Camille Larue

Accumulation, translocation and impact of TiO2 nanoparticles in wheat (Triticum aestivum spp.): Influence of diameter and crystal phase

Intensive production of TiO(2) nanoparticles (TiO(2)-NPs) would lead to their release in the environment. Their ecotoxicological impact is still poorly documented, while their use in commercial goods is constantly increasing. In this study we compare root accumulation and root-to-shoot translocation in wheat of anatase and rutile TiO(2)-NPs with diameters ranging from 14 nm to 655 nm, prepared in water. NP distribution in plant tissues was mapped by synchrotron-radiation micro-X-ray fluorescence, observed by transmission electron microscopy and quantified in the different compartments of plant roots by micro-particle-induced X-ray emission. Our results provide evidence that the smallest TiO(2)-NPs accumulate in roots and distribute through whole plant tissues without dissolution or crystal phase modification. We suggest a threshold diameter, 140 nm, above which NPs are no longer accumulated in wheat roots, as well as a threshold diameter, 36 nm, above which NPs are accumulated in wheat root parenchyma but do not reach the stele and consequently do not translocate to the shoot. This accumulation does not impact wheat seed germination, biomass and transpiration. It does not induce any modification of photosynthesis nor induce oxidative stress. However exposure of wheat plantlets to the smallest NPs during the first stages of development causes an increase of root elongation. Collectively, these data suggest that only the smallest TiO(2)-NPs may be accumulated in wheat plants, although in limited amounts and that their impact is moderate.

Foliar exposure of the crop Lactuca sativa to silver nanoparticles: Evidence for internalization and changes in Ag speciation

The impact of engineered nanomaterials on plants, which act as a major point of entry of contaminants into trophic chains, is little documented. The foliar pathway is even less known than the soil-root pathway. However, significant inputs of nanoparticles (NPs) on plant foliage may be expected due to deposition of atmospheric particles or application of NP-containing pesticides. The uptake of Ag-NPs in the crop species Lactuca sativa after foliar exposure and their possible biotransformation and phytotoxic effects were studied. In addition to chemical analyses and ecotoxicological tests, micro X-ray fluorescence, micro X-ray absorption spectroscopy, time of flight secondary ion mass spectrometry and electron microscopy were used to localize and determine the speciation of Ag at sub-micrometer resolution. Although no sign of phytotoxicity was observed, Ag was effectively trapped on lettuce leaves and a thorough washing did not decrease Ag content significantly. We provide first evidence for the entrapment of Ag-NPs by the cuticle and penetration in the leaf tissue through stomata, for the diffusion of Ag in leaf tissues, and oxidation of Ag-NPs and complexation of Ag(+) by thiol-containing molecules. Such type of information is crucial for better assessing the risk associated to Ag-NP containing products.

Quantitative evaluation of multi-walled carbon nanotube uptake in wheat and rapeseed

Environmental contamination with carbon nanotubes would lead to plant exposure and particularly exposure of agricultural crops. The only quantitative exposure data available to date which can be used for risk assessment comes from computer modeling. The aim of this study was to provide quantitative data relative to multi-walled carbon nanotube (MWCNT) uptake and distribution in agricultural crops, and to correlate accumulation data with impact on plant development and physiology. Roots of wheat and rapeseed were exposed in hydroponics to uniformly (14)C-radiolabeled MWCNTs. Radioimaging, transmission electron microscopy and raman spectroscopy were used to identify CNT distribution. Radioactivity counting made it possible absolute quantification of CNT accumulation in plant leaves. Impact of CNTs on seed germination, root elongation, plant biomass, evapotranspiration, chlorophyll, thiobarbituric acid reactive species and H(2)O(2) contents was evaluated. We demonstrate that less than 0.005‰ of the applied MWCNT dose is taken up by plant roots and translocated to the leaves. This accumulation does not impact plant development and physiology. In addition, it does not induce any modifications in photosynthetic activity nor cause oxidative stress in plant leaves. Our results suggest that if environmental contamination occurs and MWCNTs are in the same physico-chemical state than the ones used in the present article, MWCNT transfer to the food chain via food crops would be very low.

Comparative uptake and impact of TiO₂ nanoparticles in wheat and rapeseed.

Up to 2 million tons per year of titanium dioxide (TiO2) nanoparticles (NP) are produced worldwide. This extensive production is postulated to result in release into the environment with subsequent contamination of soils and plants; however, few studies have examined TiO2-NP uptake and impact on plants. In this study, wheat and rapeseed plantlets were exposed to 14 nm or 25 nm anatase TiO2-NP in hydroponics conditions, either through root or leaf exposure. Microparticle-induced x-ray emission (μPIXE) coupled with Rutherford backscattering spectroscopy (RBS) was used to quantify absorbed titanium (Ti). Micro x-ray fluorescence (μXRF) based on synchrotron radiation was used to evaluate Ti distribution in roots and leaves. Our results show that both TiO2-NP are accumulated in these plantlets upon root exposure and that Ti content is higher in rapeseed than wheat. Ti distribution in root cross sections depended on NP agglomeration state. NP are also accumulated in plantlets upon leaf exposure. Finally, it was found that TiO2-NP exposure induced increased root elongation but did not affect germination, evapotranspiration, and plant biomass. Taken together, these results confirm that TiO2-NP may be accumulated in plant crops but may only moderately impact plant development.

Investigation of titanium dioxide nanoparticles toxicity and uptake by plants

Nanoparticles (NP) are introduced in a growing number of commercial products and their production may lead to their release in the environment. Plants may be a potential entry point for NP in the food chain. Up to now, results describing NP phytotoxical effects and plant accumulation are scarce and contradictory. To increase knowledge on titanium dioxide NP (TiO2-NPs) accumulation and impact on plants, we designed a study on three plant species, namely wheat (Triticum aestivum), oilseed rape (Brassica napus) and Arabidopsis thaliana. These plants were exposed in hydroponics to a panel of well-characterized TiO2-NPs, with diameters ranging from 12 to 140 nm, either anatase or rutile. Their accumulation in plant tissues is currently being assessed by complementary imaging techniques: scanning electron microscopy (SEM), transmission electron microscopy (TEM), micro-X-ray fluorescence (SR-μ-XRF) imaging and micro-particle induced X-ray emission (μ-PIXE) imaging. Moreover, the impact of TiO2-NP exposure on germination rate, root elongation, dry biomass and evapotranspiration is evaluated. Preliminary results are presented here, with data collected on wheat plants exposed to 12 nm and 25 nm anatase TiO2-NPs. These results show that TiO2-NPs are taken up by plants, and do not significantly alter their germination and root elongation. These results underline the necessity of deeper evaluation of nanoparticle ecotoxicity, and particularly on their interaction with plants.

Lung distribution, quantification, co-localization and speciation of silver nanoparticles after lung exposure in mice

Large knowledge gaps still exist on the toxicological mechanisms of silver (Ag) engineered nanoparticles (ENPs); a comprehensive understanding of the sources, biodistribution, toxicity and transformation of Ag ENPs along their life cycle and after transfer in living organisms is needed. In a previous study, mice were pulmonary exposed to Ag ENPs and local (lung) and systemic toxic effects together with biodistribution to organs including heart, liver, spleen and kidney were investigated. Here, Ag lung distribution, local concentration, co-localization with other elements such as Fe, Cu and S, and speciation were determined after lung exposure to Ag ENPs using micro X-ray fluorescence (μXRF), micro X-ray absorption near edge structure spectroscopy (μXANES) and micro proton-induced X-ray emission (μPIXE) techniques. We found that approximately a quarter of all macrophages in the lumen of the airways contained ENPs. High local concentrations of Ag were also detected in the lung tissue, probably phagocytized by macrophages. The largest part of the ENPs was dissolved and complexed to thiol-containing molecules. Increased concentrations of Fe and Cu observed in the Ag-rich spots suggest that these molecules are metallothioneins (MTs). These results give more insights on the behavior of Ag ENPs in the lung in vivo and will help in the understanding of the toxicological mechanisms of Ag ENPs.

Mother-plant-mediated pumping of zinc into the developing seed

Insufficient intake of zinc and iron from a cereal-based diet is one of the causes of ‘hidden hunger’ (micronutrient deficiency), which affects some two billion people1,2. Identifying a limiting factor in the molecular mechanism of zinc loading into seeds is an important step towards determining the genetic basis for variation of grain micronutrient content and developing breeding strategies to improve this trait3. Nutrients are translocated to developing seeds at a rate that is regulated by transport processes in source leaves, in the phloem vascular pathway, and at seed sinks. Nutrients are released from a symplasmic maternal seed domain into the seed apoplasm surrounding the endosperm and embryo by poorly understood membrane transport processes4–6. Plants are unique among eukaryotes in having specific P1B-ATPase pumps for the cellular export of zinc7. In Arabidopsis, we show that two zinc transporting P1B-ATPases actively export zinc from the mother plant to the filial tissues. Mutant plants that lack both zinc pumps accumulate zinc in the seed coat and consequently have vastly reduced amounts of zinc inside the seed. Blockage of zinc transport was observed at both high and low external zinc supplies. The phenotype was determined by the mother plant and is thus due to a lack of zinc pump activity in the seed coat and not in the filial tissues. The finding that P1B-ATPases are one of the limiting factors controlling the amount of zinc inside a seed is an important step towards combating nutritional zinc deficiency worldwide.

Etiolated Seedling Development Requires Repression of Photomorphogenesis by a Small Cell-Wall-Derived Dark Signal

Etiolated growth in darkness or the irreversible transition to photomorphogenesis in the light engages alternative developmental programs operating across all organs of a plant seedling. Dark-grown Arabidopsis de-etiolated by zinc (dez) mutants exhibit morphological, cellular, metabolic, and transcriptional characteristics of light-grown seedlings. We identify the causal mutation in TRICHOME BIREFRINGENCE encoding a putative acyl transferase. Pectin acetylation is decreased in dez, as previously found in the reduced wall acetylation2-3 mutant, shown here to phenocopy dez. Moreover, pectin of dez is excessively methylesterified. The addition of very short fragments of homogalacturonan, tri-galacturonate, and tetra-galacturonate, restores skotomorphogenesis in dark-grown dez and similar mutants, suggesting that the mutants are unable to generate these de-methylesterified pectin fragments. In combination with genetic data, we propose a model of spatiotemporally separated photoreceptive and signal-responsive cell types, which contain overlapping subsets of the regulatory network of light-dependent seedling development and communicate via a pectin-derived dark signal.

Negative Bias Temperature Instability Effect on the Single Event Transient Sensitivity of a 65 nm CMOS Technology

Impact of NBTI degradation on the SET sensitivity of 65 nm CMOS test structures is investigated. Pre- and post-aging SET laser thresholds measurements on chains of gates indicate a decrease of SET sensitivity due to NBTI.

Imaging the Single Event Burnout sensitive volume of vertical power MOSFETs using the laser Two-Photon Absorption technique

The Single Event Burnout sensitive volume of power MOSFETs is investigated using the laser Two-Photon Absorption Technique. A first discussion about the efficiency of this technique is given.