| MS Description
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A total of 200 seeds of each independent m … A total of 200 seeds of each independent mutant and accession were homogenized using a mixer mill MM 200 (Retsch) in 80 μL of extraction buffer [40% acetonitrile in H2O with 25 μM hydroxyphenyl–glucosinolate (GSL) and 50 μM norleucine as internal standards].The extracts were diluted with 500 lL of LC–MS grade H2O and centrifuged (1,000 g) for 5 min. The supernatants were filtered through a CAPTIVA 0.45μlm filter (Varian).<br />
Thirty-six metabolites (17 amino acids, 18 GSLs, and 1 flavonoid; Table S1) were separated on UPLC through a reverse phase column (50 9 2.1 mm,HSS T3 1.8 μm; Waters) (Table S1) and detected using ZQ mass spectrometers (Waters) with an electrospray ionization (ESI) interface (positive mode for amino acid analysis; negative mode for glucosinolates and flavonoid analysis; capillary voltage, +3.0 and -3.0 keV; cone voltage, +25 and -40 V; source temperature, 120℃; desolvation temperature, 350℃; cone gas flow, 50 L/h; desolvation gas flow, 600 L/h). w, 50 L/h; desolvation gas flow, 600 L/h).
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