Metabolite analysis was performed by gas chromatography mass spectrometry (GC–MS). Lyophilized samples (10 mg), the BS, and the DE were extracted with 0.8 mL of acetone:water:acetic acid (80:19:1 v:v:v) containing ribitol at 4 µg/mL, followed by shaking for 10 min at 4°C at 1400 rpm. After centrifugation (20,000 ×g, 5 min), 100 µL of supernatant were collected and dried for 5 h in a SpeedVac vacuum centrifuge.
Analysis and data processing were performed as previously described (Moret et al., 2018 (link)). Samples were derivatized and analyzed using an Agilent 7890A gas chromatograph coupled to an Agilent 5975C mass spectrometer, as previously described (Fiehn, 2006 (link); Fiehn et al., 2008 (link)). For processing, data files were converted to NetCDF format and analyzed with AMDIS (http://chemdata.nist.gov/mass-spc/amdis/ ). A homemade retention indices and mass spectra library built from the NIST, Golm (http://gmd.mpimp-golm.mpg.de/ ), and Fiehn databases and standard compounds was used to identify metabolites. Peak areas were determined with Targetlynx software (Waters) after conversion of the NetCDF file to Masslynx format. AMDIS and Target Lynx in splitless and split 30 mode data were compiled in a single Excel file for comparison. After blank mean subtraction, peak areas were normalized to ribitol and fresh weight.
Analysis and data processing were performed as previously described (Moret et al., 2018 (link)). Samples were derivatized and analyzed using an Agilent 7890A gas chromatograph coupled to an Agilent 5975C mass spectrometer, as previously described (Fiehn, 2006 (link); Fiehn et al., 2008 (link)). For processing, data files were converted to NetCDF format and analyzed with AMDIS (
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