Ion Profiling of Arabidopsis Mutants

Ion profiling was determined by following a previous publication with slight modification (Wu et al., 2019 (link)). In brief, 18-day-old seedlings from Col-0 and G protein mutants from treatment plates (Yamagami medium with 1% purified agar) were dried at 60 °C for 3 d. A 25 mg aliquot of ground leaf samples was boiled in 10 ml of 65% (v/v) HNO₃ solution at 120 °C for 90 min. A 3 ml volume of 30% (v/v) H₂O₂ was subsequently added and continuously boiled at 120 °C for 90 min. Metal concentrations were determined using inductively coupled plasma-optical emission spectroscopy (ICP-OES) (Varian Vista-MPX CCD Simultaneous ICP-OES) and expressed per gram of shoot dry weight. Data were analyzed using Student’s t-test. The criterion of P<0.05 was used to determine statistically significant differences between Col-0 and mutant lines.