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negative - reverse-phaseLiquid Chromatography MS - positive - reverse-phaseChromatographic separation of metabolites was performed on the Thermo UHPLC Vanquish Horizon system equipped with an Accucore C30 column (2.6 μm, 2.1 mm × 100 mm). The separation was achieved at 0.4 mL/min flow rate with a mobile phase as a gradient consisted of 10 mM ammonium acetate in acetonitrile:water (1:1, v/v), containing 0.1% formic acid (solvent A) and 2 mM ammonium acetate in acetonitrile:isopropanol:water (10:88:2, v/v/v), containing 0.1% formic acid (solvent B).Interspecific variation in symbiosis with the swainsonine-producing fungus decouples the chemical defense strategies of Oxytropis locoweeds versus nontoxic congeners.Hao ZhouWuhan Universityleafmass spectrometry assayThe leaf materials (50 mg) were accurately weighted and then washed by sterilized 1× PBS, and a 6-mm grinding bead was added to each tube. Leaf materials were mixed with 280 μL methanol:water (2:5, v/v) solution containing 0.02 mg/mL L-2-chlorophenylalanine as an internal standard. extraction, and then mixed with 400 μL MTBE. Mixture was treated by a high-throughput tissue crusher Wonbio-96c (Wanbo Biotechnology Co., LTD., Shanghai, China) at 50 Hz and −10℃ for 6 minutes, followed by ultrasound treatment at 40 kHz and 5℃ for 30 minutes. After centrifugation (at 12,000 rpm for 15 minutes), supernatants were transferred to a 2 mL autosampler vial for LC-MS/MS analysis.blankOxytropis microphyllaOxytropis falcataOxytropis sericopetalaOxytropis bifloraOxytropis glacialishttps://www.ebi.ac.uk/metabolights/MTBLS15472Hao Zhou. Wuhan University. hao-zhou@whu.edu.cn.After analysis, all raw data were imported into the LipidSearch (Thermo, CA) for peak detection, alignment and identification. The preprocessing results generated a data matrix that consisted of the metabolite class, retention time (RT), mass-to-charge ratio (m/z) values, and peak intensity. Features detected at least 80 % in any set of samples were retained. After filtering, minimum metabolite values were imputed for specific samples in which the metabolite levels fell below the lower limit of quantitation and features were normalized by sum. In order to reduce the errors caused by sample preparation and instrument instability, the response intensity of the sample mass spectrum peaks was normalized by the sum normalization method, and the normalized data matrix was obtained. At the same time, the variables with relative standard deviation (RSD) > 30% of QC samples were removed, and log10 logarithmization was performed to obtain the final data matrix for subsequent analysis.Specieshao-zhou@whu.edu.cnSamples were collected from the leaves of Oxytropis plants.MetabolomicsMetabolomicsOxytropis microphyllablankuntargeted analysisOxytropis sericopetalaOxytropis falcataLipidSearchWuhan Universitysecondary metabolite biosynthetic processOxytropis glacialisleafThermo Scientific Vanquish UHPLC SystemThermo Scientific Q Exactive HF-XOxytropis bifloraexperimental blankMetabolomicsblankOxytropis microphyllauntargeted analysisOxytropis falcataOxytropis sericopetalaLipidSearchWuhan Universitysecondary metabolite biosynthetic processOxytropis glacialisleafThermo Scientific Vanquish UHPLC SystemThermo Scientific Q Exactive HF-XOxytropis bifloraexperimental blankMass spectrometric data was collected using a Thermo UHPLC-Q-Exactive HF-X Benchtop Orbitrap Mass Spectrometer equipped with heated-electrospray ionization (HESI) source operating in positive and negative ion mode. The detection was carried out over a mass range of 200-2000 m/z.falseMetabolomics data of leaf samples of Oxytropis plantsThis experiment aimed to analyze the leaf metabolite profiles of Oxytropis plants.2026-09-102026-08-27MTBLS15472