Jason C. White

Assay-Dependent Phytotoxicity of Nanoparticles to Plants

The effects of five nanomaterials (multiwalled carbon nanotubes [MWCNTs], Ag, Cu, ZnO, Si) and their corresponding bulk counterparts on seed germination, root elongation, and biomass of Cucurbita pepo (zucchini) were investigated. The plants were grown in hydroponic solutions amended with nanoparticles or bulk material suspensions at 1000 mg/L. Seed germination was unaffected by any of the treatments, but Cu nanoparticles reduced emerging root length by 77% and 64% relative to unamended controls and seeds exposed to bulk Cu powder, respectively. During a 15-day hydroponic trial, the biomass of plants exposed to MWCNTs and Ag nanoparticles was reduced by 60% and 75%, respectively, as compared to control plants and corresponding bulk carbon and Ag powder solutions. Although bulk Cu powder reduced biomass by 69%, Cu nanoparticle exposure resulted in 90% reduction relative to control plants. Both Ag and Cu ion controls (1-1000 mg/L) and supernatant from centrifuged nanoparticle solutions (1000 mg/L) indicate that half the observed phytotoxicity is from the elemental nanoparticles themselves. The biomass and transpiration volume of zucchini exposed to Ag nanoparticles or bulk powder at 0-1000 mg/mL for 17 days was measured. Exposure to Ag nanoparticles at 500 and 100 mg/L resulted in 57% and 41% decreases in plant biomass and transpiration, respectively, as compared to controls or to plants exposed to bulk Ag. On average, zucchini shoots exposed to Ag nanoparticles contained 4.7 greater Ag concentration than did the plants from the corresponding bulk solutions. These findings demonstrate that standard phytotoxicity tests such as germination and root elongation may not be sensitive enough or appropriate when evaluating nanoparticle toxicity to terrestrial plant species.

Concurrent plant uptake of heavy metals and persistent organic pollutants from soil

The extent of anthropogenic environmental pollution—in the United States (Black, 1999; Glass, 1999), in the European Union (Chaudhry et al., 2002), and in the third world is well documented. For example, the Food and Agriculture Organization of the United Nations has estimated that at a minimum 10 t of unwanted pesticides are in ‘‘storage’’ in undeveloped countries, with at least 2 10 t in African countries (Chaudhry et al., 2002). The potential for negative impacts of these stocks on humans and the environment is of major concern. In situ remediation techniques, such as phytoremediation— the attenuation of pollution through the use of plants—which impose minimal environmental disturbance, offer economic, agronomic, and societal benefits to all countries. Up to the present time phytoremediation of soilborne heavy metals and of organic contaminants has been pursued as two distinct disciplines. This compartmentalized approach applies to fundamental studies of the mechanisms of action, as well as to the development of remediation technologies. Based on data from the authors’ laboratories over the preceding several years we propose that far more convergence as opposed to divergence in the underlying plant physiology and soil science impacts the soil/vegetation microcosm to attenuate both soil-borne heavy metals and organic pollutants. For example, our published research has established that zucchini (Cucurbita pepo L.) and spinach (Spinacia oleracea) bioaccumulate soil-bound persistent organic pollutants (POPs) (Mattina et al., 2002). Other published reports have shown that spinach bioaccumulates heavy metals from soil (Romer and Keller, 2001). The data which are presented here demonstrate that these two plants simultaneously bioconcentrate and translocate both categories of weathered, soil-bound pollutant. Such simultaneous uptake and translocation of heavy metal and organic pollutants, if confirmed and optimized, could have enormous implications for plant/soil interaction mechanisms, and impact on practical remediation approaches, and ultimately on risk to human health.

Graphene in the Aquatic Environment: Adsorption, Dispersion, Toxicity and Transformation

Graphene-family nanomaterials (GFNs) including pristine graphene, reduced graphene oxide (rGO) and graphene oxide (GO) offer great application potential, leading to the possibility of their release into aquatic environments. Upon exposure, graphene/rGO and GO exhibit different adsorption properties toward environmental adsorbates, thus the molecular interactions at the GFN-water interface are discussed. After solute adsorption, the dispersion/aggregation behaviors of GFNs can be altered by solution chemistry, as well as by the presence of colloidal particles and biocolloids. GO has different dispersion performance from pristine graphene and rGO, which is further demonstrated from surface properties. Upon exposure in aquatic environments, GFNs have adverse impacts on aquatic organisms (e.g., bacteria, algae, plants, invertebrates, and fish). The mechanisms of GFNs toxicity at the cellular level are reviewed and the remaining unclear points on toxic mechanisms such as membrane damage are presented. Moreover, we highlight the transformation routes of GO to rGO. The degradation of GFNs upon exposure to UV irradiation and/or biota is also reviewed. In view of the unanswered questions, future research should include comprehensive characterization of GFNs, new approaches for explaining GFNs aggregation, environmental behaviors of metastable GO, and the relationship between dispersion of GFNs and the related adsorption properties.

Xylem- and Phloem-Based Transport of CuO Nanoparticles in Maize (Zea mays L.)

This work reports on the toxicity of CuO nanoparticles (NPs) to maize (Zea mays L.) and their transport and redistribution in the plant. CuO NPs (100 mg L(-1)) had no effect on germination, but inhibited the growth of maize seedlings; in comparison the dissolved Cu(2+) ions and CuO bulk particles had no obvious effect on maize growth. CuO NPs were present in xylem sap as examined by transmission electron microscopy (TEM) and energy dispersive spectroscopy (EDS), showing that CuO NPs were transported from roots to shoots via xylem. Split-root experiments and high-resolution TEM observation further showed that CuO NPs could translocate from shoots back to roots via phloem. During this translocation, CuO NPs could be reduced from Cu (II) to Cu (I). To our knowledge, this is the first report of root-shoot-root redistribution of CuO NPs within maize. The current study provides direct evidence for the bioaccumulation and biotransformation of CuO NPs (20-40 nm) in maize, which has significant implications on the potential risk of NPs and food safety.

Trophic transfer, transformation, and impact of engineered nanomaterials in terrestrial environments.

Engineered nanomaterials (ENMs) are released into the environment with unknown implications in the food chain. Recent findings demonstrate that ENMs may accumulate and/or increase concentrations of the component metal or carbon nanomaterials in the fruits/grains of agricultural crops, have detrimental or beneficial effects on the agronomic traits, yield, and productivity of plants, induce modifications in the nutritional value of food crops, and transfer within trophic levels. Given this information, important questions needed to be resolved include a determination of actual or predicted concentrations of ENMs through the development of new and perhaps hybridized analytical tools, assessment of the nutritional content modifications and/or accumulation of ENMs, component metal, and cocontaminants in edible plants and their implications on human diet, nutrition, and health, assessment of the consequences of ENM-induced changes in soil health, physiological process, and yield on agricultural production and food security, and transfer of ENMs in trophic levels. Given the significant implications of ENMs exposure and the rather large knowledge gaps that exist, it will be prudent to observe judicious and targeted use of ENMs so as to minimize environmental release until a comprehensive environmental fate and effects assessment can be undertaken.

Identification and avoidance of potential artifacts and misinterpretations in nanomaterial ecotoxicity measurements.

Novel physicochemistries of engineered nanomaterials (ENMs) offer considerable commercial potential for new products and processes, but also the possibility of unforeseen and negative consequences upon ENM release into the environment. Investigations of ENM ecotoxicity have revealed that the unique properties of ENMs and a lack of appropriate test methods can lead to results that are inaccurate or not reproducible. The occurrence of spurious results or misinterpretations of results from ENM toxicity tests that are unique to investigations of ENMs (as opposed to traditional toxicants) have been reported, but have not yet been systemically reviewed. Our objective in this manuscript is to highlight artifacts and misinterpretations that can occur at each step of ecotoxicity testing: procurement or synthesis of the ENMs and assessment of potential toxic impurities such as metals or endotoxins, ENM storage, dispersion of the ENMs in the test medium, direct interference with assay reagents and unacknowledged indirect effects such as nutrient depletion during the assay, and assessment of the ENM biodistribution in organisms. We recommend thorough characterization of initial ENMs including measurement of impurities, implementation of steps to minimize changes to the ENMs during storage, inclusion of a set of experimental controls (e.g., to assess impacts of nutrient depletion, ENM specific effects, impurities in ENM formulation, desorbed surface coatings, the dispersion process, and direct interference of ENM with toxicity assays), and use of orthogonal measurement methods when available to assess ENMs fate and distribution in organisms.

A review of the use of engineered nanomaterials to suppress plant disease and enhance crop yield

Nanotechnology has the potential to play a critical role in global food production, food security, and food safety. The applications of nanotechnology in agriculture include fertilizers to increase plant growth and yield, pesticides for pest and disease management, and sensors for monitoring soil quality and plant health. Over the past decade, a number of patents and products incorporating nanomaterials into agricultural practices (e.g., nanopesticides, nanofertilizers, and nanosensors) have been developed. The collective goal of all of these approaches is to enhance the efficiency and sustainability of agricultural practices by requiring less input and generating less waste than conventional products and approaches. This review evaluates the current literature on the use of nanoscale nutrients (metals, metal oxides, carbon) to suppress crop disease and subsequently enhance growth and yield. Notably, this enhanced yield may not only be directly linked to the reduced presence of pathogenic organisms, but also to the potential nutritional value of the nanoparticles themselves, especially for the essential micronutrients necessary for host defense. We also posit that these positive effects are likely a result of the greater availability of the nutrients in the “nano” form. Last, we offer comments on the current regulatory perspective for such applications.

Toxicity of silver and copper to Cucurbita pepo: Differential effects of nano and bulk‐size particles

The phytotoxicity of bulk and nanoparticle Cu and Ag was directly compared. NP Ag reduced biomass and transpiration by 66–84% when compared with bulk Ag. The Ag ion concentration was 4.4–10‐times greater in NP than bulk particle solutions. The Cu ion concentration was 1.4–4.4‐times greater in bulk than NP amended solutions. Humic acid (50 mg/L) decreased the ion content of bulk Cu solution by 38–42% but increased ion Cu content of NP solutions by 1.4–2.9 times. Bulk and NP Cu were highly phytotoxic; growth and transpiration were reduced by 60–70% relative to untreated controls. NP Cu phytotoxicity was unaffected by solution type, but humic acid (50 mg/L) completely alleviated phytotoxicity caused by bulk Cu. The data demonstrate differential toxicity of Ag NP relative to bulk Ag. The finding that humic acid and solution chemistry differentially impact bulk and NP behavior highlights the importance of evaluating nanoparticles under environmentally relevant conditions.

Multiwalled Carbon Nanotubes and C60 Fullerenes Differentially Impact the Accumulation of Weathered Pesticides in Four Agricultural Plants

The effect of multiwalled carbon nanotubes (MWCNT) or C60 fullerenes on the uptake of weathered chlordane or DDx (DDT + metabolites) by Cucurbita pepo (zucchini), Zea mays (corn), Solanum lycopersicum (tomato), and Glycine max (soybean) was investigated. The plants were grown in 50 g of soil with weathered chlordane (2150 ng/g) and DDx (118 ng/g) that was amended with 0, 500, 1000, or 5000 mg/kg MWCNT or C60. After 28 d, the root and shoot content of chlordane components and DDx was determined by GC-MS. Zucchini and tomato growth were unaffected by carbon nanomaterial coexposure, although C60 at 500 mg/kg reduced corn and soybean biomass by 36.5-45.0%. Total chlordane content ranged from 1490 (tomato) to 4780 (zucchini) ng; DDx amounts ranged from 77.8 (corn) to 395 ng (zucchini). MWCNT coexposure decreased chlordane and DDx accumulation 21-80% across all crops, depending on species and nanotube concentration. Conversely, C60 had species- and contaminant-specific effects on pesticide uptake, ranging from complete suppression of DDx uptake (corn/tomato) to 34.9% increases in chlordane accumulation (tomato/soybean). The data show that pesticide accumulation varies greatly with crop species and carbon nanomaterial type/concentration. These findings have implications for food safety and for the use of engineered nanomaterials in agriculture.

Metal-based nanotoxicity and detoxification pathways in higher plants.

The potential risks from metal-based nanoparticles (NPs) in the environment have increased with the rapidly rising demand for and use of nanoenabled consumer products. Plants central roles in ecosystem function and food chain integrity ensure intimate contact with water and soil systems, both of which are considered sinks for NPs accumulation. In this review, we document phytotoxicity caused by metal-based NPs exposure at physiological, biochemical, and molecular levels. Although the exact mechanisms of plant defense against nanotoxicity are unclear, several relevant studies have been recently published. Possible detoxification pathways that might enable plant resistance to oxidative stress and facilitate NPs detoxification are reviewed herein. Given the importance of understanding the effects and implications of metal-based NPs on plants, future research should focus on the following: (1) addressing key knowledge gaps in understanding molecular and biochemical responses of plants to NPs stress through global transcriptome, proteome, and metablome assays; (2) designing long-term experiments under field conditions at realistic exposure concentrations to investigate the impact of metal-based NPs on edible crops and the resulting implications to the food chain and to human health; and (3) establishing an impact assessment to evaluate the effects of metal-based NPs on plants with regard to ecosystem structure and function.