Examining a synchrotron-based approach for in situ analyses of Al speciation in plant roots.

Aluminium (Al) K- and L-edge X-ray absorption near-edge structure (XANES) has been used to examine Al speciation in minerals but it remains unclear whether it is suitable for in situ analyses of Al speciation within plants. The XANES analyses for nine standard compounds and root tissues from soybean (Glycine max), buckwheat (Fagopyrum tataricum), and Arabidopsis (Arabidopsis thaliana) were conducted in situ. It was found that K-edge XANES is suitable for differentiating between tetrahedral coordination (peak of 1566 eV) and octahedral coordination (peak of 1568 to 1571 eV) Al, but not suitable for separating Al binding to some of the common physiologically relevant compounds in plant tissues. The Al L-edge XANES, which is more sensitive to changes in the chemical environment, was then examined. However, the poorer detection limit for analyses prevented differentiation of the Al forms in the plant tissues because of their comparatively low Al concentration. Where forms of Al differ markedly, K-edge analyses are likely to be of value for the examination of Al speciation in plant tissues. However, the apparent inability of Al K-edge XANES to differentiate between some of the physiologically relevant forms of Al may potentially limit its application within plant tissues, as does the poorer sensitivity at the L-edge.

Understanding the delayed expression of Al resistance in signal grass (Urochloa decumbens).

BACKGROUND AND AIMS: Signal grass (Urochloa decumbens) is a widely used pasture grass in tropical and sub-tropical areas due to its high aluminiun (Al) resistance. However, the underlying mechanisms conferring this resistance are not clearly understood. METHODS: The Al concentrations of bulk root tissues and the intracellular compartment were examined, including the impact of a metabolic inhibitor, carbonyl cyanide m-chlorophenyl hydrazone (CCCP). Next, we examined changes in the properties of signal grass root tissues following exposure to toxic levels of Al, including the cell wall cation exchange capacity (CEC), degree of methylation and concentrations of cell wall fractions. KEY RESULTS: Although signal grass was highly resistant to Al, there was a delay of 24-48 h before the expression of this resistance. We found that this delay in the expression of Al resistance was not related to the total Al concentration in the bulk apical root tissues, nor was it related to changes in the Al bound to the cell wall. We also examined changes in other properties of the cell wall, including the CEC, degree of methylation and changes in the concentration of pectin, hemicellulose and cellulose. We noted that concentrations of intracellular Al decreased by approx. 50 % at the same time that the root elongation rate improved after 24-48 h. Using CCCP as a metabolic inhibitor, we found that the intracellular Al concentration increased approx. 14-fold and that the CCCP prevented the subsequent decrease in intracellular Al. CONCLUSIONS: Our results indicate that the delayed expression of Al resistance was not associated with the Al concentration in the bulk apical root tissues or bound to the cell wall, nor was it associated with changes in other properties of the cell wall. Rather, signal grass has an energy-dependent Al exclusion mechanism, and this mechanism requires 24-48 h to exclude Al from the intracellular compartment.

Silver Sulfide Nanoparticles Reduce Nitrous Oxide Emissions by Inhibiting Denitrification in the Earthworm Gut.

The accumulation of Ag2S in agricultural soil via application of Ag-containing sludge potentially affects the functioning of soil microorganisms and earthworms (EWs) due to the strong antimicrobial properties of Ag. This study examined the effects of Ag2S nanoparticles (Ag2S-NPs) on the EW-mediated (Eisenia fetida and Pontoscolex corethrurus) soil N cycle. We used 16S rRNA gene-based sequencing and quantitative polymerase chain reaction to examine the bacterial community and nitrification/denitrification-related gene abundance. The presence of either EWs or Ag significantly increased denitrification and N2O emissions. However, the addition of Ag2S to EW-inhabited soil reduced N2O emissions by 14-33%. Furthermore, Ag2S caused a low-dose stimulation but a high-dose inhibition to N2O flux from the EW gut itself. Accordingly, an increase in Ag in the EW gut caused a decrease in the relative abundance of denitrifiers in both the soil and the gut, especially for the dominant genus Bacillus. Ag2S also decreased the copy numbers of nitrification gene (nxrB) and denitrification genes (napA, nirS, and nosZ) in EW gut, leading to the observed decrease in N2O emissions. Collectively, applying Ag2S-containing sludge disturbs the denitrification function of the EW gut microbiota and the cycling of N in soil-based systems.

Minimizing experimental artefacts in synchrotron-based X-ray analyses of Fe speciation in tissues of rice plants.

Iron (Fe) plays an important role within environmental systems. Synchrotron-based X-ray approaches, including X-ray absorption spectroscopy (XAS), provide powerful tools for in situ analyses of Fe speciation, but beam damage during analysis may alter Fe speciation during its measurement. XAS was used to examine whether experimental conditions affect the analysis of Fe speciation in plant tissues. Even when analyzed in a cryostat at 12 K, it was found that FeIII can rapidly (within 0.5-1 min) photoreduce to FeII, although the magnitude of photoreduction varied depending upon the hydration of the sample, the coordination chemistry of the Fe, as well as other properties. For example, photoreduction of FeIII was considerably higher for aqueous standard compounds than for hydrated plant-root tissues. The use of freeze-dried samples in the cryostat (12 K) markedly reduced the magnitude of this FeIII photoreduction, and there was no evidence that the freeze-drying process itself resulted in experimental artefacts under the current experimental conditions, such as through the oxidation of FeII, although some comparatively small differences were observed when comparing spectra of hydrated and freeze-dried FeII compounds. The results of this study have demonstrated that FeIII photoreduction can occur during X-ray analysis, and provides suitable conditions to preserve Fe speciation to minimize the extent of beam damage when analyzing environmental samples. All studies utilizing XAS are encouraged to include a preliminary experiment to determine if beam damage is occurring, and, where appropriate, to take the necessary steps (such as freeze drying) to overcome these issues.

Chemical and physical influence of sodic soils on the coleoptile length and root growth angle of wheat genotypes.

BACKGROUND AND AIMS: High exchangeable sodium percentage (ESP) and bulk density of sodic soils can reduce seedling emergence. This study examined variation in seedling coleoptile length and seminal root angle of wheat (Triticum aestivum. L) genotypes to determine whether these traits vary between genotypes that differ in their tolerance to sodic soils. METHODS: Wheat genotypes were grown in three different experiments. First, four wheat genotypes were grown using soils of three ESPs (4, 10 and 17 %) and secondly in soils of three different bulk densities (1.2, 1.4 and 1.5 g cm-3) and ESP 10 %. Thirdly, seedling coleoptile length and seminal root angle were determined for 16 genotypes grown in a soil of ESP 10 % and bulk density 1.2 g cm-2. Seminal root angle and coleoptile length measurements from the current study were compared with seedling emergence rate and force measured previously. KEY RESULTS: The seedling coleoptile length of all genotypes decreased with increasing soil ESP and bulk density, but with no significant differences between genotypes. In contrast, seminal root angles differed significantly between genotypes, but were not significantly affected by ESP or bulk density. There was an inverse relationship between the seminal root angle of the 16 genotypes and seedling emergence rate (R2 = 0.89) and also between seminal root angle and seedling emergence force (R2 = 0.61). CONCLUSIONS: Lack of significant variation in coleoptile length between genotypes suggests that this may not be a suitable characteristic to identify wheat tolerance to sodic conditions. However, a narrower seminal root angle was correlated with rate and force of seedling emergence, traits likely to improve establishment. The mechanism underlying this correlation is not yet clear. Genotypes with a narrow root angle had greater root depth. One possible mechanism might be that genotypes with narrow root angles were able to take up more soil moisture at depth, leading to a higher proportion of seedling emergence.

Absorption of foliar-applied Zn in sunflower (Helianthus annuus): importance of the cuticle, stomata and trichomes.

Background and Aims: The pathways whereby foliar-applied nutrients move across the leaf surface remain unclear. The aim of the present study was to examine the pathways by which foliar-applied Zn moves across the sunflower (Helianthus annuus) leaf surface, considering the potential importance of the cuticle, stomata and trichomes. Methods: Using synchrotron-based X-ray florescence microscopy and nanoscale secondary ion mass spectrometry (NanoSIMS), the absorption of foliar-applied ZnSO4 and nano-ZnO were studied in sunflower. The speciation of Zn was also examined using synchrotron-based X-ray absorption spectroscopy. Key Results: Non-glandular trichomes (NGTs) were particularly important for foliar Zn absorption, with Zn preferentially accumulating within trichomes in ≤15 min. The cuticle was also found to have a role, with Zn appearing to move across the cuticle before accumulating in the walls of the epidermal cells. After 6 h, the total Zn that accumulated in the NGTs was approx. 1.9 times higher than in the cuticular tissues. No marked accumulation of Zn was found within the stomatal cavity, probably indicating a limited contribution of the stomatal pathway. Once absorbed, the Zn accumulated in the walls of the epidermal and the vascular cells, and trichome bases of both leaf sides, with the bundle sheath extensions that connected to the trichomes seemingly facilitating this translocation. Finally, the absorption of nano-ZnO was substantially lower than for ZnSO4, with Zn probably moving across the leaf surface as soluble Zn rather than nanoparticles. Conclusions: In sunflower, both the trichomes and cuticle appear to be important for foliar Zn absorption.

Manganese distribution and speciation help to explain the effects of silicate and phosphate on manganese toxicity in four crop species.

Soil acidity and waterlogging increase manganese (Mn) in leaf tissues to potentially toxic concentrations, an effect reportedly alleviated by increased silicon (Si) and phosphorus (P) supply. Effects of Si and P on Mn toxicity were studied in four plant species using synchrotron-based micro X-ray fluorescence (µ-XRF) and nanoscale secondary ion mass spectrometry (NanoSIMS) to determine Mn distribution in leaf tissues and using synchrotron-based X-ray absorption spectroscopy (XAS) to measure Mn speciation in leaves, stems and roots. A concentration of 30 µM Mn in solution was toxic to cowpea and soybean, with 400 µM Mn toxic to sunflower but not white lupin. Unexpectedly, µ-XRF analysis revealed that 1.4 mM Si in solution decreased Mn toxicity symptoms through increased Mn localization in leaf tissues. NanoSIMS showed Mn and Si co-localized in the apoplast of soybean epidermal cells and basal cells of sunflower trichomes. Concomitantly, added Si decreased oxidation of Mn(II) to Mn(III) and Mn(IV). An increase from 5 to 50 µM P in solution changed some Mn toxicity symptoms but had little effect on Mn distribution or speciation. We conclude that Si increases localized apoplastic sorption of Mn in cowpea, soybean and sunflower leaves thereby decreasing free Mn2+ accumulation in the apoplast or cytoplasm.

Absorption of foliar-applied Zn fertilizers by trichomes in soybean and tomato.

The present study investigated the role of trichomes in absorption of foliar-applied zinc fertilizers in soybean and tomato. Using synchrotron-based X-ray fluorescence microscopy for in situ analyses of hydrated leaves, we found that upon foliar application of ZnSO4, Zn accumulated within 15 min in some non-glandular trichomes in soybean, but not in tomato. However, analyses of cross-sections of soybean leaves did not show any marked accumulation of Zn in tissues surrounding trichomes. Furthermore, when near-isogenic lines of soybean differing 10-fold in trichome density were used to compare Zn absorption, it was found that foliar Zn absorption was not related to trichome density. Therefore, it is suggested that trichomes are not part of the primary pathway through which foliar-applied Zn moves across the leaf surface in soybean and tomato. However, this does not preclude trichomes being important in other plant species, as they are known to be highly diverse. We also compared the absorption of Zn when supplied as either ZnSO4, nano-ZnO, or bulk-ZnO, and found that absorption from ZnSO4 was about 10-fold higher than from nano- and bulk-ZnO, suggesting that it was mainly absorbed as soluble Zn. This study improves our understanding of the absorption of foliar-applied nutrients.

Nitrogen-rich microbial products provide new organo-mineral associations for the stabilization of soil organic matter.

Understanding the cycling of C and N in soils is important for maintaining soil fertility while also decreasing greenhouse gas emissions, but much remains unknown about how organic matter (OM) is stabilized in soils. We used nano-scale secondary ion mass spectrometry (NanoSIMS) to investigate the changes in C and N in a Vertisol and an Alfisol incubated for 365 days with 13 C and 15 N pulse labeled lucerne (Medicago sativa L.) to discriminate new inputs of OM from the existing soil OM. We found that almost all OM within the free stable microaggregates of the soil was associated with mineral particles, emphasizing the importance of organo-mineral interactions for the stabilization of C. Of particular importance, it was also found that 15 N-rich microbial products originating from decomposition often sorbed directly to mineral surfaces not previously associated with OM. Thus, we have shown that N-rich microbial products preferentially attach to distinct areas of mineral surfaces compared to C-dominated moieties, demonstrating the ability of soils to store additional OM in newly formed organo-mineral associations on previously OM-free mineral surfaces. Furthermore, differences in 15 N enrichment were observed between the Vertisol and Alfisol presumably due to differences in mineralogy (smectite-dominated compared to kaolinite-dominated), demonstrating the importance of mineralogy in regulating the sorption of microbial products. Overall, our findings have important implications for the fundamental understanding of OM cycling in soils, including the immobilization and storage of N-rich compounds derived from microbial decomposition and subsequent N mineralization to sustain plant growth.

Risk of Silver Transfer from Soil to the Food Chain Is Low after Long-Term (20 Years) Field Applications of Sewage Sludge.

The increasingly widespread usage of silver (Ag) nanoparticles has raised concerns regarding their environmental risk. The behavior of Ag and its transfer risk to the food chain were investigated using a long-term field experiment that commenced in 1942 in which Ag-containing sewage sludge was repeatedly applied to the soil (25 applications during 20 years). The speciation of the Ag in both the sludge and the soils retrieved from the long-term experimental archive was examined using synchrotron-based X-ray absorption spectroscopy, and extractable Ag concentrations from soils were determined using 0.01 M Ca(NO3)2 and 0.005 M DTPA. The total Ag in the sludge during the time period of 1942-1961 ranged from 155 to 463 mg kg-1. These values are 1-2 orders of magnitude higher than those in currently produced sludge (ca. 0.5-20 mg kg-1). Long-term repeated applications of these sludges resulted in an increase of Ag in soils from 1.9 mg kg-1 in the control to up to 51 mg kg-1. The majority (>80%) of the Ag in both the sludge and the sludge-treated soils was present as insoluble Ag2S, thereby markedly reducing the bioavailability of this Ag. Concentrations of Ag in the archived crop samples were generally <0.70 mg kg-1 in edible tissues, much less than those in diets that may cause an adverse effects in animals and humans (>100 mg kg-1). These data indicate that the transfer of Ag (derived from both traditional Ag industry and current nano Ag industry) to the terrestrial food chain is limited.

Soil Organic Carbon Stabilization: Mapping Carbon Speciation from Intact Microaggregates.

The clearing of land for agricultural production depletes soil organic carbon (OC) reservoirs, yet despite their importance, the mechanisms by which C is stabilized in soils remain unclear. Using synchrotron-based infrared microspectroscopy, we have for the first time obtained in situ, laterally resolved data regarding the speciation of C within sections taken from intact free microaggregates from two contrasting soils (Vertisol and Oxisol, 0-20 cm depth) impacted upon by long-term (up to 79 y) agricultural production. There was no apparent gradient in the C concentration from the aggregate surface to the interior for any of the three forms of C examined (aliphatic C, aromatic C, and polysaccharide C). Rather, organo-mineral interactions were of critical importance in influencing overall C stability, particularly for aliphatic C, supporting the hypothesis that microaggregates form through organo-mineral interactions. However, long-term cropping substantially decreased the magnitude of the organo-mineral interactions for all three forms of C. Thus, although organo-mineral interactions are important for OC stability, C forms associated with the mineral phases are not entirely resistant to degradation. These results provide important insights into the underlying mechanisms by which microaggregates form and the factors influencing the persistence of OC in soils.

Defining appropriate methods for studying toxicities of trace metals in nutrient solutions.

The use of inappropriate experimental conditions for examining trace metal phytotoxicity results in data of questionable value. The present study aimed to identify suitable parameters for study of phytotoxic metals in nutrient solutions. First, the literature was reviewed to determine the concentration of six metals (Cd, Cu, Hg, Ni, Pb, and Zn) from solution of contaminated soils. Next, the effects of pH, P, Cl, NO3, and four Fe-chelators were investigated by using thermodynamic modelling and by examining changes in root elongation rate of soybean (Glycine max cv. Bunya). The literature review identified that the solution concentrations of metals in soils were low, ranging from (µM) 0.069-11Cd, 0.19-15.8 Cu, 0.000027-0.000079 Hg, 1.0-8.7 Ni, 0.004-0.55 Pb, and 0.4-36.3 Zn. For studies in nutrient solution, pH should generally be low given its effects on solubility and speciation, as should the P concentration due to the formation of insoluble phosphate salts. The concentrations of Cl, NO3, and various chelators also influence metal toxicity through alteration of metal speciation. The nutrient solutions used to study metal toxicity should consider environmentally-relevant conditions especially for metal concentrations, with concentrations of other components added at levels that do not substantially alter metal toxicity.

Global changes in soil stocks of carbon, nitrogen, phosphorus, and sulphur as influenced by long-term agricultural production.

Quantifying changes in stocks of C, N, P, and S in agricultural soils is important not only for managing these soils sustainably as required to feed a growing human population, but for C and N, they are also important for understanding fluxes of greenhouse gases from the soil environment. In a global meta-analysis, 102 studies were examined to investigate changes in soil stocks of organic C, total N, total P, and total S associated with long-term land-use changes. Conversion of native vegetation to cropping resulted in substantial losses of C (-1.6 kg m-2 , -43%), N (-0.15 kg m-2 , -42%), P (-0.029 kg m-2 , -27%), and S (-0.015 kg m-2 , -33%). The subsequent conversion of conventional cropping systems to no-till, organic agriculture, or organic amendment systems subsequently increased stocks, but the magnitude of this increase (average of +0.47 kg m-2 for C and +0.051 kg m-2 for N) was small relative to the initial decrease. We also examined the conversion of native vegetation to pasture, with changes in C (-11%), N (+4.1%), and P (+25%) generally being modest relative to changes caused by conversion to cropping. The C:N ratio remained relatively constant irrespective of changes in land use, whilst in contrast, the C:S ratio decreased by 21% in soils converted to cropping - this suggesting that biochemical mineralization is of importance for S. The data presented here will assist in the assessment of different agricultural production systems on soil stocks of C, N, P, and S - this information assisting not only in quantifying the effects of existing agricultural production on these stocks, but also allowing for informed decision-making regarding the potential effects of future land-use changes.

Effects of changes in leaf properties mediated by methyl jasmonate (MeJA) on foliar absorption of Zn, Mn and Fe.

Background and Aims: Foliar fertilization to overcome nutritional deficiencies is becoming increasingly widespread. However, the processes of foliar nutrient absorption and translocation are poorly understood. The present study aimed to investigate how cuticular leaf properties affect the absorption of foliar-applied nutrients in leaf tissues. Methods: Given that methyl jasmonate (MeJA) can cause alterations in leaf properties, we applied 1 mm MeJA to sunflower (Helianthus annuus), tomato (Solanum lycopersicum) and soybean (Glycine max) to assess changes in leaf properties. Using traditionally analytical approaches and synchrotron-based X-ray fluorescence microscopy, the effects of these changes on the absorption and translocation of foliar-applied Zn, Mn and Fe were examined. Key Results: The changes in leaf properties caused by the application of MeJA increased foliar absorption of Zn, Mn and Fe up to 3- to 5-fold in sunflower but decreased it by 0·5- to 0·9-fold in tomato, with no effect in soybean. These changes in the foliar absorption of nutrients could not be explained by changes in overall trichome density, which increased in both sunflower (86%) and tomato (76%) (with no change in soybean). Similarly, the changes could be not attributed to changes in stomatal density or cuticle composition, given that these properties remained constant. Rather, the changes in the foliar absorption of Zn, Mn and Fe were related to the thickness of the cuticle and epidermal cell wall. Finally, the subsequent translocation of the absorbed nutrients within the leaf tissues was limited (<1·3mm) irrespective of treatment. Conclusions: The present study highlights the potential importance of the combined thickness of the cuticle and epidermal cell wall in the absorption of foliar-applied nutrients. This information will assist in increasing the efficacy of foliar fertilization.

Evaluation of pyritic mine tailings as a plant growth substrate.

At the Kidston gold mine, Australia, the direct establishment of vegetation on tailings was considered as an alternative to the use of a waste rock cover. The tailings acid/base account was used to predict plant growth limitation by acidity, and thus methods capable of identifying tailings that would acidify to pH 4.5 or lower were sought. Total S was found to be poorly correlated with acid-generating sulfide, and total C was poorly correlated with acid-neutralizing carbonate, precluding the use of readily determined total S and C as predictors of net acid generation. Therefore, the selected approach used assessment of sulfide content as a predictor of acid generation, and carbonate content as a measure of the acid-neutralizing capacity available at pH 5 and above. Using this approach, the majority of tailings (67%) were found to be non-acid generating. However, areas of potentially acid-generating tailings were randomly distributed across the dam, and could only be located by intensive sampling. The limitations imposed by the large sample numbers, and costly analysis of sulfide and carbonate, make it impractical to identify and ameliorate acid-generating areas prior to vegetation establishment. However, as only a small proportion of the tailings will acidify, a strategy of re-treating acid areas following oxidation is suggested. The findings of the present study will assist in the selection of appropriate methods for the prediction of net acid generation, particularly where more conservative measurements are required to allow vegetation to be established directly in tailings.

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.

Kinetics and nature of aluminium rhizotoxic effects: a review.

Acid soils with elevated levels of soluble aluminium (Al) comprise ~40% of the world's arable land, but there remains much uncertainty regarding the mechanisms by which Al is rhizotoxic. This review examines the kinetics of the toxic effects of Al on the root elongation rate (RER), its effects on root tissues, and its location at a subcellular level. Depending upon the concentration and plant species, soluble Al decreases the RER in a median time of 73min, but in as little as 5min in soybean. This is initially due to a decreased rate at which cells expand anisotropically in the elongation zone. Thereafter, rhizodermal and outer cortical cells rupture through decreased cell wall relaxation. It is in this region where most Al accumulates in the apoplast. Subsequently, Al impacts root growth at a subcellular level through adverse effects on the plasma membrane (PM), cytoplasm, and nucleus. At the PM, Al alters permeability, fluidity, and integrity in as little as 0.5h, whilst it also depolarizes the PM and reduces H(+)-ATPase activity. The Al potentially crosses the PM within 0.5h where it is able to bind to the nucleus and inhibit cell division; sequestration within the vacuole is required to reduce the toxic effects of Al within the cytoplasm. This review demonstrates the increasing evidence of the importance of the initial Al-induced inhibition of wall loosening, but there is evidence also of the deleterious effects of Al on other cellular processes which are important for long-term root growth and function.

Aluminium effects on mechanical properties of cell wall analogues.

Aluminium (Al) toxicity adversely impacts plant productivity in acid soils by restricting root growth and although several mechanisms are involved the physiological basis of decreased root elongation remains unclear. Understanding the primary mechanisms of Al rhizotoxicity is hindered due to the rapid effects of soluble Al on root growth and the close proximity of many cellular components within the cell wall, plasma membrane, cytosol and nucleus with which Al may react. To overcome some of these difficulties, we report on a novel method for investigating Al interactions with Komagataeibacter xylinus bacterial cellulose (BC)-pectin composites as cell wall analogues. The growth of K. xylinus in the presence of various plant cell wall polysaccharides, such as pectin, has provided a unique in vitro model system with which to investigate the interactions of Al with plant cell wall polysaccharides. The BC-pectin composites reacted in a similar way with Al as do plant cell walls, providing insights into the effects of Al on the mechanical properties of the BC-pectin composites as cell wall analogues. Our findings indicated that there were no significant effects of Al (4-160 µM) on the tensile stress, tensile strain or Young's modulus of the composites. This finding was consistent with cellulose, not pectin, being the major load bearing component in BC-pectin composites, as is also the case in plant cell walls.

Silver Nanoparticles Entering Soils via the Wastewater-Sludge-Soil Pathway Pose Low Risk to Plants but Elevated Cl Concentrations Increase Ag Bioavailability.

The widespread use of silver nanoparticles (Ag-NPs) results in their movement into wastewater treatment facilities and subsequently to agricultural soils via application of contaminated sludge. On-route, the chemical properties of Ag may change, and further alterations are possible upon entry to soil. In the present study, we examined the long-term stability and (bio)availability of Ag along the "wastewater-sludge-soil" pathway. Synchrotron-based X-ray absorption spectroscopy (XAS) revealed that ca. 99% of Ag added to the sludge reactors as either Ag-NPs or AgNO3 was retained in sludge, with ≥79% of this being transformed to Ag2S, with the majority (≥87%) remaining in this form even after introduction to soils at various pH values and Cl concentrations for up to 400 days. Diffusive gradients in thin films (DGT), chemical extraction, and plant uptake experiments indicated that the potential (bio)availability of Ag in soil was low but increased markedly in soils with elevated Cl, likely due to the formation of soluble AgClx complexes in the soil solution. Although high Cl concentrations increased the bioavailability of Ag markedly, plant growth was not reduced in any treatment. Our results indicate that Ag-NPs entering soils through the wastewater-sludge-soil pathway pose low risk to plants due to their conversion to Ag2S in the wastewater treatment process, although bioavailability may increase in saline soils or when irrigated with high-Cl water.

Synchrotron-based X-ray absorption near-edge spectroscopy imaging for laterally resolved speciation of selenium in fresh roots and leaves of wheat and rice.

Knowledge of the distribution of selenium (Se) species within plant tissues will assist in understanding the mechanisms of Se uptake and translocation, but in situ analysis of fresh and highly hydrated plant tissues is challenging. Using synchrotron-based fluorescence X-ray absorption near-edge spectroscopy (XANES) imaging to provide laterally resolved data, the speciation of Se in fresh roots and leaves of wheat (Triticum aestivum L.) and rice (Oryza sativa L.) supplied with 1 µM of either selenate or selenite was investigated. For plant roots exposed to selenate, the majority of the Se was efficiently converted to C-Se-C compounds (i.e. methylselenocysteine or selenomethionine) as selenate was transported radially through the root cylinder. Indeed, even in the rhizodermis which is exposed directly to the bulk solution, only 12-31% of the Se was present as uncomplexed selenate. The C-Se-C compounds were probably sequestered within the roots, whilst much of the remaining uncomplexed Se was translocated to the leaves-selenate accounting for 52-56% of the total Se in the leaves. In a similar manner, for plants exposed to selenite, the Se was efficiently converted to C-Se-C compounds within the roots, with only a small proportion of uncomplexed selenite observed within the outer root tissues. This resulted in a substantial decrease in translocation of Se from the roots to leaves of selenite-exposed plants. This study provides important information for understanding the mechanisms responsible for the uptake and subsequent transformation of Se in plants.