Measurement of SA and JA content
SA and JA content was measured using high-performance liquid
chromatography-tandem mass spectrometry
(HPLC-MS/MS).
The leaves of A. thaliana before and after inoculation withS. sclerotiorum were obtained. 0.5-1 g of homogenized frozen
leaves were suspended in 10 mL extraction solvent
(2-propanol/H2O/HCl), and then add 8
μL
internal standard (1 μg/mL). Samples were shaken
for
30 min at 4°C with a speed of 100 rpm, and add 5 mL dichloromethane to
the samples and shake for 30 min at
4°C.
The organic phase was obtained in a centrifuge at 4°C for 5 min at
13,000×g . The
organic
phase was transferred to a new fresh tube and evaporated with the
nitrogen flow. Pellets were resuspended in 0.4 mL of methanol. The
extraction was analyzed using HPLC-MS/MS in Nanjing Convinced-test
Technology Co., Ltd (Zang et al. 2021).