Search Results (273)

Microbial remediation serves as a cost-effective and eco-friendly tactic for petroleum-contaminated sites. In real-world practices, petroleum-degrading microbial consortia display prominent superiority over single strains. To tap into their intrinsic synergistic degradation capacity, it is essential to dissect microbial commonalities, individual traits and functional complementarity for complicated bioremediation scenarios. This study applied bibliometric approaches to collect 15 well-documented representative petroleum-hydrocarbon-degrading strains, and summarized research progress via comparative genomic analysis. Pseudomonas aeruginosa, Bacillus subtilis and Rhodococcus erythropolis are the most frequently reported degraders with remarkable petroleum removal performance. Moreover, co-occurrence network analysis identifies Pseudomonas, Bacillus, Acinetobacter and Rhodococcus as core co-existing genera sustaining natural pollutant-degrading communities. Genomic evidence indicates these strains harbour abundant functional elements encoding diverse oxygenases, alcohol dehydrogenases, cytochrome P450 monooxygenases and key LuxR-, AraC- and GntR-family regulatory factors. P. aeruginosa has the highest copy numbers of catabolic enzyme genes, consistent with its outstanding degradation phenotype. Degraders from different genera exhibit high genetic heterogeneity, with accessory gene clusters significantly enriched in hydrocarbon degradation pathways. Six strains including P. aeruginosa and R. erythropolis assemble a complete gene cascade targeting recalcitrant polycyclic aromatic hydrocarbons. This work provides reliable genomic support for rational strain screening and synthetic-consortium optimization, facilitating knowledge-driven strategies for efficient petroleum bioremediation. Full article

Spodoptera frugiperda (J.E. Smith) is a notoriously destructive and invasive agricultural pest, causing substantial economic losses worldwide. Research has shown that cytochrome P450s play an important role in detoxifying insecticides. However, the precise molecular mechanism of P450-mediated detoxification of pyrethroids in S. frugiperda remains largely unknown. Enzyme activity assays demonstrated the involvement of P450 monooxygenases in pyrethroid detoxification in S. frugiperda. Subsequently, the results of induction experiments revealed that CYP9A58 and CYP9A59 were significantly upregulated by bifenthrin (up to 4.59- and 5.54-fold) and lambda-cyhalothrin (up to 4.17- and 3.28-fold). Stage- and tissue-specific expression analyses suggested that CYP9A59 exhibited predominant expression in sixth-instar larvae, and CYP9A58 was primarily expressed in the moth stage, with both P450 genes showing high transcript levels in the midgut. The functional role of P450s was confirmed by RNAi, where knockdown of either CYP9A58 or CYP9A59 significantly increased larval susceptibility to lambda-cyhalothrin, raising mortality by 45.71% and 57.68%, respectively. Furthermore, structural modeling demonstrated the capacity of both P450 enzymes—CYP9A58 and CYP9A59—to form stable complexes with pyrethroid insecticides. Collectively, these results confirm that CYP9A58 and CYP9A59 are key mediators of pyrethroid detoxification, conferring insecticide tolerance in S. frugiperda. Full article

Huperzine A (HupA) is a natural Lycopodium alkaloid known for its potent neuroprotective properties through the inhibition of acetylcholinesterase. Nevertheless, the limited understanding of its biosynthesis restricts its broader application. This study integrates full-length and second-generation transcriptomes with the quantification of HupA and its precursor, huperzine B, across various tissues of Huperzia serrata, the primary source plant. By employing phylogenetic clustering, expression profiling, and correlation analysis between gene expression and metabolite abundance, we identified 71 candidate genes from seven enzyme families potentially involved in the synthesis of the HupA backbone, including lysine/ornithine decarboxylases, copper amine oxidases (CAOs), chalcone synthases, and cytochrome P450 monooxygenases. Additionally, 28 genes from two families were identified for modification reactions, specifically 2-oxoglutarate/Fe(II)-dependent dioxygenases and caffeoyl shikimate esterases. Comparative analysis between young and mature leaves revealed 3801 genes with higher expression in young leaves, with 84 showing a high correlation with HupA content across seven families. Protein–protein interaction network analysis indicated possible interactions with transcription factors from the MYB, NF-YC, GRAS, ERF, BHLH, and SAP families. Functional validation of two candidate CAOs in planta confirmed their catalytic roles in amine/alkaloid metabolism. This study provides a theoretical foundation and a set of candidate genes for elucidating the biosynthetic pathway of HupA and related alkaloids in H. serrata. Full article

Plant diterpenoids are a diverse class of natural products with important ecological roles and wide applications in the pharmaceutical, agricultural, food additive, and chemical industries. Biosynthesis represents a primary strategy for accessing these valuable compounds. However, the identification of downstream tailoring enzymes (hereafter referred to as tailoring enzymes) involved in diterpenoid biosynthetic pathways remains a major bottleneck, particularly in non-model plant species with limited genomic resources. This review summarizes current strategies for discovering plant diterpenoid biosynthetic pathways and recent advances in elucidating their metabolic routes. We further highlight the lineage-biased distribution of diterpene scaffolds across plant taxa. We propose that scaffold enrichment in specific evolutionary lineages, when integrated with enzyme family expansion and functional divergence, may provide a complementary framework for prioritizing candidate tailoring enzymes. Importantly, scaffold enrichment alone cannot establish enzyme function or evolutionary causality; rather, it provides a complementary layer of evidence that can guide future experimental investigation. Future perspectives include predictive substrate–enzyme mapping, computational and generative design of cytochrome P450 enzymes, and the integration of enzyme discovery, structural modeling, and heterologous chassis engineering. Full article

Cytochrome P450 monooxygenases (CYPs/P450s) are versatile heme-containing enzymes involved in drug metabolism, steroidogenesis, secondary metabolite biosynthesis, and biotechnology. Although African researchers have contributed to P450 research for nearly four decades, no comprehensive assessment of African-led research has been conducted. This study presents a historical and bibliometric analysis of African-led P450 research published between 1987 and 2025 using PubMed and Web of Science. Of 1382 retrieved articles, 558 had African corresponding authors, and 149 included African co-authors. Research output increased markedly after 2011, with South Africa contributing 64% of African-led publications. Corresponding authors represented 31 African countries, while collaborations extended to 54 non-African countries, demonstrating strong international integration. Drug metabolism, pharmacology, and toxicology dominated the research landscape, followed by molecular biology, genomics, enzymology, and structural biology. African researchers have made important contributions to pharmacogenomics, steroidogenesis, insecticide resistance, microbial biotechnology, biosensors, and natural-product metabolism. Collectively, African-led publications have received more than 20,500 citations on Google Scholar, highlighting their global scientific impact. Despite this progress, important gaps remain in large-scale pharmacogenomics, structural biology, systems biology, industrial biotechnology, and omics-based studies. Overall, African researchers have made substantial contributions to global cytochrome P450 research and have created opportunities for future scientific growth and collaboration. Full article

Since its first detection in 2010, Aedes albopictus has spread across northern Algeria, where vector control relies on the use of chemical insecticides. This study aimed to evaluate the susceptibility of Ae. albopictus populations from Algiers to commonly used larvicides and adulticides, as well as to characterize the underlying resistance mechanisms. Eggs were collected from three sites in Algiers. The susceptibility of larvae to temephos and Bacillus thuringiensis israelensis (Bti), as well as that of adults to permethrin, deltamethrin, malathion, and bendiocarb was evaluated using WHO bioassays. Genotyping of knockdown resistance (kdr) mutations was performed via PCR and sequencing. Metabolic resistance mechanisms were investigated using CDC bottle bioassays. The larvae were found to be susceptible to temephos and Bti. Bioassays on adults demonstrated susceptibility to deltamethrin, suspected resistance to permethrin, and resistance to malathion and bendiocarb. Genotyping revealed low frequencies of heterozygous kdr mutations (V1016G, I1532T, F1534C/S). Synergist assays highlighted the key role of esterases in malathion resistance, a minimal involvement of glutathione S-transferases and an unexpected antagonistic effect of cytochrome P450 monooxygenases. Although larvicides remain effective, resistance to organophosphates, carbamates, as well as suspected resistance to permethrin, has been detected. Esterase-mediated metabolic resistance and kdr mutations may contribute to this profile. These findings highlight the need for resistance monitoring and integrated vector control to ensure sustainable control of Ae. albopictus. Full article

Plant cytochrome P450 monooxygenases (CYPs) are valuable biocatalysts for the regioselective hydroxylation of aromatic compounds. However, their expression in bacterial hosts is hampered by poor solubility, membrane anchoring and the requirement for redox partners. In this work, we report the design and characterization of modular expression systems that enable the functional production of SbCYP82D1.1 from Scutellaria baicalensis (SbF6H) in Escherichia coli. Both independent expression and synthetic fusion systems were evaluated by combining a CYP with a compatible reductase (ATR2_tr from Arabidopsis thaliana) to catalyze the conversion of chrysin into baicalein. A combinatorial library of N-terminal variants, host strains, media, and induction strategies was constructed and screened. Among the tested host, E. coli DH 10-beta provided the highest product titers, particularly when cultures were supplemented with 5-aminolevulinic acid. Truncation of the native transmembrane anchor significantly improved catalytic performance, whereas the addition of the heterologous MALLLAVF tag decreased activity. Fusion systems outperformed separate expression formats, showing approximately two-fold higher activity, with the flexible glycine–serine linker (L_GS) supporting the highest hydroxylation product formation. The corresponding fusion construct showed an apparent conversion of 0.1 mM chrysin to baicalein of up to 90% under the applied whole-cell reaction and analytical conditions, although this value should be interpreted with caution due to the concurrent instability of baicalein observed in all reactions and culture conditions. This result nevertheless indicates a marked improvement in whole-cell baicalein formation compared with previously reported bacterial systems. Together, these results demonstrate that rational N-terminal engineering combined with fusion protein design can enable efficient bacterial expression of plant CYPs, representing a promising step toward scalable production of hydroxylated flavonoids. Full article

Thiamethoxam is the main neonicotinoid insecticide used for controlling Riptortus pedestris (Fabricius) (Hemiptera: Alydidae). However, sublethal concentration stress may induce intergenerational transcriptional memory, leading to transcriptional patterns that may contribute to the intergenerational accumulation of metabolic tolerance, and evaluating only the toxicity of the current generation would underestimate the long-term risk. Therefore, this study investigated the effect of parental exposure on the expression of detoxification enzyme genes in offspring. Using transcriptome sequencing, we systematically identified three detoxification enzyme gene families (cytochrome P450 monooxygenases (CYPs), carboxylesterases (CCEs), and glutathione S-transferases (GSTs)) in R. pedestris and compared their differential expression patterns between the parental and filial generations after thiamethoxam treatment at three sublethal concentrations (LC₁₀, LC₃₀, and LC₅₀). In the parental generation, a Theta family GST was consistently upregulated, while in the filial generation, detoxification genes were predominantly downregulated, and the genes upregulated in the parents were not also upregulated in the offspring. Comparisons of parents and offspring at the same concentration revealed that the medium concentration induced the highest number of intergenerationally upregulated genes, exhibiting a non-linear response pattern. These results indicate that parental exposure to sublethal thiamethoxam leaves an intergenerational transcriptional imprint in the offspring, and the transmission pattern involves transcriptional reprogramming rather than simple replication of the parental response, the mechanism of which remains to be determined. This study provides transcriptomic evidence for understanding the metabolic adaptation and intergenerational resistance evolution of R. pedestris to thiamethoxam, offering important reference value for field resistance monitoring and rational insecticide application. Full article

Plagiodera versicolora (Coleoptera: Chrysomelidae), the willow leaf beetle, is a leaf-eating pest that generally occurs on salicaceous trees. However, there is a blank of identification and phylogenetic relationship of the detoxification genes in P. versicolora. Here, we identified four detoxification gene families (glutathione S-transferases: GSTs, UDP-glycosyltransferases: UGTs, cytochrome P450 monooxygenases: CYPs and carboxylesterases: COEs) from the adult antennal transcriptome data. In all, 146 candidate detoxification genes including 22 GSTs, 20 UGTs, 60 CYPs, and 44 COEs were identified. We used quantitative real-time PCR technology to explore the tissue expression patterns of 12 PvGSTs in P. versicolora. The results showed that 7 PvGSTs have significantly high expression in antennae, indicating these PvGSTs may play an important role in degrade and/or inactivate the sex pheromones and host volatiles. The identification and phylogenetics of the detoxification genes in P. versicolora extended the database in Coleoptera and contributed to the subsequent in-depth research for function about detoxification genes. Full article

Kadsura heteroclita (Roxb.) Craib, commonly known as “Xuetong”, is a traditional Tujia ethnomedicine with anti-rheumatoid arthritis (RA) activity, and schizanlactone E (Xuetongsu) is its major bioactive component whose biosynthetic pathway remains uncharacterized. As a cycloartane-type tetracyclic triterpenoid, Xuetongsu’s biosynthesis is likely to involve multiple oxidation steps. Cytochrome P450 (CYP450) is a versatile monooxygenase encoded by a large and diverse gene superfamily and plays a critical role in various oxidation reactions in plants’ secondary metabolism. In this study, 367 KhCYP450s were identified and systematically analyzed for their physicochemical properties, phylogenetic analysis, conserved motifs, gene structures, collinearity, and cis-acting elements. Weighted gene co-expression network analysis (WGCNA) revealed a turquoise module strongly associated with Xuetong root tissue, which had the highest Xuetongsu accumulation; 32 candidate KhCYP450s within this module were screened via correlation analysis between gene expression and xuetongsu content and partially validated by qRT-PCR. Five of these candidates showed significant homology with known triterpenoid biosynthetic genes via protein structure analyses. This study deepened our comprehension of the CYP450 superfamily in Xuetong and provided a valuable reference for further research on the biosynthesis of Xuetongsu. Full article

The rapid evolution of metabolic resistance to chemical insecticides and the adaptation to plant allelochemicals in insect pests have become major challenges in global pest management. While the overexpression of cytochrome P450 monooxygenases (P450s) is a well-recognized classic detoxification mechanism, the upstream epigenetic and post-transcriptional regulatory networks governing this process have only recently been elucidated. In this narrative review, the latest research progress on microRNAs (miRNAs) as crucial “fine-tuners” in insect detoxification networks is systematically summarized. The classic regulatory model is highlighted: the induced or constitutive downregulation of specific miRNAs relieves the translational repression of their target P450 genes, thereby contributing to metabolic resistance to major insecticide classes, including neonicotinoids, diamides, and pro-insecticides. Furthermore, the evolutionary recruitment mechanisms of conserved miRNAs in host plant adaptation are explored, and how endocrine signals, such as juvenile hormone (JH) and 20-hydroxyecdysone (20E), synergistically regulate the miRNA–P450 axis is analyzed. The “sponge effect”, wherein highly expressed P450 mRNAs act as competitive endogenous RNAs (ceRNAs) to sequester miRNAs, and the consequent physiological trade-offs (fitness costs) resulting from the prioritization of metabolic resources toward the detoxification system are comprehensively discussed. Finally, the current core methodologies for miRNA functional validation are critically evaluated, and the application potential and ecological safety prerequisites of miRNA-based nanobiopesticides for targeted and sustainable pest management are discussed. By integrating mechanistic insights with translational perspectives, this review highlights miRNA–P450 regulatory networks as key determinants of insecticide resistance evolution and as promising targets for developing more precise, environmentally compatible pest-management strategies. Full article

Background/Objectives: Hepatocellular carcinoma (HCC) is challenging to treat with chemotherapy. Immunotherapy has shown moderate responses in inflammatory and immunosuppressive tumor environments. Hepatic cytochrome P450 monooxygenases (CYPs) play a crucial role in xenobiotic and drug metabolism, as well as lipid and steroid metabolism. We aimed to investigate whether CYP expression and various parameters of the innate and adaptive immune system are prognostic factors for the survival of HCC patients. Methods: HCC biopsies (n = 370) from The Cancer Genome Atlas (TCGA) database were analyzed using Kaplan–Meier statistics and the KMPlotter algorithm. Parameters such as immune cell infiltration, DNA mutation rates, and neoantigen load were selected for survival analysis and subjected to hierarchical cluster analysis. The expression of candidate CYP genes in tumors was compared to that in normal liver tissues. Furthermore, tumor infiltration of innate immune cells (basophilic and eosinophilic granulocytes, natural killer cells), adaptive immune cells (CD4+ memory and CD8+ cytotoxic T cells, regulatory T cells, type 1 and type 2 helper T cells), and mesenchymal stem cells was examined. Results: High expression of CYP19A1 and CYP26B1 was associated with shorter survival, whereas high expression of CYP3A5, CYP3A43, CYP7A1, and CYP27A1 was linked to longer survival. Mutation rates combined with CYP expression showed a correlation with five out of six CYP genes, while a high neoantigen load produced less definitive results. A specific cluster exhibiting high CYP expression and immune cell counts or mutation/neoantigen rates was associated with shorter survival, while another cluster was linked to longer survival. Conclusions: CYPs involved in the metabolic regulation of HCC, including CYP3A5, CYP3A43, CYP7A1, CYP19A1, CYP26B1, and CYP27A1, were found to have prognostic value for patient survival. Combined signatures that include CYP expression, mutational rates, and immune cell infiltration into tumors further enhanced the prognostic value for patient survival. This suggests that CYPs may influence the creation of a tumor-specific metabolic microenvironment that impacts immune functions. These combined signatures could be utilized for patient stratification to personalize tumor treatment and develop novel combination therapies aimed at optimizing treatment outcomes, such as combining transarterial chemoembolization (TACE) with immune checkpoint inhibitors. Full article

The Colorado potato beetle, Leptinotarsa decemlineata, is a major insect pest of potatoes. Our previous studies have demonstrated that two cytochrome P450 monooxygenase (P450s) genes, CYP9Z140 and CYP9AY1, and a uridine diphosphate–glycosyltransferase (UGT) gene, UGT321AP1, play important roles in thiamethoxam resistance to L. decemlineata. However, the related upstream regulatory mechanism remains unclear. In this study, we first monitored the resistance of L. decemlineata field populations to thiamethoxam in Xinjiang to determine the resistance ratios. The predicted results demonstrated that four transcription factors (TFs), CncC/Maf, Abd-B, FoxO, and Ptx1, may bind to the core regions of three gene promoters. The qRT-PCR results revealed that the TFs were significantly upregulated by thiamethoxam and exhibited specific spatiotemporal expression patterns. Dual-luciferase reporter assays indicated that the CncC pathway could regulate the expression of three detoxification genes, whereas Abd-B and FoxO only regulate CYP9Z140 and UGT321AP1 expressions, respectively. Ptx1 could regulate the expression of both CYP9AY1 and UGT321AP1. Furthermore, knockdown of several TFs through RNA interference significantly reduced expression of the corresponding detoxification genes, consistent with the dual-luciferase reporter assay results, and increased the thiamethoxam susceptibility of test adults. These findings aid in gaining a deeper understanding of the transcriptional regulation mechanisms of insecticide resistance in insects. Full article

The tomato borer, Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae), is a highly destructive invasive pest of tomato, and its ability to develop resistance to insecticides is well known. The susceptibility of three field populations from Egypt (Luxor, Assuit, and Giza) to the insecticides chlorantraniliprole, chlorfenapyr, indoxacarb, emamectin benzoate, and spinetoram was evaluated by leaf-dip bioassays. The second-instar larvae were exposed, and mortality was noted 24, 48, and 72 h later. Probit analysis was used to calculate lethal concentrations (LC₅₀ and LC₉₀) and resistance ratios (RR₅₀). Furthermore, key detoxification enzymes (α-esterases, cytochrome P450 monooxygenases, and glutathione S-transferases) were also measured to identify possible metabolic resistance mechanisms. Considerable inter-population differences were observed. Detoxification enzymes (α-esterases, cytochrome P450 monooxygenases, and glutathione S-transferases) were measured. The Luxor population showed the highest LC₅₀ values and resistance ratios, especially to chlorfenapyr and indoxacarb, suggesting low-to-moderate resistance, while the Giza population was the most susceptible. Emamectin benzoate and spinetoram provided excellent control of all laboratory and field strains. Full article

Maize ear rot, primarily caused by Fusarium verticillioides (Fusarium ear rot, FER) and Fusarium graminearum (Gibberella ear rot, GER), is a devastating disease that causes significant yield losses and mycotoxin contamination. Breeding resistant varieties is the most effective control strategy, but this requires the identification of stable genetic loci for resistance. In this study, we employed bulked segregant analysis (BSA) on two F₂ mapping populations to identify quantitative trait loci (QTLs) conferring resistance to FER and GER. We identified five and eleven QTLs for FER and GER, respectively. Notably, chromosome 4 was identified as a major hotspot for resistance to both diseases, and there was a co-localization of the FER QTL (qFER4.05) and GER QTL (qGER4.05-1) within a 58.58–71.34 Mb interval on bin 4.05, suggesting a potential locus for broad-spectrum resistance. Within this overlapping region, we identified 18 high-confidence candidate genes, including genes encoding leucine-rich repeat receptor-like kinases (LRR-RLKs), remorin, cytochrome P450 monooxygenases, and wall-associated receptor kinase-like (WAKL) protein, all with established roles in plant defense. These findings advance the understanding of the genetic architecture of ear rot resistance and provide critical resources for marker-assisted breeding to develop maize hybrids with durable resistance to both FER and GER. Full article