1290 high performance liquid chromatograph

Lipidomic analysis was performed on a 6460 triple quadruple electrospray ionization mass spectrometer (ESI/MS) coupled with a 1290 high-performance liquid chromatograph (Agilent, USA). The specific methods were previously described (Li et al., 2008 (link); Gwak et al., 2014 (link)). The detailed method can be found in Method S1. For lipidomics analysis, six biological replicates were included. For each biological replicate, lipids were extracted in duplicate for independent LC/MS analysis.

The endogenous hormones IAA, ABA, JA, SA, GA₁, GA₃, and zeatin were analyzed by HPLC–MS/MS according to the method by Pan et al. (2010) [58 (link)]. The 7 analytes were separated using a Poroshell 120 SB-C18 column (2.1 × 150 mm, 2.7 μm) with the mobile phase containing water and methanol in gradient elution mode on an Agilent 1290 high-performance liquid chromatograph (Agilent, Santa Clara, CA, USA). The flow rate was 0.3 mL/min, and the column temperature was 30 °C. Then, a quantitative analysis was performed in multiple-reaction monitoring (MRM) mode with electrospray ionization (ESI) on an AB SCIEX QTRAP 6500+LC-MS/MS system (AB, Fairfield, OH, USA).