Graphene Quantum Dots Nonmonotonically Influence the Horizontal Transfer of Extracellular Antibiotic Resistance Genes via Bacterial Transformation.

Graphene quantum dots (GQDs) coexist with antibiotic resistance genes (ARGs) in the environment. Whether GQDs influence ARG spread needs investigation, since the resulting development of multidrug-resistant pathogens would threaten human health. This study investigates the effect of GQDs on the horizontal transfer of extracellular ARGs (i.e., transformation, a pivotal way that ARGs spread) mediated by plasmids into competent Escherichia coli cells. GQDs enhance ARG transfer at lower concentrations, which are close to their environmental residual concentrations. However, with further increases in concentration (closer to working concentrations needed for wastewater remediation), the effects of enhancement weaken or even become inhibitory. At lower concentrations, GQDs promote the gene expression related to pore-forming outer membrane proteins and the generation of intracellular reactive oxygen species, thus inducing pore formation and enhancing membrane permeability. GQDs may also act as carriers to transport ARGs into cells. These factors result in enhanced ARG transfer. At higher concentrations, GQD aggregation occurs, and aggregates attach to the cell surface, reducing the effective contact area of recipients for external plasmids. GQDs also form large agglomerates with plasmids and thus hindering ARG entrance. This study could promote the understanding of the GQD-caused ecological risks and benefit their safe application.

Estrogens and xenoestrogen residues in manure-based fertilizers and their potential ecological risks.

Optimal manure treatment aimed at usage as agricultural soil fertilizers is a prerequisite ecological pollution control strategy. In this work, livestock manure-based fertilizers were collected from 71 animal farms across 14 provinces in China. The contamination levels and potential ecotoxicological risks of residual steroid estrogens (SEs): estrone (E1), estriol (E3), 17α-estradiol (17α-E2), 17ß-estradiol (17ß-E2) and xenoestrogen (XE) bisphenol A (BPA), were investigated. The results showed that the occurrence frequencies for SEs and XE ranged from 66.67% to 100%, and the mean concentration varied considerably across the study locations. The total content of SEs and XE in Hebei province was the highest, and swine manure-based fertilizers concentrations were higher than the levels reported in other animal fertilizers. Compared with farm level manure, manure-based fertilizers are processed by composting, and the micropollutants quantities are significantly reduced (mean: 87.65 - 534.02 µg/kg). The total estradiol equivalent quantity (EEQ) that might migrate to the soil was estimated to be 1.23 µg/kg. Based on the estimated application rate of manure, 38% of the fertilizers risk quotients exceeded 0.1, indicating medium to high risks pressure on terrestrial organisms. Nonetheless, the estrogenic risk was lower in manure-based fertilizers than in manure. This study highlights the significance of proper treatment of livestock manure and designing an optimal manure fertilization strategy to mitigate the risks posed by SEs and XEs to the agroecosystems.

Identification of novel catabolic genes involved in 17ß-estradiol degradation by Novosphingobium sp. ES2-1.

Novosphingobium sp. ES2-1 is an efficient 17ß-estradiol (E2)-degrading bacterium, which can convert E2 to estrone (E1), then to 4-hydroxyestrone (4-OH-E1) for subsequent oxidative cracking. In this study, the molecular bases for this process were elucidated. Two novel monooxygenase systems EstP and EstO were shown to catalyse the oxygenation of E1 and 4-OH-E1, respectively. EstP was a three-component cytochrome P450 monooxygenase system consisting of EstP1 (P450 monooxygenase), EstP2 (ferredoxin) and EstP3 (ferredoxin reductase). Ultraperformance liquid chromatography-high resolution mass spectrometry (UPLC-HRMS) analysis revealed that EstP catalysed the 4-hydroxylation of E1 to produce 4-OH-E1. The resultant 4-OH-E1 was further oxidized by a two-component monooxygenase system EstO consisting of EstO1 (flavin-dependent monooxygenases) and EstO2 (flavin reductase). UPLC-HRMS combined with 1 H-nuclear magnetic resonance analysis demonstrated that EstO catalysed the breakage of C9-C10 to yield a ring B-cleavage product. In addition, the oxygenase component genes estP1 and estO1 exhibited contrary inductive behaviours when exposed to different steroids, suggesting that EstP1-mediated 4-hydroxylation was E2-specific, whereas EstO1-mediated monooxygenation might be involved in the degradation of testosterone, androstenedione, progesterone and pregnenolone. This also implied that the mechanisms of the catabolism of different steroids by the same microorganism might be partially interlinked.

Endophytic Bacteria in in planta Organopollutant Detoxification in Crops.

Microbe-assisted organopollutant removal, or in planta crop decontamination, is based on an interactive system between organopollutant-degrading endophytic bacteria (DEBOP) and crops in alleviating organic toxins in plants. This script focuses on the fast-growing body of literature that has recently bloomed in organopollutant control in agricultural plants. The various facets of DEBOP under study include their colonization, distribution, plant growth-promoting mechanisms, and modes of action in the detoxification process in plants. Also, an assessment of the biotechnological advances, advantages, and bottlenecks in accelerating the implementation of this decontamination strategy will be undertaken. The highlighted key research directions from this review will shape the future of agro-environmental sustainability and preservation of human health.

Inoculation of a phenanthrene-degrading endophytic bacterium reduces the phenanthrene level and alters the bacterial community structure in wheat.

Colonization by polycyclic aromatic hydrocarbon (PAH)-degrading endophytic bacteria (PAHDEB) can reduce the PAH contamination risk in plant. However, little information is available on the impact of PAHDEB colonization on the endophytic bacterial community of inner plant tissues. A phenanthrene-degrading endophytic bacterium (PDEB), Massilia sp. Pn2, was inoculated onto the roots of wheat and subjected to greenhouse container experiments. The endophytic bacterial community structure in wheat was investigated using high-throughput sequencing technology. The majority of endophytic bacteria in wheat were Proteobacteria, and the dominant genus was Pseudomonas. Phenanthrene contamination clearly increased the diversity of endophytic bacteria in wheat. The cultivable endophytic bacteria counts in wheat decreased with increasing the level of phenanthrene contamination; the endophytic bacterial community structure changed correspondingly, and the bacterial richness first increased and then decreased. Inoculation of strain Pn2 reduced the phenanthrene contamination in wheat, enlarged the biomass of wheat roots, changed the bacterial community structure and enhanced the cell counts, diversity and richness of endophytic bacteria in phenanthrene-contaminated wheat in a contamination level-dependent manner. The findings of this investigation provide insight into the responses of endophytic bacterial community in plant to external PAH contamination and PAHDEB colonization.

Inoculating wheat (Triticum aestivum L.) with the endophytic bacterium Serratia sp. PW7 to reduce pyrene contamination.

This research was conducted to find an optimal inoculation way for a pyrene-degrading endophytic Serratia sp. PW7 to colonize wheat for reducing pyrene contamination. Three inoculation ways, which are soaking seeds in inocula (TS), dipping roots of seedlings in inocula (TR), and spraying inocula on leaves of seedlings (TL), were used in this study. Inoculated seedlings and noninoculated seedlings (CK) were, respectively, cultivated in Hoagland solutions supplemented with pyrene in a growth chamber. The results showed that strain PW7 successfully colonized the inoculated seedlings in high numbers, and significantly promoted the growth of seedlings (TS and TR). More importantly, strain PW7 reduced pyrene levels in the seedlings and the Hoagland solutions. Compared to the noninoculated seedlings, the pyrene contents of the inoculated seedlings were decreased by 35.7-86.3% in the shoots and by 26.8-60.1% in the roots after 8-day cultivation. By comparing the efficiencies of decreasing pyrene residues, it can be concluded that TR was an optimal inoculation way for endophytic strains to colonize the inoculated plants and to reduce the pyrene contamination. Our findings provide an optimized inoculation way to reduce organic contamination in crops by inoculating plants with functional endophytic bacteria.

Micro-interfacial behavior of antibiotic-resistant bacteria and antibiotic resistance genes in the soil environment: A review.

Overutilization and misuse of antibiotics in recent decades markedly intensified the rapid proliferation and diffusion of antibiotic resistance genes (ARGs) within the environment, thereby elevating ARGs to the status of a global public health crisis. Recognizing that soil acts as a critical reservoir for ARGs, environmental researchers have made great progress in exploring the sources, distribution, and spread of ARGs in soil. However, the microscopic state and micro-interfacial behavior of ARGs in soil remains inadequately understood. In this study, we reviewed the micro-interfacial behaviors of antibiotic-resistant bacteria (ARB) in soil and porous media, predominantly including migration-deposition, adsorption, and biofilm formation. Meanwhile, adsorption, proliferation, and degradation were identified as the primary micro-interfacial behaviors of ARGs in the soil, with component of soil serving as significant determinant. Our work contributes to the further comprehension of the microstates and processes of ARB and ARGs in the soil environments and offers a theoretical foundation for managing and mitigating the risks associated with ARG contamination.

A novel remediation strategy of mixed calcium peroxide and degrading bacteria for polycyclic aromatic hydrocarbon contaminated water.

Polycyclic aromatic hydrocarbons (PAHs) are a class of toxic organic pollutants commonly detected in the aqueous phase. Traditional biodegradation is inefficient and advanced oxidation technologies are expensive. In the current study, a novel strategy was developed using calcium peroxide (CP) and PAH-degrading bacteria (PDB) to effectively augment PAH degradation by 28.62-59.22%. The PDB consisted of the genera Acinetobacter, Stenotrophomonas, and Comamonas. Applying the response surface model (RSM), the most appropriate parameters were identified, and the predictive degradation rates of phenanthrene (Phe), pyrene (Pyr), and ΣPAHs were 98%, 76%, and 84%, respectively. The constructed mixed system could reduce 90% of Phe and more than 60% of ΣPAHs and will perform better at pH 5-7 and lower salinity. Because PAHs tend to bind to dissolved organic matter (DOM) with larger molecular weights, humic acid (HA) had a larger negative effect on the PAH-degradation efficiency of the CP-PDB mixed system than fulvic acid (FA). The proposed PAH-degradation pathways in the mixed system were based on the detection of intermediates at different times. The investigation constructed and optimized a novel environmental PAH-degradation strategy. The synergistic application of PDB and oxidation was extended for organic contaminant degradation in aqueous environments.

Remediation potential of an immobilized microbial consortium with corn straw as a carrier in polycyclic aromatic hydrocarbons contaminated soil.

Polycyclic aromatic hydrocarbons (PAHs) are widespread in soils and threaten human health seriously. The immobilized microorganisms (IM) technique is an effective and environmentally sound approach for remediating PAH-contaminated soil. However, the knowledge of the remedial efficiency and the way IM operates using natural organic materials as carriers in complex soil environments is limited. In this study, we loaded a functional microbial consortium on corn straw to analyze the effect of IM on PAH concentration and explore the potential remediation mechanisms of IM in PAH-contaminated soil. The findings revealed that the removal rate of total PAHs in the soil was 88.25% with the application of IM after 20 days, which was 39.25% higher than the control treatment, suggesting that IM could more easily degrade PAHs in soil. The findings from high-throughput sequencing and quantitative PCR revealed that the addition of IM altered the bacterial community structure and key components of the bacterial network, enhanced cooperative relationships among bacteria, and increased the abundance of bacteria and functional gene copies such as nidA and nahAc in the soil, ultimately facilitating the degradation of PAHs in the soil. This study enhances our understanding of the potential applications of IM for the treatment of PAH-contaminated soil.

Biodegradation of PAEs in contaminated soil by immobilized bacterial agent and the response of indigenous bacterial community.

Phthalic acid esters (PAEs) are common hazardous organic contaminants in agricultural soil. Microbial remediation is an effective and eco-friendly method for eliminating PAEs. Nevertheless, the operational mode and potential application of immobilized microorganisms in PAEs-contaminated soil are poorly understood. In this study, we prepared an immobilized bacterial agent (IBA) using a cedar biochar carrier to investigate the removal efficiency of PAEs by IBA in the soil. We found that IBA degraded 88.35% of six optimal-control PAEs, with 99.62% biodegradation of low-molecular-weight PAEs (DMP, DEP, and DBP). The findings demonstrated that the IBA achieved high efficiency and a broad-spectrum in degrading PAEs. High-throughput sequencing revealed that IBA application altered the composition of the soil bacterial community, leading to an increase in the relative abundance of PAEs-degrading bacteria (Rhodococcus). Furthermore, co-occurrence network analysis indicated that IBA promoted microbial interactions within the soil community. This study introduces an efficient method for the sustainable remediation of PAEs-contaminated soil.

Dissemination of antibiotic resistance genes is regulated by iron oxides: Insight into the influence on bacterial transformation.

The transportation of antibiotic resistance genes (ARGs) in manure-soil-plant continuums poses risks to human health. Horizontal gene transfer, particularly for bacterial transformation, is an important way for ARG dissemination. As crucial components in soils, iron oxides impacted the fates of various abiotic and biotic contaminants due to their active properties. However, whether they can influence the transformation of ARGs is unknown, which waits to be figured out to boost the assessment and control of ARG spread risks. In this study, we have investigated the effects of goethite, hematite, and magnetite (0-250 mg/L, with sizes < 100 nm and > 100 nm) on the transfer of ampicillin resistance genes to Escherichia coli cells. At lower iron oxide concentrations, the transformation of ARGs was first facilitated (transformation frequency reached up to 3.38-fold higher), but the facilitating effects gradually weakened and eventually disappeared as concentrations further increased. Particle size and iron oxide type were not the universal determinants controlling the transformation. At lower concentrations, iron oxides interacted with proteins and phospholipids in E. coli envelope structures, and induced the overgeneration of intracellular reactive oxygen species. Consequently, they led to pore formation and permeability enhancement on the cell membrane, thus promoting the transformation. The facilitation was also associated with the carrier-like effect of iron oxides for antibiotic resistance plasmids. At higher concentrations, the weakened facilitations were attributed to the aggregation of iron oxides. In this study, we highlight the crucial roles of the concentrations (contents) of iron oxides on the dissemination of ARGs in soils; this study may serve as a reference for ARG pollution control in future agricultural production.

Dissolved organic matter influences the indigenous bacterial community and polycyclic aromatic hydrocarbons biodegradation in soils.

In polycyclic aromatic hydrocarbon (PAH) contaminated soils, bioremediation is superior to other strategies owing to its low cost and environmental friendliness. However, dissolved organic matter (DOM) and indigenous bacterial communities can affect the efficiency of PAH-degrading bacteria (PDB). This study found that exogenous PDB (C1) including the genera Acinetobacter, Stenotrophomonas, and Comamonas, decreased the bacterial diversity of Alfisol, Ultisol, Inceptisol, and Mollisol, and DOM enhanced the diffusion of PDB and the bioavailability of PAH. In addition, bacteria preferred to ingest low molecular weight DOM fractions, and the abundances of lipid-like and protein-like substances decreased by 0.12-3.03 % and 1.73-4.60 %. The DOM fractions had a more marked influence on the indigenous bacteria than the exogenous PDB, and PDB dominated the PAH biodegradation process in the soils. More COO functional groups promoted the utilization of higher molecular weight-related homologue fractions by bacteria, and lower molecular weight fractions carrying more CH2 functional groups declined during biodegradation. This study investigated the variations in bacterial communities during biodegradation and revealed the effects of DOM fractions on biodegradation in PAH-contaminated soils at the molecular level. These results will promote the development of bioremediation strategies for organics-contaminated soil and provide guidance for prediction models of soil biodegradation kinetics.

Research progress and prospect of glomalin-related soil protein in the remediation of slightly contaminated soil.

Soil pollution caused by organic pollutants and potentially toxic elements poses a serious threat to sustainable agricultural development, global food security and human health. Therefore, strategies for reducing soil pollution are urgently required. Arbuscular mycorrhizal fungi (AMF)-assisted phytoremediation is widely recognized for its ability to remediate slightly-contaminated soil. Glomalin-related soil protein (GRSP) production by AMF is considered a vital mechanism of AMF-assisted phytoremediation. GRSP is widespread in soils and may contribute to the remediation of slightly contaminated soils. GRSP facilitates stabilization of pollutants in soils by interacting with pollutants owing to its abundant functional groups, recalcitrance, and long turnover time. It also enhances soil bioremediation and phytoremediation by stimulating soil microbial activity, improving soil structure, and providing nutrients for plants. However, research on GRSP is still in its early stages, and studies on contaminated soil remediation are limited. The effectiveness of GRSP in situ remediation remains to be proved. This review summarizes current knowledge regarding the GRSP distribution and its contribution to the remediation of slightly contaminated soils. Additionally, we present strategies to increase the GRSP content in contaminated soils, as well as prospects for future studies on the use of GRSP in contaminated soil remediation. This study focuses on recent developments that aim to improve awareness of the role of GRSP in soil remediation and relevant future directions.

Potential interaction mechanisms between PAHs and glomalin related-soil protein (GRSP).

Glomalin-related soil protein (GRSP) has gained widespread attention because of its benefits to carbon sequestration, improving soil quality and fixing heavy metals. However, studies on how GRSP affects the environmental fate of organic contaminants are scarce. In this study, different types of GRSPs were isolated from forest soils and characterized to study the binding of GRSPs and PAHs under different environmental conditions. The results indicated that GRSPs contain abundant functional groups (such as -NH, -COOH, and CO) and material composition, like humic acid, proteins, and lipids. For the tested GRSPs, EE-GRSP has lower DOC, SUVA260 and SUVA280 values, as well as higher E2/E3 values, indicating that EE-GRSP has lower hydrophobicity and molecular weight. These properties can lead to strong interactions between GRSP and PAHs, especially with benzopyrene, which has a high Kow and Ksw and a large molecular size, with binding constants ranging from 16,119 to 163,697 L·kg-1. Furthermore, low pH (pH = 3) and temperature (15 °C) could increase GRSP's aggregation, enhance the GRSP binding ability with PAHs, whose binding constants were 11,595 and 5067.3 L·kg-1. Therefore, the binding between GRSP and PAHs may lead to changes in the fate of PAHs in the soil and affect the environmental risk of PAHs. The results presented here will deepen our understanding of the environmental function of GRSPs and provide a theoretical basis to further elucidate the mechanisms of GRSPs and organic pollutants.

Safety of benzophenone-type UV filters: A mini review focusing on carcinogenicity, reproductive and developmental toxicity.

Consumer products containing benzophenone-type ultraviolet (UV) filters (BPs) have been widely accepted by the public, resulting in the widely existence of various BPs in the human body and environment. In recent years, more and more evidences show that BPs are endocrine disruptors. In view of the continuous exposure risk of BPs and their endocrine disrupting characteristics, the carcinogenicity of BPs and their effects on reproduction and development are of particular concern. However, due to the wide varieties of BPs and the scattered toxicity studies on BPs, people have a limited understanding on the safety of BPs. Therefore, this paper systematically reviews the carcinogenicity, reproductive and developmental toxicity of BPs in order to expand people's knowledge on the health risks of BPs and screen for more safe BPs. Although existing toxicological studies are insufficient, it can be determined that BPs have different potentials for carcinogenicity, and reproductive and developmental toxicity. Among those BPs, 2-hydroxyl-4-methoxyl benzophenone needs to be used with caution due to its adverse effects on cancer cell proliferation and migration, reproductive ability, sex differentiation, neurodevelopment, and so on. It is worth noting that functional group substitutions significantly affect the interaction between BPs and biomolecules such as DNA, cancer cells, and seminal fluid, resulting in different levels of toxicity. In short, it is very necessary to evaluate the carcinogenicity, reproductive and developmental toxicity of BPs, which is of great significance for establishing reasonable BPs content standards in cosmetics, water quality treatment standards for BPs, and so on.

Variations of different PAH fractions and bacterial communities during the biological self-purification in the soil vertical profile.

Wild PAH-contaminated sites struggle to provide continuous and stable monitoring, resulting in the potential risks of contaminated soil utilization could not be evaluated effectively. This work provided a 9-months laboratory simulation which was close to the natural ecological process. These results believed that PAH-degrading bacteria (PDB) preferred to degrade organic extracted PAH (PAH_OS) and fresh bound-PAH (79.36-99.97%). The formation and migration efficiency of PAH binding with HA humic acid (HA) (PAH_HA) was lower than that of PAH binding with fulvic acid (FA) and humin (HM) (PAH_FA and PAH_HM), leading to PAH_HA had more persistent retention and influenced bacterial communities in shallow soils. Besides, phylum Proteobacteria gradually dominated the bacterial community and decreased 12.05-20.48% diversity at all depths during the biological self-purification process. Although the effect of this process enhanced the abundance of 28 genes 16 s rRNA and three PAH-degrading genes (PDGs) by 5.91-2047.34 times (phe, nahAc and nidA), the top 30 genera maintained their ecological characteristics. This study provided insights into the important influencing factor and mechanism of the biological self-purification processes and discerned the linkages between bacterial communities and environmental variables in the vertical profile, which is important to the isolation and application of PDB and ecological risk assessment.

Distribution of bound-PAH residues and their correlations with the bacterial community at different depths of soil from an abandoned chemical plant site.

The situ pollutant residue and microbial characteristics in contaminated environments are crucial for ecological restoration and soil utilization. This work reported the variation of polycyclic aromatic hydrocarbon (PAH) residues and the bacterial community at different depths in an aged-abandoned site. These results unveiled that over 90% of low molecular weight (LMW) and medium molecular weight (MMW), 52.84-76.88% of high molecular weight (HMW) bound-PAH (BP) residues were sequestrated in humin (HM). The stresses of PAH and soil depth enhanced the frequency of bacteria associations, especially positive associations. We enriched and cultured PAH degradation bacteria (PDB) from the sampling site mainly consisting of Pseudomonas and Acinetobacter, which were originally 0.39-0.52% abundant in the sampling site. The abundances of PDB and PAH-degradation genes (PDGs) were higher at shallower depths and increased with high PAH concentration. Simultaneously, Pearson correlation analysis and experimental verification found that the process of PAH binding with SOM limited the further increase of PDB and PDGs in PAH-contaminated sites. These findings may illustrate possible ecological risks of contaminated soils and provide guidance for the isolation and application of PDB.

Environmental health risks induced by interaction between phthalic acid esters (PAEs) and biological macromolecules: A review.

As a kind of compounds abused in industry productions, phthalic acid esters (PAEs) cause serious problems in natural environment. PAEs pollution has penetrated into environmental media and human food chain. This review consolidates the updated information to assess the occurrence and distribution of PAEs in each transmission section. It is found that micrograms per kilogram of PAEs are exposed to humans through daily diets. After entering the human body, PAEs often undergo the metabolic process of hydrolysis to monoesters phthalates and conjugation process. Unfortunately, in the process of systemic circulation, PAEs will interact with biological macromolecules in vivo under the action of non-covalent binding, which is also the essence of biological toxicity. The interactions usually operate in the following pathways: (a) competitive binding; (b) functional interference; and (c) abnormal signal transduction. While the non-covalent binding forces mainly contain hydrophobic interaction, hydrogen bond, electrostatic interaction, and π interaction. As a typical endocrine disruptor, the health risks of PAEs often start with endocrine disorder, further leading to metabolic disruption, reproductive disorders, and nerve injury. Besides, genotoxicity and carcinogenicity are also attributed to the interaction between PAEs and genetic materials. This review also pointed out that the molecular mechanism study on biological toxicity of PAEs are deficient. Future toxicological research should pay more attention to the intermolecular interactions. This will be beneficial for evaluating and predicting the biological toxicity of pollutants at molecular scale.

Genotoxicity of organic contaminants in the soil: A review based on bibliometric analysis and methodological progress.

Organic contaminants (OCs) are ubiquitous in the environment, posing severe threats to human health and ecological balance. In particular, OCs and their metabolites could interact with genetic materials to induce genotoxicity, which has attracted considerable attention. In this review, bibliometric analysis was executed to analyze the publications on the genotoxicity of OCs in soil from 1992 to 2021. The result indicated that significant contributions were made by China and the United States in this field and the research hotspots were biological risks, damage mechanisms, and testing methods. Based on this, in this review, we summarized the manifestations and influencing factors of genotoxicity of OCs to soil organisms, the main damage mechanisms, and the most commonly utilized testing methods. OCs can induce genotoxicity and the hierarchical response of soil organisms, which could be influenced by the physicochemical properties of OCs and the properties of soil. Specific mechanisms of genotoxicity can be classified into DNA damage, epigenetic toxicity, and chromosomal aberrations. OCs with different molecular weights lead to genetic material damage by inducing the generation of ROS or forming adducts with DNA, respectively. The micronucleus test and the comet test are the most commonly used testing methods. Moreover, this review also pointed out that future studies should focus on the relationships between bioaccessibilities and genotoxicities, transcriptional regulatory factors, and potential metabolites of OCs to elaborate on the biological risks and mechanisms of genotoxicity from an overall perspective.

Polyphenol oxidase activity in subcellular fractions of tall fescue contaminated by polycyclic aromatic hydrocarbons.

Understanding enzyme responses to contamination with persistent organic pollutants (POPs) is a key step in the elucidation of POP metabolic mechanisms in plants. However, there is little information available on enzyme activity in subcellular fractions of POP-contaminated plants. To our knowledge, this is the first study to investigate the activities of polyphenol oxidase (PPO) in cell fractions of plants under contamination stress from polycyclic aromatic hydrocarbons (PAHs) using a greenhouse batch technique. Three parameters, E(cell), E(cell-n), and P(cell), denoting the amount of PPO activity, cell fraction content-normalized PPO activity, and proportion of PPO activity in each cell fraction, respectively, were used in this study. Contamination with phenanthrene, as a representative PAH, at a relatively high level (>0.23 mg L⁻¹) in culture solution generally stimulated PPO activity in tall fescue (Festuca arundinacea Schreb.) roots and shoots and their cellular fractions. The amount and distribution proportion of PPO activity in each cell fraction of phenanthrene-contaminated roots and shoots were (in descending order): cell solution > > cell wall > cell organelles. Cell solution was the dominant storage domain of PPO activity and contributed 84.0 and 82.8% of PPO activity in roots and shoots, respectively. The cell wall had the highest density of PPO activity in roots and shoots, based on the highest cell fraction content normalized PPO activity in this cell fraction. Our results provide new information on enzyme responses in plant intracellular fractions to xenobiotic POPs and fundamental information on within-plant POP metabolic mechanisms.