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30 pages, 15411 KB  
Article
Selenium Nanobiostimulants Attenuate Copper-Induced Oxidative Damage in Brassica napus Through Genotype-Specific Antioxidant and Metabolic Adaptation
by Sundas Fatima, Muhammad Arslan Yousaf, Saba Yaseen, Muhammad Kamran, Basharat Ali, Yingying Zhou, Asad Ullah, Fangbin Cao, Skhawat Ali and Weijun Zhou
Plants 2026, 15(9), 1333; https://doi.org/10.3390/plants15091333 - 27 Apr 2026
Abstract
Copper (Cu) contamination poses severe threats to agricultural productivity and food safety, particularly affecting economically important crops such as rapeseed (Brassica napus L.). This study investigated the protective effects of selenium nanoparticles (SeNPs) against Cu toxicity in four B. napus cultivars. Exposure [...] Read more.
Copper (Cu) contamination poses severe threats to agricultural productivity and food safety, particularly affecting economically important crops such as rapeseed (Brassica napus L.). This study investigated the protective effects of selenium nanoparticles (SeNPs) against Cu toxicity in four B. napus cultivars. Exposure to Cu (200 μM) caused severe reductions in growth and photosynthetic efficiency while significantly elevating oxidative stress markers across all cultivars. Application of SeNPs (25 μM) effectively mitigated these adverse effects, improving biomass, restoring chlorophyll content, and enhancing photosynthetic performance compared to Cu-stressed plants. SeNP treatment significantly enhanced antioxidant enzyme activities, with corresponding upregulation of antioxidant gene expression. Secondary metabolite profiling revealed cultivar-specific responses, with sensitive cultivar Zheda 622 exhibiting metabolic adaptation and higher volatile organic compound (VOC) accumulation, while tolerant cultivar Zheda 635 maintained metabolic stability. PCA analysis demonstrated distinct metabolic clustering patterns, reflecting differential stress-responsive strategies. The study demonstrates that SeNPs attenuate Cu-induced toxicity through integrated mechanisms encompassing diminished Cu acquisition, augmented antioxidant defense systems, and comprehensive metabolic reprogramming. Cultivar-specific responses highlighted substantial genetic variation in tolerance mechanisms across B. napus genotypes. These findings substantiate SeNPs as a viable and efficacious nanomaterial for sustainable agronomic management in Cu-contaminated edaphic environments. The approach offers dual benefits of improved crop productivity and reduced Cu accumulation, ensuring enhanced food safety. Full article
(This article belongs to the Special Issue Nanobiotechnology in Plant Health and Stress Resilience)
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17 pages, 2140 KB  
Article
Lysolecithin Improves Lipid Metabolism and Gut Microbiota: An Integrated Transcriptome and Microbiome Analysis in Largemouth Bass (Micropterus salmoides) Fed Stearin-Based High-Lipid Diets
by Yuexing Zhang, Tianyu Feng, Zhiyong Dong, Tianhong Ke, Trond Storebakken, Wanjie Cai, Bo Shi and Liying Huang
Metabolites 2026, 16(5), 297; https://doi.org/10.3390/metabo16050297 (registering DOI) - 27 Apr 2026
Abstract
Background: Supplementing aquafeeds with emulsifiers can enhance lipid utilization, yet the physiological effects of lysolecithin, derived from enzymatic lecithin conversion, remain under-explored. Objectives: This study examined the effects of lysolecithin supplementation on hepatopancreatic transcriptome and gut microbiota in largemouth bass (Micropterus salmoides [...] Read more.
Background: Supplementing aquafeeds with emulsifiers can enhance lipid utilization, yet the physiological effects of lysolecithin, derived from enzymatic lecithin conversion, remain under-explored. Objectives: This study examined the effects of lysolecithin supplementation on hepatopancreatic transcriptome and gut microbiota in largemouth bass (Micropterus salmoides) fed stearin-based high-lipid diets. Methods: Two diets were formulated: a control containing 130 g kg−1 stearin fish oil (SO), and in the experimental diet (SL), 3.1 g kg−1 rapeseed oil was replaced with 3.1 g kg−1 lysolecithin oil. Each diet was fed to three replicate groups for 56 days. Hepatopancreas and distal intestine were sampled for transcriptome profiling, and gut microbiota were characterized at 28 and 56 days. Results: Lysolecithin supplementation resulted in 424 differentially expressed genes compared with the control (322 up- and 102 downregulated). KEGG enrichment indicated major effects on lipid metabolic processes, notably activation of the PI3K-AKT signaling pathway, enhanced adipocyte lipolysis, and modulation of adipocytokine signaling, suggesting improved insulin sensitivity and lipid mobilization. Histological analysis showed mild distal intestinal inflammation in the SO group. Gut microbiota composition shifted over time; lysolecithin increased the relative abundance of Cetobacterium and reduced potential opportunistic taxa compared with the control. Conclusions: Overall, dietary inclusion of lysolecithin improved lipid utilization in largemouth bass, likely by enhancing lipid metabolism and promoting beneficial gut microbial profiles. These findings support lysolecithin as a promising feed additive for optimizing high-lipid aquafeeds. Full article
(This article belongs to the Special Issue Metabolism and Nutrition in Fish)
16 pages, 2355 KB  
Article
Ginkgo biloba DFR2 Gene Remodels the Flavonoid Metabolic Network in Transgenic Nicotiana benthamiana
by Xinru Sun, Cheng Ji, Pengfei Yu, Guibin Wang and Jing Guo
Plants 2026, 15(9), 1331; https://doi.org/10.3390/plants15091331 - 27 Apr 2026
Abstract
Dihydroflavonol 4-reductase (DFR) plays a pivotal role in regulating flavonoid and anthocyanin biosynthesis, governing the accumulation of plant secondary metabolites. This study aimed to characterize the DFR gene family in Ginkgo biloba and elucidate the function of the predominant gene GbDFR2 in the [...] Read more.
Dihydroflavonol 4-reductase (DFR) plays a pivotal role in regulating flavonoid and anthocyanin biosynthesis, governing the accumulation of plant secondary metabolites. This study aimed to characterize the DFR gene family in Ginkgo biloba and elucidate the function of the predominant gene GbDFR2 in the flavonoid metabolic network. Through transcriptome analysis, three differentially expressed GbDFR genes were identified. Bioinformatic analysis revealed that all three GbDFR proteins are hydrophilic and acidic and belong to the NADB_Rossmann superfamily. RT-qPCR analysis of different tissues of ginkgo revealed that all three GbDFR genes exhibited the highest expression levels in the leaves. An overexpression vector of GbDFR2 was constructed and stably transformed into Nicotiana benthamiana. Metabolomic and qPCR analyses showed that heterologous GbDFR2 expression significantly remodeled the flavonoid profile, upregulating sakuranetin and 3,7-Di-O-methylquercetin while downregulating narcissin and naringenin chalcone. Additionally, it upregulated endogenous NbCHI and NbDFR, and suppressed the transcription factors NbMYL2b and NbERF4a. These findings suggest that GbDFR2 can act as a regulator of flavonol biosynthesis and provide a candidate gene for the metabolic engineering of flavonoids in woody plants. Full article
19 pages, 2112 KB  
Article
A Comprehensive Larval microRNA Atlas of Hyphantria cunea Identifies Candidate miRNAs and Potential Molecular Targets for Green Pest Management
by Yanqin Zhu, Kai Tang, Mao Lin, Shuaishuai Fanji and Shouke Zhang
Int. J. Mol. Sci. 2026, 27(9), 3884; https://doi.org/10.3390/ijms27093884 (registering DOI) - 27 Apr 2026
Abstract
Hyphantria cunea (Drury) causes extensive ecological damage primarily during its larval stages, characterized by voracious feeding and rapid dispersal. Given that conventional dsRNA-mediated RNA interference (RNAi) is generally recalcitrant in Lepidoptera, endogenous microRNAs (miRNAs) may represent an additional class of regulatory molecules worthy [...] Read more.
Hyphantria cunea (Drury) causes extensive ecological damage primarily during its larval stages, characterized by voracious feeding and rapid dispersal. Given that conventional dsRNA-mediated RNA interference (RNAi) is generally recalcitrant in Lepidoptera, endogenous microRNAs (miRNAs) may represent an additional class of regulatory molecules worthy of systematic investigation. In this study, we utilized high-throughput sequencing to construct nine comprehensive miRNA libraries across three critical developmental milestones (three biological replicates per instar): the 1st, 4th, and 7th instars (L1, L4, and L7). A total of 1667 miRNA entries were catalogued, including 1080 known and 587 bioinformatically predicted, as yet unvalidated novel miRNA candidates. Comparative transcriptomic analysis revealed 52 differentially expressed miRNAs with significant stage-dependent profiles, with the most pronounced divergence observed between the L1 and L7 groups. Bioinformatic prediction identified 16,784 non-redundant target genes. GO and KEGG enrichment analyses indicated that the predicted target genes of these differentially expressed miRNAs were enriched in developmental and metabolic categories, including cellular development, protein digestion, and nutrient absorption, suggesting that these miRNAs may be associated with tissue remodeling and larval developmental transitions. Collectively, our findings expand the currently available miRNA resource for H. cunea and define stage-associated miRNA expression patterns during larval development. Rather than establishing direct functional roles, this work provides a framework and candidate molecules for future design of RNAi-based biopesticides. Full article
(This article belongs to the Section Molecular Plant Sciences)
26 pages, 2961 KB  
Review
The Muco-Microbiotic Layer in Respiratory Pathophysiology: Integrating Transcriptomics, Inflammation Phenotypes, and Clinical Biomarkers in Precision Pulmonology
by Claudio Candia, Adelaide Carista, Melania Ionelia Gratie, Domiziana Picone, Giuseppa D’Amico, Celeste Caruso Bavisotto, Fabio Bucchieri, Silvestro Ennio D’Anna, Alessandro Pitruzzella, Mauro Maniscalco, Giuseppe Bonaventura, Stefano Burgio and Francesco Cappello
Biology 2026, 15(9), 684; https://doi.org/10.3390/biology15090684 (registering DOI) - 27 Apr 2026
Abstract
The lower airways are a dynamic environment where physical, microbial, and molecular factors intersect to regulate respiratory health and disease. The muco-microbiotic (MuMi) layer, composed of mucus, resident microbes, and extracellular vesicles (EVs), is not just a passive barrier but also an active [...] Read more.
The lower airways are a dynamic environment where physical, microbial, and molecular factors intersect to regulate respiratory health and disease. The muco-microbiotic (MuMi) layer, composed of mucus, resident microbes, and extracellular vesicles (EVs), is not just a passive barrier but also an active site for host–microbe communication. This layer integrates epithelial cell biology, microbial activity, and immune responses within the bronchial environment. New transcriptomic and metatranscriptomic technologies show that it is not only which microbes are present but also their gene activity that closely links to airway inflammation and disease. EV-associated RNAs from both host and microbial cells act as key messengers, influencing epithelial responses, immune activity, mucus properties, and microbial behaviour. This review highlights evidence that positions the MuMi layer as central to understanding lower airway disease, particularly asthma and chronic obstructive pulmonary disease (COPD). Distinct gene expression programs and biomarker profiles, such as exhaled nitric oxide, may reflect different disease mechanisms even in cases with similar clinical features, such as eosinophilia. Multi-omic approaches focused on the MuMi layer enable better disease classification, biomarker discovery, and therapy selection. By putting the MuMi interface at the core of precision pulmonology, we provide a framework for advancing personalised care in chronic respiratory diseases. Full article
21 pages, 2542 KB  
Article
Integrated Metabolomics and Transcriptomics Reveal the Influence of Natural and Cultivation-Managed Habitats on Metabolic Divergence and Flavonoid Enrichment in Anoectochilus roxburghii
by Erli Wang, Weicheng Gao, Peng Wang and Xiaoping Wang
Metabolites 2026, 16(5), 294; https://doi.org/10.3390/metabo16050294 (registering DOI) - 27 Apr 2026
Abstract
Background/Objectives: Environmental conditions in natural and cultivation-managed habitats strongly influence plant physiology and medicinal quality. However, the molecular mechanisms underlying metabolic differentiation in Anoectochilus roxburghii remain poorly understood. This study aimed to elucidate the metabolic and transcriptional differences between wild and cultivated [...] Read more.
Background/Objectives: Environmental conditions in natural and cultivation-managed habitats strongly influence plant physiology and medicinal quality. However, the molecular mechanisms underlying metabolic differentiation in Anoectochilus roxburghii remain poorly understood. This study aimed to elucidate the metabolic and transcriptional differences between wild and cultivated A. roxburghii and to identify the regulatory mechanisms driving habitat-associated variation in metabolite profiles. Methods: We applied integrated non-targeted metabolomics and transcriptomics to compare metabolic profiles and gene expression in the leaves and stems of 15-month-old wild and cultivated A. roxburghii plants. Gene–metabolite correlation analysis was performed to identify coordinated correlation networks associated with key biosynthetic pathways. Results: Our analyses revealed clear differences in metabolite composition and transcriptional patterns between habitat types, suggesting distinct strategies of metabolic resource allocation. Wild plants showed significant enrichment of amino acids and other primary metabolites, whereas cultivated plants accumulated higher levels of flavonoids. Gene–metabolite correlation analysis indicated that multiple flavonoid metabolites were closely associated with key structural genes, including F3H, C12RT1, and HHT1, forming a tightly connected correlation network. In addition, several transcription factor families, including MYB, bHLH, WRKY, and AP2/ERF, showed strong correlations with genes involved in the flavonoid pathway, suggesting that flavonoid accumulation in cultivated plants may be associated with coordinated transcriptional control. Conclusions: Taken together, these findings suggest that habitat conditions are associated with differences in metabolic networks and resource allocation in A. roxburghii. This work provides new insight into the metabolic plasticity of this medicinal plant and highlights potential factors associated with molecular mechanisms that may contribute to variation in medicinal quality. Full article
(This article belongs to the Section Pharmacology and Drug Metabolism)
18 pages, 7901 KB  
Article
Genome-Wide Identification and Expression Analysis of Starch Biosynthesis-Related Gene Families in Wheat
by Qinlong Zhao, Longjiao Hu, Xinye Wu, Bo Ma, Weining Song, Xiaojun Nie and Shuzuo Lv
Int. J. Mol. Sci. 2026, 27(9), 3876; https://doi.org/10.3390/ijms27093876 (registering DOI) - 27 Apr 2026
Abstract
Starch synthesis is critical for crop yield and quality and is regulated and coordinated by a series of key enzymes encoded by starch synthesis-related genes (SSRGs). Although this process is well characterized in many crops, the genomic location and expression patterns of SSRGs [...] Read more.
Starch synthesis is critical for crop yield and quality and is regulated and coordinated by a series of key enzymes encoded by starch synthesis-related genes (SSRGs). Although this process is well characterized in many crops, the genomic location and expression patterns of SSRGs in wheat remain unclear. Here, we performed a genome-wide analysis and identified 78 SSRGs in wheat, classified into the AGPase, SSS, GBSS, SBE, and DBE subfamilies. SSRGs within each subfamily showed conserved motifs and domain organization. RNA-seq analysis indicated that most SSRGs are expressed during early grain development. We further examined genetic variation in SSRGs across wheat and its progenitors using re-sequencing data. Diploid wheat showed greater genetic differentiation and diversity than tetraploid and hexaploid wheat. Five SSRGs exhibited significant haplotype differences between emmer wheat and common wheat; emmer wheat displayed diverse haplotypes, whereas common wheat showed a single dominant haplotype. Finally, starch characteristics differed between emmer wheat and common wheat in amylose content and thermodynamic properties, while viscosity, crystal structure, and morphology were largely similar. Overall, this study systematically characterizes SSRGs in wheat and provides insights for improving starch quality. Full article
(This article belongs to the Special Issue Molecular Characterization and Utilization of Plant Genetic Resources)
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26 pages, 1379 KB  
Review
Epigenetic Variation in Plant Populations: DNA Methylation as a Driver of Phenotypic Diversity and Adaptation
by Jakub Sawicki, Wiktoria Czochór, Aniela Garbowska, Kamil Koczwara, Jerzy Andrzej Przyborowski, Natan Pupek, Paweł Sulima, Joanna Szablińska and Monika Szczecińska
Diversity 2026, 18(5), 259; https://doi.org/10.3390/d18050259 (registering DOI) - 27 Apr 2026
Abstract
DNA methylation constitutes a primary layer of epigenetic regulation in plants, operating across three sequence contexts (CG, CHG, and CHH) through distinct enzymatic pathways. Over the past fifteen years, accumulating evidence has shown that DNA methylation varies substantially among individuals and populations of [...] Read more.
DNA methylation constitutes a primary layer of epigenetic regulation in plants, operating across three sequence contexts (CG, CHG, and CHH) through distinct enzymatic pathways. Over the past fifteen years, accumulating evidence has shown that DNA methylation varies substantially among individuals and populations of wild plants, sometimes independently of underlying genetic polymorphism. This variation can influence gene expression, transposable element activity, and phenotypic traits relevant to ecological adaptation. Population epigenetics, the study of methylation variation at the population scale, has matured from initial surveys using methylation-sensitive amplified fragment length polymorphism (MS-AFLP) into a discipline increasingly reliant on reduced-representation bisulfite sequencing (epiGBS, bsRADseq), whole-genome bisulfite sequencing (WGBS), enzymatic methyl-seq (EM-seq), and direct long-read detection by nanopore sequencing. These methodological advances are opening population epigenetics to non-model organisms across the full breadth of the plant phylogeny, from angiosperms and gymnosperms to ferns and bryophytes. We cover (i) the molecular machinery underlying plant DNA methylation, including the debated status of N6-methyladenine (6mA); (ii) empirical evidence for natural epigenetic variation in plant populations, spanning clonal, invasive, and outcrossing species; (iii) the methodological toolkit available for population-scale methylation profiling, with emphasis on approaches suitable for non-model taxa; and (iv) the ecological and evolutionary significance of population epigenetic variation, including transgenerational inheritance, stress memory, epigenetic clocks, conservation applications, and the emerging integration of epigenetics into the extended evolutionary synthesis. We identify critical knowledge gaps, particularly the near-complete absence of population-level epigenetic data for bryophytes, ferns, and lycophytes, and outline priorities for future research. Full article
(This article belongs to the Special Issue 2026 Feature Papers by Diversity's Editorial Board Members)
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22 pages, 6391 KB  
Article
Differential Expression and Target Gene Analysis of PBMC-Derived microRNAs as Prognostic Biomarkers in Acute Lymphoblastic Leukemia
by Fatemah S. Basingab, Hadil Alahdal, Deemah Alwadaani, Ghaida Almuneef, Ahmed S. Barefah, Ali H. Algiraigri, Rawan Hammad, Mohamed Elnakeeb, Jehan S. Alrahimi, Kawther A. Zaher and Alia M. Aldahlawi
Int. J. Mol. Sci. 2026, 27(9), 3868; https://doi.org/10.3390/ijms27093868 (registering DOI) - 27 Apr 2026
Abstract
Acute lymphoblastic leukemia (ALL) is a clinically diverse cancer in which microRNA (miRNA)-mediated post-transcriptional regulation contributes to leukemogenesis and subtype heterogeneity. In this study, miRNA expression profiling by microarray was performed on ALL cases (B-ALL and T-ALL) and healthy controls. Data were normalized [...] Read more.
Acute lymphoblastic leukemia (ALL) is a clinically diverse cancer in which microRNA (miRNA)-mediated post-transcriptional regulation contributes to leukemogenesis and subtype heterogeneity. In this study, miRNA expression profiling by microarray was performed on ALL cases (B-ALL and T-ALL) and healthy controls. Data were normalized and analyzed for differential expression using false discovery rate (FDR)-adjusted p-values. Differentially expressed miRNAs were further examined using unsupervised visualization to assess overall disease-related expression patterns. To explore their biological significance, experimentally validated miRNA–target interactions were obtained using multiMiR, limited to validated databases (miRTarBase, TarBase, and miRecords) and summarized via target-burden ranking, miRNA–target network analysis, and Circos–style interaction mapping. A unique miRNA expression signature was identified in ALL. Upregulated miRNAs included miR-106a-5p, miR-106b-5p, miR-17-5p, miR-20a-5p, miR-20b-5p, miR-181b-5p, and miR-128-3p, while miR-127-3p, miR-139-5p, miR-433-3p, and miR-584-5p were downregulated. Validated targets concentrated on key leukemia-related genes like PTEN, BCL2L11, CDKN1A, CCND1, RB1, E2F1, and TGFBR2. KEGG pathway analysis highlighted pathways associated with leukemic cell survival and growth, including MAPK, cell cycle, autophagy, Hippo, ubiquitin-mediated proteolysis, and mTOR signaling pathways. These findings reveal a concise ALL-associated miRNA panel predominantly comprising the miR-17/20/106 family and provide a prioritized set of candidate regulatory networks for subtype-specific validation and functional follow-up studies. Full article
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31 pages, 16448 KB  
Article
Punicic Acid in Ovarian Cancer: Anticancer Activity and Mechanistic Insights
by Jingjia Mo, Isabella Mendieta, Alexander J. Adams, Katherine Wiest, Hannah Lee, Victoria Gorman, Rachel Koo, Santiago Garcia, Ethan Nguyen, Aaron Lee, Jihua Feng and Zhiqing Huang
Cells 2026, 15(9), 792; https://doi.org/10.3390/cells15090792 (registering DOI) - 27 Apr 2026
Abstract
Ovarian cancer (OC) remains the deadliest gynecological malignancy, with aged tumor microenvironments linked to poorer outcomes. Our prior work identified reduced levels of free fatty acids (FFAs) within tumor-surrounding adipose tissue of aged OC xenograft rats compared to younger counterparts. In this study, [...] Read more.
Ovarian cancer (OC) remains the deadliest gynecological malignancy, with aged tumor microenvironments linked to poorer outcomes. Our prior work identified reduced levels of free fatty acids (FFAs) within tumor-surrounding adipose tissue of aged OC xenograft rats compared to younger counterparts. In this study, we investigated the therapeutic potential of one such FFA, punicic acid (PunA). We evaluated PunA’s effects on OC and normal cell viability and compared its activity with that of its structural isomer, α-eleostearic acid (α-ESA). Both compounds decreased OC cell viability; however, α-ESA was cytotoxic to normal cells, whereas PunA selectively impaired OC cell viability while sparing normal cells. Additionally, PunA enhanced cisplatin efficacy, demonstrating its potential for use in combination therapy to reduce cisplatin dosage and toxicity without compromising antitumor activity. Mechanistically, PunA induced ferroptosis in OC cells while sparing normal cells by differently modulating lipid peroxidation, fatty acid oxidation, and mitochondrial function. Transcriptomic profiling further revealed coordinated gene expression changes associated with oxidative stress and ferroptosis in PunA-treated OC and normal cells. In a preliminary C57BL/6J-ID8 OC mouse model, PunA suppressed tumor growth. Collectively, these findings identify PunA as a promising therapeutic candidate for OC, acting through ferroptosis and mitochondrial dysfunction, and enhancing cisplatin efficacy while sparing normal cells. Full article
(This article belongs to the Special Issue From Molecular Mechanisms to Treatment Progress of Ovarian Cancer)
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16 pages, 2797 KB  
Article
Synergistic Effects of Amino Acids and Bacillus velezensis N35 on Suppressing Phelipanche aegyptiaca Parasitism and Modulating Tomato Growth: Insights from Transcriptomic Profiling
by Wei He, Yiguang Wang, Siqiong Tang, Wenfang Luo, Xin Huang, Junhui Zhou, Xiang Zhang and Jianjun Xu
Plants 2026, 15(9), 1327; https://doi.org/10.3390/plants15091327 - 27 Apr 2026
Abstract
Phelipanche aegyptiaca is a root parasitic weed that causes severe yield losses in tomato production. Current control methods are constrained by limited efficacy and environmental concerns. Although biocontrol microbes and amino acids have each been reported to suppress broomrape parasitism individually, their synergistic [...] Read more.
Phelipanche aegyptiaca is a root parasitic weed that causes severe yield losses in tomato production. Current control methods are constrained by limited efficacy and environmental concerns. Although biocontrol microbes and amino acids have each been reported to suppress broomrape parasitism individually, their synergistic effects and underlying molecular mechanisms remain largely unexplored. This study evaluated the biocontrol performance of Bacillus velezensis strain N35, applied alone or in combination with five amino acids (methionine, isoleucine, valine, histidine, and proline), against P. aegyptiaca parasitism in tomato using pot experiments coupled with transcriptomic profiling of host roots. Both individual and combined treatments significantly reduced the number and fresh weight of P. aegyptiaca parasitic tubercles. Notably, the combinations of methionine + N35 and isoleucine + N35 achieved near-complete suppression of parasitism. Transcriptomic analysis revealed extensive reprogramming of gene expression in tomato roots, with significant enrichment in pathways associated with plant hormone signal transduction, MAPK signaling, phenylpropanoid biosynthesis, and carotenoid biosynthesis. The synergistic treatments coordinately activated ethylene, jasmonic acid, and salicylic acid-mediated signaling, while suppressing auxin and abscisic acid signaling. Moreover, key strigolactone biosynthesis genes (CCD7 and CCD8) were strongly downregulated, and specific genes involved in the biosynthesis of defense-related secondary metabolites were selectively upregulated. Collectively, these findings demonstrate a pronounced synergy between B. velezensis N35 and specific amino acids in suppressing P. aegyptiaca parasitism. This enhanced host resistance is achieved through the coordinated reprogramming of hormonal and metabolic networks, particularly via interference with strigolactone-mediated germination signal secretion. This study provides a theoretical basis for the development of microbe–metabolite synergistic strategies as sustainable and environmentally benign alternatives for broomrape management. Full article
(This article belongs to the Section Plant Protection and Biotic Interactions)
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23 pages, 8570 KB  
Article
The Heterogeneous Nuclear Ribonucleoprotein K (hnrnpk) Gene Targeted by miR-460a-5p Functions in the Gonadal Differentiation and Development in Chinese Tongue Sole (Cynoglossus semilaevis)
by Kaimin Li, Haipeng Yan, Qi Liu, Wenjie Li, Chengbin Gao and Songlin Chen
Animals 2026, 16(9), 1327; https://doi.org/10.3390/ani16091327 - 27 Apr 2026
Abstract
Chinese tongue sole (Cynoglossus semilaevis), an economically important mariculture species in China, exhibits pronounced sexual dimorphism in growth, underscoring the importance of elucidating sex regulatory mechanisms for aquaculture development. Heterogeneous nuclear ribonucleoprotein K (hnrnpk) critically regulates mammalian reproductive development, [...] Read more.
Chinese tongue sole (Cynoglossus semilaevis), an economically important mariculture species in China, exhibits pronounced sexual dimorphism in growth, underscoring the importance of elucidating sex regulatory mechanisms for aquaculture development. Heterogeneous nuclear ribonucleoprotein K (hnrnpk) critically regulates mammalian reproductive development, yet its role in fish sex regulation remains elusive. Here, we systematically investigated the underlying function and mechanisms of hnrnpk in C. semilaevis through integrated molecular cloning, expression profiling, upstream regulatory analysis, functional assays, and transcriptome sequencing. We found that hnrnpk was highly expressed in the gonad and liver, with female-biased expression during gonadal development. Promoter activity assays revealed that sox2 and c-Jun enhanced hnrnpk transcription, whereas foxl2 and ar suppressed it. Additionally, hnrnpk was directly targeted by miR-460a-5p in C. semilaevis, revealing multi-level transcriptional and post-transcriptional regulation. Functional analyses showed that hnrnpk regulated cyp19a1a in a cell type-dependent and dose-sensitive manner: the expression of cyp19a1a was both upregulated in hnrnpk-knockdown ovarian cells and hnrnpk-overexpression testicular cells. Interestingly, foxl2 was upregulated in hnrnpk-knockdown ovarian cells but suppressed in hnrnpk-overexpression testicular cells, which showed the distinct regulation mechanisms in the different sexual programs. Transcriptomic analyses further revealed that several sex-related genes (sox9a with downregulation, etc.) were significantly regulated, and cell development and cycle pathways were dramatically enriched in functional enrichment analyses. This might indicate that hnrnpk overexpression drives C. semilaevis testis (CSTE) toward feminization reprogramming through sox9 switching and multi-pathway perturbations. Overall, our findings might reveal that hnrnpk, a female-biased gene regulated by miR-460a-5p and transcription factors, influences sex-related gene expression through sox9 switching. This study will offer new insights for C. semilaevis hnrnpk into sex determination and also provide a potential target for monosex breeding in aquaculture. Full article
(This article belongs to the Special Issue Morphological and Physiological Research on Fish: Second Edition)
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27 pages, 34800 KB  
Article
Antiviral and Anticancer Activities of Stingless Bee Propolis from Tetragonula drescheri and Tetragonula pagdeni: Toward Development of Prototype Healthcare Pharmaceuticals
by Karnjanee Greegrainuch, Wiratchanee Kansandee, Chamsai Pientong, Tipaya Ekalaksananan and Jureeporn Chuerduangphui
Int. J. Mol. Sci. 2026, 27(9), 3855; https://doi.org/10.3390/ijms27093855 (registering DOI) - 27 Apr 2026
Abstract
Honey and propolis from the stingless bees Tetragonula drescheri and Tetragonula pagdeni remain underexplored for their health-promoting application. This study investigated the bioactive compounds, and antiviral and anticancer activities of honey and propolis extracts against herpes simplex virus (HSV), and human papillomavirus (HPV-16/18)-positive [...] Read more.
Honey and propolis from the stingless bees Tetragonula drescheri and Tetragonula pagdeni remain underexplored for their health-promoting application. This study investigated the bioactive compounds, and antiviral and anticancer activities of honey and propolis extracts against herpes simplex virus (HSV), and human papillomavirus (HPV-16/18)-positive cervical cancer cells. Water and ethanol extracts were prepared and evaluated for anti-HSV activity using plaque assay, and for anticancer effects on CaSki and HeLa cells using apoptosis, colony formation, cell migration, and candidate gene expression analysis. Propolis water extract most potentially inhibited HSV wild-type and drug-resistant strains. Propolis ethanol extract from T. drescheri markedly suppressed CaSki and HeLa cell growth, induced apoptosis, downregulated HPV-16/18 E6, and upregulated BAX expression. Chemical profiles were identified by electrospray ionization quadrupole time-of-flight mass spectrometry. Most candidate compounds displayed preferable drug-likeness properties. Prototype herbal soup formulations containing selected extracts significantly inhibited HSV-1 drug-resistant strain and HPV-16 E6 expression. These findings demonstrated the high antiviral and anticancer potential of the extracted compounds from T. drescheri and T. pagdeni propolis, supporting their application in health-promoting products against HSV and HPV infection. Full article
(This article belongs to the Section Bioactives and Nutraceuticals)
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21 pages, 9783 KB  
Article
Leucine-Rich Repeat Extension 7 Gene Confers Cotton Resistance to Verticillium Wilt
by Xue Du, Yanfang Li, Wankui Gong, Zhen Wei, Qiankun Liu, Aiming Zhang, Yuting Ge, Yangyang Wei, Yuling Liu, Quanwei Lu, Xianghui Xiao, Pengtao Li, Juwu Gong and Renhai Peng
Int. J. Mol. Sci. 2026, 27(9), 3852; https://doi.org/10.3390/ijms27093852 (registering DOI) - 26 Apr 2026
Abstract
Leucine-rich repeat extensins (LRXs) are essential regulators of plant development, cell wall integrity, and stress responses. However, genome-wide LRX studies in cotton are limited. Analysis of four Gossypium species identified 29, 28, 16, and 16 LRX genes in G. hirsutum, G [...] Read more.
Leucine-rich repeat extensins (LRXs) are essential regulators of plant development, cell wall integrity, and stress responses. However, genome-wide LRX studies in cotton are limited. Analysis of four Gossypium species identified 29, 28, 16, and 16 LRX genes in G. hirsutum, G. barbadense, G. arboreum, and G. raimondii, respectively. Phylogenetic analysis resolved these 89 genes into four subfamilies (I–IV). Structural annotation revealed that cotton LRX family members exhibit conserved domain architectures. This finding was corroborated by motif analysis, which revealed notable conservation in the motif compositions of most cotton LRX proteins, suggesting functional conservation across evolutionary lineages. Distinct spatiotemporal expression patterns were uncovered between G. hirsutum and G. barbadense. Prolonged exposure to extreme temperatures induced widespread down-regulation of most GhLRX genes, whereas genes in subgroup IV were significantly up-regulated under salt and drought stress conditions, respectively. Notably, GhLRX7 showed a more proactive responding profile to Verticillium wilt (VW) infection, which was therefore selected for functional validation employing virus-induced gene silencing in the cotton cultivars MBI9626 and CCRI36. Phenotypic analysis of silenced plants revealed exacerbated disease symptoms compared to wild-type controls, providing direct evidence implicating GhLRX7 as a key contributor to defense against VW. Full article
(This article belongs to the Section Molecular Plant Sciences)
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Article
Kratom (Mitragyna speciosa) as a Phytochemical-Based Natural Product Exhibiting Opioid-like Analgesic Effects with Reduced Tolerance and Dependence Liability via TLR4-Associated Neuroimmune Modulation
by Fajar Prasetya, Niken Indriyanti, Nurul Muhlisa Mus, Mentarry Bafadal, Raisa Fadilla, Yuli Widiyastuti, Chaidir Chaidir, Hadi Kuncoro, Sofa Fajriah, Rudi Heryanto, Angga Cipta Narsa, Onny Ziasti Fricillia, Yurika Sastyarina, Victoria Yulita Fitriani, Siti Rouchmana, Nurus Sobah, Zulhaerana Bahar, Nur Rezky Khairun Nisaa, Helmi Helmi and Hady Anshory
Molecules 2026, 31(9), 1428; https://doi.org/10.3390/molecules31091428 (registering DOI) - 26 Apr 2026
Abstract
Kratom (Mitragyna speciosa) is a botanical candidate for pain management with potentially reduced opioid-related risks, partly through modulation of neuroimmune pathways involving Toll-Like Receptor 4 (TLR4). This study aimed to characterize the phytochemical profile of kratom ethanol extract and evaluate its [...] Read more.
Kratom (Mitragyna speciosa) is a botanical candidate for pain management with potentially reduced opioid-related risks, partly through modulation of neuroimmune pathways involving Toll-Like Receptor 4 (TLR4). This study aimed to characterize the phytochemical profile of kratom ethanol extract and evaluate its effects on TLR4 signalling, neuroinflammatory cytokines, analgesic activity, withdrawal behaviours, and organ safety in morphine-dependent mice. Metabolite profiling was conducted using UHPLC–Q-Exactive Orbitrap HRMS, followed by molecular docking of major constituents to the TLR4 complex. In vivo assessments included flow cytometry and gene expression analyses of TLR4-mediated cytokines (NF-κB, IL-1β, IL-6), behavioural assays for antinociception, endurance, and withdrawal symptoms, and histopathological and biochemical evaluation of liver, kidney, and spleen tissues. More than 100 metabolites were identified, including mitragynine and flavonoids such as rutin and isoquercetin, which showed interactions with key TLR4 residues. Selected fractions suppressed pro-inflammatory cytokine expression, increased tail-pinch latency comparable to morphine, reduced withdrawal manifestations, and demonstrated nephroprotective and immunomodulatory effects, although mild reversible hepatic alterations were observed in specific fractions. Overall, kratom ethanol extract exhibited fraction-dependent analgesic and anti-neuroinflammatory activities associated with TLR4 modulation, supporting its potential as a botanical analgesic candidate while emphasizing the importance of safety optimization and standardized fraction development. Full article
(This article belongs to the Special Issue Redox-Active Molecules as Key Players for Inflammatory Diseases)
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