Three-dimensional architecture of ESCRT-III flat spirals on the membrane.

The endosomal sorting complexes required for transport (ESCRTs) are responsible for membrane remodeling in many cellular processes, such as multivesicular body biogenesis, viral budding, and cytokinetic abscission. ESCRT-III, the most abundant ESCRT subunit, assembles into flat spirals as the primed state, essential to initiate membrane invagination. However, the three-dimensional architecture of ESCRT-III flat spirals remained vague for decades due to highly curved filaments with a small diameter and a single preferred orientation on the membrane. Here, we unveiled that yeast Snf7, a component of ESCRT-III, forms flat spirals on the lipid monolayers using cryogenic electron microscopy. We developed a geometry-constrained Euler angle-assigned reconstruction strategy and obtained moderate-resolution structures of Snf7 flat spirals with varying curvatures. Our analyses showed that Snf7 subunits recline on the membrane with N-terminal motifs α0 as anchors, adopt an open state with fused α2/3 helices, and bend α2/3 gradually from the outer to inner parts of flat spirals. In all, we provide the orientation and conformations of ESCRT-III flat spirals on the membrane and unveil the underlying assembly mechanism, which will serve as the initial step in understanding how ESCRTs drive membrane abscission.

High-nitrogen fertilizer alleviated adverse effects of drought stress on the growth and photosynthetic characteristics of Hosta 'Guacamole'.

BACKGROUND: Several plants are facing drought stress due to climate change in recent years. In this study, we aimed to explore the effect of varying watering frequency on the growth and photosynthetic characteristics of Hosta 'Guacamole'. Moreover, we investigated the effect of high-nitrogen and -potassium fertilizers on alleviating the impacts of drought stress on the morphology, photosynthetic characteristics, chlorophyll fluorescence, fast chlorophyll a fluorescence transient, JIP-test parameters, and enzymatic and non-enzymatic scavenging system for reactive oxygen species (ROS) in this species. RESULTS: Leaf senescence, decreased chlorophyll contents, limited leaf area, and reduced photosynthetic characteristics and oxygen-evolving complex (OEC) activity were observed in Hosta 'Guacamole' under drought stress. However, high-nitrogen fertilizer (30-10-10) could efficiently alleviate and prevent the adverse effects of drought stress. High-nitrogen fertilizer significantly increased chlorophyll contents, which was higher by 106% than drought stress. Additionally, high-nitrogen fertilizer significantly improved net photosynthetic rate and water use efficiency, which were higher by 467% and 2900% than those under drought stress. It attributes that high-nitrogen fertilizer could reduce transpiration rate of leaf cells and stomatal opening size in drought stress. On the other hand, high-nitrogen fertilizer enhanced actual photochemical efficiency of PS II and photochemical quenching coefficient, and actual photochemical efficiency of PS II significantly higher by 177% than that under drought stress. Furthermore, high-nitrogen fertilizer significantly activated OEC and ascorbate peroxidase activities, and enhanced the performance of photosystem II and photosynthetic capacity compared with high-potassium fertilizers (15-10-30). CONCLUSIONS: High-nitrogen fertilizer (30-10-10) could efficiently alleviate the adverse effects of drought stress in Hosta 'Guacamole' via enhancing OEC activity and photosynthetic performance and stimulating enzymatic ROS scavenging system.

Changes in carbohydrate distribution in cotton photosynthetic organs and increase in boll weight reduce yield loss under high temperature.

Yield of cotton (Gossypium hirsutum) does not always fall with high temperature (HT) even though this induces significant reductions in fruit retention. To investigate the underlying mechanisms, a greenhouse experiment was conducted with two temperature regimes [control treatment, 28 °C; high temperature (HT), 34 °C] for 7 d. Results showed HT did not significantly influence cotton yield, but reduced boll number and increased boll weight. The 13C distribution ratio of the leaf subtending the cotton boll (LSCB) decreased while that of the cotton boll increased under HT. Transcriptomic and proteomic analyses of the LSCB revealed up-regulated genes involved in cytokinin and jasmonic acid synthesis, as well as SWEET15 (GH_D01G0218), which positively regulated photosynthesis and transport photosynthate, ultimately leading to increased boll weight. After 7 d recovery from HT, the 13C distribution ratio of the LSCB increased while that of the cotton boll decreased. However, boll weight still increased, which was related to increased amylase and sucrose phosphate synthase activities and up-regulated sucrose transport genes in the main-stem leaf and capsule wall. Thus, both accelerated sucrose synthesis and transport in the LSCB under HT and increased sucrose supply ability of the main-stem leaf and capsule wall after recovery from HT contributed to an increased boll weight, which finally maintained cotton yield.

Stimulated Raman Scattering Microscopy Reveals Bioaccumulation of Small Microplastics in Protozoa from Natural Waters.

Microplastics (MPs) are pollutants of global concern, and bioaccumulation determines their biological effects. Although microorganisms form a large fraction of our ecosystem's biomass and are important in biogeochemical cycling, their accumulation of MPs has never been confirmed in natural waters because current tools for field biological samples can detect only MPs > 10 µm. Here, we show that stimulated Raman scattering microscopy (SRS) can image and quantify the bioaccumulation of small MPs (<10 µm) in protozoa. Our label-free method, which differentiates MPs by their SRS spectra, detects individual and mixtures of different MPs (e.g., polyethylene, polypropylene, polyvinyl chloride, polyethylene terephthalate, polystyrene, and poly(methyl methacrylate)) in protozoa. The ability of SRS to quantify cellular MP accumulation is similar to that of flow cytometry, a fluorescence-based method commonly used to determine cellular MP accumulation. Moreover, we discovered that protozoa in water samples from Yangtze River, Xianlin Wastewater Treatment Plant, Lake Taihu and the Pearl River Estuary accumulated MPs < 10 µm, but the proportion of MP-containing cells was low (∼2-5%). Our findings suggest that small MPs could potentially enter the food chain and transfer to organisms at higher trophic levels, posing environmental and health risks that deserve closer scrutiny.

In situ biosynthesized polyphosphate nanoparticles/reduced graphene oxide composite electrode for highly sensitive detection of heavy metal ions.

The development of an effective sensing platform is critical for the electrochemical detection of heavy metal ions (HMIs) in water. In this study, we fabricated a newly designed sensor through the in situ assembly of reduced graphene oxide (rGO) and polyphosphate nanoparticles (polyP NPs) on a carbon cloth electrode via microorganism-mediated green biochemical processes. The characterization results revealed that the rGO produced via microbial reduction had a three-dimensional porous structure, serving as an exceptional scaffold for hosting polyP NPs, and the polyP NPs were evenly distributed on the rGO network. In terms of detecting HMIs, the numerous functional groups of polyP NPs play a major role in the coordination with the cations. This electrochemical sensor, based on polyP NPs/rGO, enabled the individual and simultaneous determination of lead ion (Pb2+) and copper ion (Cu2+) with detection limits of 1.6 nM and 0.9 nM, respectively. Additionally, the electrode exhibited outstanding selectivity for the target analytes in the presence of multiple interfering metal ions. The fabricated sensor was successfully used to determine Pb2+/Cu2+ in water samples with satisfactory recovery rates ranging from 92.16% to 104.89%. This study establishes a facile, cost-effective, and environmentally friendly microbial approach for the synthesis of electrode materials and the detection of environmental pollutants.

The response of structure and nitrogen removal function of the biofilm on submerged macrophytes to high ammonium in constructed wetlands.

The ammonium exceedance discharge from sewage treatment plants has a great risk to the stable operation of subsequent constructed wetlands (CWs). The effects of high ammonium shocks on submerged macrophytes and epiphytic biofilms on the leaves of submerged macrophytes in CWs were rarely mentioned in previous studies. In this paper, the 16S rRNA sequencing method was used to investigate the variation of the microbial communities in biofilms on the leaves of Vallisneria natans plants while the growth characteristics of V. natans plants were measured at different initial ammonium concentrations. The results demonstrated that the total chlorophyll and soluble sugar synthesis of V. natans plants decreased by 51.45% and 57.16%, respectively, and malondialdehyde content increased threefold after 8 days if the initial NH4+-N concentration was more than 5 mg/L. Algal density, bacterial quantity, dissolved oxygen, and pH increased with high ammonium shocks. The average removal efficiencies of total nitrogen and NH4+-N reached 73.26% and 83.94%, respectively. The heat map and relative abundance analysis represented that the relative abundances of phyla Proteobacteria, Cyanobacteria, and Bacteroidetes increased. The numbers of autotrophic nitrifiers and heterotrophic nitrification aerobic denitrification (HNAD) bacteria expanded in biofilms. In particular, HNAD bacteria of Flavobacterium, Hydrogenophaga, Acidovorax, Acinetobacter, Pseudomonas, Aeromonas, and Azospira had higher abundances than autotrophic nitrifiers because there were organic matters secreted from declining leaves of V. natans plants. The analysis of the nitrogen metabolic pathway showed aerobic denitrification was the main nitrogen removal pathway. Thus, the nitrification and denitrification bacterial communities increased in epiphytic biofilms on submerged macrophytes in constructed wetlands while submerged macrophytes declined under ammonium shock loading.

Label-Free Imaging of Humic Substance Bioaccumulation by Pump-Probe Microscopy.

Humic substances (HS) are the most abundant forms of natural organic matter on the earth surface. Comprised of decomposed plant and animal materials rich in carbon, oxygen, hydrogen, nitrogen, and sulfur complexes, HS facilitate global carbon and nitrogen cycling and the transport of anthropogenic contaminants. While it is known that HS also interact with organisms at different trophic levels to produce beneficial and harmful effects whether HS exert these biological effects through accumulation remains unknown. Current radiolabeling techniques, which only detect the amount of accumulated radiolabels, cannot visualize the transport and accumulation behavior of HS. Here, using a label-free method based on pump-probe microscopy, we show HS entered the protozoan Tetrahymena thermophila, zebrafish embryos, and human cells and exerted direct effects on these organisms. HS accumulated in the nucleus of T. thermophila, chorion pore canals of zebrafish embryos, and nucleus of intestinal and lung cells in a concentration- and time-dependent way. Epigenetic and transcriptomics assays show HS altered chromatin accessibility and gene transcription in T. thermophila. In zebrafish larvae, HS induced neurotoxicity, altering spontaneous muscle contraction and locomotor activity. Detailed images showing HS accumulation in our study reveal new insights on the ecological and environmental behavior of HS.

Pre-exposure to Fe2O3 or TiO2 Nanoparticles Inhibits Subsequent Biological Uptake of 55Fe-Labeled Fe2O3 Nanoparticles.

Aquatic organisms are frequently exposed to various nanoparticles (NPs) in the natural environment. Thus, studies of NP bioaccumulation should include organisms that have been previously exposed to NPs. Our study investigated the effects of pre-exposure of Tetrahymena thermophila (T. thermophila) to Fe2O3 or TiO2 NPs on the protozoan's subsequent uptake of 55Fe-labeled Fe2O3 (55Fe2O3) NPs. Molecular mechanisms underlying the pre-exposure effects were explored in transcriptomic and metabolomic experiments. Pre-exposure to either NPs inhibited the subsequent uptake of 55Fe2O3 NPs. The results of the transcriptomic experiment indicated that NP pre-exposure influenced the expression of genes related to phagosomes and lysosomes and physiological processes such as glutathione and lipid metabolism, which are closely associated with the endocytosis of 55Fe2O3 NPs. The differentially expressed metabolites obtained from the metabolomic experiments showed an enrichment of energy metabolism and antioxidation pathways in T. thermophila pre-exposed to NPs. Together, these results demonstrate that the pre-exposure of T. thermophila to Fe2O3 or TiO2 NPs inhibited the protozoan's subsequent uptake of 55Fe2O3 NPs, possibly by mechanisms involving the alteration of endocytosis-related organelles, the induction of oxidative stress, and a lowering of the intracellular energy supply. Thus, NP pre-exposure represents a scenario which can inform increasingly realistic estimates of NP bioaccumulation.

Development of EST-SSR markers based on transcriptome sequencing for germplasm evaluation of 65 lilies (Lilium).

BACKGROUND: Lilium genus consists of approximately 100 species and numerous varieties, many of which are interspecific hybrids, which result in a complicated genetic background. The germplasm identification, genetic relationship analysis, and breeding of Lilium rely on exploiting genetic information among different accessions. Hence, an attempt was made to develop new EST-SSR markers and study the molecular divergence among 65 genotypes of Lilium. METHODS AND RESULTS: A total of 5509 EST-SSRs were identified from the high-throughput sequencing database of L. 'Elodie'. After primer screening, six primer pairs with the most abundant polymorphic bands were selected from 100 primer pairs. Combined with the other 10 reported SSR primers, a total of 16 pairs detected genetic information with an average PIC value of 0.7583. The number of alleles per locus varied from four to 33, the expected heterozygosity varied from 0.3289 to 0.9231, and the observed heterozygosity varied from 0.2857 to 0.5122. Based on the phylogenic results, 22 Asiatic hybrids (A), seven Longiflorum × Asiatic hybrids (LA), as well as two native species were grouped. Eighteen Oriental hybrids (O) and nine Oriental × Trumpet (OT) hybrids, four native species, one LO, and one LL (L. pardalinum × L. longiflorum) variety were grouped. CONCLUSIONS: Two major clusters were reported and a large number of genotypes were grouped based on UPGMA and STRUCTURE analysis methods. The PIC value as well as other parameters revealed that the EST-SSR markers selected were informative. In addition, the clustering pattern displayed better agreement with the cultivar's pedigree. The newly identified SSRs in this study provides molecular markers for germplasm characterization and genetic diversity for Lilium.

The ppk-expressing transgenic rice floating bed improves P removal in slightly polluted water.

With significant economic advantages, the plant floating bed has been widely utilized in the ecological remediation of eutrophic water because of the excessive phosphorus (P) and nitrogen discharge in China. Previous research has demonstrated that polyphosphate kinase (ppk)-expressing transgenic rice (Oryza sativa L. ssp. japonica) (ETR) can increase the P absorption capacity to support rice growth and boost rice yield. In this study, the floating beds of ETR with single copy line (ETRS) and double copy line (ETRD) are built to investigate their capacity to remove aqueous P in slightly polluted water. Compared with the wild type Nipponbare (WT) floating bed, the ETR floating beds greatly reduce the total P concentration in slightly polluted water though the ETR floating beds have the same removal rates of chlorophyll-a, NO3--N, and total nitrogen in slightly polluted water. The P uptake rate of ETRD on the floating bed is 72.37% in slightly polluted water, which is higher than that of ETRS and WT on the floating beds. Polyphosphate (polyP) synthesis is a critical factor for the excessive phosphate uptake of ETR on the floating beds. The synthesis of polyP decreases the level of free intracellular phosphate (Pi) in ETR on the floating beds, simulating the phosphate starvation signaling. The OsPHR2 expression in the shoot and root of ETR on the floating bed increased, and the corresponding P metabolism gene expression in ETR was changed, which promoted Pi uptake by ETR in slightly polluted water. The Pi accumulation further promoted the growth of ETR on the floating beds. These findings highlight that the ETR floating beds, especially ETRD floating bed, have significant potential for P removal and can be exploited as a novel method for phytoremediation in slightly polluted water.

Molecular and Metabolic Insights into Anthocyanin Biosynthesis for Spot Formation on Lilium leichtlinii var. maximowiczii Flower Petals.

Plants exhibit remarkable diversity in their petal colors through biosynthesis and the accumulation of various pigments. Lilium, an important cut and potted flower, has many coloring pattern variations, including bicolors and spots. To elucidate the mechanisms regulating spot formation in Lilium leichtlinii var. maximowiczii petals, we used multiple approaches to investigate the changes in petal carotenoids, spot anthocyanins, and gene expression dynamics. This included green petals without spots (D1-Pe and D1-Sp), yellow-green petals with purple spots (D2-Pe and D2-Sp), light-orange petals with dark-purple spots (D3-Pe and D3-Sp), and orange petals with dark-purple spots (D4-Pe and D4-Sp). D3-Pe and D4-Pe contained large amounts of capsanthin and capsorubin and small amounts of zeaxanthin and violaxanthin, which contributed to the orange color. In addition to cyanidin-3-O-glucoside, pelargonidin-3-O-rutinoside, cyanidin-3-O-rutinoside, and peonidin-3-O-rutinoside may also contribute to L. leichtlinii var. maximowiczii's petal spot colors. KEGs involved in flavonoid biosyntheses, such as CHS, DFR, and MYB12, were significantly upregulated in D2-Sp and D3-Sp, compared with D1-Sp, as well as in spots, compared with petals. Upregulated anthocyanin concentrations and biosynthesis-related genes promoted spot formation and color transition. Our results provide global insight into pigment accumulation and the regulatory mechanisms underlying spot formation during flower development in L. leichtlinii var. maximowiczii.

SWATH-MS-Based Proteomics Reveals the Regulatory Metabolism of Amaryllidaceae Alkaloids in Three Lycoris Species.

Alkaloids are a class of nitrogen-containing alkaline organic compounds found in nature, with significant biological activity, and are also important active ingredients in Chinese herbal medicine. Amaryllidaceae plants are rich in alkaloids, among which galanthamine, lycorine, and lycoramine are representative. Since the difficulty and high cost of synthesizing alkaloids have been the major obstacles in industrial production, particularly the molecular mechanism underlying alkaloid biosynthesis is largely unknown. Here, we determined the alkaloid content in Lycoris longituba, Lycoris incarnata, and Lycoris sprengeri, and performed a SWATH-MS (sequential window acquisition of all theoretical mass spectra)-based quantitative approach to detect proteome changes in the three Lycoris. A total of 2193 proteins were quantified, of which 720 proteins showed a difference in abundance between Ll and Ls, and 463 proteins showed a difference in abundance between Li and Ls. KEGG enrichment analysis revealed that differentially expressed proteins are distributed in specific biological processes including amino acid metabolism, starch, and sucrose metabolism, implicating a supportive role for Amaryllidaceae alkaloids metabolism in Lycoris. Furthermore, several key genes collectively known as OMT and NMT were identified, which are probably responsible for galanthamine biosynthesis. Interestingly, RNA processing-related proteins were also abundantly detected in alkaloid-rich Ll, suggesting that posttranscriptional regulation such as alternative splicing may contribute to the biosynthesis of Amaryllidaceae alkaloids. Taken together, our SWATH-MS-based proteomic investigation may reveal the differences in alkaloid contents at the protein levels, providing a comprehensive proteome reference for the regulatory metabolism of Amaryllidaceae alkaloids.

Comprehensive Exposure Studies of Per- and Polyfluoroalkyl Substances in the General Population: Target, Nontarget Screening, and Toxicity Prediction.

Novel per- and polyfluoroalkyl substances (PFASs) in the environment and populations have received extensive attention; however, their distribution and potential toxic effects in the general population remain unclear. Here, a comprehensive study on PFAS screening was carried out in serum samples of 202 individuals from the general population in four cities in China. A total of 165 suspected PFASs were identified using target and nontarget analysis, including seven identified PFAS homolog series, of which 16 PFASs were validated against standards, and seven PFASs [4:2 chlorinated polyfluorinated ether sulfonate (4:2 Cl-PFESA), 7:2 chlorinated polyfluorinated ether sulfonate (7:2 Cl-PFESA), hydrosubstituted perfluoroheptanoate (H-PFHpA), chlorine-substituted perfluorooctanoate (Cl-PFOA), chlorine-substituted perfluorononanate (Cl-PFNA), chlorine-substituted perfluorodecanoate (Cl-PFDA), and perfluorodecanedioic acid (PFLDCA n = 8)] were reported for the first time in human serum. The Tox21-GCN model (a graph convolutional neural network model based on the Tox21 database) was established to predict the toxicity of the discovered PFASs, revealing that PFASs containing sulfonic acid groups exhibited multiple potential toxic effects, such as estrogenic effects and stress responses. Our study indicated that the general population was exposed to various PFASs, and the toxicity prediction results of individual PFASs suggested potential health risks that could not be ignored.

Novel simulation of aqueous total nitrogen and phosphorus concentrations in Taihu Lake with machine learning.

This study demonstrates the utility of internal nutrient loads as an additional parameter to improve the performance of machine learning models in predicting the temporal variations of aqueous TN and TP concentrations in Taihu Lake, a large shallow lake. Internal loads, as a potential input parameter for machine learning models, were estimated using a mass balance calculation. The results showed that between 2011 and 2018 the maximum monthly internal loads of nitrogen and phosphorus in Taihu Lake were 4200 t and 178 t, respectively. Monthly changes in the aqueous TN and TP concentrations of Taihu Lake did not correlate significantly with inflow loads whereas the correlations with estimated internal loads were positive and significant. Long short-term memory (LSTM), random forest (RF), and gradient boosting regression tree (GBRT) models were built, and for all of them the inclusion of internal loads in the input parameters improved their performance. LSTM model III, whose input parameters included both inflow loads and internal loads, had the best performance, based on a testing root mean square error of 0.11 mg TN/L and 0.017 mg TP/L. A 28 % decrease in the annual aqueous TP concentration in Taihu Lake in 2018 simulated by LSTM model III was achieved by lowering the average water level from 3.29 m to 2.99 m, suggesting a possible strategy to control the TP concentration in the lake. In summary, our study showed that aqueous TN and TP concentrations in shallow lakes can be simulated using machine learning, with LSTM models outperforming RF and GBRT models; in these models, internal loads should be included as an input parameter. Additionally, our study identified the water level as an important factor affecting the aqueous TP concentration in Taihu Lake.

Changes in carbohydrate metabolism and endogenous hormone regulation during bulblet initiation and development in Lycoris radiata.

BACKGROUND: Lycoris species have great ornamental and medicinal values; however, their low regeneration efficiency seriously restricts their commercial production. Understanding the mechanism of bulblet propagation in this genus, which has remained underexplored to date, could provide a theoretical basis for improving the reproductive efficiency. Therefore, we studied the bulblet initiation and developmental processes in Lycoris radiata. RESULTS: We found that bulblets are formed on the junctions of the innermost layers of scales and the basal plate, and initially present as an axillary bud and gradually develop into a bulblet. We also determined the changes in carbohydrate and endogenous hormone contents during bulblet initiation and development, as well as the expression patterns of genes involved in carbohydrate metabolism and hormone biosynthesis and signaling through transcriptome analysis. Soluble sugars derived from starch degradation in the outer scales are transported to and promote bulblet initiation and development through starch synthesis in the inner scales. This process is mediated by several genes involved in carbohydrate metabolism, especially genes encoding ADP glucose pyrophosphorylase, a crucial starch synthesis enzyme. As for hormones, endogenous IAA, GA, and ABA content showed an increase and decrease during bulblet initiation and development, respectively, which were consistent with the expression patterns of genes involved in IAA, GA, and ABA synthesis and signal transduction. In addition, a decrease in ZR content may be down- and up-regulated by CK biosynthesis and degradation related genes, respectively, with increasing auxin content. Furthermore, expression levels of genes related to BR, JA, and SA biosynthesis were increased, while that of ethylene biosynthesis genes was decreased, which was also consistent with the expression patterns of their signal transduction genes. CONCLUSIONS: The present study provides insights into the effect of carbohydrate metabolism and endogenous hormone regulation on control of L. radiata bulblet initiation and development. Based on the results, we propose several suggestions to improve L. radiata propagation efficiency in production, which will provide directions for future research.

Unbound Natural Organic Matter Competes with Nanoparticles for Internalization Receptors During Cell Uptake.

Natural organic matter (NOM) that forms coronas on the surface of engineered nanoparticles (NPs) affects their stability, bio-uptake, and toxicity. After corona formation, a large amount of unbound NOM remains in the environment and their effects on organismal uptake of NPs remain unknown. Here, the effects of unbound NOM on the uptake of polyacrylate-coated hematite NPs (HemNPs) by the protozoan Tetrahymena thermophila were examined. HemNPs were well-dispersed without any detectable NOM adsorption. Kinetics experiments showed that unbound NOM decreased the uptake of HemNPs with greater inhibition at lower concentrations of the particles in the presence of NOM of higher molecular weight. The unbound NOM suppressed clathrin-mediated endocytosis but not the phagocytosis of HemNPs. Confirmation of these events was obtained using label-free hyperspectral stimulated Raman spectroscopy imaging and dissipative particle dynamics simulation. Overall, the present study demonstrates that unbound NOM can compete with HemNPs for internalization receptors on the surface of T. thermophila and inhibit particle uptake, highlighting the need to consider the direct effects of unbound NOM in bioapplication studies and in safety evaluations of NPs.

Combined Toxicity of Silver Nanoparticles with Hematite or Plastic Nanoparticles toward Two Freshwater Algae.

In the natural environment, the interactions of different types of nanoparticles (NPs) may alter their toxicity, thus masking their true environmental effects. This study investigated the toxicity of silver NPs (AgNPs) combined with hematite (HemNPs) or polystyrene (PsNPs) NPs toward the freshwater algae Chlamydomonas reinhardtii and Ochromonas danica. The former has a cell wall and cannot internalize these NPs, while the latter without a cell wall can. Therefore, the toxicity of AgNPs toward C. reinhardtii was attributed to the released Ag ions, while AgNPs had direct toxic effects on O. danica. Moreover, nontoxic HemNPs ameliorated AgNP toxicity toward C. reinhardtii, by decreasing the bioavailability of Ag ions through adsorption. Despite their role as Ag-ion carriers, HemNPs still reduced the toxicity of AgNPs toward O. danica by competitively inhibiting AgNP uptake. In both algae, Ag accumulation fully accounted for the combined toxicity of AgNPs and HemNPs. However, the combined toxicity of AgNPs and PsNPs was complicated by their significant individual toxicities and the synergistic interactions of these particles with the algae, regardless of differences in Ag accumulation. Overall, in environmental assessments, considerations of the combined toxicity of dissimilar NPs will allow more accurate assessments of their environmental risks.

Algal Foods Reduce the Uptake of Hematite Nanoparticles by Downregulating Water Filtration in Daphnia magna.

Rapid developments in nanotechnology have led to the release of substantial amounts of nanoparticles (NPs) into aquatic environments, where many types of biotic particles are present and could potentially interact with the NPs. Nevertheless, how biotic particles may affect the bioaccumulation and toxicity of NPs remains largely unknown. In the present study, we investigated the effects of the green alga Chlamydomonas reinhardtii on the accumulation kinetics (uptake, assimilation, efflux) and toxicity of polyacrylate-coated hematite NPs (HemNPs), using Daphnia magna as the test organism. As a biotic particle and daphnid food, C. reinhardtii reduced the accumulation and toxicity of HemNPs in D. magna. The HemNPs were well-dispersed with little adsorption to the alga. Their decreased accumulation could thus be partly explained by their low trophic transfer from the alga to the daphnid and by the inductive effects of the alga on HemNP efflux. However, the main cause was the direct inhibition of HemNP uptake from the water phase as a result of the reduced water-filtration activity of D. magna in the presence of C. reinhardtii. Overall, in bioaccumulation studies, the inhibitory effects of biotic particles on NP uptake from the water phase should be paid attention.

Label-Free Imaging of Nanoparticle Uptake Competition in Single Cells by Hyperspectral Stimulated Raman Scattering.

Imaging and quantification of nanoparticles in single cells in their most natural condition are expected to facilitate the biotechnological applications of nanoparticles and allow for better assessment of their biosafety risks. However, current imaging modalities either require tedious sample preparation or only apply to nanoparticles with specific physicochemical characteristics. Here, the emerging hyperspectral stimulated Raman scattering (SRS) microscopy, as a label-free and nondestructive imaging method, is used for the first time to investigate the subcellular distribution of nanoparticles in the protozoan Tetrahymena thermophila. The two frequently studied nanoparticles, polyacrylate-coated α-Fe2 O3 and TiO2 , are found to have different subcellular distribution pattern as a result of their dissimilar uptake routes. Significant uptake competition between these two types of nanoparticles is further discovered, which should be paid attention to in future bioapplications of nanoparticles. Overall, this study illustrates the great promise of hyperspectral SRS as an analytical imaging tool in nanobiotechnology and nanotoxicology.

Waterborne and Dietborne Toxicity of Inorganic Arsenic to the Freshwater Zooplankton Daphnia magna.

Waterborne and dietborne exposure are both important sources for the accumulation of inorganic arsenic (iAs) in aquatic organisms. Although the waterborne toxicity of iAs has been extensively investigated, its dietborne toxicity has received little attention. The present study examined the acute and chronic toxicity of arsenate (iAsV) and arsenite (iAsIII) to the freshwater zooplankton species Daphnia magna under both waterborne and dietborne exposure scenarios. The bioaccumulation, speciation, and tissue and subcellular distributions of arsenic were analyzed to understand the mechanisms accounting for differences in toxicity related to different arsenic species, exposure scenarios, and exposure duration. The toxicity of iAs increased with exposure time, and iAsIII was more toxic than iAsV. Moreover, although dietborne iAs had no acute effect on D. magna, it incurred significant toxicity in the chronic-exposure experiment. Nevertheless, the toxicity of dietborne iAs was still lower than that of waterborne iAs regardless of the exposure duration. This difference was found to be caused by the lower bioaccumulation of dietborne iAs, its higher distribution in the gut and in the biologically detoxified subcellular fraction, and greater transformation to the less toxic dimethylarsinic acid. Overall, the dietborne toxicity of iAs should be considered when evaluating the environmental risks posed by arsenic.