend in the localization of Cd in young or mature leaves. At the leaf scale, an enrichment of the petiole, central vein and trichomes was observed after 3 weeks. After 9 weeks, leaf edges were the most Cd-enriched tissues, and regions along leaf vascular bundles appeared less concentrated. Bulk Cd K-edge extended X-ray absorption fine structure (EXAFS) spectroscopy showed that Cd was predominantly bound to COOH/OH groups belonging to organic acids and/or cell wall components. Cd bound to thiol groups was found as a secondary species (less than 25%). Thiols ligands might correspond, at least partly, to glutathione found in significant amount in aerial parts, but phytochelatins were not detected. These results show that the mechanisms of Cd storage and detoxification in A. halleri differ from what was previously found for Zn.     

Ultrastructural and metabolic disorders induced by short‐term cadmium exposure in Avicennia schaueriana plants and its excretion through leaf salt glands

• Environmental cadmium (Cd) sources have increased in mangrove sediments in recent decades, inducing cellular damage to many plants. Avicennia schaueriana is abundant in mangrove sites and has been subject to Cd contamination. The possible effects of Cd toxicity and the structural and physiological disturbances to this plant were studied. Can this plant express early cellular tolerance mechanisms to such metal contamination?
• Seedlings of A. schaueriana were collected from sites of their natural occurrence, placed in plastic pots containing nutrient solution for 60 days, and subsequently exposed to increasing Cd concentrations for 5 days under experimental conditions. The anatomical, ultrastructural and physiological changes induced by Cd were analysed.
• Cd accumulated mainly in the root system and in pneumatophores, stems and leaves, induced differential accumulation of mineral nutrients, but did not induce necrosis or changes in leaf anatomy. However, there was a decrease in starch grains and an increase in deposited electron‐dense material in the cortex and vascular bundles. Cd induced both increases in calcium (Ca) content in shoots and Ca oxalate crystal precipitation in leaf mesophyll and was detected in crystals and in the secretion of salt glands.
• Our observations and experimental results provide evidence of Cd tolerance in A. schaueriana. As a new feature, despite the clear cellular physiological disorders, this plant is able to eliminate Cd through leaf salt glands and immobilise it in Ca crystals, representing fast mechanisms for Cd exclusion and complexation in leaves in heavy metal coastal polluted marine ecosystems.

     

Bioaccumulation and Physiological Response of Five Willows to Toxic Levels of Cadmium and Zinc

We evaluated the phytoremediation potential of Salix spp. exposed to high cadmium (Cd) and zinc (Zn) concentrations to select feasible plant materials for restoration and revegetation of mining soil contaminated by heavy metals on the basis of their Cd and Zn accumulation, Cd-Zn interaction on bioaccumulation, and the changes of photosynthetic parameters. The Cd and Zn concentrations were in the order of root > leaf > stem, regardless of the species. In the combined Cd and Zn treatment, the leaf and stem Cd concentration in all species were higher relative to Cd-alone treatment. In contrast, the Zn concentration in plant tissues when exposed to the combined Cd + Zn treatment decreased relative to the Zn-alone treatment. The translocation factor (TF) of Cd and Zn from root to leaf was generally higher compared to TF from root to stem than those in the single treatment. The Cd + Zn treatments resulted in enhanced translocation of Cd from root to aboveground tissue (synergistic), while the same treatment suppressed the Zn translocation from root to leaf and stem (antagonistic). The reduction of photosynthetic parameters in Zn alone and Cd + Zn treatments was generally higher than that of Cd-alone treatment. Among the different species, S. caprea and P. alba×glandulosa have the lowest photosynthetic reduction relative to the control. Overall, S. caprea could be a potential candidate for phytoremediation of Cd- and Zn-contaminated sites.     

Cadmium biosorption properties of the metal‐resistant Ochrobactrum cytisi Azn6.2

The aim of this work was to establish the conditions for using Ochrobactrum cytisi Azn6.2 as a metal biosorbent. Azn6.2 is a novel strain from the legume symbiont O. cytisi that has been isolated from nodules of Medicago polymorpha plants grown on heavy metal‐polluted soils. Compared with the strain ESC1, Azn6.2 showed some biochemical differences, as well as antibiotic susceptibility, Azn6.2 was multi‐resistant to heavy metals, such as Cu, Cd and Zn, and bacterial pellets were able to biosorb high amounts of Cd and Zn. As shown by scanning electron microscopy coupled to energy dispersive X‐ray, most of Cd was attached to the cell surface. Optimal conditions for Cd biosorption were established, being 1 mM Cd ions in solution and 2 h of contact with the biosorbent at room temperature. At these conditions, maximal Cd loading capacity reached 32–34 mg/g. Cd desorption from bacterial pellets was achieved after washing with EDTA or, at higher efficiency, at pH 1.0. These results indicated that biosorption/desorption on O. cytisi Azn6.2 biomass should be a cost‐effective method for Cd recovery from contaminated solutions.     

Cadmium reduces zinc uptake but enhances its translocation in the cadmium-accumulator, <Emphasis Type="Italic">Carpobrotus rossii</Emphasis>, without affecting speciation

Background and aims

Interactions between Cd and Zn occur in soils and plants but are inconsistent. This study examined how Cd/Zn interactions influence the growth of Carpobrotus rossii (Haw.) and the accumulation of Cd and Zn in plants.

Methods

Plants were grown in nutrient solutions containing 5–100 μM Zn and 0, 5 or 15 μM Cd. Plant growth and tissue concentrations were measured, and the speciation of Zn within the plant tissues determined using synchrotron-based X-ray absorption spectroscopy.

Results

There was an additive negative interaction between Cd and Zn on root growth. Only the highest level of Zn (100 μM) decreased Cd concentrations in root and shoot tissues (by 40–64%), whilst 100 μM Zn enhanced Cd translocation at 5 μM Cd but decreased it at 15 μM Cd. In contrast, both 5 and 15 μM Cd decreased Zn concentrations in root and shoot tissues but increased Zn translocation by 30–90%. This interaction was not associated with changes in Zn speciation within the plants, with most Zn associated with oxalate (48–87%).

Conclusions

The presence of Zn and Cd resulted in an additive negative effect on root growth, but an antagonistic pattern in their accumulation in shoots of C. rossii.

     

Tolerance and potential for bioaccumulation of <Emphasis Type="Italic">Alternanthera tenella</Emphasis> Colla to cadmium under in vitro conditions

High concentrations of cadmium (Cd) in the environment can threaten the local biota and one of its main sources is anthropic activities such as zinc (Zn) mining. Some plant species are able to tolerate high Cd concentrations, using anatomical and physiological strategies to avoid the absorption or accumulation of this element in their biomass. The in vitro assessment of these strategies is an efficient way to control variables external to the experiment. We aimed to investigate the anatomical and physiological changes in Alternanthera tenella exposed to Cd and its potential for accumulation in controlled microenvironmental conditions. We evaluated changes in the leaf and root anatomy, antioxidant system, and biomass of A. tenella grown in a culture medium containing increasing Cd concentrations (0, 50, 100, and 150 μM), in the presence of 1500 μM Zn. Alternanthera tenella was able to accumulate Cd and Zn and these elements competed for absorption by the species. Increase in Cd in the medium led to a progressive thickening of the root tissues, which was also observed on the leaves, albeit only at concentrations below 100 μM Cd. The concentration of 150 μM Cd was toxic to the leaf tissue and stimulated the formation of hydrogen peroxide, interfering with the antioxidant system and reducing plant biomass and the chlorophyll levels. Therefore, in vitro cultivated A. tenella can accumulate Cd and tolerate up to 100 μM Cd by modifying its anatomy and physiology in order to cope with Cd stress.