Synchrotron-based X-ray Fluorescence Microscopy Research Articles

Synchrotron-Based Imaging Reveals the Fate of Selenium in Striped Marsh Frog Tadpoles.

Synchrotron-based X-ray fluorescence microscopy (XFM) coupled with X-ray absorption near-edge structure (XANES) imaging was used to study selenium (Se) biodistribution and speciation in Limnodynastes peronii tadpoles. Tadpoles were exposed to dissolved Se (30 μg/L) as selenite (SeIV) or selenate (SeVI) for 7 days followed by 3 days of depuration. High-resolution elemental maps revealed that Se partitioned primarily in the eyes (specifically the eye lens, iris, and retinal pigmented epithelium), digestive and excretory organs of SeIV-exposed tadpoles. Speciation analysis confirmed that the majority of accumulated Se was converted to organo-Se. Multielement analyses provided new information on Se colocalization and its impact on trace element homeostasis. New insights into the fate of Se on a whole organism scale contribute to our understanding of the mechanisms and risks associated with Se pollution.

Metal ion fluxes controlling amphibian fertilization.

Mammalian oocytes undergo major changes in zinc content and localization in order to be fertilized, the most striking being the rapid exocytosis of over ten billion zinc ions, known as zinc sparks. Here we report that fertilization of amphibian Xenopus laevis eggs also initiates a zinc spark that progresses across the cell surface in coordination with dynamic calcium waves. This zinc exocytosis is accompanied by a newly recognized loss of intracellular manganese: synchrotron-based X-ray fluorescence and analytical electron microscopy reveal that zinc and manganese are sequestered in a system of cortical granules that are abundant at the animal pole. Through Electron-Nuclear Double-Resonance (ENDOR) studies, we rule out Mn2+ complexation with phosphate or nitrogenous ligands in intact eggs but the data are consistent with a carboxylate coordination environment. Our observations suggest that zinc and manganese fluxes are a conserved feature of fertilization in vertebrates and that they function as part of a physiological block to polyspermy.

Biologically facilitated precipitation of metals in low-Fe waters at the sulphidic Mount Chalmers mine, Queensland, Australia

Biofilms enriched in copper and zinc have formed at the aqueous-air interface and in subaqueous environments at the abandoned Mount Chalmers volcanic-hosted massive sulphide mine in Queensland, Australia. Remedial activity on site has limited acid mine drainage and associated iron-rich discharges, and surface waters leave the site at pH ~ 7. Unlike in many other oxygenated metal-sulphide environments, (bio)precipitates at this site conspicuously lack iron and manganese. Hence, this site provides an opportunity to observe the interactions of microorganisms with other metals, unobscured by the usually dominant Fe and Mn processes. Small subunit rRNA gene sequencing analyses suggest that biofilms are dominated by neutrophilic species of moss, diatoms, fungi and rotifera, as well as minor cyanobacteria and photosynthesising microorganisms adapted to high metal loads (e.g., up to ~ 8 mg/L Zn and ~ 2 mg/L Cu) in surface waters. Microbial communities formed metal-bearing biofilms on a time-scale of months. Investigations of these biofilms using scanning electron microscopy with electron dispersive x-ray spectroscopy and synchrotron-based X-ray fluorescence microscopy indicate that biofilms are intimately associated with metals, especially Cu and Zn, with Cu concentrations of up to ~ 20 wt%. Microorganisms on site help to catalyse dissolution and precipitation processes, although abiotic precipitation and anthropogenic intervention (raised pH) are the main geochemical drivers of remediation. Our observations suggest that microorganisms catalysed metal accumulation processes by (1) providing a substrate to which metals can attenuate during wetting and drying cycles and (2) micro-fossilisation induced by microorganism-metal interactions. This site provides a unique view of the impacts of rehabilitation processes on the microbial community, and vice versa. However, small numbers of bacteria that can contribute to acid mine drainage remain in the biome, ready to proliferate should remediation activities wane. Processes identified at this site are also relevant for potentially economic, enhanced metal extraction from legacy mine sites.

High phosphorus fertilization changes the speciation and distribution of manganese in wheat grains grown in a calcareous soil

The physiological disorders in humans resulting from the excess dietary intake of manganese (Mn) via whole-grain food has attracted considerable attention. However, the speciation and bioavailability of Mn in wheat grains and their response to different phosphorus (P) fertilization rates are still unclear. In the current study, using a long-term field trial with P application rates of 0, 21.8, 43.6, 65.5 and 87.3 kg/ha, we examined changes in the concentration, distribution, and speciation of Mn of wheat grains using synchrotron-based X-ray fluorescence microscopy and X-ray absorption spectroscopy. The total Mn concentration in grains was found to be increased by phosphorus fertilization, especially in embryo in the form of Mn(II), but this phosphorus fertilization also decreased Mn concentrations in the nucellar projection. In this study, the speciation of Mn in different wheat grain tissues was examined, and results indicate that in calcareous soils, high rates of P fertilizers can increase Mn concentrations in wheat grain, including Mn which is likely to be of high bioavailability, and thus may increase the risk for human to expose to high Mn intake via whole-grain food.

Non-glandular trichomes of sunflower are important in the absorption and translocation of foliar-applied Zn.

Trichomes are potentially important for absorption of foliar fertilizers. A study has shown that the non-glandular trichromes (NGTs) of sunflower (Helianthus annuus) accumulated high concentrations of foliar-applied zinc (Zn); however, the mechanisms of Zn accumulation in the NGTs and the fate of this Zn are unclear. Here we investigated how foliar-applied Zn accumulates in the NGTs and the subsequent translocation of this Zn. Time-resolved synchrotron-based X-ray fluorescence microscopy and transcriptional analyses were used to probe the movement of Zn in the NGTs, with the cuticle composition of the NGTs examined using confocal Raman microscopy. The accumulation of Zn in the NGTs is both an initial preferential absorption process and a subsequent translocation process. This preferred absorption is likely because the NGT base has a higher hydrophilicity, whilst the subsequent translocation is due to the presence of plasmodesmata, Zn-chelating ligands, and Zn transporters in the NGTs. Furthermore, the Zn sequestered in the NGTs was eventually translocated out of the trichome once the leaf Zn concentration had decreased, suggesting that the NGTs are also important in maintaining leaf Zn homeostasis. This study demonstrates for the first time that trichomes have a key structural and functional role in the absorption and translocation of foliar-applied Zn.

Synchrotron-based X-ray fluorescence microscopy reveals accumulation of polymyxins in single human alveolar epithelial cells.

Intravenous administration of the last-line polymyxins results in poor drug exposure in the lungs and potential nephrotoxicity; while inhalation therapy offers better pharmacokinetics/pharmacodynamics for pulmonary infections by delivering the antibiotic to the infection site directly. However, polymyxin inhalation therapy has not been optimized and adverse effects can occur. This study aimed to quantitatively determine the intracellular accumulation and distribution of polymyxins in single human alveolar epithelial A549 cells. Cells were treated with an iodine-labeled polymyxin probe FADDI-096 (5.0 and 10.0 μM) for 1, 4, and 24 h. Concentrations of FADDI-096 in single A549 cells were determined by synchrotron-based X-ray fluorescence microscopy. Concentration- and time-dependent accumulation of FADDI-096 within A549 cells was observed. The intracellular concentrations (mean ± SEM, n ≥ 189) of FADDI-096 were 1.58 ± 0.11, 2.25 ± 0.10, and 2.46 ± 0.07 mM following 1, 4 and 24 h of treatment at 10 μM, respectively. The corresponding intracellular concentrations following the treatment at 5 μM were 0.05 ± 0.01, 0.24 ± 0.04, and 0.25 ± 0.02 mM (n ≥ 189). FADDI-096 was mainly localized throughout the cytoplasm and nuclear region over 24 h. The intracellular zinc concentration increased in a concentration- and time-dependent manner. This is the first study to quantitatively map the accumulation of polymyxins in human alveolar epithelial cells and provides crucial insights for deciphering the mechanisms of their pulmonary toxicity. Importantly, our results may shed light on the optimization of inhaled polymyxins in patients and the development of new-generation safer polymyxins.

Incidence of hyperaccumulation and tissue-level distribution of manganese, cobalt, and zinc in the genus Gossia (Myrtaceae)

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 these elemental distributions at the macro (organ) and micro (cellular) levels with laboratory- and synchrotron-based X-ray Fluorescence Microscopy (XFM). Gossia pubiflora and G. hillii were newly discovered to be Mn hyperaccumulator plants. In the dosing trial, G. fragrantissima accumulated 17400µg g-1 Mn, 545µg g-1 Co and 13 000µg g-1Zn. 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, and in the cortex and phloem of roots. This study combined novel analytical approaches with controlled experimentation to examine metal hyperaccumulation in slow-growing woody species, thereby enabling insight into the phenomenon not possible through field studies.

Zinc from foliar-applied nanoparticle fertiliser is translocated to wheat grain: A 65Zn radiolabelled translocation study comparing conventional and novel foliar fertilisers

Foliar zinc (Zn) fertilisers can be used to supplement or replace soil applications of Zn in situations where soil properties may decrease the plant bioavailability of Zn. However, conventional foliar Zn formulations such as zinc sulfate can cause leaf damage due to the rapid release of high amounts of Zn2+ into leaf tissue which can be locally phytotoxic. Zinc oxide nanoparticles (ZnO-NPs) offer an alternative approach by providing a more sustained release of Zn into leaf tissue, and potentially avoiding the need for multiple applications. We compared the efficacy of ZnO-NPs and microparticles (ZnO-MPs) to that of conventional formulations (ZnCl2 and ZnEDTA) in wheat. This is the first study to use 65Zn radiolabelled formulations and gamma spectrometry to determine the translocation of Zn to the grains and subsequent efficiency of foliar-applied ZnO-NP fertilisers. We found that ZnEDTA was the most efficient fertiliser in terms of the proportion of applied Zn translocated to wheat grain. We also investigated the effect of Zn application rate on fertiliser efficiency. For all forms of Zn, when plants were treated with Zn at 750 mg/L or 75 mg/L, there were no significant differences in the concentration of applied Zn translocated to the grain. This suggests that current Zn application rates could be decreased while still maintaining the nutritional quality of grain. Finally, using photo-stimulated luminescence (PSL) autoradiography and synchrotron-based X-ray fluorescence microscopy (XFM) we showed that the grain distribution of foliar-applied Zn mirrors that of Zn derived from root uptake.

Penetration of foliar-applied Zn and its impact on apple plant nutrition status: in vivo evaluation by synchrotron-based X-ray fluorescence microscopy

The absorption of foliar fertilizer is a complex process and is poorly understood. The ability to visualize and quantify the pathway that elements take following their application to leaf surfaces is critical for understanding the science and for practical applications of foliar fertilizers. By the use of synchrotron-based X-ray fluorescence to analyze the in vivo localization of elements, our study aimed to investigate the penetration of foliar-applied Zn absorbed by apple (Malus domestica Borkh.) leaves with different physiological surface properties, as well as the possible interactions between foliar Zn level and the mineral nutrient status of treated leaves. The results indicate that the absorption of foliar-applied Zn was largely dependent on plant leaf surface characteristics. High-resolution elemental maps revealed that the high binding capacity of the cell wall for Zn contributed to the observed limitation of Zn penetration across epidermal cells. Trichome density and stomatal aperture had opposite effects on Zn fertilizer penetration: a relatively high density of trichomes increased the hydrophobicity of leaves, whereas the presence of stomata facilitated foliar Zn penetration. Low levels of Zn promoted the accumulation of other mineral elements in treated leaves, and the complexation of Zn with phytic acid potentially occurred owing to exposure to high-Zn conditions. The present study provides direct visual evidence for the Zn penetration process across the leaf surface, which is important for the development of strategies for Zn biofortification in crop species.

Cobalt hyperaccumulation in Rinorea cf. bengalensis (Violaceae) from Sabah: accumulation potential and tissue and cellular-level distribution of cobalt

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. 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. The maximum foliar cobalt concentration reached 1200 μg 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. 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.

Zinc Dynamics during Drosophila Oocyte Maturation and Egg Activation.

SummaryTemporal fluctuations in zinc concentration are essential signals, including during oogenesis and early embryogenesis. In mammals, zinc accumulation and release are required for oocyte maturation and egg activation, respectively. Here, we demonstrate that zinc flux occurs in Drosophila oocytes and activated eggs, and that zinc is required for female fertility. Our synchrotron-based X-ray fluorescence microscopy reveals zinc as the most abundant transition metal in Drosophila oocytes. Its levels increase during oocyte maturation, accompanied by the appearance of zinc-enriched intracellular granules in the oocyte, which depend on transporters. Subsequently, in egg activation, which mediates the transition from oocyte to embryo, oocyte zinc levels decrease significantly, as does the number of zinc-enriched granules. This pattern of zinc dynamics in Drosophila oocytes follows a similar trajectory to that in mammals, extending the parallels in female gamete processes between Drosophila and mammals and establishing Drosophila as a model for dissecting reproductive roles of zinc.

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.

Spatially Resolved Localization of Lanthanum and Cerium in the Rare Earth Element Hyperaccumulator Fern Dicranopteris linearis from China.

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.

Zinc Dynamics During <i>Drosophila</i> Oocyte Maturation and Egg Activation

Temporal fluctuations in zinc concentration are emerging as essential signaling events in the regulation of meiosis during oogenesis and early embryogenesis. For example, accumulation and release of zinc in mammalian gametes are required for oocyte maturation and egg activation, respectively. Here, we report that significant zinc fluctuations occur in Drosophila melanogaster oocytes and activated eggs, and that zinc is required for female fertility. Using synchrotron-based X-ray fluorescence microscopy (XFM), we mapped the subcellular localization of essential transition metals during fly oogenesis and egg activation. Quantitative analysis of the XFM data reveals that zinc is the most abundant transition metal in oocytes and that its levels increase during oocyte maturation. This increase is accompanied by appearance of zinc-enriched intracellular granules in the oocyte. The zinc transporter Znt35C is required for maintenance of these zinc-enriched granules in mature oocytes. During egg activation, the process that follows oocyte maturation and mediates the transition of the egg to an embryo, oocyte zinc levels decrease significantly, accompanied by a decrease in the number of zinc-enriched granules. This essential pattern of zinc dynamics in Drosophila oocytes follows a similar trajectory to that in mammals, rising during oogenesis and decreasing during egg activation. This further extends the parallels in female gamete processes between Drosophila and mammals, suggesting that the former may provide an important genetically tractable model for studying the role of zinc in female reproduction.

Zinc and lead encapsulated in amorphous ferric cements within hardpans in situ formed from sulfidic Cu-Pb-Zn tailings

Hardpans are massively indurated layers formed at the top layer of sulfidic tailings dams, which develop cementation structures and result in heavy metal immobilization. However, the micro-structural and complex forms of the cementing materials are not fully understood, as well as the mechanisms by which Zn and Pb are stabilized in the hardpans. The present study deployed synchrotron-based X-ray fluorescence microscopy (XFM) to have characterized the cementing structures, examined the distribution of Fe, Zn and Pb, and obtained laterally-resolved speciation of Zn within the hardpans using fluorescence X-ray absorption near-edge structure (XANES) imaging. The XFM analyses revealed that the Fe-rich cement layers consisted of Fe (oxyhydr)oxides coupled with amorphous Si materials, immobilizing Zn and Pb. Through laterally-resolved XANES imaging analyses, Zn-ferrihydrite-like precipitates were predicted to account for >76% of the total Zn within the Fe-rich cement layers. In contrast, outside of the cement layers, 9–63% of the Zn was estimated as labile ZnSO4.7H2O, with the remainder in the form of Zn-sulfide. These findings demonstrated that the Fe-rich cement layers were critical in immobilizing Zn and Pb within hardpans via mineral passivation and encapsulation, as the basis for long-term geochemical stability in the hardpan layer of sulfidic mine tailings.

Copper distribution and oxidation states near corroded fasteners in treated wood

Metal fasteners are used to hold wood structures together. In outdoor applications, these fasteners are subject to corrosion when the wood is treated with certain preservative treatments. Typically, these treatments contain copper. Prior work has hypothesized that the mechanism of corrosion in treated wood involves reduction of copper ions from the wood treatments. However, copper was rarely detected in the corrosion products of metals embedded in treated wood, which contradicts the hypothesized mechanism. This present work utilizes synchrotron based X-ray fluorescence microscopy (XFM) and X-ray absorption near edge spectroscopy to examine the corrosion mechanism in treated wood by looking at the spatial distribution and oxidation states of copper in the treated wood near the fastener and in the corrosion products removed from the fastener surface. The samples were obtained after a 1-year corrosion test. In the wood cell walls, the oxidation state of the copper treatment did not change in the immediate vicinity of the fastener, although there was a depletion of copper near the fastener. Furthermore, copper was detected in the corrosion products in trace amounts using XFM. Together, these techniques confirm that the corrosion mechanism involves transport of the cupric ions to the fastener surface, where they are reduced and suggest that previous attempts to detect copper were unsuccessful because the concentration of copper in the corrosion products was below the level of detection of the previously used techniques.

Investigating the foliar uptake of zinc from conventional and nano-formulations: a methodological study

Environmental contextZinc, an essential micronutrient often applied to crops as a fertiliser, can be difficult to analyse in plants due to limitations of conventional techniques. Here, we use radiotracers and a non-destructive imaging technique to visualise how zinc applied as a nanofertiliser moves within wheat plants over time. This is an important step towards developing cost-effective fertilisers to help solve one of the world’s most widespread plant deficiencies. AbstractZinc (Zn) deficiency affects half of the world’s arable soil and one-third of the world’s human population. Application of Zn foliar fertilisers to cereal crops can be an effective way to increase grain Zn content; however, commonly used formulations can scorch the leaf (e.g. soluble Zn salts) or are prohibitively expensive (e.g. chelated Zn, ZnEDTA). Zinc oxide nanoparticles (ZnO-NPs) may offer an efficient and cost-effective alternative, but little is known regarding the mechanisms of Zn uptake and translocation within the plant. Foliar-applied Zn is analytically challenging to detect, locate and quantify, as it is omnipresent. Furthermore, any single analytical technique does not have the detection limit or spatial resolution required. In this study, the uptake and mobility of foliar-applied ZnEDTA, ZnO-NPs and ZnO microparticles (ZnO-MPs) to wheat (Triticum aestivum L.) were investigated using inductively coupled plasma mass spectroscopy (ICP-MS), synchrotron-based X-ray fluorescence microscopy (XFM) and radiotracing techniques using 65Zn-labelled formulations. The three techniques were compared to highlight limitations and advantages of each. We also report, for the first time, a novel time-resolved invivo autoradiography imaging technique that can be used to visualise 65Zn in live plants treated with foliar applications of 65ZnO-NPs and MPs. The images were supplemented by gamma spectroscopy analysis for quantification. The results of this study provide important insights into the analytical challenges faced when investigating foliar-applied Zn nanofertilisers in plants. Potential solutions using nuclear techniques are also discussed, which in turn may ultimately lead to the development of more efficient foliar fertilisers.

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.

Absorption of foliar applied Zn is decreased in Zn deficient sunflower (Helianthus annuus) due to changes in leaf properties

Despite the importance of foliar Zn fertilisation, it remains unclear how the Zn status of the plant itself influences the efficacy of foliar applied Zn, with this forming the focus of the present study. Sunflower (Helianthus annuus) was grown in nutrient solutions with either 0.5 (sufficient) or 0 μM Zn (deficient) to relate the absorption of foliar applied Zn to leaf properties. The distribution and speciation of Zn within the leaf were examined in situ using synchrotron-based X-ray fluorescence microscopy and X-ray absorption spectroscopy. Zinc deficiency decreased foliar absorption of Zn as ZnSO4 by 50–66% compared to Zn sufficient sunflower, despite μ-XRF analysis showing the pattern of Zn absorption was similar for both plants. Rather, Zn deficiency decreased the leaf trichome density and likely altered the adaxial leaf surface composition and structure, with these presumably being the main causes for the reduced Zn absorption. The Zn status of the plant also influenced the speciation of the absorbed Zn, with 37–53% as Zn phytate in Zn sufficient leaves but 55% as Zn phosphate in Zn deficient leaves. Zinc status of the plants influenced leaf surface properties, with Zn deficiency leading to decreased absorption of foliar applied Zn.

In situ analyses of inorganic nutrient distribution in sweetcorn and maize kernels using synchrotron-based X-ray fluorescence microscopy.

Understanding the spatial distribution of inorganic nutrients within edible parts of plant products helps biofortification efforts to identify and focus on specific uptake pathways and storage mechanisms. Kernels of sweetcorn (Zea mays) variety 'High zeaxanthin 103146' and maize inbred line 'Thai Floury 2' were harvested at two different maturity stages, and the distributions of K, P, S, Ca, Zn, Fe and Mn were examined in situ using synchrotron-based X-ray fluorescence microscopy. The distribution of inorganic nutrients was largely similar between maize and sweetcorn, but differed markedly depending upon the maturity stage after further embryonic development. The micronutrients Zn, Fe and Mn accumulated primarily in the scutellum of the embryo during early kernel development, while trace amounts of these were found in the aleurone layer at the mature stage. Although P accumulated in the scutellum, there was no direct relationship between the concentrations of P and those of the micronutrients, compared with the linear trend between Zn and Fe concentrations. This study highlights the important role of the embryo as a micronutrient reserve for sweetcorn and maize kernels, and the need to understand how biofortification efforts can further increase the inorganic nutrient concentration of the embryo for human consumption.