Metabolomic and Lipidomic Analysis of Mycelia

Sample preparation and metabolites/lipid extraction and identification were performed as described previously (44 (link), 45 (link)). For metabolomic analysis, fresh mycelia were extracted using MeOH:water (1:1). Extracts were dried in a vacuum centrifuge, resuspended in MeOX-pyridine and N-methyl-N-(trimethylsilyl)trifluoroacetamide (MSTFA) with 1% trimethylsilyl chloride (TMCS) for derivatization, and analyzed by GC-MS (Thermo Fisher Scientific, Boston, MA, USA) equipped with an RTX-5MS column (30 m by 0.25 mm by 0.25 μm, Restek, Bellefonte, PA, USA). For lipidomic analysis, freeze-dried mycelia were extracted with methyl tert-butyl ether. The lipidomic analysis was conducted using a Dionex UltiMate 3000 UPLC system (Santa Clara, CA, USA) coupled to a HESI probe with a Q-Exactive Orbitrap mass spectrometer (Thermo Fisher, CA, USA). The “raw” format files were converted to “abf” format using the ABF converter. The MSDIAL4.20 equipped with the DB_FiehnBinbase-FiehnRI and LipidMsmsBinaryDB-VS46-FiehnO database was used for metabolite and lipid molecule identification, respectively (46 (link)). All annotations were manually checked again. The mass spectrometry data have been deposited to MetaboLights with the data set identifier MTBLS4103.