Arabidopsis Seedling Growth Protocol

Arabidopsis seedlings were grown as described in Jia et al. (2019) (link). Briefly, 25 Arabidopsis seeds per bundle were vapor-phase sterilized (90% sodium hypochlorite 13 and 10% HCl 36%) and sown on plates which were layered with a thin nylon mesh on top of 1/2 MS media (Murashige and Skoog salt + Gamborg B5 vitamin mixture [Duchefa] supplemented with 0.5 g/liter MES and 1% sucrose, pH 5.7) plates (with 1% plant agar [Duchefa]) followed by 3 d of vernalization at 4°C in dark. Vernalized seeds were grown at 21°C under LEDs with 85 µM/m²/s with a 14 h light/10 h dark photoperiod for 7 d. 7-d-old seedlings were subsequently transferred to 1/2 MS media (CaissonLabs) or nitrogen-deficient 1/2 MS media (Murashige and Skoog salt without nitrogen [CaissonLabs] + Gamborg B5 vitamin mixture [Duchefa] supplemented with 0.5 g/liter MES and 1% sucrose, pH 5.7) plates (+0.8% plant agar [Duchefa]) and grown for 2 wk.