MetaboLights

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.ac.uk/pub/databases/metabolights/studies/public/MTBLS14580/FILES/RAW_FILES/117bRF-2.D.zipftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14580/FILES/RAW_FILES/135RF-2.D.zipprimaryOK200ftp://ftp.ebi.ac.uk/pub/databases/metabolights/studies/public/MTBLS14580Metabolites were annotated by comparing spectra with the NIST commercial library (Ausloos et al., 1999) and manually validated against the Golm Metabolome Database (GMD; (Kopka, 2006)).MetaboLightsPublicGas Chromatography MS - positive - low-polarityGC-MS analyses were performed on an Agilent 5977A mass spectrometer with a quadrupole analyzer and an HP-5MS UI capillary column (30 m × 0.25 mm × 0.25 mm; Agilent 19091S- 433UI). Helium was used as the carrier gas (1 ml/min, constant flow). The injector was operated in splitless mode (79.6 ml/min, 1 min), and the injection volume was 2 ml. The temperature program was as follows: 60 °C (5 min), ramped at 10 °C/min to 250 °C (5 min), then 20 °C/min to 300 °C (10 min). The transfer line, ion source, and quadrupole temperatures were set to 280 °C, 230 °C, and 150 °C, respectively. Mass spectra were acquired in electron impact mode (70 eV), with a scan range of 30–650 m/z and a solvent delay of 8 min. Each sample was injected in duplicate as technical replicates.Poikilohydria, polyols, and homeoviscosity: lichen metabolomic remodeling across environmental gradients. 10.3389/fpls.2026.1792494.Universitat de ValÃ¨nciaFrancisco MarcoThallusmass spectrometry assayMetabolites were extracted and derivatized according to a protocol for plant samples (Lisec et al., 2006), with major modifications. First, thallus fragments were randomly pooled per lichen specimen to obtain 15 mg dry weight (DW), weighed on a precision balance. Then, lichen material was finely ground with approximately 300 mg glass beads (500-750 mm diameter) using a vortex with a bead-beating attachment (Vortex-Genie 2) at maximum speed for 15 min. A volume of 1.4 ml of HPLC-grade methanol (pre-cooled to -20 °C) and 350 ml of antipyrine stock solution (10 mM in methanol, used as the internal standard, IS) were added to the ground material. Samples were incubated at 70 °C with shaking (950 rpm, 10 min), centrifuged (11,000 g, 10 min), and 1.2 ml of the supernatant was transferred to PCR clean purity grade tubes. A 400 ml aliquot of the extract was then dried in a SpeedVac for chemical derivatization, while the remainder volume was stored at -20 °C for future use.Derivatization involved methoxyamination followed by silylation of the dried samples, with an empty tube processed in parallel as a blank control. For methoxyamination, 40 ml of freshly prepared methoxyamine hydrochloride (20 mg/ml in pyridine) were added, and samples were incubated at 37 °C (650 rpm, 2 h). For silylation, 70 ml of N-methyl -N-(trimethylsilyl) trifluoroacetamide (MSTFA) were added, along with 5 ml of C7-C30 saturated alkanes (1 mg/ml) to the first and last samples of each batch for retention index calibration (van Den Dool and Kratz, 1962). Samples were incubated at 37 °C (650 rpm, 30 min) and transferred to glass-insert vials for Gas Chromatography-Mass Spectrometry (GC-MS) analyses.Ramalina farinaceahttps://www.ebi.ac.uk/metabolights/MTBLS14580Francisco Marco. Universitat de València. Instituto de Investigación en Biotecnología y Biomedicina (BIOTECMED), E-46110 Burjassot, Spain. francisco.marco@uv.es.Marta Pérez-Rodrigo. Universitat de València. Instituto de Investigación en Biotecnología y Biomedicina (BIOTECMED), Facultad de Ciencias Biológicas, E-46100 Burjassot, Spain. marta.perez-rodrigo@uv.es.Pedro Carrasco. Universitat de València. Instituto de Investigación en Biotecnología y Biomedicina (BIOTECMED), Facultad de Ciencias Biológicas, E-46100 Burjassot, Spain. Pedro.Carrasco@uv.es.Peak deconvolution, spectral matching, and integration were performed using MassHunter GC/MS Acquisition and MassHunter Qualitative software (Agilent Technologies). C7-C30 alkanes were used for peak alignment. Metabolites were annotated by comparing spectra with the NIST commercial library (Ausloos et al., 1999) and manually validated against the Golm Metabolome Database (GMD; (Kopka, 2006)). Abundance values were normalized to the IS by applying a scaling factor to each sample, calculated as fj = median(IS)=ISj and multiplying it by all peak areas in that sample. Pure standards of selected metabolites were analyzed for method validation, confirming annotations and supporting relative abundance assessment.CountrySeasonFrancisco.Marco@uv.esSpecimens fitting the morphological concept of Ramalina farinacea were collected from six sites (countries) across a geographical gradient in Europe: Czech Republic, Estonia, Finland, Norway, Spain, and Sweden. Sampling was carried out in winter (January-March 2023) and summer (June-July 2023), with six individuals collected per site and season. Fresh, medium-sized (≤5 cm in length) thalli were processed immediately after collection. Each thallus was lyophilized overnight and stored in the dark with silica gel for metabolite analysis.MetabolomicsMetabolomicsgeographicalClimateAgilent 5977A MSDMassHunter Data Acquisitionuntargeted analysisThallusEuropegradientwinterlichensummerMassHunter Qualitative Analysisexperimental sampleUniversitat de ValènciaRamalina farinacea5977A MSDMetabolomicsgeographicalClimateAgilent 5977A MSDMassHunter Data Acquisitionuntargeted analysisThallusEuropegradientwinterlichensummerMassHunter Qualitative Analysisexperimental sampleUniversitat de ValènciaRamalina farinacea5977A MSDGC-MS analyses were performed on an Agilent 5977A mass spectrometer with a quadrupole analyzer and an HP-5MS UI capillary column (30 m × 0.25 mm × 0.25 mm; Agilent 19091S- 433UI). Helium was used as the carrier gas (1 ml/min, constant flow). The injector was operated in splitless mode (79.6 ml/min, 1 min), and the injection volume was 2 ml. The temperature program was as follows: 60 °C (5 min), ramped at 10 °C/min to 250 °C (5 min), then 20 °C/min to 300 °C (10 min). The transfer line, ion source, and quadrupole temperatures were set to 280 °C, 230 °C, and 150 °C, respectively. Mass spectra were acquired in electron impact mode (70 eV), with a scan range of 30–650 m/z and a solvent delay of 8 min. Each sample was injected in duplicate as technical replicates. falseIn-situ metabolome of Ramalina farinacea from thalli collected across six European regions during winter and summerSamples of the model lichen Ramalina farinacea were collected from six sites (countries) across a geographical gradient in Europe: Czech Republic, Estonia, Finland, Norway, Spain, and Sweden. Sampling was carried out in winter (January-March 2023) and summer (June-July 2023), with six individuals collected per site and season. Metabolites were extracted and derivatized according to a protocol for plant samples (Lisec et al., 2006), with major modifications. Briefly, methanol extracts from dry samples were obtained by grinding and 70ºC incubation. Extracts were vacuum dried and derivatized by methoxyamination followed by silylation. Derivatized samples were analyzed by GC-MS. GC-MS analysis was performed on an Agilent 5977A mass spectrometer with a quadrupole analyzer and an HP-5MS UI capillary column (30 m × 0.25 mm × 0.25 mm; Agilent 19091S-433UI). Mass spectra were acquired in electron impact mode (70 eV), with a scan range of 30–650 m/z and a solvent delay of 8 min. Each sample was injected in duplicate as technical replicates. Peak deconvolution, spectral matching, and integration were performed using MassHunter GC/MS Acquisition and MassHunter Qualitative software (Agilent Technologies). C7-C30 alkanes were used for peak alignment. Metabolites were annotated by comparing spectra with the NIST commercial library (Ausloos et al., 1999) and manually validated against the Golm Metabolome Database (GMD; (Kopka, 2006)).2026-07-222026-05-25MTBLS14580