Non-targeted metabolite profiling of plants

Six independent plants were used as biological replicates, and about five leaves were sampled from each plant in primary metabolic profiling. Non-targeted metabolite profiling was carried out by GC-MS using a modified method described by Yun et al. [37 (link)]. A total of 200 mg ground leaf samples were extracted in 2,700 μl methanol and ribitol solution (300 μl, 0.2 mg ml⁻¹) was added as an internal standard. The samples were centrifuged, dried and derivatized. GC-MS analysis was performed by using a Thermo Trace GC Ultra, coupled with Thermo Fisher a DSQ II mass spectrometer (Thermo Fisher Scientific, Waltham, MA, USA). Metabolites were identified by using an available chromatogram library and PCA analysis was performed by using the software Simca-P (Ver 11, Umetrics, Umea, Sweden).