2.2.5 Fatty acid profile
Seed fatty acid contents were determined using gas chromatography (GC) following trans- methylation of fatty acids and production of fatty acid methyl ester (FAME) by the modified method of Xue et al., 2013. In short, dried seeds (300 mg) in 500 μl of methanol containing H2SO4 (49:1 v,v) were heated at 80°C for 2h. After the mixture was cooled at room temperature, FAMEs separated with 150 μl hexane and 100 μl %0.9 NaCl. After vortexing and centrifuging at 3000rpm for 5min, supernatants (hexane phase) were injected to GC analyzer using a DB-225 column (30m × 250µm × 0.25 µm) on Agilent 7890B.