Transport proteins and their differential roles in the accumulation of phenanthrene in wheat.

The study focuses on the uptake, accumulation, and translocation of polycyclic aromatic hydrocarbons (PAHs) in cereals, specifically exploring the role of peroxidase (UniProt accession: A0A3B5XXD0, abbreviation: PX1) and unidentified protein (UniProt accession: A0A3B6LUC6, abbreviation: UP1) in phenanthrene solubilization within wheat xylem sap. This research aims to clarify the interactions between these proteins and phenanthrene. Employing both in vitro and in vivo analyses, we evaluated the solubilization capabilities of recombinant transport proteins for phenanthrene and examined the relationship between protein expression and phenanthrene concentration. UP1 displayed greater transport efficiency, while PX1 excelled at lower concentrations. Elevated PX1 levels contributed to phenanthrene degradation, marginally diminishing its transport. Spectral analyses and molecular dynamics simulations validated the formation of stable protein-phenanthrene complexes. The study offers crucial insights into PAH-related health risks in crops by elucidating the mechanisms of PAH accumulation facilitated by transport proteins.

Photoaging process and mechanism of four commonly commercial microplastics.

Microplastics (MPs) are the widespread emerging pollutants in the terrestrial systems, and photo-oxidation is an effective process for aging MPs on land. Here, four common commercial MPs were exposed to ultraviolet (UV) light to simulate the photo-aging of MPs on soil, and the changes in surface properties and eluates of photoaging MPs were studied. Results revealed that polyvinyl chloride (PVC) and polystyrene (PS) exhibited more pronounced physicochemical changes than polypropylene (PP) and polyethylene (PE) during photoaging on the simulated topsoil, due to the dechlorination of PVC and the debenzene ring of PS. Oxygenated groups accumulated in aged MPs were strongly correlated with dissolved organic matters (DOMs) leaching. Through analysis of the eluate, we found that photoaging altered the molecular weight and aromaticity of DOMs. PS-DOMs showed the greatest increase in humic-like substances after aging, whereas PVC-DOMs exhibited the highest amount of additive leaching. The chemical properties of additives explained their differences in photodegradation responses, which also accounted for the greater importance of chemical structure of MPs to their structural stability. These findings demonstrate that the extensive presence of cracks in aged MPs facilitates DOMs formation and the complexity of DOMs composition poses a potential threat to soil and groundwater safety.

New mechanistic insights into PAHs transport across wheat root cell membrane: Evidence for ABC transporter mediation.

Polycyclic aromatic hydrocarbons (PAHs) are a class of highly carcinogenic organic pollutants. Our previous results revealed that the active uptake of PAHs by plant roots is performed through H+/PAHs co-transport. However, the proteins and mechanisms of co-transport of PAHs remain unknown. We hypothesized that ABC transporters are involved in PAHs co-transport via the roots. We found a total of 47 ABC transporters with alkalinity and hydrophobicity which were up-regulated or newly expressed in the wheat roots after phenanthrene exposure. And the concentration of ABC transporters rose. There was a positive relationship between the concentration of phenanthrene and ABC transporter expression in the wheat roots. Additionally, the trend observed in the ABC transporters expression was also found in the gene expression. With energies below -6 kcal mol-1, a stable docking conformation formed between ABC transporters and PAHs. π-π stacking and van der Waals force bound PAHs to ABCB or ABCG. The binding strength of ABCB subfamily proteins with homodimers is stronger than that of ABCG subfamily proteins with single molecules. ABC transporters may transport PAHs by forming a dimer-shaped pocket, translocating it into cells, then opening it within the cells, to release the bound PAHs. These results contributed to our understanding of how ABC transporters aid plant root uptake of PAHs.

Mechanistic insights into phenanthrene acropetal translocation via wheat xylem: Separation and identification of transfer proteins.

Polycyclic aromatic hydrocarbons (PAHs) have the potential to cause cancer, teratogenicity, and mutagenesis in humans. Long-term plant safe production relies on how PAHs are transported and coordinated across organs. However, the acropetal transfer mechanism of PAHs in staple crop stems, particularly in xylem, a critical path, is unknown. Herein, we first confirmed the presence of specific interaction between the proteins and phenanthrene by employing the magnetic phenanthrene-bound bead immunoassay and label free liquid chromatograph mass spectrometer (LC-MS/MS), suggesting that peroxidase (uniprot accession: A0A3B5XXD0) and unidentified proteins (uniprot accession: A0A3B6LUC6) may function as the carriers to load and acropetally translocate phenanthrene (a model PAH) in wheat xylem. This specified binding of protein-phenanthrene may form through hydrophobic interactions in the conservative binding region, as revealed by protein structural investigations and molecular docking. To further investigate the role of these proteins in phenanthrene solubilization, phenanthrene exposure was conducted: a substantial quantity of peroxidase was produced; an unusually high expression of uncharacterized proteins was observed, indicating their positive effects in the acropetal transfer of phenanthrene in wheat xylem. These data confirmed that the two proteins are crucial in the solubilization of phenanthrene in wheat xylem sap. Our findings provide fresh light on the molecular mechanism of PAH loading in plant xylem and techniques for ensuring the security of staple crops and improving the efficacy of phytoremediation in a PAH-contaminated environment.

Role of miR164 in the growth of wheat new adventitious roots exposed to phenanthrene.

Polycyclic aromatic hydrocarbons (PAHs), ubiquitous organic pollutants in the environment, can accumulate in humans via the food chain and then harm human health. MiRNAs (microRNAs), a kind of non-coding small RNAs with a length of 18-30 nucleotides, regulate plant growth and development and respond to environmental stress. In this study, it is demonstrated that miR164 can regulate root growth and adventitious root generation of wheat under phenanthrene exposure by targeting NAC (NAM/ATAF/CUC) transcription factor. We observed that phenanthrene treatment accelerated the senescence and death of wheat roots, and stimulated the occurrence of new roots. However, it is difficult to compensate for the loss caused by old root senescence and death, due to the slower growth of new roots under phenanthrene exposure. Phenanthrene accumulation in wheat roots caused to generate a lot of reactive oxygen species, and enhanced lipoxygenase activity and malonaldehyde concentration, meaning that lipid peroxidation is the main reason for root damage. MiR164 was up-regulated by phenanthrene, enhancing the silence of NAC1, weakening the association with auxin signal, and inhibiting the occurrence of adventitious roots. Phenanthrene also affected the expression of CDK (the coding gene of cyclin-dependent kinase) and CDC2 (a gene regulating cell division cycle), the key genes in the cell cycle of pericycle cells, thereby affecting the occurrence and growth of lateral roots. In addition, NAM (a gene regulating no apical meristem) and NAC23 may also be related to the root growth and development in wheat exposed to phenanthrene. These results provide not only theoretical basis for understanding the molecular mechanism of crop response to PAHs accumulation, but also knowledge support for improving phytoremediation of soil or water contaminated by PAHs.

Acropetal translocation of phenanthrene in wheat seedlings: Xylem or phloem pathway?

Due to the potential toxicity of polycyclic aromatic hydrocarbons (PAHs) to humans, the uptake and translocation of PAHs in food crops have gained much attention. However, it is still unclear whether phloem participates in the acropetal translocation of PAHs in plants. Herein, the evidence for acropetal translocation of phenanthrene (a model PAH) via phloem is firstly tested. Wheat (Triticum aestivum L.) new leaves contain significantly higher phenanthrene concentration than old leaves (P < 0.05), and the inhibitory effect on phenanthrene translocation is stronger in old leaves after abscisic acid and polyvinyl alcohol (two common transpiration inhibitors) application. Phenanthrene concentration in xylem sap is slightly higher than in phloem sap. Ring-girdling treatment can significantly reduce phenanthrene concentration in castor bean (Ricinus communis L.) leaves. Two-photon fluorescence microscope images indicate a xylem-to-phloem and acropetal phloem translocation of phenanthrene in castor bean stem. Therefore, phloem is involved in the acropetal translocation of phenanthrene in wheat seedlings, especially when the xylem is not mature enough in scattered vascular bundle plants. Our results provide a deeper understanding of PAH translocation in plants, which have significant implications for food safety and phytoremediation enhancement of PAH-contaminated soil and water.

miR398 is involved in the relief of phenanthrene-induced oxidative toxicity in wheat roots.

Polycyclic aromatic hydrocarbons (PAHs) are ubiquitous environmental pollutants and could produce oxidative toxicity to plants. Our previous study has shown that miR398 is involved in response to phenanthrene treatment by targeting CSD1 and CSD2. However, it is not clear which is essential for CSD1 and CSD2 and how miR398 changes. In this study, we performed discontinuous PAGE to separate superoxide dismutase (SOD) isozymes and found that two bands of the cytosolic Cu/Zn-SOD are induced by phenanthrene at day 5 and 7. Low expression of pri-miR398 and high expression of pre-miR398 indicate that the conversion process from pri-miR398 to pre-miR398 is impeded, which causes decrease in mature miR398. The relative expression of CSD1 is entirely up-regulated, further confirming the important role of CSD1 in response to phenanthrene exposure. Besides, the overexpression of WRKY implies its potential function in answering the call from phenanthrene stress. Therefore, it is concluded that the gene silencing of CSD1 recedes due to the biosynthesis inhibition of miR398, causing the increase of SOD activity in response to phenanthrene exposure in wheat roots. Our results are useful not only for better understanding miRNAs regulation in detoxication of reactive oxygen species, but also for alleviating the toxicity to crops caused by PAHs.