Toxicity Mechanisms of Nanoplastics on Crop Growth, Interference of Phyllosphere Microbes, and Evidence for Foliar Penetration and Translocation.

Despite the increasing prevalence of atmospheric nanoplastics (NPs), there remains limited research on their phytotoxicity, foliar absorption, and translocation in plants. In this study, we aimed to fill this knowledge gap by investigating the physiological effects of tomato leaves exposed to differently charged NPs and foliar absorption and translocation of NPs. We found that positively charged NPs caused more pronounced physiological effects, including growth inhibition, increased antioxidant enzyme activity, and altered gene expression and metabolite composition and even significantly changed the structure and composition of the phyllosphere microbial community. Also, differently charged NPs exhibited differential foliar absorption and translocation, with the positively charged NPs penetrating more into the leaves and dispersing uniformly within the mesophyll cells. Additionally, NPs absorbed by the leaves were able to translocate to the roots. These findings provide important insights into the interactions between atmospheric NPs and crop plants and demonstrate that NPs' accumulation in crops could negatively impact agricultural production and food safety.

Role of Nanoscale Hydroxyapatite in Disease Suppression of Fusarium-Infected Tomato.

The present study investigated the mechanisms by which large- and small-sized nanoscale hydroxyapatite (nHA) suppressed Fusarium-induced wilt disease in tomato. Both nHA sizes at 9.3 mg/L (low) and 46.5 mg/L (high dose) phosphorus (P) were foliar-sprayed on Fusarium-infected tomato leaf surfaces three times. Diseased shoot mass was increased by 40% upon exposure to the low dose of large-sized nHA compared to disease controls. Exposure to both nHA sizes significantly elevated phenylalanine ammonialyase activity and total phenolic content in Fusarium-infected shoots by 30-80% and 40-68%, respectively. Shoot salicylic acid content was also increased by 10-45%, suggesting the potential relationship between antioxidant and phytohormone pathways in nHA-promoted defense against fungal infection. Exposure to the high dose of both nHA sizes increased the root P content by 27-46%. A constrained analysis of principal coordinates suggests that high dose of both nHA sizes significantly altered the fatty acid profile in diseased tomato. Particularly, the diseased root C18:3 content was increased by 28-31% in the large-sized nHA treatments, indicating that nHA remodeled the cell membrane as part of defense against Fusarium infection. Taken together, our findings demonstrate the important role of nHA in promoting disease suppression for the sustainable use of nHA in nanoenabled agriculture.

Dose-Dependent Physiological and Transcriptomic Responses of Lettuce (Lactuca sativa L.) to Copper Oxide Nanoparticles-Insights into the Phytotoxicity Mechanisms.

Understanding the complex mechanisms involved in plant response to nanoparticles (NPs) is indispensable in assessing the environmental impact of nano-pollutants. Plant leaves can directly intercept or absorb NPs deposited on their surface; however, the toxicity mechanisms of NPs to plant leaves are unclear. In this study, lettuce leaves were exposed to copper oxide nanoparticles (CuO-NPs, 0, 100, and 1000 mg/L) for 15 days, then physiological tests and transcriptomic analyses were conducted to evaluate the negative impacts of CuO-NPs. Both physiological and transcriptomic results demonstrated that CuO-NPs adversely affected plant growth, photosynthesis, and enhanced reactive oxygen species (ROS) accumulation and antioxidant system activity. The comparative transcriptome analysis showed that 2270 and 4264 genes were differentially expressed upon exposure to 100 and 1000 mg/L CuO-NPs. Gene expression analysis suggested the ATP-binding cassette (ABC) transporter family, heavy metal-associated isoprenylated plant proteins (HIPPs), endocytosis, and other metal ion binding proteins or channels play significant roles in CuO-NP accumulation by plant leaves. Furthermore, the variation in antioxidant enzyme transcript levels (POD1, MDAR4, APX2, FSDs), flavonoid content, cell wall structure and components, and hormone (auxin) could be essential in regulating CuO-NPs-induced stress. These findings could help understand the toxicity mechanisms of metal NPs on crops, especially NPs resulting from foliar exposure.

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 1000 mg L-1) for 5, 10, and 15 days. 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 6350 mg kg-1), which was one magnitude greater than that in the roots (up to 525 mg kg-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.

Tracing isotopically labeled selenium nanoparticles in plants via single-particle ICP-mass spectrometry.

Agronomic biofortification using selenium nanoparticles (SeNPs) shows potential for addressing selenium deficiency but further research on SeNPs-plants interaction is required before it can be effectively used to improve nutritional quality. In this work, single-particle inductively coupled plasma-mass spectrometry (SP-ICP-MS) was used for tracing isotopically labeled SeNPs (82SeNPs) in Oryza sativa L. tissues. For this purpose, SeNPs with natural isotopic abundance and 82SeNPs were synthesized by a chemical method. The NPs characterization by transmission electron microscopy (TEM) confirmed that enriched NPs maintained the basic properties of unlabeled NPs, showing spherical shape, monodispersity, and sizes in the nano-range (82.8 ± 6.6 nm and 73.2 ± 4.4 nm for SeNPs and 82SeNPs, respectively). The use of 82SeNPs resulted in an 11-fold enhancement in the detection power for ICP-MS analysis, accompanied by an improvement in the signal-to-background ratio and a reduction of the size limits of detection from 89.9 to 39.9 nm in SP-ICP-MS analysis. This enabled 82SeNPs to be tracked in O. sativa L. plants cultivated under foliar application of 82SeNPs. Tracing studies combining SP-ICP-MS and TEM-energy-dispersive X-ray spectroscopy data confirmed the uptake of intact 82SeNPs by rice leaves, with most NPs remaining in the leaves and very few particles translocated to shoots and roots. Translocation of Se from leaves to roots and shoots was found to be lower when applied as NPs compared to selenite application. From the size distributions, as obtained by SP-ICP-MS, it can be concluded that a fraction of the 82SeNPs remained within the same size range as that of the applied NP suspension, while other fraction underwent an agglomeration process in the leaves, as confirmed by TEM images. This illustrates the potential of SP-ICP-MS analysis of isotopically enriched 82SeNPs for tracing NPs in the presence of background elements within complex plant matrices, providing important information about the uptake, accumulation, and biotransformation of SeNPs in rice plants.

Unveiling of interactions between foliar-applied Cu nanoparticles and barley suffering from Cu deficiency.

The objective of this study was to evaluate nano-Cu-plant interactions under Cu deficiency. Nano-Cu at rates of 100 and 1000 mg L-1 was applied as foliar spray to Hordeum vulgare L. during increased demand for nutrients at tillering stage. Corresponding treatment with CuSO4 was used to exam the nano-specific effects. Cu compounds-plant leaves interactions were analyzed with spectroscopic and microscopic methods (ICP-OES, FTIR/ATR, SEM-EDS). Moreover, the effect of Cu compounds on plants in terms of biomass, pigments content, lipid peroxidation, antiradical properties, the activity of enzymes involved in plant defense against stress (SOD, CAT, POD, GR, PAL, PPO) and the content of non-enzymatic antioxidants (GSH, GSSG, TPC) was determined after 1 and 7 days of exposure. Cu loading to plant leaves increased over time, but the content of Cu under treatment with nano-Cu at 100 mg L-1 was lower by 76% than CuSO4 at 7th day of exposure. The changes induced by applied Cu compounds in biochemical traits were mostly observed after 1 day. Our data showed that CuSO4 exposure induce oxidative stress (increased MDA level and GSSG content) when compared to control and nano-Cu treated plants. Noteworthy, nano Cu at 100 mg L-1 demonstrated enhanced stress tolerance as indicated by boosted GSH content. After 7 days, the antioxidant response was almost same compared to control sample. However, based on other indicators (pigment content, chlorosis sign, biomass), it should be noted that CuSO4 caused serve oxidative burst of plant which may resulted in damage of defense system. Nano-Cu, especially at 100 mg L-1, showed promising effect on plant health, and obtained results may be useful for optimizing of nano-Cu application as fertilizer agent.

Co-exposure of polystyrene microplastics influence cadmium trophic transfer along the "lettuce-snail" food chain: Focus on leaf age and the chemical fractionations of Cd in lettuce.

Cadmium (Cd) and polystyrene microplastics (PS) co-contamination always occurs in environment; however, the trophic transfer of Cd and PS is still poorly understood. A hydroponic experiment was conducted to investigate the behavior of Cd in lettuce, together with the root or foliar exposure of different sized PS. Accumulation and chemical form distributions of Cd in leaves were distinguished into young and mature leaves. Subsequently, a 14-day snail feeding experiment was performed. Data showed that Cd accumulation in roots, rather than in leaves, are significantly affected by PS coexistence. However, mature leaves had a higher Cd content than young leaves under the root exposure of PS, while a reverse effect was observed in the foliar exposure. There existed a positive correlation between the food-chain transfer associated Cd (CdFi+Fii+Fiii) in mature leaves and Cd content in snail soft tissue (r = 0.705, p < 0.001), but not in young leaves. Though no bio-amplification of Cd in food chain was observed, an increase of Cd transfer factor (TF) from lettuce to snail was noted in the root exposure of 5 µm PS and the foliar exposure of 0.2 µm PS. Moreover, we observed a highest increase rate of 36.8 % in TF values from lettuce to snail viscera, and a chronic inflammatory response in snail stomach tissue. Therefore, more attentions should be paid to study the ecological risks of heavy metals and microplastics co-contamination in environment.

Uptake and accumulation of naphthalene, phenanthrene, and benzo(b)fluoranthene in winter wheat affected by foliar exposure at different growth stages.

Foliar uptake, as an important pathway of polycyclic aromatic hydrocarbons (PAHs) accumulation in winter wheat, has a great contribution to wheat PAHs, which mainly depends on atmospheric PAHs level. An indoor simulation experiment was conducted to explore the effects of foliar exposure to PAHs at different growth stages on PAHs uptake in wheat. Three levels (0, 0.75, 4.5 mg L-1) of mixed solution of three PAHs (Σ3PAHs) including naphthalene (NAP), phenanthrene (PHE), and benzo(b)fluoranthene (BbF) were sprayed on leaves of two varieties (Yunong, YN; Xiaoyan, XY) of winter wheat (Triticum aestivum L.) during the booting, heading, pre-filling, and post-filling stage. The results showed that the sprayed PAHs exhibited little effects on the growth of the two varieties except the stem biomass of YN was significantly (p < 0.05) reduced when high concentration of PAHs was applied at the post-filling stage. PAHs concentration in winter wheat grain was highest under foliar exposure at the pre-filling stage, while the lowest was found under foliar exposure at the post-filling stage. Transfer factor of PAHs from stem to root (TFroot/stem) of three PAHs when foliar exposure to PAHs at the booting and heading stage was significantly (p < 0.001) higher than that at the pre-filling and post-filling stage, while TFgrain/glume of three PAHs when foliar exposure to PAHs at pre-filling stage was significantly (p < 0.01) higher than that at the other three stages. These results indicated that foliar exposure to PAHs during the vegetative growth stage was transferred and distributed to the root, while PAHs are mainly transferred and accumulated to the grain during the grain filling stage. Additionally, the higher lipophilic PAHs showed a lower ability to transfer from the glume to grain, and larger flag leaf area had the potential to promote the enrichment of PAHs in grain. This study indicated that the health risk of PAHs in winter wheat could be effectively reduced by controlling atmospheric PAHs level during pre-filling stage.

Foliar-applied polystyrene nanoplastics (PSNPs) reduce the growth and nutritional quality of lettuce (Lactuca sativa L.).

Currently, there is a lack of information about the influence of foliar-applied nanoplastics on crop growth and nutritional quality. To fill the knowledge gap, soil-grown lettuces (Lactuca sativa L.) were foliar-exposed to polystyrene nanoplastics (PSNPs) at 0, 0.1 and 1 mg/L for one month. Foliar exposure to PSNPs significantly decreased the dry weight, height, and leaf area of lettuce by 14.3%-27.3%, 24.2%-27.3%, and 12.7%-19.2%, respectively, compared with the control. Similarly, plant pigment content (chlorophyll a, b and carotenoid) was considerably reduced (9.1%, 8.7%, 12.5%) at 1 mg/L PSNPs. However, the significant increase in electrolyte leakage rate (18.6%-25.5%) and the decrease in total antioxidant capacity (12.4%-26%) were the key indicators of oxidative stress in lettuce leaves, demonstrating the phytotoxicity of PSNPs by foliar exposure. In addition, the remarkable reduction in micronutrients and essential amino acids demonstrated a decrease in nutritional quality of lettuce caused by PSNPs. Besides, SEM and TEM analysis indicated the possible absorption of PSNPs through leaves stoma and the translocation downwards to plant roots. This study provides new information about the interaction of airborne NPs with plants. It also warns against atmospheric NPs pollution that the adverse effects of airborne NPs on crop production and food quality should be assessed as a matter of urgency.

Foliar spray of TiO2 nanoparticles prevails over root application in reducing Cd accumulation and mitigating Cd-induced phytotoxicity in maize (Zea mays L.).

Cadmium (Cd) pollution is considered one of the global environmental issues due to its adverse effects on plant and human health. With the rapid development of nanotechnology and the practical application of engineered nanoparticles (ENPs) in agriculture, the mechanisms underlying the interactions between NPs and heavy metal on their uptake, accumulation, and phytotoxicity in crops are still not fully understood. Therefore, the impact of TiO2 NPs (0, 100, 250 mg/L) and Cd (0, 50 µM) co-exposure on hydroponic maize (Zea mays L.) was determined under two exposure modes. Results showed that root co-exposure to TiO2 NPs and 100 mg/L Cd significantly enhanced Cd uptake and produced greater phytotoxicity in maize than foliar exposure to TiO2 NPs. Meanwhile, plant dry weight and chlorophyll content showed a reduction of 45.3% and 50.5%, respectively, when compared with single Cd treatment. In addition, the accumulation of Ti in shoots and roots increased by 1.61 and 4.29 times, respectively when root exposure to 250 mg/L TiO2 NPs. By contrast, foliar exposure of TiO2 NPs could markedly decrease shoot Cd contents from 15.2% to 17.8% and had a stronger influence on alleviating Cd-induced toxicity via increasing superoxide dismutase (SOD) and glutathione S-transferase (GST) activities and upregulating several metabolic pathways, including galactose metabolism and citrate cycle, alanine, aspartate and glutamate metabolism, as well as glycine, serine and threonine metabolism. This study provides a new strategy for the application of TiO2 NPs in crop safety production in Cd contaminated soils.

Integration of subcellular partitioning and chemical forms to understand silver nanoparticles toxicity to lettuce (Lactuca sativa L.) under different exposure pathways.

The current understanding of the biological impacts of silver nanoparticles (AgNPs) is restricted to the direct interactions of the particles with biota. Very little is known about their intracellular fate and subsequent toxic consequences. In this research we investigated the uptake, internal fate (i,e., Ag subcellular partitioning and chemical forms), and phytotoxicity of AgNPs in lettuce following foliar versus root exposure. At the same AgNP exposure concentrations, root exposure led to more deleterious effects than foliar exposure as evidenced by a larger extent of reduced plant biomass, elevated oxidative damage, as well as a higher amount of ultrastructural injuries, despite foliar exposure leading to 2.6-7.6 times more Ag bioaccumulation. Both Ag subcellular partitioning and chemical forms present within the plant appeared to elucidate this difference in toxicity. Following foliar exposure, high Ag in biologically detoxified metals pool (29.2-53.0% by foliar exposure vs. 12.8-45.4% by root exposure) and low Ag proportion in inorganic form (6.1-11.9% vs. 14.1-19.8%) potentially associated with AgNPs tolerance. Silver-containing NPs (24.8-38.6 nm, 1.5-2.3 times larger than the initial size) were detected in lettuce plants exposed to NPs and to dissolved Ag+, suggesting possible transformation and/or aggregation of AgNPs in the plants. Our observations show that the exposure pathway significantly affects the uptake and internal fate of AgNPs, and thus the associated phytotoxicity. The results are an important contribution to improve risk assessment of NPs, and will be critical to ensure food security.

Uptake of iron oxide nanoparticles inhibits the photosynthesis of the wheat after foliar exposure.

Iron oxide nanoparticles (nFe2O3)-filled materials have been widely employed in various products and their effects on plants have attracted considerable attention because of their potential release into the environment. Currently, numerous studies reporting the influences of iron-bearing nanoparticles on plants are focused on root or seed exposure. However, plants exposed to atmospheric iron-bearing nanoparticles through the leaves and their impacts on plants are still not well understood. This study focused on the uptake, translocation, and effects of foliar exposure of nFe2O3 on wheat seedlings. Wheat seedlings were foliar applied to various concentrations of nFe2O3 (0, 60 and 180 µg per plant) for 1, 7, 14 or 21 d. Our results demonstrated that after exposure for 21 d, the concentrations of Fe in leaves, stems, and roots were 1100, 280 and 160 µg kg-1, respectively. Scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS), as well as the backscattered electron (BSE) images, revealed the stomatal opening was likely the pathway for nFe2O3 uptake. Analysis of the transfer rate, translocation of Fe from leaves to stems and roots, suggested the involvement of plant Fe regulation processes. Particularly, the antioxidant enzymatic activities and malondialdehyde levels in leaves were modified, which was ascribed to the excessive hydroxyl radical (OH) generated via the Fenton-like reaction mediated by nFe2O3. Finally, the OH facilitated the degradation of chlorophyll, posting a negative impact on the photosynthesis, and thus inhibited the biomass production. These findings are meaningful to understand the fate and physiological effects of atmospheric nFe2O3 in crops.

Effects of exposure pathways on the accumulation and phytotoxicity of silver nanoparticles in soybean and rice.

The widespread use of silver nanoparticles (AgNPs) raises concerns both about their accumulation in crops and human exposure via crop consumption. Plants take up AgNPs through their leaves and roots, but foliar uptake has been largely ignored. To better understand AgNPs-plant interactions, we compared the uptake, phytotoxicity and size distribution of AgNPs in soybean and rice following root versus foliar exposure. At similar AgNP application levels, foliar exposure led to 17-200 times more Ag bioaccumulation than root exposure. Root but not foliar exposure significantly reduced plant biomass, while root exposure increased the malondialdehyde and H2O2 contents of leaves to a larger extent than did foliar exposure. Following either root or foliar exposure, Ag-containing NPs larger (36.0-48.9 nm) than the originally dosed NPs (17-18 nm) were detected within leaves. These particles were detected using a newly developed macerozyme R-10 tissue extraction method followed by single-particle inductively coupled plasma mass spectrometry. In response to foliar exposure, these NPs were stored in the cell wall and plamalemma of leaves. NPs were also detected in planta following Ag ion exposure, indicating their in vivo formation. Leaf-to-leaf and root-to-leaf translocation of NPs in planta was observed but the former did not alter the size distribution of the NPs. Our observations point to the possibility that fruits, seeds and other edible parts may become contaminated by translocation processes in plants exposed to AgNPs. These results are an important contribution to improve the risk assessment of NPs under environmental exposure scenarios.