Search Results (2,807)

Background/Objectives: Due to the numerical dominance of environmental and commensal strains, understanding antimicrobial resistance (AMR) transmission in Escherichia coli requires consideration of non-clinical as well as pathogenic isolates. In this cross-sectional study, associations between the genetic context of non-clinical E. coli and AMR carriage are examined in isolates sampled from different niches within a One Health continuum. Methods: Two hundred eighty-eight E. coli isolates collected in Alberta, Canada (2018–2019) from wastewater, well water, feces of broiler chickens and feedlot cattle, and retail beef and chicken meat were selected from existing surveillance collections using a stratified random sampling structure. Using short-read whole genome assemblies, phylogenetic relationships were inferred from pan-genome multiple sequence alignments. Principal coordinate analysis and permutational analysis of variance (PERMANOVA) of a Jaccard dissimilarity matrix derived from gene presence/absence data were used to investigate contributions of source and AMR strata to observe genetic dissimilarity. Population clustering and gene under- or over-representation by source and cluster were also explored. Results: Minimal phylogenetic segregation of isolates was noted based on source or AMR strata, and both contributed significant but small proportions of observed genetic dissimilarity, with the largest proportion attributed to phylogroup. There was notable diversity of E. coli within and between sources; however, in some larger clusters, differential gene presence/absence was potentially linked to ecological niche rather than source of isolation. Conclusions: This study highlights the ecological complexity of AMR in E. coli in non-clinical contexts, offering a novel lens on how niche-specific factors can influence population structure and AMR carriage. It also provides insight into apparent discrepancies in the literature regarding clustering of E. coli by source. These findings support a more integrative One Health approach to AMR surveillance, emphasizing the need to account for microbial diversity and niche-specific adaptation across interconnected systems. Full article

Roselle (Hibiscus sabdariffa L.) is a plant rich in bioactive constituents, serving as a unique material for the food and beverage industry and therapeutic applications. Despite its significant utility, few studies have focused on the molecular breeding of the plant. Chloroplasts are organelles in plant cells with independent genetic information, making them ideal for investigating plant phylogeny and genetic evolution. In this study, the roselle breeding material ‘Zhe Xiao Luo 1’ was selected to assemble and analyze the entire chloroplast genome using the Illumina NovaSeq X Plus platform. The phylogenetic relationships between roselle and other species within Malvaceae family, particularly within the genus Hibiscus, were clarified. The results showed that the complete chloroplast genome of roselle was 162,428 bp in length, with nucleotide proportions of 31.14% (A), 18.73% (C), 18.01% (G), 32.12% (T), and 36.74% (GC). It exhibited a typical tetrad structure consisting of four segments: the large single copy (LSC) region (90,327 bp), the small single-copy (SSC) region (19,617 bp), and two inverted repeat sequences (IRa and IRb, each 26,242 bp). A total of 130 genes were identified, including 37 tRNA genes, 8 rRNA genes, and 85 mRNA genes, and no pseudogenes were detected. Phylogenetic analysis using 23 revealed a clear phylogenetic relationship between H. sabdariffa and H. esculentus (okra) among all tested species. Building on previous research, this study further explored the functional annotation of genes in the roselle chloroplast genome, as well as its codon preference, repetitive sequences, simple sequence repeats (SSR), Ka/Ks ratio, nucleotide diversity (pi) analysis, and boundary analysis. The complete gene sequences have been uploaded to the NCBI database (accession number PX363576). This study provides evidence for elucidating the phylogenetic relationships and taxonomic status of H. sabdariffa, laying a theoretical foundation for studies on molecular mechanism resolution and cultivar development. Full article

The Influenza A Virus (IAV) is known to hijack cellular proteins during its replication. IAV infection increases the expression of Heat-shock-protein family A (Hsp70) member 5 (HSPA5) in human cells, but its specific function in the viral life cycle remains unclear. This study aims to elucidate the function of HSPA5 in IAV replication, by implementing HSPA5 knockdown (KD) in A549 cells and assessing its impact on IAV’s viral protein translation, genomic RNA transcription, and the host cellular proteome. HSPA5 KD significantly reduced progeny virus release, although viral RNA levels were unaffected. Interestingly, levels of viral structural proteins increased in HSPA5 KD cells after infection. Treatment with HSPA5 inhibitor also suppressed IAV replication, confirming its role as a host dependency factor. Proteomic profiling revealed 116 proteins altered in wild-type cells and 223 in HSPA5 KD cells, with 32 uniquely dysregulated in wild-type and 139 unique to HSPA5 KD cells. In HSPA5 knockdown cells, the altered proteins were linked to pathways such as EIF2, EGF, PEDF, CNTF, IL-13, and G-protein receptor signaling, as well as to cellular processes like lymphocyte activation and regulation of immune and blood cell death, which were not affected in wild-type cells after IAV infection. Overall, this study suggests that HSPA5 contributes to late stages of IAV replication, likely assembly or maturation, and represents a promising target for antiviral drug development. Full article

Polymyxin antibiotics are often the last line of defense against multidrug-resistant Gram-negative pathogens. A key resistance mechanism involves the addition of 4-amino-4-deoxy-L-arabinose (L-Ara4N) to lipid A, mediated by the bifunctional enzyme ArnA. However, the evolutionary rationale and structural basis for ArnA’s domain fusion, hexameric assembly, and catalytic coordination remain mechanistically unresolved. Here, we integrate evolutionary genomics, high-resolution cryo-electron microscopy (cryo-EM), and computational protein design to provide a comprehensive mechanistic analysis of ArnA. Our evolutionary analysis reveals that the dehydrogenase (DH) and formyltransferase (TF) domains evolved independently and were selectively fused in Gammaproteobacteria, suggesting an adaptive advantage. A 2.89 Å cryo-EM structure of apo-ArnA resolves the flexible interdomain linker and reveals a DH-driven hexameric architecture essential for enzymatic activity. 3D variability analysis captures intrinsic conformational dynamics, indicating a molecular switch that may coordinate sequential catalysis and substrate channeling. Structure-based peptide inhibitors targeting the hexamerization and predicted ArnA–ArnB interaction interfaces were computationally designed, offering a novel strategy for disrupting L-Ara4N biosynthesis. These findings illuminate a previously uncharacterized structural mechanism of antimicrobial resistance and lay the groundwork for therapeutic intervention. Full article

The expansion of genomes is a major evolutionary force, yet its role in facilitating adaptation to extreme environments remains enigmatic. Here, we investigate alpine Parnassius butterflies, a rare genus characterized by exceptionally large genomes, to unravel the interplay between genome architecture and high-altitude colonization. We present a new, 1.46 Gb draft genome assembly for Parnassius epaphus and perform a comparative analysis across six species. Our findings reveal a massive 3- to 5-fold genome expansion driven predominantly by Long Interspersed Nuclear Elements (LINEs). Counterintuitively, we discover that larger genomes possess a proportionally smaller fraction of young, active transposable elements (TEs), challenging the prevailing paradigm that recent TE proliferation is the primary driver of genome size. Instead, our temporal analysis demonstrates that this expansion is a legacy of two ancient TE waves (~8 and ~14 Mya), which remarkably coincide with major uplift phases of the Tibetan Plateau. We propose a model where the selective retention of these ancient TEs, mechanistically linked to major geological upheavals, provided the crucial genomic plasticity for colonizing Earth’s most extreme terrestrial habitats. This study re-frames TEs not merely as genomic parasites but as pivotal architects of adaptive genome evolution in response to profound environmental change. Full article

To enhance the L-threonine synthesis level in Escherichia coli, this study constructed screening markers rich in L-threonine rare codons. By replacing all the threonine codons in the protein sequences with a high proportion of threonine with L-threonine rare codons and linking them to the fluorescent proteins with the same replacement, high-throughput screening of L-threonine production mutant strains was achieved. To address the metabolic imbalance caused by overexpression of a single enzyme, an artificial multi-enzyme complex system was constructed based on the principle of cellulosome self-assembly. By co-locating ThrC-DocA and ThrB-CohA, the substrate transfer path was shortened, achieving a 31.7% increase in L-threonine production. Furthermore, combined with multi-copy chromosomal integration technology via CRISPR-associated transposase (MUCICAT) technology, the thrC-docA-thrB-cohA gene cluster was integrated into the genome of the high-yield strains obtained through screening, eliminating the plasmid-dependent metabolic burden and significantly enhancing genetic stability. The modular assembly of metabolic pathways by using cellulosome elements provides a new paradigm for the optimization of complex pathways and lays a theoretical and technical foundation for the efficient production of L-threonine. Full article

Background/Objectives: Bacteroides species are key members of the human gut microbiota but can act as opportunistic pathogens. This study investigated the genomic features of clinical Bacteroides isolates from southern Thailand. Methods: Sixteen isolates were collected from body fluids, tissues, and pus at Songklanagarind Hospital (2022–2024). Whole-genome sequencing was performed on the BGI platform, followed by genome assembly, annotation, average nucleotide identity (ANI), pairwise single-nucleotide polymorphism (SNP) analysis, antimicrobial resistance (AMR) gene profiling, plasmid prediction, virulence screening, and phylogenetic analysis. Results: ANI and SNP analysis revealed two clusters: one comprising B. ovatus, B. intestinigallinarum, and B. thetaiotaomicron, and another mainly B. fragilis with one B. hominis isolate. All isolates were resistant to ampicillin, cephalothin, and penicillin; six B. fragilis strains were resistant to all tested antibiotics. The β-lactamase gene cepA was detected in all B. fragilis isolates, and plasmids were predicted in two genomes. Three virulence types (capsule formation, lipopolysaccharide modification, and stress response) were identified. Phylogenomic analysis confirmed species-level assignments and revealed underrecognized lineages, emphasizing the value of genome-based approaches for accurate classification. Conclusions: Clinical Bacteroides isolates display diverse resistance and virulence profiles, highlighting the importance of strain-level genomic surveillance. Full article

This study examines the genomic composition and resistance potential of eight putative plasmid-derived contig assemblies reconstructed from marine Enterobacterales isolated in the central Adriatic Sea. Using a combination of Illumina-based whole genome sequencing, de novo assembly, and a multi-tool bioinformatics pipeline, we annotated antimicrobial resistance genes (ARGs), insertion sequences (ISs), and plasmid replicon types. Clinically significant resistance markers such as bla_KPC, bla_TEM, aacA4, tetA, and folP were identified, frequently co-localised with mobile genetic elements including IS110, IS4, and IS1182. The plasmid-associated contigs were assigned to MOBP and MOBQ types and contained replicon markers (IncP6, IncA/C2) characteristic of broad-host-range plasmids. Our findings provide valuable insight into the plasmidome of environmental Enterobacterales, emphasising the role of coastal pollution in shaping the distribution and potential mobility of antimicrobial resistance genes. This supports the One Health framework by linking environmental reservoirs to clinically relevant resistance mechanisms. Full article

Diospyros, the most species-rich woody plant genus in Ebenaceae, has attracted significant academic interest due to its ecological and economic importance. This study presented the first complete assembly and annotation of the chloroplast genome of Diospyros tsangii. The chloroplast genome measured 157,445 bp, with a typical quadripartite circular structure and 132 annotated coding genes. A comprehensive analysis of evolutionary traits and codon usage preferences across chloroplast genomes of 15 Diospyros species were conducted. The main objective was to provide a theoretical basis for understanding phylogenetic relationships and assessing genetic diversity within Diospyros. Our findings showed that genetic diversity in the IR regions of the chloroplast genomes is notably lower than that in the LSC and SSC regions. The boundary regions exhibited high conservation with minimal variation. Selected pressure analysis indicated that most coding genes are under purifying selection. Phylogenetic analysis showed that D. tangii was sister to Diospyros oleifera, and Diospyros kaki was closely related to Diospyros vaccinioides with high supporting values. The examination of codon usage patterns showed that the GC content at the first, second, and third codon positions of 52 protein-coding sequences followed the order GC1 > GC2 > GC3, with a preference for A or U bases at the third position. The effective number of codons ranged from 45.13 to 45.43, which indicated the weak codon bias. The neutral-plot, ENC-plot, and PR2-plot analysis suggested that natural selection predominantly influences the codon usage patterns in Diospyros plants. These results would be vital to understand the evolutionary dynamics of the genus Diospyros. Full article

Lycopene, a natural carotenoid with antioxidant and health-promoting properties, has attracted attention as a valuable compound for the food, pharmaceutical, and cosmetic industries. Conventional production methods based on plant extraction or chemical synthesis are limited by low yields, high costs, and environmental concerns. In this study, the oleaginous yeast Yarrowia lipolytica was engineered as an alternative microbial cell factory for sustainable lycopene biosynthesis using short-chain fatty acids (SCFAs)—such as acetate, butyrate, and propionate—as inexpensive, renewable carbon sources. Four heterologous genes from Pantoea agglomerans (crtI, crtB, crtE, and idi) were codon-optimized and integrated into the Y. lipolytica genome using different expression systems, including the Golden Gate Assembly strategy. Among the tested strains, PS05/4lyc/GGA, characterized by enhanced phospholipid biosynthesis, demonstrated the highest lycopene yield of 462.9 mg/g dry cell weight and a titer of 3.41 g/L on butyrate medium—values comparable to or exceeding those reported for bioreactor-scale fermentations. The results indicate that co-activation of phospholipid and carotenoid biosynthesis pathways creates favorable intracellular conditions for hydrophobic pigment accumulation. Moreover, the use of SCFAs improved acetyl-CoA availability and redirected carbon flux through the mevalonate pathway, enhancing productivity. Strains with elevated membrane lipid biosynthesis also exhibited higher metabolic stability and stress tolerance. Full article

Docosahexaenoic acid (DHA), an omega-3 polyunsaturated fatty acid essential for human health, is primarily produced at scale using Schizochytrium sp. Mutagenesis-based strain improvement has increased DHA yields, but the genetic and metabolic mechanisms underlying high productivity remain poorly understood. Here, we conducted the comparative whole-genome sequencing and transcriptomic profiling of a high-DHA-yielding mutant strain (HS01) and its parental strain (GS00). The GS00 genome assembly spans 62.4 Mb and encodes 14,886 predicted genes. Functional annotation highlighted pathways involved in central metabolism, saturated fatty acid (SFA) synthesis, and polyunsaturated fatty acid (PUFA)/DHA biosynthesis. Comparative genomics identified 40 insertions/deletions and 396 single-nucleotide polymorphisms between HS01 and GS00, including mutations in the coding and regulatory regions of key metabolic genes. Transcriptomic analysis revealed extensive metabolic reprogramming in HS01, including the upregulation of glycolysis and tricarboxylic acid (TCA) cycle genes, along with a distinct fatty acid profile and the altered expression of fatty acid metabolism genes compared with GS00. Collectively, the integrated genomic and transcriptomic analyses not only pinpointed specific mutations potentially associated with the HS01 high-DHA phenotype but also revealed substantial transcriptional and metabolic remodeling, providing valuable insights into the mechanisms that drive enhanced DHA biosynthesis. Full article

Claviceps purpurea is a specialized phytopathogenic fungus that infects grasses and produces pharmacologically active compounds, attracting considerable interest in genomic research. In this study, we assembled and annotated the complete mitogenomes of 15 C. purpurea strains isolated from different host plants, including seven newly sequenced isolates from China. Analysis of the C. purpurea pan-mitogenome demonstrated that the accessory regions, with an average proportion of 48.23%, are the main contributor to mitogenome variation. Analysis of the 14 protein-coding genes revealed Ka/Ks ratios below 1, indicating strong purifying selection. Notably, the atp9 gene was absent in all strains, suggesting a potential adaptive gene loss. Structural variations were predominantly located in the intergenic region between rns and rnl. Phylogenetic analysis based on concatenated mitochondrial genes placed Claviceps as most closely related to the genus Epichloë. The 15 C. purpurea strains grouped into five well-supported subclades, with Chinese and non-Chinese isolates forming distinct lineages. Among these, the Chinese strains ACCC 37001 and KUNCC 11030 represented the earliest diverging lineages. This study elucidates the intraspecific variation and evolutionary patterns of the mitogenome in C. purpurea and highlights the value of mitogenome in resolving phylogenetic relationships. Full article

Grapes (Vitis spp.) are a globally significant fruit crop with a long history of cultivation and substantial cultivar diversity. Their high genetic differentiation and complex evolutionary history make them a valuable system for studying plant evolution. The chloroplast genome, known for its structural conservation and uniparental inheritance, offers a reliable molecular marker for phylogenetic reconstruction. In this study, we sequenced and assembled the complete chloroplast genomes of nine representative grape cultivars, analyzed their phylogenetic relationships, and compared structural variations. All chloroplast genomes displayed a typical quadripartite structure, with high conservation in genomic architecture, gene order and content, codon usage, and simple sequence repeats (SSRs). However, additional sequence comparisons revealed seven regions with high variation, including the genes rbcL and ndhF, and the intergenic regions rps16-trnQ, ndhC-trnV, accD-psaI, ndhF-rpl32, and trnL-ccsA. At the same time, seven natural variation sites were identified in the amino acid sequences of rbcL and ndhF. Additionally, the study’s maximum likelihood (ML) phylogenetic trees and photosynthetic index measurements suggest that developmental characteristics of grape photosynthesis may be related to the evolutionary origins of different populations. This phylogenetic classification not only elucidates the evolutionary origins of these germplasm resources but also provides a foundation for molecular-assisted breeding by identifying distinct genetic groups. Full article

Gene duplication, a major source of new genes in evolution, often occurs via reverse transcription of mRNA, leading to the integration of a retrocopy into a new genomic locus. Here, we performed an in-depth analysis of the evolutionary history of the e(y)2 gene in Metazoa. The E(y)2 protein is a shared subunit of two highly conserved complexes involved in transcription regulation (the DUB module of the SAGA complex) and mRNA transport (TREX-2). In Deuterostomes, the e(y)2 gene has undergone multiple independent retropositions, often giving rise to functional retrogenes. In contrast, among Protostomes, duplications of e(y)2 were identified only in Drosophilinae and a member of the Lepidoptera family (Manduca sexta). In Drosophila, the retrocopy e(y)2 acquired an almost ubiquitous expression pattern and compensates for the function of the parental gene in all tissues except the testes. The parental gene, e(y)2b, evolved a testis-specific expression pattern, lost the ability to incorporate into the DUB module, but retained nuclear envelope localization and the capacity to assemble into the TREX-2 complex. Knockout of the D. melanogaster e(y)2b gene resulted in reduced male fertility. Overall, our study highlights distinct evolutionary trajectories of the e(y)2 gene in Deuterostomes and Protostomes. Full article

Stream-dwelling fishes face diverse hydrological pressures, making the broadly distributed Opsariichthys bidens an ideal model for analyzing adaptive evolution. To elucidate its adaptation to a high-dissolved-oxygen and high-flow-velocity stream environment, a high-quality genome with comprehensive annotation is essential. In this study, we present the first telomere-to-telomere (T2T) reference genome for O. bidens, constructed using PacBio HiFi, Oxford Nanopore Ultra-long, and Hi-C technologies. The assembled genome spans 841.96 Mb, comprising 38 chromosomes, each in a single contig (contig N50 = 22.42 Mb, 2.5-fold higher than the previous version), achieving a gap-free standard with 99.34% BUSCO completeness. Additionally, 38 centromeric sequences, 37 double-telomeric sequences, and 1 single-telomeric sequence were successfully identified, providing essential molecular markers. Phylogenetic analysis revealed a divergence time of 13.5 million years between O. bidens and its closely related species Z. platypus, with collinearity analysis confirming their high genomic conservation. Gene family analysis revealed 350 expanded families enriched in pathways associated with adaptation to high-dissolved-oxygen environments (e.g., antioxidant defense, oxidative phosphorylation, mitochondrial electron transport chain) and high-flow-velocity environments (e.g., exercise endurance, myocardial contraction, actin binding). Positive selection analysis further identified multiple pathways and key genes involved in mitochondrial optimization, oxygen utilization, and metabolic regulation. The T2T assembly greatly improves assembly continuity and enabling precise identification of centromeres and telomeres for O. bidens. These results provide a robust foundation for studying its adaptive evolution to stream environment. Full article