Dicyclohexyl phthalate (DCHP) is a widely used plasticizer, yet its potential toxicity remains insufficiently characterized. In this study, Caenorhabditis elegans was employed to investigate the reproductive and neurotoxic effects of DCHP, along with its transgenerational impacts. Parental exposure to DCHP significantly impaired head thrashing and body bending, accompanied by morphological abnormalities in dopaminergic neurons. Additionally, DCHP exposure led to a reduction in both the number of eggs laid and the number of fertilized eggs, which was associated with the upregulation of apoptosis-related genes (cep-1, ced-3, ced-4 and ced-9). Elevated levels of reactive oxygen species and oxidative stress were also observed in C. elegans. Notably, these toxic effects were transmitted to the F1 generation but were alleviated in the F2 generation, potentially due to increased expression of skn-1 and gst-4. This study addresses critical gaps in the toxicological profile of DCHP and provides a scientific foundation for evaluating its ecological and human health risks.

Although recent advancements have shed light on the crucial role of coordinated evolution among cell subpopulations in influencing disease progression, the full potential of these insights has not yet been fully harnessed in the clinical application of personalized precision medicine for prostate cancer (PCa). In this study, we utilized single-cell sequencing to identify the evolutionary characteristics of tumoral cell states and employed comprehensive bulk RNA sequencing to evaluate their potential as prognostic indicators and therapeutic targets. Leveraging advancements in artificial intelligence, we integrated machine learning with multi-omics to develop and validate the tumor evolutionary characteristic predictive indicator (TECPI). TECPI not only demonstrated superior prognostic performance compared to traditional clinical predictors and 81 previously published models but also improved patient outcomes by accurately identifying individuals who would benefit from immunotherapy and targeted therapies. Furthermore, we experimentally validated the critical role of AMOTL1 in PCa pharmacodynamics through its interaction with AR, pivotal for modulating the sensitivity to AR antagonist. Additionally, we demonstrated the generalizability and applicability of TECPI across pan-cancers. In summary, this study emphasizes the importance of understanding cellular diversity and dynamics within the tumor microenvironment to predict PCa progression and to guide targeted therapy effectively.

Silver nanoparticles (AgNPs), the most extensively used nanomaterials, are inevitably released into the environment, yet their toxicological effects on plants remain not fully elucidated. To address a gap in the literature regarding the effects of AgNPs on cotton (Gossypium hirsutum L. cv. TM-1), a 15-days exposure experiment was conducted using AgNPs (10 mg/L), along with Ag+ (0.4 mg/L AgNO3) serving as a mock control and water as the control (CK). Compared to the Ag+and CK groups, the roots of cotton seedlings exposed to AgNPs showed noticeable browning with significant reductions in biomass and root length. To explore the protein-level changes in cotton root response to AgNPs, a 4D label-free proteomics analysis was performed on the AgNPs, Ag+, and CK groups. Finally, 1588 differentially expressed proteins (DEPs) unique expressed in the AgNPs-treated group, which were primarily enriched in lignin and gossypol biosynthesis pathways, as revealed by GO (Gene Ontology) and KEGG (Kyoto Encyclopedia of Genes and Genomes) analyses. Subsequent phloroglucinol staining and quantification confirmed an increased lignin biosynthesis, while LC-MS examination demonstrated elevated gossypol production in cotton roots following AgNPs exposure. Furthermore, roots from a glandless cotton mutant (CCRI12gl, lacking gossypol) were significant shorter than those of its wide-type counterpart (CCRI12, glandular, containing gossypol) after AgNPs treatment, suggesting that gossypol enhances cotton tolerance to AgNPs. Additionally, parallel reaction monitoring (PRM) technology validated that the expression levels of proteins involved in lignin and gossypol biosynthesis were consistent with the findings of the 4D label-free proteomics analysis. This study provides new insights into the mechanisms of plant responses to AgNPs exposure and offers a fundamental reference for investigating the toxicity of nanometal in crops.

Amid the rapid growth of offshore aquaculture in China, a Deeper Offshore Aquaculture System (DOAS) for hybrid Jinhu groupers (Epinephelus fuscoguttatus♀× Epinephelus tukula♂) has been established in Guangdong. This study aims to elucidate the biological and metabolic differences between Jinhu groupers cultured in Recirculating Aquaculture Systems (RAS) and those in DOAS. A three-month growth comparison experiment was conducted to assess the developmental differences of Jinhu groupers. This research reveals significant variations in growth performance and flesh fatty acid composition between fish from these two systems, particularly in the content of eicosapentaenoic acid and nine other fatty acids. Through transcriptome analysis of liver and muscle tissues, 278 and 186 differentially expressed genes were identified, respectively, highlighting notable metabolic shifts. Key pathways affected include lipid metabolism and muscle fiber growth. These insights deepen the understanding of the acclimation mechanisms of Jinhu groupers to diverse aquaculture settings and offer valuable strategies for optimizing deeper offshore aquaculture practices, enhancing the sustainability and efficiency of the industry.

Metal tolerance proteins (MTPs) play central roles in metal homeostasis in plant cells and tissues. Cotton (Gossypium hirsutum) has strong tolerance to metal stresses and can be used as a phytoremediation crop for heavy metal-contaminated areas. However, the information and biological functions of the MTP gene family in cotton and their roles in metal stresses remain unclear. In this study, a comprehensive analysis of MTP genes was carried out with cotton plants. We identified a total of 23 GhMTP proteins ranged from 371 to 496 amino acids, which were predicted to be located in the cell membrane and vacuole, while most of which have 4 to 6 transmembrane domains. The 23 GhMTPs were divided into 3 clusters by phylogenetic analysis, Mn-CDF, Zn/Fe-CDF, and Zn-CDF clusters. Meanwhile, the promoter region of GhMTPs contains 4282 binding sites associated with 12 transcription factor families. The expression pattern of GhMTPs was relatively stable in anther, bract, filament, leaf, petal, pistil, root, sepal, stem, and torus, indicating that GhMTP genes were functionally conserved in cotton evolution. In addition, we found that GhMTP2, GhMTP3, GhMTP5, GhMTP8, GhMTP15, GhMTP17, GhMTP19, and GhMTP22 in the cotton leaves could be induced by specific metal ions. These results provide important information for understanding the heavy metal tolerance mechanism and genetic improvement of cotton, and will be helpful for the phytoremediation of heavy metal contaminated soils in the future.

INTRODUCTION:Prostate cancer (PCa) is one of the most commonly diagnosed cancers in men, with a high global incidence. The Meiotic Nuclear Division 1 (MND1) protein is essential for the repair of DNA double-strand breaks during meiosis, but its role in PCa remains poorly understood. This study aims to explore the function of MND1 in PCa progression and the mechanism involved. METHODS:RNA-Seq data from the TCGA and GEO databases were analyzed. Kaplan-Meier (KM) method and χ2 test examined the association between MND1 expression, prognosis, and clinical parameters. PCa cell lines (22RV1 and C4-2) were used for functional assays. CCK- 8, EdU, colony formation assay, flow cytometry analysis and xenograft model were used to evaluate the effects of MND1 on PCa cell proliferation in vitro and in vivo. RESULTS:MND1 expression was significantly upregulated in PCa tissues, particularly in cases with Gleason scores ≥8, and correlated with poorer disease-free survival (DFS) and adverse clinical features. Functionally, elevated MND1 expression promoted PCa cell proliferation both in vitro and in vivo. Mechanistically, MND1 facilitated cell cycle progression from G0/G1 to S phase via activation of the CCNB1/p53 signaling pathway. CONCLUSION:MND1 promotes prostate cancer progression by facilitating the G0/G1 to S phase transition via the CCNB1/p53 pathway, making it a promising prognostic marker and potential therapeutic target.

Ferroptosis induction has emerged as a promising therapeutic approach for prostate cancer (PCa), either as a monotherapy or in combination with hormone therapy. Therefore, identifying the mechanisms regulating ferroptosis in PCa cells is essential. Our previous study demonstrated that HJURP, an oncogene upregulated in PCa cells, plays a role in tumor proliferation. Here, we expand these findings by elucidating a novel mechanism by which HJURP inhibits sensitivity to ferroptosis inducers in PCa cells via the PRDX1/reactive oxygen species (ROS) pathway in vitro and in vivo. Mechanistically, HJURP forms disulfide-linked intermediates with PRDX1 through Cys327 and Cys457 residues. This disulfide binding promotes PRDX1 redox cycling and inhibits its hyperoxidation. As a result, HJURP enhances the peroxidase activity of PRDX1, leading to a decrease in ROS levels and subsequently suppressing lipid peroxidation induced by ferroptosis inducers. These findings reveal the potential of HJURP/PRDX1 as novel therapeutic targets and biomarkers of ferroptosis in PCa patients.

As an important endogenous cytokine in mammals, interferon-γ ( IFNγ ) primes and significantly enhances the recognition and response of TLRs to pathogen-associated molecular patterns (PAMPs) in leukocytes. Subsequently, a series of phenomena in the TLR signalling pathway are activated, including increased expression of TLR mRNA, activation of the downstream signalling pathway mediated by myeloid differentiation primary response protein 88 ( MyD88 )-tumour necrosis factor receptor-associated factor 6 ( TRAF6 ), and nuclear translocation of the nuclear transcription factor NF-κB . There is only one IFNγ isoform in mammals, while many fishes possess two IFNγ genes, IFNγ1 (also called IFNγ-rel) and IFNγ2 . The aim of this work was to explore how these two IFNγ isoforms regulate the TLR signalling pathway in green-spotted pufferfish ( Tetraodon nigroviridis ). Real-time quantitative PCR (RT-qPCR) was performed to detect the expression of TLR1 , TLR2 , TLR3 , TLR5 , TLR7 , TLR8 , TLR9 , MyD88 , and TFAF6 in T. nigroviridis after stimulation with recombinant IFNγ1 ( rIFNγ1 ) or rIFNγ2 . Western blot analysis showed that the expression of MyD88 , TFAF6, and NF-κB gradually increased between 0.5 and 8 h after stimulation with rTnIFNγ1. However, following treatment with IFNγ2 , the expression of NF-κB at the protein level initially increased rapidly, only to decline swiftly thereafter. Meanwhile, the expression of MyD88 and TFAF6 exhibited a fluctuating pattern of rapid initial decline, subsequent rapid increase, and subsequent rapid decline. Electrophoretic mobility shift assay (EMSA) analysis showed that there was a significant and rapid shift of NF-κB within 5 min soon after rIFNγ2 stimulation, while rIFNγ1 had no significant effect. In summary, we revealed that IFNγ1 and IFNγ2 modulate the innate immune response via TLRs in T. nigroviridis and their regulatory mechanisms are different and multileveled. IFNγ1 and IFNγ2 could be used as immunomodulators to fine-tune the immune response of T. nigroviridis in different ways.

IntroductionMacrophages are components of the innate immune system and can play an anti-tumor or pro-tumor role in the tumor microenvironment owing to their high heterogeneity and plasticity. Meanwhile, prostate cancer (PCa) is an immune-sensitive tumor, making it essential to investigate the value of macrophage-associated networks in its prognosis and treatment.MethodsMacrophage-related marker genes (MRMGs) were identified through the comprehensive analysis of single-cell sequencing data from GSE141445 and the impact of macrophages on PCa was evaluated using consensus clustering of MRMGs in the TCGA database. Subsequently, a macrophage-related marker gene prognostic signature (MRMGPS) was constructed by LASSO-Cox regression analysis and grouped based on the median risk score. The predictive ability of MRMGPS was verified by experiments, survival analysis, and nomogram in the TCGA cohort and GEO-Merged cohort. Additionally, immune landscape, genomic heterogeneity, tumor stemness, drug sensitivity, and molecular docking were conducted to explore the relationship between MRMGPS and the tumor immune microenvironment, therapeutic response, and drug selection.ResultsWe identified 307 MRMGs and verified that macrophages had a strong influence on the development and progression of PCa. Furthermore, we showed that the MRMGPS constructed with 9 genes and the predictive nomogram had excellent predictive ability in both the TCGA and GEO-Merged cohorts. More importantly, we also found the close relationship between MRMGPS and the tumor immune microenvironment, therapeutic response, and drug selection by multi-omics analysis.DiscussionOur study reveals the application value of MRMGPS in predicting the prognosis of PCa patients. It also provides a novel perspective and theoretical basis for immune research and drug choices for PCa.

BACKGROUND:Prostate cancer (PCa), a globally prevalent malignancy, displays intricate heterogeneity within its epithelial cells, closely linked with disease progression and immune modulation. However, the clinical significance of genes and biomarkers associated with these cells remains inadequately explored. To address this gap, this study aimed to comprehensively investigate the roles and clinical value of epithelial cell-related genes in PCa.METHODS:Leveraging single-cell sequencing data from GSE176031, we conducted an extensive analysis to identify epithelial cell marker genes (ECMGs). Employing consensus clustering analysis, we evaluated the correlations between ECMGs, prognosis, and immune responses in PCa. Subsequently, we developed and validated an optimal prognostic signature, termed the epithelial cell marker gene prognostic signature (ECMGPS), through synergistic analysis from 101 models employing 10 machine learning algorithms across five independent cohorts. Additionally, we collected clinical features and previously published signatures from the literature for comparative analysis. Furthermore, we explored the clinical utility of ECMGPS in immunotherapy and drug selection using multi-omics analysis and the IMvigor cohort. Finally, we investigated the biological functions of the hub gene, transmembrane p24 trafficking protein 3 (TMED3), in PCa using public databases and experiments.RESULTS:We identified a comprehensive set of 543 ECMGs and established a strong correlation between ECMGs and both the prognostic evaluation and immune classification in PCa. Notably, ECMGPS exhibited robust predictive capability, surpassing traditional clinical features and 80 published signatures in terms of both independence and accuracy across five cohorts. Significantly, ECMGPS demonstrated significant promise in identifying potential PCa patients who might benefit from immunotherapy and personalized medicine, thereby moving us nearer to tailored therapeutic approaches for individuals. Moreover, the role of TMED3 in promoting malignant proliferation of PCa cells was validated.CONCLUSIONS:Our findings highlight ECMGPS as a powerful tool for improving PCa patient outcomes and supply a robust conceptual framework for in-depth examination of PCa complexities. Simultaneously, our study has the potential to develop a novel alternative for PCa diagnosis and prognostication.

As a new source of natural fibers, the Bombax ceiba tree can provide thin, light, extremely soft and warm fiber material for the textile industry. Natural fibers are an ideal model system for studying cell growth and differentiation, but the molecular mechanisms that regulate fiber initiation are not fully understood. In B. ceiba, we found that fiber cells differentiate from the epidermis of the inner ovary wall. Each initiated cell then divides into a cluster of fiber cells that eventually develop into mature fibers, a process very different from the classical fiber initiation process of cotton. We used high-throughput single-cell RNA sequencing (scRNA-seq) to examine the special characteristics of fiber initiation in B. ceiba. A total of 15 567 high-quality cells were identified from the inner wall of the B. ceiba ovary, and 347 potential marker genes for fiber initiation cell types were identified. Two major cell types, initiated fiber cells and epidermal cells, were identified and verified by RNA in situ hybridization. A developmental trajectory analysis was used to reconstruct the process of fiber cell differentiation in B. ceiba. Comparative analysis of scRNA-seq data from B. ceiba and cotton (Gossypium hirsutum) confirmed that the additional cell division process in B. ceiba is a novel species-specific mechanism for fiber cell development. Candidate genes and key regulators that may contribute to fiber cell differentiation and division in B. ceiba were identified. This work reveals gene expression signatures during B. ceiba fiber initiation at a single-cell resolution, providing a new strategy and viewpoint for investigation of natural fiber cell differentiation and development.

Abscisic acid (ABA) is a plant hormone that plays an important role in cotton fiber development. In this study, the physiological changes and proteomic profiles of cotton ( Gossypium hirsutum ) ovules were analyzed after 20 days of ABA or ABA inhibitor (ABAI) treatment. The results showed that compared to the control (CK), the fiber length was significantly decreased under ABA treatment and increased under ABAI treatment. Using a tandem mass tags-based quantitative technique, the proteomes of cotton ovules were comprehensively analyzed. A total of 7321 proteins were identified, of which 365 and 69 differentially accumulated proteins (DAPs) were identified in ABA versus CK and ABAI versus CK, respectively. Specifically, 345 and 20 DAPs were up- and down-regulated in the ABA group, and 65 and 4 DAPs were up- and down-regulated in the ABAI group, respectively. The DAPs in the ABA group were mainly enriched in the biosynthesis of secondary metabolites, phenylpropanoid biosynthesis and flavonoid secondary metabolism, whereas the DAPs in the ABAI group were mainly enriched in the indole alkaloid biosynthesis and phenylpropanoid biosynthesis pathways. Moreover, 9 proteins involved in phenylpropanoid biosynthesis were upregulated after ABA treatment, suggesting that this pathway might play important roles in the response to ABA, and 3 auxin-related proteins were upregulated, indicating that auxin might participate in the regulation of fiber development under ABAI treatment.

Background: Prostate cancer (PCa) ranks as the second most prevalent malignancy among males on a global scale. Accumulating evidence suggests that inflammation has an intricate relationship with tumorigenesis, tumor progression and tumor immune microenvironment. However, the overall impact of inflammation-related genes on the clinical prognosis and tumor immunity in PCa remains unclear. Methods: Machine learning methods were utilized to construct and validate a signature using The Cancer Genome Atlas (TCGA) for training, while the Memorial Sloan Kettering Cancer Center (MSKCC) and GSE70769 cohorts for independent validation. The efficacy of the signature in predicting outcomes and its clinical utility were assessed through a series of investigations encompassing in vitro experiments, survival analysis, and nomogram development. The association between the signature and precision medicine was explored via tumor immunity, genomic heterogeneity, therapeutic response, and molecular docking analyses, using bulk and single-cell RNA-sequencing data. Results: We identified 7 inflammation-related genes with prognostic significance and developed an inflammation-related prognostic signature (IRPS) with 6 genes. Furthermore, we demonstrated that both the IRPS and a nomogram integrating risk score and pathologic T stage exhibited excellent predictive ability for the survival outcomes in PCa patients. Moreover, the IRPS was found to be significantly associated with the tumor immune, genomic heterogeneity, therapeutic response, and drug selection. Conclusion: IRPS can serve as a reliable predictor for PCa patients. The signature may provide clinicians with valuable information on the efficacy of therapy and help personalize treatment for PCa patients.

Grass carp (Ctenopharyngodon idella) is among the most important freshwater fish species in China. However, it remained unclear how salinity could affect grass carp. Two experiments were performed. The first experiment was a 4-day acute salt tolerance experiment with six salinities (0, 4, 8, 12, 16, and 20 ppt). The second experiment was an 8-week chronic salt stress experiment with three salinities (0, 2 and 6 ppt). To investigate the intestinal bacterial community of grass carp from three salinities (0, 2, and 6 ppt), the 16S rDNA sequencing was performed. The results showed that grass carp exhibited great adaptability to low salinity (2 ppt), with no significant difference in growth and maintained stable physiological and immune status. However, exposed to high salinity (6 ppt) caused significant deleterious effects on grass carp, including growth inhibition as well as physiological and immune-related changes. The gut microbiota in grass carp changed with salinity. With the increase of salinity, the proportion of beneficial bacteria in the gut of grass carp gradually decreased, while some harmful bacteria gradually occupied the dominant position. Changes in gut microbial composition ultimately affected the growth of grass carp. This study helps further clarify the effects of salinity on grass carp.

Objective:To investigate the expression of G2 and S phase-expressed-1 (GTSE1) in prostate cancer(PCa) tissues and its regulation in PCa metastasis.Methods:GEPIA, UALCAN and FireBrowse databases were analyzed to determine the correlation between GTSE1 expression and clinical indexes and metastasis in patients with PCa. The co-expression genes and possible pathways of GTSE1 were further analyzed. Wound healing and transwell invasion assays were used to detect the effect of GTSE1 on the migration and invasion of PCa cells in vitro. Western blotting assay was used to analyze the effects of GTSE1 on epithelial-to-mesenchymal transition (EMT) markers.Results:The expression of GTSE1 was significantly increased in PCa tissues, especially in metastatic PCa, and was closely related to the disease progression and lymph node metastasis. Overexpression of GTSE1 enhanced the ability of migration and invasion of PCa cells, and promoted the formation of EMT, which showed the opposite effects with GTSE1 knockdown. GTSE1 may act on the microtubules of PCa cells and cooperate with TPX2, TOP2A and CENPF to drive EMT and metastasis of PCa. Conclusion:GTSE1 is highly expressed in PCa tissues and can promote the invasion and migration of PCa cells, which may be developed as a new molecular target for the treatment of metastatic PCa.

Breast cancer is the most prevalent malignancy and the first leading cause of cancer-related mortality among the female population worldwide. Approaches for precise and reliable detection of breast cancer cells, particularly in the nascent state, are desperately needed for elevating the survival rate of patients bearing the breast tumor. In this work, we successfully performed the sensitive, precise, and reliable breast cancer cell detection using facilely fabricated bovine serum albumin-gold nanocluster (BSA-AuNCs) protein corona stabilized, epithelial cell adhesion molecule (EpCAM) aptamer linked fluorescent polystyrene nanoparticle (PS NP), termed as PS-BSA-AuNCs-Apt. The rapidly adsorbed BSA-AuNCs hard protein corona without complicated covalent conjugation not only imparted excellent colloidal stability to the PS nanoparticles, but also offered numerous active anchors for the targeted EpCAM aptamers to locate. With the remarkable aid of the aptamers specifically targeting the EpCAM-positive breast cancer cells, the PS-BSA-AuNCs-Apt emitted strong and photostable dual-color fluorescent signals for precise and reliable cancer cell detection by overcoming the false signals. The specific identification potency of the PS-BSA-AuNCs-Apt system was further verified by successfully detecting the xenografted breast tumor tissue. Notably, to the best of our knowledge, the protein corona formed nanoprobes was exploited for direct tumor cell and tissue detection with high efficacy for the first time, demonstrating their promising potential in clinical tumor detection.

Nucleotide-binding oligomerization domain-like receptor protein 3 (NLRP3) inflammasome is an important inflammasome in mammals, which is of great significance to eliminate pathogens. However, the research of the NLRP3 inflammasome in teleost is limited. Tetraodon nigroviridis has the characteristics of small genome and easy feeding, which can be used as a model for the study of fish immune function. In present study, three NLRP3 inflammasome component genes (NLRP3, ASC and caspase-1) in T. nigroviridis has been cloned. Real-time fluorescence quantitative PCR showed that TnNLRP3 (T. nigroviridis NLRP3), TnASC (T. nigroviridis ASC) and Tncaspase-1 (T. nigroviridis caspase-1) mRNA in various tissues from health T. nigroviridis were highly expressed in immune-related tissues, such as spleen, gill, head kidney and intestine. After Vibrio parahemolyticus infection, the expression of TnNLRP3, TnASC and Tncaspase-1 mRNA in spleen, gill, head kidney reached a peak at 24 h, and the expression levels of these genes in intestine were the highest at 48 h. After the transfection of TnASC-pAcGFP-N1 monomer GFP plasmid into cos-7 cells, ASC specks, the activation marker of NLRP3 inflammasome, were observed. Bimolecular fluorescence complementarity and fluorescence colocation experiment showed that TnASC and Tncaspase-1 of TnNLRP3 inflammasome were co-located near the cell nucleus, and potentially interacted with each other. NLRP3 inflammasome inducer nigericin and agonist ATP could significantly induce the expression of TnNLRP3, TnASC and Tncaspase-1 mRNA, and activation of NLRP3 inflammasome could promote the generation of mature TnIL-1β (T. nigroviridis IL-1β). These results uncover that T. nigroviridis NLRP3 inflammasome could participate in the antibacterial immune response and the generation of mature TnIL-1β after activation.

In the area of climate change, nanotechnology provides handy tools for improving crop production and assuring sustainability in global agricultural system. Due to excellent physiological and biochemical properties, silver nanoparticles (AgNPs) have been widely studied for potential use in agriculture. However, there are concerns about the mechanism of the toxic effects of the accumulation of AgNPs on crop growth and development. In this study, the impacts of AgNPs on cotton ( Gossypium hirsutum ) seedlings were evaluated by integrating physiological and comprehensive metabolomic analyses. Potting-soil-grown, two-week-old cotton seedlings were foliar-exposed to 5 mg/plant AgNP or 0.02 mg/plant Ag + (equivalent to the free Ag + released from AgNPs). Primary metabolites and volatile organic compounds (VOCs) were identified by gas chromatography–mass spectrometry (GC-MS) and solid-phase microextraction (SPME) GC-MS, respectively. AgNPs inhibited the photosynthetic capacity of the cotton leaves. The metabolic spectrum analysis identified and quantified 73 primary metabolites and 45 VOCs in cotton leaves. Both treatments significantly changed the metabolite profiles of plant leaves. Among the primary metabolites, AgNPs induced marked changes in amino acids, sugars and sugar alcohols. Among the VOCs, 13 volatiles, mainly aldehydes, alkanes and terpenoids, were specifically altered only in response to AgNPs. In summary, our study showed that the comprehensive influence of AgNPs on primary metabolites and VOCs was not merely attributed to the released Ag + but was caused by AgNP-specific effects on cotton leaves. These results provide important knowledge about the physiological and chemical changes in cotton leaves upon exposure to AgNPs and offer a new insight for supporting the sustainable use of AgNPs in agriculture.

Magnesium (Mg) is an essential macronutrient for plant growth and development. Physiological and transcriptome analyses were conducted to elucidate the adaptive mechanisms to long-term Mg deficiency (MD) in banana seedlings at the 6-leaf stage. Banana seedlings were irrigated with a Mg-free nutrient solution for 42 days, and a mock control was treated with an optimum Mg supply. Leaf edge chlorosis was observed on the 9 th leaf, which gradually turned yellow from the edge to the interior region. Accordingly, the total chlorophyll content was reduced by 47.1%, 47.4%, and 53.8% in the interior, center and edge regions, respectively, and the net photosynthetic rate was significantly decreased in the 9 th leaf. Transcriptome analysis revealed that MD induced 9,314, 7,425 and 5,716 differentially expressed genes (DEGs) in the interior, center and edge regions, respectively. Of these, the chlorophyll metabolism pathway was preferentially enriched according to Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis. The expression levels of the five candidate genes in leaves were consistent with what is expected during chlorophyll metabolism. Our results suggest that changes in the expression of genes related to chlorophyll synthesis and decomposition result in the yellowing of banana seedling leaves, and these results are helpful for understanding the banana response mechanism to long-term MD.