Search Results (325)

Streptococcus cristatus (S. cristatus) belongs to the viridans group of streptococci and is a commensal of the human upper respiratory tract. With the non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase, GapN, and the oxidative part of the pentose phosphate pathway (oxPPP), S. cristatus can use two different metabolic pathways to provide reduced nicotinamide adenine dinucleotide phosphate (NADPH), an essential cofactor of anabolic reactions such as fatty acid and amino acid biosynthesis. Regarding their NADP-reducing capacity, streptococci can be categorized into three groups: those that have only GapN, those that use only the oxPPP, and those that use both pathways. Here, we report on the experimental and computational characterization of the catalytic properties of the three NADP-reducing enzymes: GapN, glucose-6-phosphate dehydrogenase (G6PDH), and 6-phosphogluconate dehydrogenase (6PGDH) of S. cristatus. Kinetic analyses showed moderate substrate and cofactor affinities, with GapN displaying the tightest substrate binding, followed by 6PGDH and G6PDH, in agreement with structural and computational predictions. All three enzymes preferentially utilized NADP⁺, with only G6PDH exhibiting limited NAD⁺ promiscuity. Growth-phase-dependent activity patterns suggest dynamic adjustment of NADPH-generating pathways, with reduced GapN contribution and sustained oxPPP activity in the stationary phase. Regulatory screening indicated limited allosteric control, though feedback inhibition by NADPH and the ATP sensitivity of G6PDH point to conserved redox regulatory mechanisms. Comparative analysis across streptococci supports the concept that the coexistence of GapN and the oxidative pentose phosphate pathway in S. cristatus may provide metabolic flexibility by offering alternative routes for NADPH generation. Full article

Background/Objective: Cinnamaldehyde (CA) is a naturally occurring bioactive compound derived from the leaves, bark, roots, and flowers of the Chinese medicinal plant Cinnamomum cassia. It exhibits a broad spectrum of pharmacological properties, encompassing antioxidant, antibacterial, anti-diabetic, antifungal, and anticancer activities. Notably, it has shown potential therapeutic benefits in the management of type 2 diabetes mellitus (T2DM) and non-alcoholic fatty liver disease (NAFLD). Ochratoxin A (OTA), a common contaminant found in foods such as cereals, coffee, and raisins, is also present in traditional Chinese medicinal materials, including Astragalus and liquorice. T2DM and NAFLD share intertwined pathophysiological pathways, including insulin resistance, dyslipidaemia, chronic low-grade inflammation and oxidative stress, with insulin resistance serving as the common pathological hub for both conditions. Consequently, they frequently co-occur and exacerbate each other. OTA exerts dual-targeted toxicity to the pancreas and liver, which may synergistically drive the development of the comorbidity of T2DM and NAFLD. These two processes are mutually causal and together constitute the pathological basis of metabolic comorbidity. Methods: Network toxicology employs toxicological data, gene expression, and protein–protein interaction (PPI) networks to predict the targets of toxins, while network pharmacology, based on systems biology principles, reveals how drugs exert regulatory effects through multiple targets and pathways. In this study, we employed an integrated network toxicology and network pharmacology approach to jointly decipher the potential mechanisms by which CA intervenes in OTA-induced comorbid T2DM-NAFLD. First, a network toxicology approach was employed to preliminarily screen for core toxicological targets responsible for OTA’s pathogenicity. Subsequently, network pharmacology was used to identify potential targets of CA-mediated intervention in the disease. Finally, the common overlap among the CA intervention targets, OTA toxicity targets, and disease targets was defined as the final set of potential targets for CA-mediated intervention in OTA-induced T2DM-NAFLD comorbidity. A PPI network was constructed using the STRING database, and topological analysis was performed with Cytoscape. Core targets were selected using the median values of six parameters—betweenness centrality, closeness centrality, degree centrality, eigenvector centrality, LAC (local average connectivity) score, and network centrality—as cut-off thresholds, and the top 10 key genes were further identified using the cytoHubba plugin. Gene Ontology (GO) functional enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analyses were conducted via the DAVID database, and the results were visualized on the CNSknowall platform. Lastly, molecular docking of the core targets was performed using the CB-DOCK2 platform to validate binding affinity. Results: Based on an integrated analysis of network toxicology, network pharmacology, and molecular docking, 10 key targets were systematically identified. These may serve as potential mediators of cinnamaldehyde in the treatment of OTA-induced T2DM-NAFLD comorbidity. Among these, six targets—albumin (ALB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), interleukin-6 (IL-6), tumor necrosis factor (TNF), actin beta (ACTB), and estrogen receptor 1 (ESR1)—possess crystal structures amenable to molecular docking. KEGG enrichment analysis revealed that CA and OTA jointly participate in key pathological processes such as the cancer pathway, the lipid and atherosclerosis pathway, the advanced glycation end-products–receptor for advanced glycation end-products (AGE-RAGE) signaling pathway, the phosphatidylinositol 3-kinase–protein kinase B (PI3K-Akt) signaling pathway, the TNF signaling pathway, and the interleukin-17 (IL-17) signaling pathway. OTA exacerbates inflammatory responses, impairs insulin signaling, promotes hepatic steatosis, and disrupts systemic metabolic homeostasis, ultimately contributing to T2DM-NAFLD comorbidity. Conversely, cinnamaldehyde counteracts these pathological processes through multiple mechanisms, including antioxidant and anti-inflammatory effects as well as regulation of glucose and lipid metabolism, thereby restoring metabolic homeostasis. Conclusions: This study has preliminarily identified the toxicological targets of OTA and the potential intervention targets of CA, offering new avenues for preventing and intervening in OTA-induced metabolic toxicity. Furthermore, it provides a theoretical basis for CA as a potential multi-target therapeutic agent and presents novel insights worthy of further investigation into the prevention of T2DM-NAFLD comorbidity. Full article

Low-frequency alternating magnetic field (LF-AMF) was applied to liquid fermentation of Pleurotus citrinopileatus for the production of extracellular polysaccharides (EPS). Two types of EPS (POL1 and POL2) were secreted by P. citrinopileatus under normal fermentation conditions, while only one type of EPS (POL3) was produced under LF-AMF-assisted fermentation. The study demonstrated that LF-AMF-assisted fermentation altered the monosaccharide composition, molar ratios of component monosaccharides, and molecular weight distribution of EPS. LF-AMF enhanced the flocculation activity of EPS in a concentration-dependent and dosage-specific manner. Transcriptomic analysis revealed a total of 32,413 differentially expressed genes (DEGs) (|log2(fold-change)| ≥ 1, p < 0.05) between P. citrinopileatus under LF-AMF-assisted fermentation and normal fermentation. Of those, 5818 genes were up-regulated and 26,595 genes were down-regulated. The DEGs were enriched in metabolic pathways and biosynthesis of secondary metabolite. Genes involved in glycolysis and gluconeogenesis, such as hexokinase, 6-phosphofructokinase, fructose-1,6-phosphate aldolase, glyceraldehyde 3-phosphate dehydrogenase and glucose-1,6-diphosphatase were up-regulated. Additionally, genes in the tricarboxylic acid (TCA) cycle, including citrate synthase, isocitrate dehydrogenase and α-ketoglutarate dehydrogenase were also up-regulated. These findings suggest that LF-AMF-induced transcriptional alterations in glycolysis may modulate the metabolic supply of precursors for EPS biosynthesis, thereby affecting the biosynthetic process and altering the compositional characteristics of EPS. Full article

Reliable reference genes are critical for ensuring the accuracy of RT-qPCR-based gene expression analysis, especially under environmental stress conditions. In this study, Leymus chinensis was used as the experimental material, and eight candidate reference genes—alpha-tubulin (TUA), beta-tubulin (TUB), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), elongation factor 1-alpha (EF1α), 18S ribosomal RNA (18S rRNA), adenylyl cyclase-associated protein (CAP), adenine phosphoribosyl transferase (APRT), and actin (ACT)—were selected to evaluate their expression stability under cold, drought, heat, NaCl, high pH, wounding, abscisic acid (ABA) and jasmonic acid (JA) treatments. Primer specificity and amplification efficiency were first assessed, and the candidate genes were then comprehensively analyzed using geNorm, NormFinder, BestKeeper, and RefFinder. The results showed that the amplification efficiencies of all primers ranged from 95.0% to 107.2%, and the Ct values of the candidate genes ranged from 17.31 to 30.99. Comprehensive analysis using RefFinder showed that ACTIN was the most stable gene under ABA and NaCl treatments, EF1α under heat and wounding treatments, CAP under JA and high pH treatments, APRT under cold treatment, and TUB under drought treatment. geNorm analysis indicated that two reference genes were sufficient for accurate normalization under each treatment condition. The reliability of the screening results was further confirmed by expression-level validation of LcbZIP46, LcWRKY5, and LcFIN1. This study provides a stable reference gene system for RT-qPCR-based expression analysis in Leymus chinensis under environmental stress conditions. Full article

Background/Objectives: Early childhood caries (ECC) remains a major public health concern, with Streptococcus mutans as a primary etiological agent. Current treatments rely on broad-spectrum antimicrobials, which can disrupt the oral microbiome and promote resistance. This study applied a structure-based in silico pipeline to identify molecule modulators of Toll-like receptor 2 (TLR2), a key host receptor implicated in ECC, and to explore their binding potential against major S. mutans proteins. Methods: ECC-related genes were collected from public databases and analyzed by functional enrichment and protein–protein interaction (PPI) network analysis. Hub genes were ranked using centrality algorithms. Virtual screening on TLR2 (DrugCLIP) was followed by molecular docking of selected compounds against the TLR1/TLR2 heterodimer and 50 S. mutans proteins, complemented by in silico absorption, distribution, metabolism, excretion, and toxicity (ADMET) profiling. Results: Fifty-four ECC-related genes and nine hub genes were identified, with TLR2 and cathelicidin antimicrobial peptide (CAMP) as central nodes. Virtual screening yielded five lead compounds fulfilling drug-likeness and toxicity criteria. Docking to TLR1/TLR2 showed favorable binding energies, with Z7684613096 showing the most consistent binding. V026-2549 displayed the highest number of strong interactions with S. mutans targets, including dTDP-glucose 4,6-dehydratase (rmlB), NADP-dependent glyceraldehyde-3-phosphate dehydrogenase (gapN), glucosyltransferase C (gtfC), and 5-methyltetrahydropteroyltriglutamate-homocysteine methyltransferase (metE). Conclusions: Five candidate compounds with promising dual activity against TLR1/TLR2 and S. mutans proteins were prioritized for experimental validation, including TLR2 functional assays and in vitro anti-biofilm studies. Full article

Water deficit (WD) irrigation has garnered considerable attention for its potential to enhance crop water use efficiency. However, limited research has been conducted on its ability to improve tomato fruit sweetness-related traits without significantly affecting yield. In this study, we investigated the effects of varying WD levels [T1–T4: 80%, 65%, 55%, and 45% of field capacity (FC)] compared with full irrigation (CK: 90% FC) on tomato fruits from the mature green to red-ripe stages, aiming to evaluate yield, textural attributes, and sugar composition. Notably, the fruit yield per plant under T2 treatment was not significantly different from that under CK. Compared with CK, T2 significantly reduced fruit water content by 2.39% while markedly increasing individual fruit dry weight by 13.61%. At 44 days after flowering, fruit firmness under T2 showed no substantial difference from CK, whereas adhesiveness was markedly elevated by 33.85%. Furthermore, T2 substantially boosted the activities of key Calvin cycle enzymes (ribulose-1,5-bisphosphate carboxylase/oxygenase, glyceraldehyde-3-phosphate dehydrogenase, fructose-1,6-bisphosphatase, fructose-1,6-bisphosphate aldolase, and transketolase) in tomato leaves, thereby increasing the photosynthetic rate and consequently elevating the total sugar content in tomato fruits. Additionally, T2 stimulated the activities of sucrose-hydrolyzing enzymes (acid invertase and neutral invertase) in fruits, leading to increased fructose and glucose accumulation at the red-ripening stage, with respective increases of 69.60% and 34.67% relative to CK. Multivariate classification based on principal component analysis and hierarchical cluster analysis revealed that T2 and T3 were distinctly separated from other treatments in terms of yield, texture parameters, and sugar profiles. These findings provide a valuable strategy for applying WD to improve fruit sugar composition without compromising yield. Full article

Iodoacetic acid (IAA), a highly cytotoxic disinfection byproduct commonly detected in drinking water, poses a potential risk to female reproductive health. The direct molecular mechanisms underlying its effects on the reproductive system epithelium remain unclear. This study demonstrates that IAA induces glycational stress in primary porcine uterine (UECs) and oviduct epithelial cells (OECs), representing an early event contributing to extensive cellular toxicity. IAA exposure inhibited Glyceraldehyde-3-Phosphate Dehydrogenase (GAPDH) enzymatic activity and promoted the accumulation of advanced glycation end products (AGEs) Nε-(carboxymethyl)lysine (CML), triggering mitochondrial dysfunction, redox imbalance, calcium dyshomeostasis, and endoplasmic reticulum stress. These disturbances activated a dysregulated signaling network involving the p38 MAPK, AKT, and NF-κB pathways, ultimately causing G1/S cell cycle arrest and apoptosis. Notably, pretreatment with the AGE inhibitor pyridoxamine reduced CML accumulation, restored mitochondrial function, and alleviated apoptotic cell death. These findings identify glycational stress as a key initiating mechanism for IAA-induced reproductive epithelial toxicity, providing mechanistic insight into the potential health risks of environmental disinfection byproducts. Full article

Enterocytozoon hepatopenaei (EHP) is a specialized parasitic microsporidian that causes significant economic losses to the shrimp farming industry. The glycolysis pathway plays an important role in the survival of EHP spores in vitro. In this study, key enzyme genes involved in the glycolysis pathway of EHP were analyzed, and purified spores were treated with KOH and a low temperature (−20 °C) to promote or inhibit germination. Quantitative analysis and enzyme activity of the initiating key gene hexokinase (HK) and the core link gene glyceraldehyde 3-phosphate dehydrogenase (GAPDH) were conducted to explore the energy response characteristics of germinating spores. The results showed that HK and GAPDH genes had significant differences from the host, based on phylogenetic analysis. The expression of HK gene and enzyme activity increased after promoting germination treatment. The expression of the GAPDH gene was stable, but the activity of the GAPDH enzyme increased significantly after germination promotion. These findings indicate that the inhibition of the HK gene expression level and GAPDH protein level can block spore germination and invasion in vitro, which could be used as potential control targets for EHP. Full article

Reactive oxygen species (ROS) are an essential component for the maintenance of cellular function. However, if produced in excess, ROS can drive cellular dysfunction and compromise cell viability. Indeed, uncontrolled ROS production plays a pivotal role in the pathogenesis of type 2 diabetes (T2D), contributing to the loss of β-cell function and the impairment in insulin signalling, as well as driving the development of diabetic complications, which can severely compromise quality of life. T2D is characterised by persistent hyperglycaemia, which is a leading contributor to ROS overproduction in this disease state. This enhanced, almost uncontrolled, increase in glucose metabolism upregulates several ROS-producing pathways, including the hexosamine pathway, protein kinase C, NADPH oxidase and the mitochondrial electron transport chain. There is accumulating evidence to suggest that in a bid to preserve redox homeostasis, ROS acts to suppress glucose metabolism by inactivating several enzymes involved in the regulation of glycolytic flux, including glucokinase, glyceraldehyde 3-phosphate dehydrogenase, phosphofructokinase-1 and pyruvate kinase. Nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) is a multi-faceted transcription factor, with a central role in ROS signalling and redox homeostasis. Whilst NF-κB mediates the transcriptional regulation of many pro-oxidants, NF-κB activity is also regulated by the oxidative status, with ROS having both inhibitory and stimulatory roles in these signalling pathways. Interestingly, NF-κB is also involved in controlling the delicate balance between glycolytic flux and mitochondrial respiration. This review will summarise the interplay linking hyperglycaemia with ROS formation, emphasising the role of glucose metabolism in the process, and the crosstalk of these pathways with NF-κB. Full article

Phosphofructokinase 1 (PfkA) mediates the ATP-dependent phosphorylation of fructose-6-phosphate and is a key, controlling enzyme in glycolysis for Escherichia coli and other organisms. In this study, 22 chromosomally expressed PfkA variants were constructed in E. coli C. These variants, the wild-type strain, and the ∆pfkA strain were compared for growth rates using glucose as the sole carbon source. The majority of variants (14 of 22) attained a growth rate less than 20% of the growth rate of the wild-type strain (0.94 h⁻¹) and thus similar to the knockout strain (0.12 h⁻¹). Three variants (R171S, F76Y, and R77A), representing a range of growth phenotypes, and strains expressing the wild-type PfkA and the ∆pfkA deletion strain were additionally examined for key intracellular metabolites and gene expression under nitrogen-limited steady-state conditions. These five strains could be distinguished by two groupings: strains with relatively high growth rates under batch conditions (wild-type and R77A variant) showed the greatest glucose consumption rate and formed acetate, whereas strains with low growth rates (F76Y, R77A, and ∆pfkA) exhibited low glucose consumption and did not accumulate acetate. As the PfkA mutation severity increased, the intracellular concentrations of acetyl-CoA and fructose-1,6-bisphosphate and the sum of dihydroxyacetone and glyceraldehyde-3-phosphate greatly decreased. Although the mutation severity had a limited effect on the expression of maeB and icd genes expressing malic enzyme and isocitrate dehydrogenase, it correlated with reduced expression of zwf and pta genes expressing glucose-6P-dehydrogenase and phosphotransacetylase, respectively. The results highlight the great sensitivity of the enzyme to substitutions and the key role it plays in controlling glycolytic flux. Full article

Targeting the Warburg effect (aerobic glycolysis) in tumor cells represents a promising metabolic therapeutic strategy in cancer research. This review analyzes the regulatory mechanisms and therapeutic potential of key glycolysis pathway components, including glucose transporters (GLUTs) and glycolytic enzymes such as hexokinase 2 (HK2), phosphofructokinase (PFK), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), pyruvate kinase M2 (PKM2), and lactate dehydrogenase A (LDHA). We evaluate the molecular mechanisms of various inhibitors and the current clinical development landscape, noting that limitations of monotherapy stem not only from tumor metabolic plasticity but also largely from the unacceptable toxicity of many inhibitors due to the essential role of glycolysis in normal cell metabolism. Furthermore, we explore the molecular basis of synergistic interactions between glycolysis inhibitors and chemotherapy, radiotherapy, immunotherapy, photothermal therapy, and targeted therapy, proposing that rational combination strategies may help overcome resistance and improve therapeutic efficacy. Finally, the review outlines future challenges and directions, emphasizing that the primary obstacle in metabolic treatments is achieving selective inhibition of glycolytic enzymes in cancer cells while sparing normal cells. To address this challenge, the development of high-selectivity agents, cancer-specific nanodelivery systems, precise biomarker identification, and innovative combination regimens based on metabolic-immune regulation is crucial for advancing glycolysis-targeted therapy toward clinical translation. Full article

Fallopia multiflora (Thunb.) Harald is a valuable medicinal plant with substantial economic and therapeutic value, widely cultivated in southern China. In 2025, a leaf disease outbreak occurred in an F. multiflora plantation in Tongnan District, Chongqing, China. Diseased samples were collected for pathogen isolation, and seven representative strains were selected from 50 pure isolates via preliminary pathogenicity tests. Species identification was performed using a combination of morphological characterization and multi-locus phylogenetic analysis, targeting the Internal Transcribed Spacer (ITS), Translation Elongation Factor 1-alpha (TEF1), RNA Polymerase II Second Largest Subunit (RPB2), Beta-tubulin (β-TUB2), Heat Shock Protein 60 (HSP60), and Glyceraldehyde-3-Phosphate Dehydrogenase (G3PDH) gene regions. The isolates were identified as Alternaria alternata, Botrytis cinerea, Botrytis polygoni, Epicoccum mackenziei, and Lasiodiplodia citricola. Pathogenicity assays on living F. multiflora leaves confirmed that all identified species could induce disease symptoms, with distinct interspecific differences. This study verifies that multiple pathogenic fungi can infect F. multiflora, with potential co-infection. It improves our understanding of the pathogenic fungal community associated with this medicinal plant and lays a foundation for subsequent disease management in its cultivation. Full article

Accurate normalization of RT-qPCR data requires selecting stable internal control genes, particularly in models characterized by dynamic metabolic transitions, such as 3T3-L1 adipocytes. The current study compares the expression stability of nine widely used housekeeping genes (HKGs) (peptidylprolyl isomerase A (Ppia), glyceraldehyde-3-phosphate dehydrogenase (Gapdh), beta-2 microglobulin (B2M), ribosomal protein, large, P0 (36b4), hydroxymethylbilane synthase (Hmbs), hypoxanthine guanine phosphoribosyl transferase (Hprt), tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein, zeta polypeptide (Ywhaz), 18S ribosomal RNA (18S), and β-actin (Actb)) across key stages of differentiation (days 0, 9, and 18) and under treatments with palmitic acid and docosahexaenoic acid. Stability was assessed using four classical algorithms—geNorm, NormFinder, BestKeeper, and RefFinder—supplemented by the ΔCt method, conventional statistical testing, correlation, and regression analysis relative to two target genes, fatty acid-binding protein 4 (Fabp4) and sterol regulatory element binding transcription factor 1 (Srebf1). The obtained data indicate that no single HKG remains universally stable across these experimental conditions, and the expression of traditionally used reference genes (Gapdh, Actb, Hprt, 18S) is highly influenced by both the stage of adipogenesis and exposure to lipid-modulating factors. In contrast, Ppia, 36b4, and B2M—despite some of them being underestimated in use as references—consistently display the lowest variability across most analytical tools, forming a reliable and functionally diverse normalization panel. It should be noted that our initial stability assessment revealed apparent discrepancies among mathematical evaluation methods, emphasizing the need for a holistic, multiple-level approach strategy. The applied combination of algorithmic and statistical methods provides a more rigorous and objective framework for assessing the stability of reference genes, which is highly recommended in such a complex adipocyte-based model. Full article

Quantitative real-time PCR (qPCR) remains a cornerstone method for analyzing gene expression due to its high sensitivity, specificity, and reproducibility. However, for reliable results in relative quantification studies, the choice of an appropriate reference gene is critical to ensure accurate normalization. The expression of commonly used reference genes can vary depending on developmental stage and experimental conditions, making their validation essential. To date, no validated reference genes have been reported for Agrostemma githago L. (corn cockle, Caryophyllaceae). To facilitate research on genes involved in natural product biosynthesis and specialized metabolism regulation, we aimed to identify the most stable reference genes across various plant organs and cultivation conditions of this species. Drawing on previous literature, we have selected seven housekeeping genes widely used for evaluation: actin, β-tubulin, elongation factor 1α, glyceraldehyde-3-phosphate dehydrogenase, histone H3, translation elongation factor 1, and eukaryotic translation initiation factor 5A1 (for which two primer sets were tested). The nucleotide sequences of these potential reference genes were identified from the A. githago transcriptome. Using qRT-PCR, transcript levels of seven potential reference genes were estimated in 40 different A. githago samples, including 25 in vitro samples under various treatment conditions and 15 soil-grown samples representing A. githago organs in different developmental stages. Expression stability of candidate reference genes was assessed using the RefFinder platform, which combines four commonly applied statistical algorithms: geNorm, NormFinder, BestKeeper, and the comparative Δ-Ct method. The results revealed that the selection of optimal reference genes varied based on the particular organ, developmental stage and condition being examined. TIF5A1-2 (one of the two primer pairs tested) and GAPHD consistently exhibited the most stable expression under various conditions in vitro. EF1α and H3 exhibited superior performance across different organs of soil-grown plants. Moreover, our integrated analysis enabled the identification of the two most stable, universal reference genes suitable for normalization in A. githago under all tested conditions—H3 and TIF5A1-2. Our work provides a robust foundation for future transcriptomic and functional studies of the specialized metabolism of A. githago and other related species. Full article

Cornus officinalis is a valuable traditional Chinese medicinal (TCM) plant species with both therapeutic and ornamental attributes. It is widely used in TCM prescriptions to nourish the liver and kidneys and constitutes a critical component of numerous classical formulas. In recent years, the large-scale cultivation of this medicinal plant has been expanded in Xixia County, Henan Province, China. Field investigations have revealed widespread brown leaf spot, accompanied by reductions in yield and quality. In this study, symptomatic leaves were collected for pathogen isolation. Tissue isolations consistently yielded a Colletotrichum fungus, and morphology combined with multi-locus phylogenetic analyses (the internal transcribed spacer, glyceraldehyde-3-phosphate dehydrogenase, chitin synthase, actin, and β-tubulin) identified the pathogen as Colletotrichum siamense. Pathogenicity assays (conducted by either wounding and inoculating detached leaves with a mycelium plug or spraying a conidium suspension on healthy potted plants) reproduced field symptoms, and the pathogen was re-isolated, thereby fulfilling Koch’s postulates. In vitro fungicide assays showed that carbendazim, tebuconazole, and prochloraz were highly effective against the pathogen, providing preliminary information for chemical management. This is the first documentation of C. siamense causing leaf anthracnose on C. officinalis and provides a basis for developing targeted control strategies to mitigate disease impacts and preserve yield and quality. Full article