Phytotoxicity Of Nanoparticles Research Articles

Effects of nZnS vs. nZnO and ZnCl2 on mungbean [Vigna radiata (L.) R. Wilczek] plant and Bradyrhizobium symbiosis: A life cycle study

Scarce of knowledge of using Zinc (Zn) nanoparticles (NPs) to augment plant growth, Zn availability to plants and its potential toxicity warrants more NPs–plant life cycle studies. The main objectives of this study were to compare nano zinc sulphide (nZnS) with nano zinc oxide (nZnO) and ionic Zn i.e., ZnCl2, as a source of Zn, as well as to establish physiological impact of NPs on growth, yield and symbiosis of mungbean [Vigna radiata (L.) R. Wilczek] plants at different concentrations (0, 0.01, 0.1, 1 and 10 mg kg−1 of soil). In this study, mungbean plants were grown for 60 days (life cycle study) in natural soil infested with Bradyrhizobium. Effects of Zn compounds (nZnS, nZnO and ZnCl2) on plant height, dry biomass, number of nodules per plant, yield and fruit agronomical parameters along with micronutrient assessment were determined. Impact of Zn compounds on Bradyrhizobium–mungbean symbiosis was also unravelled. Results showed that both the NPs, (nZnS and nZnO) were more effective than ZnCl2 in promoting growth and yield up to a critical concentration and above which phytotoxic effects were observed. Both the NPs were more effective than ZnCl2 at increasing fruit Zn content also. Whereas, nZnS treatment was found to be better than nZnO in improving overall plant growth. Bradyrhizobium–mungbean symbiosis was not affected at lower NPs concentrations, while higher concentration revealed toxicity by damaging bacterial morphology and nodule formation. There was no nano specific toxicity found while, ZnCl2 showed relatively more toxicity than both the NPs. The present investigation demonstrated the concept of nano-micronutrient as well as NPs phytotoxicity by understanding NPs–plant interactions in the soil environment.

Integrating transcriptome and physiological analyses to elucidate the molecular responses of buckwheat to graphene oxide

With the increasing application of nanomaterials, evaluation of the phytotoxicity of nanoparticles has attracted considerable interest. Buckwheat is an economically pseudocereal crop, which is a potential model for investigating the response of plants to hazardous materials. In this study, the response of buckwheat to graphene oxide (GO) was investigated by integrating physiological and transcriptome analysis. GO can penetrate into buckwheat root and stem, and high concentrations of GO inhibited seedlings growth. High concentration of GO improved ROS production and regulated the activities and gene expression of oxidative enzymes, which implying GO may affect plant growth via regulating ROS detoxification. Root and stem exhibit distinct transcriptomic responses to GO, and the GO-responsive genes in stem are more enriched in cell cycle and epigenetic regulation. GO inhibited plant hormone biosynthesis and signaling by analyzing the expression data. Additionally, 97 small secreted peptides (SSPs) encoding genes were found to be involved in GO response. The gene expression of 111 transcription factor (TFs) and 43 receptor-like protein kinases (RLKs) were regulated by GO, and their expression showed high correlation with SSPs. Finally, the TFs-SSPs-RLKs signaling networks in regulating GO response were proposed. This study provides insights into the molecular responses of plants to GO.

Integrating transcriptome and physiological analyses to elucidate the essential biological mechanisms of graphene phytotoxicity of alfalfa (Medicago sativa L.)

The phytotoxicity of nanoparticles has attracted considerable interest, given the broad applications of nanomaterials in different fields. Alfalfa (Medicago sativa L.) is a major forage crop grown worldwide with a high protein content. The molecular regulation mechanisms involved in nanomaterial-treated alfalfa were examined in this research. In our lab, 18 cDNA libraries of Golden Empress (GE) and Bara 310SC (SC) under control (CK), middle (10 g kg−1)- and high (20 g kg−1)-graphene stress treatments were constructed in 2019. All clean reads were matched to the reference Medicago_truncatula genome, the mapping ratio was higher than 50%, and a total of 3946 differentially expressed genes (DEGs) were obtained. The number of DEGs that reflect transcriptional activity is proportional to the degree of stress. For example, 1241/610 and 1794/1422 DEGs were identified as significant in the leaves of GE/SC under mid- and high-graphene treatment, respectively. Furthermore, GO analysis of the DEGs annotated in some significant biochemical process terms included ‘response to abiotic stimulus’, ‘oxidation-reduction process’, ‘protein kinase activity’, and ‘oxidoreductase activity’. KEGG pathway analysis of the DEGs revealed strongly mediated graphene-responsive genes in alfalfa mainly linked to the ‘biosynthesis of amino acids’, ‘isoflavonoid biosynthesis’, ‘linoleic acid metabolism’, and ‘phenylpropanoid biosynthesis’ pathways. In addition, hundreds of DEGs, including photosynthetic, antioxidant enzyme, nitrogen metabolism, and metabolic sucrose and starch genes, have been identified as potentially involved in the response to graphene. Physiological findings revealed that enzymes related to the metabolism of nitrogen play a crucial role in the adaptation of graphene stress to alfalfa. Ultimately, in response to graphene stress, a preliminary regulatory mechanism was proposed for the self-protective mechanism of alfalfa, which helps to explain the phytotoxicity of the molecular mechanism of nanoparticle-treated crops.

Metal nanoparticles: Phytotoxicity on tomato and effect on symbiosis with the Fusarium solani FsK strain

The effect of copper (Cu-NPs, CuO-NPs), silver (Ag-NPs) and zinc oxide (ZnO-NPs) nanoparticles (NPs) on plant growth, physiological properties of tomato plants and their symbiotic relationships with the endophytic Fusarium solani FsK strain was investigated. Fungitoxicity tests revealed that the FsK strain was significantly more sensitive to Cu-NPs and ZnO-NPs than CuO-NPs and Ag-NPs both in terms of mycelial growth and spore germination. All NPs were more toxic to FsK compared to their bulk counterparts except for AgNO3, which was 8 to 9-fold more toxic than Ag-NPs. Apart from AgNO3, NPs and bulk counterparts did not affect the number of germinated tomato seeds even in higher concentrations, while root length was significantly reduced in a dose dependent way in most cases. Dry weight of tomato plants was also significantly reduced upon treatment with NPs and counterparts with most pronounced effects in the cases of AgNO3, Cu-NPs, ZnO-NPs, and ZnSO4. Root and shoot length of grown tomato plants was also affected by treatments while differences between NPs and bulk counterparts varied. A marked oxidative stress response was recorded in all cases of NPs/bulk counterparts as indicated by increased MDA and H2O2 levels of treated plants. Treated plants had significantly reduced chlorophyl-a and carotenoid levels compared to the untreated control. NPs and counterparts did not affect FsK colonization of roots indicating a possible shielding effect of tomato plants once the endophyte was established inside the roots. Vice versa, a possible alleviation of CuO-NPs, ZnO-NPs, and ZnSO4 toxicity was observed in the presence of FsK inside tomato roots in terms of plant dry weight. The results suggest that phytotoxicity of NPs in tomato treated plants should be considered before application and while both FsK and tomato are sensitive to NPs, their reciprocal benefits may extent to resistance towards these toxic agents.

Blocking effect of fullerene nanoparticles (nC60) on the plant cell structure and its phytotoxicity

Blockage of nanoparticles on plant pore structures might produce phytotoxicity and affect plant uptake indirectly. This study examined the blocking and phytotoxic effects of fullerene nanoparticles (nC60) on plants at the cellular level. The malondialdehyde content in plant was normal during nC60 exposure, implying that nC60 caused no acute phytotoxicity, while the normalized relative transpiration significantly decreased, showing that the pore structure of roots was seriously blocked by nC60. High power optical microscopy and transmission electron microscope showed that root endothelial cells were squeezed, and inner wall structures were damaged by the extrusion of nanoparticles. Low nC60 concentrations inhibited root uptake of lindane, whereas high nC60 concentrations promoted root uptake of lindane, indicating that serious pore blocking by nC60 damaged root cell structure and hence ready transport of lindane from roots to shoots. Significant alterations of fatty acid (FA) saturation degree of root cell membrane indicated that nC60 led to phytotoxicity in the root cell membrane after long-term exposure and nC60 produced phytotoxicity in the process of blocking root pore structures and interfering with cell membrane fluidity. Moreover, the plant cell structures under phytotoxicity were more likely to be damaged mechanically by the extrusion of nanoparticles. These findings may be helpful to better understand the transport pathways of nanoparticles in plants, the phytotoxicity of nanoparticles and the potential risks of nanomaterials used in agriculture.

Evaluation of the phytotoxicity of nano-particles on mung beans by internal extractive electrospray ionization mass spectrometry.

The wide application of nano-particles (NPs) has raised a serious concern over their impact on plants. However, evaluation of the effects of NPs on plant metabolism by direct detection of chemicals inside solid tissues presents a challenge. In this study, we report on a direct ionization method in mass spectrometry, internal extractive electrospray ionization (iEESI), for the direct evaluation of phytotoxicity of three different NPs (including CdTe quantum dots (CdTe QDs), gold nano-particles (Au NPs), and silver nano-particles (Ag NPs)) both on surfaces and inside solid tissues from the mung bean seeds (Vigna radiata) that were cultured in aqueous solutions of three NPs at 50 μg mL-1. The results showed that NPs could stimulate the biological accumulation of trigonelline and the decomposition of polysaccharides/oligosaccharides to glucose and maltose within 21 h of culture. To the best of our knowledge, this is the first study to apply internal extractive electrospray ionization mass spectrometry (iEESI-MS) for the direct measurement of solid tissue samples to evaluate the phytotoxicity of NPs on mung bean sprouts. Our study lays a solid foundation for further examination of other NPs-induced damaging effects such as apoptosis/necrosis, helping us to understand the phytotoxicity of NPs on plants.

Biochemical and Physiological Toxicity of Nanoparticles in Plant.

As increasing application of nanoparticles, nanoparticles have been becoming a new emerging environmental pollution that attracts a lot of attention from the scientific community and also regulatory agents. In the past decade, studying the toxicity and environmental impacts of nanoparticles is becoming a hotresearch field and more and more researches have been published using both plant and animal system. In this chapter, using oxidized metal nanoparticles as an example, we introduce a detailed protocol for performing research on biochemical and physiological toxicity of nanoparticles in plant. We employ a hydroponics system to study phytotoxicity of nanoparticles, which makes it easier to study the impact of nanoparticles. In this chapter, we majorly focus on plant respiration and photosynthesis, root vigor as well as oxidative stress. Oxidative stress is one major physiological response to different environmental pollution, in which we present a detailed method for detecting free radical oxygen species as well as the major molecules and enzymes associating with oxidative stress, including SOD and POD. Although we introduce the methods using cotton as an example, the protocols presented in this chapter can be used almost any plant species to test the biochemical and physiological toxicity of an environmental pollution.

Foliar uptake, biotransformation, and impact of CuO nanoparticles in Lactuca sativa L. var. ramosa Hort.

Plant leaves can intercept and directly absorb nanoparticles (NPs) that deposit on their surface, which can lead severe phytotoxicity. However, there is a large blind spot when it comes to the fate and phytotoxicity of NPs after leaf exposure, even though foliar uptake is likely to occur. In this study, lettuce leaves (Lactuca sativa L. var. ramosa Hort.) were exposed to different concentrations of copper-oxide NPs (CuO-NPs, 0, 100, and 1000mg L-1) for 5, 10, and 15days. Foliar uptake, subcellular distribution, chemical forms, and impact of CuO-NPs on nutrient status, antioxidant systems, and lettuce growth were examined. Substantially elevated Cu levels were observed in lettuce leaves (up to 6350mgkg-1), which was one magnitude greater than that in the roots (up to 525mgkg-1). Cu translocation factors from leaves to roots ranged from 1.80 to 15.6%. The application of CuO-NPs severely inhibited lettuce growth and altered the nutrient status in plants (especially Mn, K, and Ca). Moreover, CuO-NPs increased H2O2 generation, malonaldehyde level (on the 5th and 10th day of exposure), and catalase activity (on the 15th day of exposure) in lettuce leaves. The Cu concentrations in subcellular fractions were ranked: cell wall ≈ organelles > soluble fraction in lettuce leaves, and organelles > cell wall > soluble fraction in lettuce roots. Undissolved Cu forms were predominant in lettuce, which may have helped to reduce the Cu's mobility and phytotoxicity in the plant. The findings of this study will be of great interest in areas with high levels of metal-NPs in the atmosphere.

Combination analysis of the physiology and transcriptome provides insights into the mechanism of silver nanoparticles phytotoxicity

Silver nanoparticles (AgNPs) have adverse impacts on plants when released into environments, but their toxic mechanism is still a matter of debate. Here we present a combined analysis of physiology and transcriptome of Arabidopsis thaliana leaves exposure to 30 mg L−1 AgNPs and Ag+ for six days to explore the toxicity mechanism of AgNPs on Arabidopsis. Both transcriptomic and physiological results showed that AgNPs induced reactive oxygen species (ROS) accumulation and damaged photosynthesis. The toxicity of AgNPs is not merely attributable to Ag+ release and much higher photosynthetic toxicity and ROS accumulation were observed in 30 mg L−1 AgNPs than that in 0.12 mg L−1 Ag+. About 60% genes were similarly up- or down-regulated at the same concentration of AgNPs and Ag+ and these genes were enriched in photosynthesis and response to the stimulus. However, 302 genes, including those involved in glucosinolates synthesis, were specifically regulated under AgNPs treatments. In conclusion, more than the released Ag+, nanoparticle-specific effects are responsible for the toxicity of AgNPs in Arabidopsis thaliana.

Nitric Oxide Ameliorates Zinc Oxide Nanoparticles Phytotoxicity in Wheat Seedlings: Implication of the Ascorbate-Glutathione Cycle.

The present study investigates ameliorative effects of nitric oxide (NO) against zinc oxide nanoparticles (ZnONPs) phytotoxicity in wheat seedlings. ZnONPs exposure hampered growth of wheat seedlings, which coincided with reduced photosynthetic efficiency (Fv/Fm and qP), due to increased accumulation of zinc (Zn) in xylem and phloem saps. However, SNP supplementation partially mitigated the ZnONPs-mediated toxicity through the modulation of photosynthetic activity and Zn accumulation in xylem and phloem saps. Further, the results reveal that ZnONPs treatments enhanced levels of hydrogen peroxide and lipid peroxidation (as malondialdehyde; MDA) due to severely inhibited activities of the following ascorbate–glutatione cycle (AsA–GSH) enzymes: ascorbate peroxidase, glutathione reductase, monodehydroascorbate reductase and dehydroascorbate reductase, and its associated metabolites ascorbate and glutathione. In contrast to this, the addition of SNP together with ZnONPs maintained the cellular functioning of the AsA–GSH cycle properly, hence lesser damage was noticed in comparison to ZnONPs treatments alone. The protective effect of SNP against ZnONPs toxicity on fresh weight (growth) can be reversed by 2-(4carboxy-2-phenyl)-4,4,5,5-tetramethyl- imidazoline-1-oxyl-3-oxide, a NO scavenger, and thus suggesting that NO released from SNP ameliorates ZnONPs toxicity. Overall, the results of the present study have shown the role of NO in the reducing of ZnONPs toxicity through the regulation of accumulation of Zn as well as the functioning of the AsA–GSH cycle.

Does seed size and surface anatomy play role in combating phytotoxicity of nanoparticles?

Rapid utilization of nano-based products will inevitably release nanoparticles into the environment with unidentified consequences. Plants, being an integral part of ecosystem play a vital role in the incorporation of nanoparticles in food chain and thus, need to be critically assessed. The present study assesses the comparative phytotoxicity of nanoparticle, bulk and ionic forms of zinc at different concentrations on selected plant species with varying seed size and surface anatomy. ZnO nanoparticles were chosen in view of their wide spread use in cosmetics and health care products, which allow their direct release in the environment. The impact on germination rate, shoot & root length and vigour index were evaluated. A concentration dependent inhibition of seed germination as well as seedling length was observed in all the tested plants. Due to the presence of thick cuticle on testa and root, pearl millet (xerophytic plant) was found to be relatively less sensitive to ZnO nanoparticles as compared to wheat and tomato (mesophytic plants) with normal cuticle layer. No correlation was observed between nanoparticles toxicity and seed size. The results indicated that variations in surface anatomy of seeds play a crucial role in determining the phytotoxicity of nanoparticles. The present findings significantly contribute to assess potential consequences of nanoparticle release in environment particularly with major emphasis on plant systems. It is the first report which suggests that variations observed in phytotoxicity of nanoparticles is mainly due to the predominant differences in size and surface anatomy of tested plant seeds and root architecture. Effect of various concentrations of nano ZnO, bulk ZnO and zinc sulphate on the growth of pearl millet (A), tomato (B) and wheat (C) seedlings.

Effect of Metal Oxides on Plant Germination: Phytotoxicity of Nanoparticles, Bulk Materials, and Metal Ions

The recent rapid expansion of nanotechnologies has increased concern over the impact of engineered nanoparticles (ENPs) on the environment and biota. Although the toxicity of ENPs has received considerable attention in the recent years, there are still gaps in our knowledge of the mechanisms responsible for their effects. In this study, we tested the toxicity of various metal oxide ENPs (Al2O3, CuO, Fe3O4, MnO, TiO2, and ZnO), including nanowires together with their bulk counter particles and soluble metal salts, on germinating seeds of Sinapis alba L. Fe3O4, TiO2, MnO2, and Al2O3 ENPs did not negatively affected seed germination at any tested concentrations. However, CuO and ZnO ENPs showed a dose-dependent inhibition of germination. Metal ions were more toxic than metal oxide particles at corresponding concentrations. The highest toxicity was exhibited by Cu, followed by Zn, Fe, Al, and Mn ions. A comparison of ENPs with bulk materials did not reveal significantly higher ENP toxicity. Similarly, nanowires showed effects similar to other nanoparticles and bulk materials. Our results indicate that the nanosize or shape of particles did not play a crucial role, whereas metal ions released into cultivation media and accumulated in seedlings contributed significantly to the phytotoxicity of metal oxides.

Effects of Stabilized Nanoparticles of Copper, Zinc, Manganese, and Iron Oxides in Low Concentrations on Lettuce (Lactuca sativa) Seed Germination: Nanotoxicants or Nanonutrients?

Information on the phytotoxicity of nanoparticles (NPs) at low concentrations (e.g., ppb to low ppm) is scarce. Therefore, this study was conducted to assess the effects of laboratory-prepared Cu, Zn, Mn, and Fe oxide NPs in low concentrations (<50 ppm) on the germination of lettuce (Lactuca sativa) seeds in a water medium. The data showed that CuO NPs were slightly more toxic than Cu ions while the toxicity of ZnO NPs was similar to that of Zn ions, and MnOx NPs and FeOx NPs were not only less toxic than their ionic counterparts but also significantly stimulated the growth of lettuce seedlings by 12–54 %. This study showed that manufactured NPs were not always more toxic than other chemical species containing the same elements. Instead, Mn or Fe NPs can significantly enhance plant growth and have the potential to be effective nanofertilizers for increasing agronomic productivity.

Plant Responses to Nanoparticle Stress.

With the rapid advancement in nanotechnology, release of nanoscale materials into the environment is inevitable. Such contamination may negatively influence the functioning of the ecosystems. Many manufactured nanoparticles (NPs) contain heavy metals, which can cause soil and water contamination. Proteomic techniques have contributed substantially in understanding the molecular mechanisms of plant responses against various stresses by providing a link between gene expression and cell metabolism. As the coding regions of genome are responsible for plant adaptation to adverse conditions, protein signatures provide insights into the phytotoxicity of NPs at proteome level. This review summarizes the recent contributions of plant proteomic research to elaborate the complex molecular pathways of plant response to NPs stress.

Influence of soil type and environmental conditions on ZnO, TiO2 and Ni nanoparticles phytotoxicity

Intensive development of nanotechnology will result in releasing nanoparticles (NPs) to the environment including soil. The objective of the study was the evaluation of phytotoxicity of inorganic nanoparticles and their bulk counterparts (ZnO, TiO2 and Ni) in various soils using Phytotoxkit F™ method. The estimation of toxicity was conducted with relation to Lepidium sativum. The toxicity of NPs was also estimated in relation to contact time between NPs and soil, effect of light and temperature and NPs–NPs interactions. In all tested variants no effect of NPs on seed germination was observed. NPs displayed varied effect on inhibition of plant root growth in relation to soil type. Only in the case of ZnO nanoparticles and their bulk counterparts a dose–effect relationship was observed. That relationship, however, was observed only in OECD soil. In a majority of cases, aging and increase of temperature caused a reduction of toxicity of NPs, while light conditions increased the toxic effect of NPs. The effect of the NPs interaction: ZnO with TiO2 or Ni had an antagonistic character, that was manifested in a reduction of the toxicity of ZnO.

The influence of ZnO and TiO2nanoparticles on the toxicity of sewage sludges

More and more often sewage sludges become the place of deposition of nanoparticles (NPs), the use of which in consumer products is increasing. In turn, the increasing amount of sewage sludges enforces the need for their utilization (e.g. through the application of sludges to the soil). Therefore, the presence of NPs in sewage sludges may create a new threat to the environment. Thus it becomes important to perform evaluation of the toxicity of sewage sludges in the context of their content of NPs. The objective of the study was to estimate the effect of nanoparticles of ZnO (nano-ZnO) and TiO2 (nano-TiO2) and their bulk counterparts (bZnO and bTiO2) on the toxicity of sewage sludges in relation to selected organisms (plants – Lepidium sativum and Sinapis alba, and microorganisms – Vibrio fischeri and 11 different strains from Microbial Assay for Risk Assessment – MARA). The study also involved the estimation of other factors that may have an effect on the phytotoxicity of NPs in sewage sludge: the size of the particles, the dose of the sewage sludge, the time of NP–sewage sludge contact and light conditions. The effect of both nano-ZnO and nano-TiO2 on the toxicity of the sludges is dependent on the kind of NPs and their concentration. Sludges containing NPs displayed a different level of toxicity from their bulk counterparts. All of the factors estimated (size of particles, dose of sludge, contact time and light conditions) had a significant effect on the phytotoxicity of NPs which was dependent both on the kind of the NPs and on that of the sewage sludge. Estimation of the leachate toxicity indicated a greater sensitivity of plants to the presence of NPs as compared to the sensitivity of microorganisms. Leachates caused a greater reduction of bioluminescence of V. fischeri in the presence of nano-TiO2 than nano-ZnO. Nano-ZnO caused a reduction of the toxicity of the sewage sludge leachates.

Environmental Nanoparticles Interactions with Plants: Morphological, Physiological, and Genotoxic Aspects

Nanoparticles (NPs) are characterized by their small size (less than 100 nm) and large surface area, which confer specific physicochemical properties as strength, electrical, and optical features. NPs can be derived from natural or anthropic sources, such as engineered or unwanted/incidental NPs. The composition, dimension, and morphology of engineered NPs enable their use in a variety of areas, such as electronic, biomedical, pharmaceutical, cosmetic, energy, environmental, catalysis, and materials science. As nanotechnology is an innovative and scientific growth area with an exponential production, more information is needed concerning the impacts of these nanomaterials (NMs) in the environment and, particularly, in animals/humans health and in plants performance. So, research on NPs as emerging contaminants is therefore a new field in environmental health. This minireview describes, briefly, the NPs characterization and their occurrence in the environment stating air, water, and soil. Finally, particular emphasis is given to the interaction of NPs with plants at different levels: morphology, physiology, and genotoxicity. By analyzing this compiled information, it is evident that research on NPs phytotoxicity is in the beginning, and more comprehensive studies are needed not only on NPs cytotoxicity and genotoxicity but also on the best and the most reliable methods of assessing NPs toxicity.

Beneficial role of carbon nanotubes on mustard plant growth: an agricultural prospect

Nowadays an increasing application of nanotechnology in different fields has arisen an extensive debate about the effect of the engineered nanoparticles on environment. Phytotoxicity of nanoparticles has come into limelight in the last few years. However, very few studies have been done so far on the beneficial aspects of nanoparticles on plants. In this article, we report the beneficial effect of multi-walled carbon nanotubes (MWCNTs) having diameter of ~30 nm on Brassica juncea (mustard) seeds. Measurements of germination rate, T50 (time taken for 50% germination), shoot and root growth have shown encouraging results using low concentration of oxidized MWCNT (OMWCNT) treated seeds as compared to non-oxidized as well as high concentration OMWCNT treated seeds. For toxicity study we measured the germination index and relative root elongation, while conductivity test and infra-red spectra were also performed to study the overall effect of oxidized and non-oxidized nanotubes on mustard seeds and seedlings.

Toxicity and bioavailability of copper nanoparticles to the terrestrial plants mung bean (Phaseolus radiatus) and wheat (Triticum aestivum): Plant agar test for water‐insoluble nanoparticles

Because of their insolubility in water, nanoparticles have a limitation concerning toxicity experiments. The present study demonstrated a plant agar test for homogeneous exposure of nanoparticles to plant species. The effect of Cu nanoparticles on the growth of a plant seedling was studied, and bioaccumulation of nanoparticles was investigated. All tests were conducted in plant agar media to prevent precipitation of water-insoluble nanoparticles in test units. The plant species were Phaseolus radiatus (mung bean) and Triticum aestivum (wheat). Growth inhibition of a seedling exposed to different concentrations of Cu nanoparticles was examined. Copper nanoparticles were toxic to both plants and also were bioavailable. The 2-d median effective concentrations for P. radiatus and T. aestivum exposed to Cu nanoparticles were 335 (95% confidence level, 251-447) and 570 (450-722) mg/L, respectively. Phaseolus radiatus was more sensitive than T. aestivum to Cu nanoparticles. A cupric ion released from Cu nanoparticles had negligible effects in the concentration ranges of the present study, and the apparent toxicity clearly resulted from Cu nanoparticles. Bioaccumulation increased with increasing concentration of Cu nanoparticles, and agglomeration of particles was observed in the cells using transmission-electron microscopy-energy-dispersive spectroscopy. The present study demonstrated that the plant agar test was a good protocol for testing the phytotoxicity of nanoparticles, which are hardly water soluble.

Phytotoxicity of nanoparticles: Inhibition of seed germination and root growth

Plants need to be included to develop a comprehensive toxicity profile for nanoparticles. Effects of five types of nanoparticles (multi-walled carbon nanotube, aluminum, alumina, zinc, and zinc oxide) on seed germination and root growth of six higher plant species (radish, rape, ryegrass, lettuce, corn, and cucumber) were investigated. Seed germination was not affected except for the inhibition of nanoscale zinc (nano-Zn) on ryegrass and zinc oxide (nano-ZnO) on corn at 2000 mg/L. Inhibition on root growth varied greatly among nanoparticles and plants. Suspensions of 2000 mg/L nano-Zn or nano-ZnO practically terminated root elongation of the tested plant species. Fifty percent inhibitory concentrations (IC 50) of nano-Zn and nano-ZnO were estimated to be near 50 mg/L for radish, and about 20 mg/L for rape and ryegrass. The inhibition occurred during the seed incubation process rather than seed soaking stage. These results are significant in terms of use and disposal of engineered nanoparticles.