The role of plants in ironstone evolution: iron and aluminium cycling in the rhizosphere.

The Carajás plateaus in Brazil host endemic epilithic vegetation ("campo rupestre") on top of ironstone duricrusts, known as canga. This capping rock is primarily composed of iron(III) oxide minerals and forms a physically resistant horizon. Field observations reveal an intimate interaction between canga's surface and two native sedges (Rhynchospora barbata and Bulbostylis cangae). These observations suggest that certain plants contribute to the biogeochemical cycling of iron. Iron dissolution features at the root-rock interface were characterised using synchrotron-based techniques, Raman spectroscopy and scanning electron microscopy. These microscale characterisations indicate that iron is preferentially leached in the rhizosphere, enriching the comparatively insoluble aluminium around root channels. Oxalic acid and other exudates were detected in active root channels, signifying ligand-controlled iron oxide dissolution, likely driven by the plants' requirements for goethite-associated nutrients such as phosphorus. The excess iron not uptaken by the plant can reprecipitate in and around roots, line root channels and cement detrital fragments in the soil crust at the base of the plants. The reprecipitation of iron is significant as it provides a continuously forming cement, which makes canga horizons a 'self-healing' cover and contributes to them being the world's most stable continuously exposed land surfaces. Aluminium hydroxide precipitates ("gibbsite cutans") were also detected, coating some of the root cavities, often in alternating layers with goethite. This alternating pattern may correspond with oscillating oxygen concentrations in the rhizosphere. Microbial lineages known to contain iron-reducing bacteria were identified in the sedge rhizospheric microbiome and likely contribute to the reductive dissolution of iron(III) oxides within canga. Drying or percolation of oxygenated water to these anaerobic niches have led to iron mineralisation of biofilms, detected in many root channels. This study sheds light on plants' direct and indirect involvement in canga evolution, with possible implications for revegetation and surface restoration of iron mine sites.

Uptake, transformation, and environmental impact of zinc oxide nanoparticles in a soil-wheat system.

Zinc oxide nanoparticles (ZnO-NPs) are metal-based nanomaterials, but their long-term effects on plant growth and the soil environment in the field remain unclear with most previous studies using short-term laboratory and glasshouse studies. In this study, we used a field experiment to examine the long-term effects of ZnO-NPs in a soil-wheat (Triticum aestivum) system. It was found that although ZnO-NPs had no significant effect on either yield or the concentration of other nutrients within the grain, the application of ZnO-NPs significantly increased Zn concentrations. Indeed, for grain, the application of ZnO-NPs to both the soil and foliage (SFZnO) (average of 33.1 mg/kg) significantly increased grain Zn concentrations compared to the the control treatment (21.7 mg/kg). Using in situ analyses, nutrients were found to accumulate primarily in the crease tissue and the aleurone layer of the grain, regardless of treatment. Specifically, the concentration of Zn in the aleurone layer for the SFZnO treatment was 2-3 times higher than that in the control, being >300 mg/kg, whilst the Zn concentration in the crease tissue was ca. 600 mg/kg in the SFZnO treatment, being two times higher than for the control. Although the application of ZnO-NPs increased the total Zn within the grain, it did not accumulate within the grain as ZnO-NPs with this being important for food safety, but rather mainly as Zn-phytate, with the remainder of the Zn complexed with either cysteine or phosphate. Finally, we also observed that ZnO-NPs caused fewer changes to the soil bacterial community structure and that it had no nano-specific toxicity.

Improved Zn bioavailability by its enhanced colocalization and speciation with S in wheat grain tissues after N addition.

Zinc bioavailability with the presence of other elements in wheat grains might be affected by fertilizers. A long-term field experiment was conducted to examine effects of N fertilizer on Zn bioavailability in wheat grain tissues, with changes in the concentrations, distribution, and speciation of Zn as well as P and sulfur S via synchrotron-based technology. Results showed that addition of N fertilizer was associated with changes in Zn concentrations and distributions in grain tissues, especially in the crease region and endosperm. Simultaneously, N addition enhanced Zn-S colocalization in the crease region and endosperm and lowered the P/Zn ratio and Zn-P colocalization. Addition of N fertilizer with P increased Zn-cysteine (9.2%) and decreased Zn-phytate (47.3%) in the crease region, leading to potentially higher grain Zn bioavailability. Thus, addition of N fertilizer improved concentrations and bioavailability of Zn, by coordinating the relationships among Zn, P and S within wheat grains.

Non-target impacts of pesticides on soil N transformations, abundances of nitrifying and denitrifying genes, and nitrous oxide emissions.

Agriculture is the leading contributor to global nitrous oxide (N2O) emissions, mostly from soils. We examined the non-target impacts of four pesticides on N transformations, N cycling genes and N2O emissions from sugarcane-cropped soil. The pesticides, including a herbicide glyphosate (GLY), an insecticide imidacloprid (IMI), a fungicide methoxy ethyl mercuric chloride (MEMC) and a fumigant methyl isothiocyanate (MITC), were added to the soil and incubated in laboratory at 25 °C. The soil microcosms were maintained at two water contents, 55 % and 90 % water holding capacity (WHC), to simulate aerobic and partly anaerobic conditions, respectively. Half of the soil samples received an initial application of KNO3 and were then maintained at 90 % WHC for 38 d, whilst the other half received (NH4)2SO4 and were maintained at 55 % WHC for 28 d followed by 10 d at 90 % WHC to favour denitrification. Responses of individual functional genes involved in nitrification and denitrification to the pesticides and their relationships to N2O emissions varied with time and soil water. Overall, MITC had pronounced repressive effects on AOA and AOB amoA gene abundances and gross nitrification. Under 55 % WHC during the initial 28 d, N2O emissions were low for all treatments (≤62 µg N kg-1 soil). However, under 90 % WHC (either during the first 28 d or the increase in water content from 55 to 90 % WHC after 28 d) the cumulative N2O emissions increased markedly. Overall, under 90 % WHC the cumulative N2O emissions were 19 (control) to 79-fold (MITC) higher than under 55% WHC; with the highest emissions observed in the MITC treatment (3140 µg N kg-1 soil). This was associated with increases in gross nitrate consumption rates and abundances of denitrifying genes (nirK, nirS and qnorB). Therefore, to minimise N2O emissions, MITC should not be applied to field under wet conditions favouring denitrification.

The Voltaic Effect as a Novel Mechanism Controlling the Remobilization of Cadmium in Paddy Soils during Drainage.

Excessive cadmium (Cd) accumulation in rice grain is a global issue that affects human health. The drainage of paddy soils during the grain filling period leads to the remobilization of Cd in soils, resulting in most of the Cd accumulated in rice grain. The rate of Cd remobilization during drainage differs markedly among soils, but the mechanisms underlying these differences remain largely unknown. Using microcosm soil incubation, electrochemical experiments, isotope labeling, and microscopic and spectroscopic analyses, here, we discover the voltaic effect as a novel mechanism controlling the remobilization of Cd during soil drainage. During soil flooding, microbial sulfate reduction results in the formation of various metal sulfides. When the soils are subsequently drained, the various metal sulfides can form within sulfide voltaic cells. The metal sulfides with a lower electrochemical potential act as anodes and are prone to oxidative dissolution, whereas the metal sulfides with a higher potential act as cathodes and are protected from oxidation. This voltaic effect explains why the presence of ZnS (with a low potential) suppresses the oxidative dissolution of Cd sulfides, whereas the presence of CuS (with a high potential) promotes the oxidative dissolution of Cd sulfides. The voltaic effect is applicable to all chalcophile trace metals coupled with the sulfur redox cycle in periodically anoxic-oxic environments, thus playing an important role in the biogeochemistry of trace metals.

Stable isotope fractionation of cadmium in the soil-rice-human continuum.

Consumption of rice (Oryza sativa) grain is a major pathway by which humans are exposed to Cd, especially in non-smoking Asian populations. Although the stable isotope signatures of Cd offer a potential tool for tracing its sources, little is known about the isotopic fractionation of Cd across the entire soil-rice-human continuum. Cadmium isotope ratios were determined in field soils, rice grain, and human urine collected from two Cd-contaminated regions in southern China. Additionally, Cd isotopic fractionation in rice plants was investigated using two transgenic plants differing in Cd uptake and accumulation. Analysis of isotope ratios revealed a preferential enrichment of the heavy Cd isotopes from soil to rice grain (δ114/110Cdgrain-soil = +0.40‰) and from grain to urine (δ114/110Cdurine-grain = +0.40‰) in both regions. The first increase was mainly caused by partitioning between the soil solid phase and the soil solution, with heavier Cd preferentially enriching in the soil solution. Within the rice plant, we identified multiple processes that alter the isotope ratio, but the net effect throughout the plant was comparatively small. Cd fractionation in humans is presumably due to the preferential enrichment of heavier Cd isotopes by metal transporters DMT1 and ZIP8 (responsible for the absorption of Cd into body from the foods). These findings provide important insights into the Cd isotopic fractionation through the soil-rice-human continuum and are helpful for tracing the sources of Cd.

Chemical Speciation and Distribution of Cadmium in Rice Grain and Implications for Bioavailability to Humans.

Consumption of rice (Oryza sativa) is the major dietary source of cadmium (Cd) for populations with rice as the staple. Little is known about the distribution and chemical speciation of Cd in rice grain, which is critical in determining the bioavailability of Cd to humans. We used synchrotron-based techniques for analyses of the speciation and distribution of Cd in rice grain. The majority of the Cd in rice grain was present as Cd-thiolate complexes (66-92%), likely in the form of Cd bound with thiol-rich proteins. The remainder was present as Cd-carboxyl compounds and Cd-histidine. Elemental mapping showed two different patterns of Cd distribution, one with an even distribution throughout the entire grain and the other with a preferential distribution in the outer tissues (aleurone layer and outer starchy endosperm). The distribution pattern is important as it affects the removal of Cd during milling. On average, milling reduced grain Cd concentrations by 23.5% (median of 27.5%), although the range varied widely from a 64.7% decrease to a 22.2% increase, depending upon the concentration of Cd in the bran. We found that the variation in the distribution pattern of Cd in the rice grain was due to a temporal change in the supply of Cd from the soil porewater during grain filling. These results have important implications for Cd bioavailability in human diets.

Increased arsenic mobilization in the rice rhizosphere is mediated by iron-reducing bacteria.

Rice (Oryza sativa) tends to accumulate elevated levels of arsenic (As) in grain, threatening food safety and human health. The rice rhizosphere has a micro-environment that differs markedly from the bulk soil. Yet, little is known about how this micro-environment influences the mobility of As in the rhizosphere. Using rhizoboxes with two rice cultivars (cv. Shenyou 957 and Yangdao 6) differing in their radial oxygen loss (ROL), we investigated the in situ transformation of As in the rhizosphere associated with changes in microbial communities and As-related functional genes. Contrary to expectation, dissolved (porewater) As concentrations within the rhizosphere increased by 1.3-2.4 fold compared to the bulk soil during the seedling stage, with the magnitude of this difference gradually decreasing over time. The increased As mobilization in the rhizosphere was associated with increased soluble Fe. This increasing trend was associated with the increased abundance of both Fe-reducing bacteria (FeRB) and As-related functional genes within the rhizosphere. Furthermore, bacterial 16S rRNA gene sequencing data showed that the abundances of Geobacter and Clostridium were 3.1 times and 12.4 times higher in the rhizosphere, respectively. The importance of FeRB was also suggested by the fact that dissolved As concentrations were highly correlated with dissolved Fe concentrations (r2 = 0.83) and also with the relative abundance of genus Clostridium_sensu_stricto_10 (r2 = 0.85). This study highlights that although the rice rhizosphere favors a more aerobic condition compared to the bulk soil, As is more mobilized in the rhizosphere, and that Geobacter and some species of Clostridium play a critical role in controlling As mobilization in the rhizosphere.

Development of ZnO Nanoparticles as an Efficient Zn Fertilizer: Using Synchrotron-Based Techniques and Laser Ablation to Examine Elemental Distribution in Wheat Grain.

Zinc (Zn) deficiency is an important problem worldwide, adversely impacting human health. Using a field trial in China, we compared the foliar application of both ZnO nanoparticles (ZnO-NPs) and ZnSO4 on winter wheat (Triticum aestivum L.) for increasing the Zn concentration within the grain. We also used synchrotron-based X-ray fluorescence microscopy and laser ablation inductively coupled plasma mass spectrometry to examine the distribution of Zn within the grain. We found that ZnO-NPs increase the Zn concentration in the wheat grain, increasing from 18 mg·kg-1 in the control up to 40 mg·kg-1 when the ZnO-NPs were applied four times. These grain Zn concentrations in the ZnO-NP-treated grains are similar to those recommended for human consumption. However, the ZnO-NPs were similar in their effectiveness to ZnSO4. When examining trace element distribution in the grain, the trace elements were found to accumulate primarily in the aleurone layer and the crease region across all treatments. Importantly, Zn concentrations in the grain endosperm increased by nearly 30-fold relative to the control, with markedly increasing Zn concentrations within the edible portion. These results demonstrate that ZnO-NPs are a suitable fertilizer for increasing Zn within wheat grain and can potentially be used to improve human nutrition.

Biogeochemical cycling of iron oxides in the rhizosphere of plants grown on ferruginous duricrust (canga).

Goethite-cemented duricrusts, also known as canga, commonly occur as a capping rock protecting underlying iron ore deposits. The processes that govern canga formation are still unclear but include recurrent partial dissolution and recrystallisation of goethite through biogeochemical cycling of iron, hypothesised to be catalysed by plants and bacteria. In the present study, the effect of plant exudates on mobilisation of iron in canga was examined using model plants grown on crushed canga in RHIZOtest devices, which separate roots from substrate by a semi-permeable membrane. Moderate plant-induced acidification of the canga was detected, however the primary driver of mineral dissolution was the synergistic effect of reductive and ligand-promoted dissolution, identified by an increase in organic acids concentration and the presence of low concentrations of free ferrous iron. Whilst organic acids exudation lasted, iron cations were stabilised in solution; once the organic acids were degraded by microorganisms, the free cations precipitated as iron oxy-hydroxides. Mineralogical analysis and high-resolution microscopy confirmed our hypothesis that plants that grow in this iron-rich substrate contribute to iron dissolution indirectly (e.g., during phosphate solubilisation), and that the resulting surplus iron not taken up by the plants is redeposited, promoting the cementation of the residual minerals. Understanding the contribution of plants to the iron cycling in canga is crucial when formulating post-mining rehabilitation strategies for iron ore sites.

Assessing radiation dose limits for X-ray fluorescence microscopy analysis of plant specimens.

BACKGROUND AND AIMS: X-ray fluorescence microscopy (XFM) is a powerful technique to elucidate the distribution of elements within plants. However, accumulated radiation exposure during analysis can lead to structural damage and experimental artefacts including elemental redistribution. To date, acceptable dose limits have not been systematically established for hydrated plant specimens. METHODS: Here we systematically explore acceptable dose rate limits for investigating fresh sunflower (Helianthus annuus) leaf and root samples and investigate the time-dose damage in leaves attached to live plants. KEY RESULTS: We find that dose limits in fresh roots and leaves are comparatively low (4.1 kGy), based on localized disintegration of structures and element-specific redistribution. In contrast, frozen-hydrated samples did not incur any apparent damage even at doses as high as 587 kGy. Furthermore, we find that for living plants subjected to XFM measurement in vivo and grown for a further 9 d before being reimaged with XFM, the leaves display elemental redistribution at doses as low as 0.9 kGy and they continue to develop bleaching and necrosis in the days after exposure. CONCLUSIONS: The suggested radiation dose limits for studies using XFM to examine plants are important for the increasing number of plant scientists undertaking multidimensional measurements such as tomography and repeated imaging using XFM.

Evaluating effects of iron on manganese toxicity in soybean and sunflower using synchrotron-based X-ray fluorescence microscopy and X-ray absorption spectroscopy.

With similar chemistry, Mn and Fe interact in their many essential roles in plants but the magnitude and mechanisms involved of these interactions are poorly understood. Leaves of soybean (a Mn-sensitive species) developed a mild chlorosis and small dark spots and distorted trifoliate leaves with 30 µM Mn and 0.6 µM Fe in nutrient solution (pH 5.6; 3 mM ionic strength). At 0.6 µM Fe, lower alternate leaves of sunflower (a Mn-tolerant species) were chlorotic at 30 µM Mn and had a pale chlorosis and necrosis at 400 µM Mn. A concentration of 30 and 300 µM Fe in solution alleviated these typical symptoms of Mn toxicity and decreased the concentration of Mn from >3000 to ca. 800 mg kg-1 dry mass (DM) in all leaf tissues. As expected, increased Fe supply increased Fe in leaves from <100 up to 1350 mg Fe kg-1 DM. In situ synchrotron-based X-ray fluorescence microscopy showed that increased Fe supply caused an overall decrease in Mn in the leaf tissue but had little effect on the pattern of its distribution. Similarly, X-ray absorption spectroscopy identified only slight effects of Fe supply on Mn speciation in leaf tissues. Thus, the results of this study indicate that increased Fe supply ameliorated Mn toxicity in soybean and sunflower largely through decreased Mn uptake and translocation to leaf tissues rather than through changes in Mn distribution or speciation within the leaves.

Cadmium contamination in agricultural soils of China and the impact on food safety.

Rapid industrialization in China during the last three decades has resulted in widespread contamination of Cd in agricultural soils. A considerable proportion of the rice grain grown in some areas of southern China has Cd concentrations exceeding the Chinese food limit, raising widespread concern regarding food safety. In this review, we summarize rice grain Cd concentrations in national Chinese markets and in field surveys from contaminated areas, and analyze the potential health risk associated with increased dietary Cd intake. For subsistence rice farmers living in some contaminated areas of southern China who mainly consume locally-produced Cd-contaminated rice, their estimated dietary Cd intake is now comparable to that for the population in the region of Japan where the Itai-Itai disease was first reported. Interventions must be taken urgently to reduce Cd intake for these farmers. We also analyze i) the main reasons causing elevated grain Cd concentrations in southern China, ii) the dominant biogeochemical processes controlling the solubility of Cd in paddy soils, and iii) molecular mechanisms for the uptake and translocation of Cd in rice plants. Based on these analyses, we propose a number of countermeasures to address soil Cd contamination, including i) mitigation of Cd transfer from paddy soils to rice grain, and ii) intervention in those farmers who consume home-grown Cd-contaminated rice. Liming to increase soil pH to 6.5 and gene editing biotechnology are effective strategies to decrease Cd accumulation in rice grain. For these local farmers with high-Cd exposure risk, local governments should monitor the Cd concentration in their home-grown rice and exchange those high-Cd rice with low-Cd rice in order to reduce their dietary Cd intake.

Speciation and accumulation of Zn in sweetcorn kernels for genetic and agronomic biofortification programs.

MAIN CONCLUSION: In sweetcorn (Zea mays L.), embryo Zn is accumulated mainly as Zn-phytate, whereas endosperm Zn is complexed with a N- or S-containing ligand. Understanding the speciation of Zn in crop plants helps improve the effectiveness of biofortification efforts. Kernels of four sweetcorn (Zea mays L.) varieties were analysed for Zn concentration and content. We also assessed the speciation of the Zn in the embryo, endosperm, and pericarp in situ using synchrotron-based X-ray absorption spectroscopy. The majority of the Zn was in the endosperm and pericarp (72%), with the embryo contributing 28%. Approximately 79% of the Zn in the embryo accumulated as Zn-phytate, whereas in the endosperm most of the Zn was complexed with a N- or S-containing ligand, possibly as Zn-histidine and Zn-cysteine. This suggests that whilst the Zn in the endosperm and pericarp is likely to be bioavailable for humans, the Zn in the embryo is of low bioavailability. This study highlights the importance of targeting the endosperm of sweetcorn kernels as the tissue for increasing bioavailable Zn concentration.

Iron-Manganese (Oxyhydro)oxides, Rather than Oxidation of Sulfides, Determine Mobilization of Cd during Soil Drainage in Paddy Soil Systems.

The preharvest drainage of rice paddy fields during the grain filling stage can result in a substantial mobilization of Cd in soil and, consequently, elevated grain Cd concentration. However, the processes controlling the mobilization of Cd remains poorly understood. Using 12 field-contaminated paddy soils, we investigated the factors controlling the temporal changes in Cd solubility in paddy soils that were incubated anaerobically for 40 d followed by a 20 d oxidation period. Soluble and extractable Cd concentrations decreased rapidly upon flooding but increased during the oxidation phase, with Cd solubility (aqueous Cd/soil Cd) largely depending upon porewater pH. Furthermore, inhibiting sulfate reduction or inhibiting oxidation dissolution of Cd-sulfides had little or no effect on the mobilization of Cd in the subsequent oxidation phase. Both sequential extraction and X-ray absorption spectroscopy (XAS) analyses revealed that changes in Cd solubility were largely dependent upon the transformation of Cd between the Fe-Mn (oxyhydro)oxide fraction and exchangeable fraction. Mobilization of Cd upon soil drainage was caused by a decrease in soil pH resulting in the release of Cd from Fe-Mn (oxyhydro)oxides. Taken together, Fe-Mn (oxyhydro)oxides play a critical (and prevalent) role in controlling the mobilization of Cd upon soil drainage in paddy systems.

Aluminum Complexation with Malate within the Root Apoplast Differs between Aluminum Resistant and Sensitive Wheat Lines.

In wheat (Triticum aestivum), it is commonly assumed that Al is detoxified by the release of organic anions into the rhizosphere, but it is also possible that detoxification occurs within the apoplast and symplast of the root itself. Using Al-resistant (ET8) and Al-sensitive (ES8) near-isogenic lines of wheat, we utilized traditional and synchrotron-based approaches to provide in situ analyses of the distribution and speciation of Al within root tissues. Some Al appeared to be complexed external to the root, in agreement with the common assumption. However, root apical tissues of ET8 accumulated four to six times more Al than ES8 when exposed to Al concentrations that reduce root elongation rate by 50% (3.5 µM Al for ES8 and 50 µM for ET8). Furthermore, in situ analyses of ET8 root tissues indicated the likely presence of Al-malate and other forms of Al, predominantly within the apoplast. To our knowledge, this is the first time that X-ray absorption near edge structure analyses have been used to examine the speciation of Al within plant tissues. The information obtained in the present study is important in developing an understanding of the underlying physiological mode of action for improved root growth in systems with elevated soluble Al.

In vivo formation of natural HgSe nanoparticles in the liver and brain of pilot whales.

To understand the biochemistry of methylmercury (MeHg) that leads to the formation of mercury-selenium (Hg-Se) clusters is a long outstanding challenge that promises to deepen our knowledge of MeHg detoxification and the role Se plays in this process. Here, we show that mercury selenide (HgSe) nanoparticles in the liver and brain of long-finned pilot whales are attached to Se-rich structures and possibly act as a nucleation point for the formation of large Se-Hg clusters, which can grow with age to over 5 µm in size. The detoxification mechanism is fully developed from the early age of the animals, with particulate Hg found already in juvenile tissues. As a consequence of MeHg detoxification, Se-methionine, the selenium pool in the system is depleted in the efforts to maintain essential levels of Se-cysteine. This study provides evidence of so far unreported depletion of the bioavailable Se pool, a plausible driving mechanism of demonstrated neurotoxic effects of MeHg in the organism affected by its high dietary intake.

Cadmium accumulation is enhanced by ammonium compared to nitrate in two hyperaccumulators, without affecting speciation.

Nitrogen fertilization could improve the efficiency of Cd phytoextraction in contaminated soil and thus shorten the remediation time. However, limited information is available on the effect of N form on Cd phytoextraction and associated mechanisms in plants. This study examined the effect of N form on Cd accumulation, translocation, and speciation in Carpobrotus rossii and Solanum nigrum Plants were grown in nutrient solution with 5-15 µM Cd in the presence of 1000 µM NH4 (+) or NO3 (-) Plant growth and Cd uptake were measured, and Cd speciation was analyzed using synchrotron-based X-ray absorption spectroscopy. Shoot Cd accumulation was 30% greater with NH4 (+) than NO3 (-) supply. Carpobrotus rossii accumulated three times more Cd than S. nigrum. However, Cd speciation in the plants was not influenced by N form, but it did vary with species and tissues. In C. rossii, up to 91% of Cd was bound to S-containing ligands in all tissues except the xylem sap where 87-95% were Cd-OH complexes. Furthermore, the proportion of Cd-S in shoots was substantially lower in S. nigrum (44-69%) than in C. rossii (60-91%). It is concluded that the application of NH4 (+) (instead of NO3 (-)) increased shoot Cd accumulation by increasing uptake and translocation, rather than changing Cd speciation, and is potentially an effective approach for increasing Cd phytoextraction.

Identification of the primary lesion of toxic aluminum in plant roots.

Despite the rhizotoxicity of aluminum (Al) being identified over 100 years ago, there is still no consensus regarding the mechanisms whereby root elongation rate is initially reduced in the approximately 40% of arable soils worldwide that are acidic. We used high-resolution kinematic analyses, molecular biology, rheology, and advanced imaging techniques to examine soybean (Glycine max) roots exposed to Al. Using this multidisciplinary approach, we have conclusively shown that the primary lesion of Al is apoplastic. In particular, it was found that 75 µm Al reduced root growth after only 5 min (or 30 min at 30 µm Al), with Al being toxic by binding to the walls of outer cells, which directly inhibited their loosening in the elongation zone. An alteration in the biosynthesis and distribution of ethylene and auxin was a second, slower effect, causing both a transient decrease in the rate of cell elongation after 1.5 h but also a longer term gradual reduction in the length of the elongation zone. These findings show the importance of focusing on traits related to cell wall composition as well as mechanisms involved in wall loosening to overcome the deleterious effects of soluble Al.

In situ distribution and speciation of toxic copper, nickel, and zinc in hydrated roots of cowpea.

The phytotoxicity of trace metals is of global concern due to contamination of the landscape by human activities. Using synchrotron-based x-ray fluorescence microscopy and x-ray absorption spectroscopy, the distribution and speciation of copper (Cu), nickel (Ni), and zinc (Zn) was examined in situ using hydrated roots of cowpea (Vigna unguiculata) exposed to 1.5 µm Cu, 5 µm Ni, or 40 µm Zn for 1 to 24 h. After 24 h of exposure, most Cu was bound to polygalacturonic acid of the rhizodermis and outer cortex, suggesting that binding of Cu to walls of cells in the rhizodermis possibly contributes to the toxic effects of Cu. When exposed to Zn, cortical concentrations remained comparatively low with much of the Zn accumulating in the meristematic region and moving into the stele; approximately 60% to 85% of the total Zn stored as Zn phytate within 3 h of exposure. While Ni concentrations were high in both the cortex and meristem, concentrations in the stele were comparatively low. To our knowledge, this is the first report of the in situ distribution and speciation of Cu, Ni, and Zn in hydrated (and fresh) plant tissues, providing valuable information on the potential mechanisms by which they are toxic.