MetaboLights

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ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14221/m_MTBLS14221_LC-MS_negative_reverse-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14221/m_MTBLS14221_LC-MS_positive_reverse-phase_v2_maf.tsvftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14221/a_MTBLS14221_LC-MS_negative_reverse-phase.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14221/a_MTBLS14221_LC-MS_positive_reverse-phase.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14221/s_MTBLS14221.txtftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14221/i_Investigation.txtprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14221Metabolite identification was performed using Progenesis QI software (version 3.0, Waters Corporation, Milford, USA). Features were identified by matching the MS and MS/MS mass spectral information against the following databases:- HMDB (Human Metabolome Database, http://www.hmdb.ca/)- Metlin (https://metlin.scripps.edu/)- Self-built database (Majorbio)The mass tolerance for MS identification was set to less than 10 ppm. Identification was based on accurate mass matching and MS/MS spectral matching scores. Metabolite identifications were reported according to the Metabolomics Standards Initiative (MSI) confidence levels.MetaboLightsPublicLiquid Chromatography MS - negative - reverse-phaseLiquid Chromatography MS - positive - reverse-phaseChromatography Instrument: Thermo Scientific Vanquish Horizon UHPLC systemAutosampler model: Thermo Scientific Vanquish autosamplerColumn model: ACQUITY UPLC HSS T3 column (100 mm × 2.1 mm i.d., 1.8 μm; Waters, Milford, USA)Column type: reverse phaseGuard column: None usedMobile phase A: 95% water + 5% acetonitrile (containing 0.1% formic acid)Mobile phase B: 47.5% acetonitrile + 47.5% isopropanol + 5% water (containing 0.1% formic acid)Flow rate: Not specified (typical UHPLC flow rate: 0.3-0.4 mL/min)Injection volume: 3 μLColumn temperature: 40°CFoHsf1 reprograms central metabolism via caffeine and vitamin D2 to modulate development and virulence of Fusarium oxysporum on Polygonatum kingianum.Lei ZhangYunnan University of Chinese Medicinemyceliamass spectrometry assayApproximately 20 ± 5 mg of frozen mycelial sample was weighed into a 2 mL centrifuge tube containing one 6 mm diameter grinding bead. Extraction was performed by adding 400 μL of extraction solvent (methanol:water = 4:1, v/v) containing four internal standards (including L-2-chlorophenylalanine at 0.02 mg/mL). The sample was ground using a frozen tissue grinder for 6 min (-10°C, 50 Hz), followed by low-temperature ultrasonic extraction for 30 min (5°C, 40 kHz). The sample was then placed at -20°C for 30 min and centrifuged at 13,000 × g for 15 min at 4°C. The supernatant was transferred to an autosampler vial for LC-MS analysis. A pooled quality control (QC) sample was prepared by mixing 20 μL of supernatant from each individual sample.Fusarium oxysporumhttps://www.ebi.ac.uk/metabolights/MTBLS14221Lei Zhang. Yunnan University of Chinese Medicine. zhlei318@163.com.Raw data files were processed using Progenesis QI software (version 3.0, Waters Corporation, Milford, USA). The processing workflow included baseline filtering, peak detection, peak integration, retention time correction, and peak alignment. Features were identified by matching MS and MS/MS mass spectral information against public databases (HMDB, Metlin) and a self-built database. The mass tolerance for MS identification was set to less than 10 ppm. The resulting data matrix containing retention time, m/z, and peak intensity information was generated for downstream statistical analysis.GenotypeStrainzhlei318@163.comMycelia of the WT, DEL, and COM strains of Fusarium oxysporum were harvested during the logarithmic growth phase by filtration, immediately frozen in liquid nitrogen, and stored at -80°C until metabolite extraction.MetabolomicsThermo Scientific Vanquish UHPLC SystemMetabolomicsmyceliaFusarium oxysporumPolygonatum kingianumuntargeted analysisThermo Scientific Q Exactive PlusHeat Shock Transcription FactorsThermo Scientific Vanquish UHPLC SystemMetabolomicsmyceliaPolygonatum kingianumFusarium oxysporumuntargeted analysisThermo Scientific Q Exactive PlusHeat Shock Transcription FactorsInstrument: Thermo Scientific Exploris 240 mass spectrometerIon source: Electrospray ionization (ESI)Scan polarity: Positive (for POS assay) / Negative (for NEG assay)Scan m/z range: 70-1050Mass analyzer: OrbitrapResolution: 60,000 (at m/z 200)Sheath gas flow rate: 40 arbAux gas flow rate: 10 arbCapillary temperature: 320°CHeater temperature: 300°CSpray voltage: 3.5 kV (positive) / 2.8 kV (negative)AGC target: 1e6Maximum injection time: 100 msfalseFoHsf1 reprograms central metabolism via caffeine and vitamin D2 to modulate development and virulence of Fusarium oxysporum on Polygonatum kingianumRoot rot has severely affected the yield and quality of Polygonatum kingianum, and Fusarium oxysporum has been identified as the primary causal agent. To explore the virulence mechanisms of this pathogen, we performed a functional characterization of FoHsf1, a gene encoding a heat shock transcription factor in F. oxysporum. Using targeted gene knockout and complementation, phenotypic assays, and virulence tests, we found that the FoHsf1 deletion mutant (DEL) exhibited significant defects in mycelial growth, conidiation, and virulence relative to the wild-type (WT) and complemented (COM) isolates. Integrated transcriptomic and metabolomic profiling demonstrated that differentially expressed genes (DEGs) and differentially accumulated metabolites (DAMs) were predominantly enriched in carbohydrate and amino acid metabolism pathways. More importantly, the metabolite abundance of caffeine and vitamin D2 was significantly reduced in DEL; exogenous application of these two metabolites partially restored fungal growth. These findings suggest that FoHsf1 regulates fungal development by modulating core metabolic pathways. Our results advance the current understanding of the molecular mechanism underlying FoHsf1-mediated virulence of F. oxysporum and offer potential targets for managing this disease.2026-04-112026-04-05MTBLS14221