Extended Data Fig. 1: Characterization of DMBQ-induced [Ca2+]cyt elevation in Arabidopsis.
From: Quinone perception in plants via leucine-rich-repeat receptor-like kinases
a–c, DICE showed different spatiotemporal dynamics to flg22-induced [Ca2+]cyt elevation in Arabidopsis. Kymographs of R-GECO1 signal in Arabidopsis in response to 10 μM DMBQ (a), 1 μM flg22 (b) or mock (c). Kymographs were extracted from time-lapse images (Supplementary Videos 1–3) along three-pixel lines as shown on the left panels. For b, two kymographs were obtained from two types of root tissues (epidermis and vasculature), which are shown in white and yellow lines, respectively. Stimuli were added at 0 min. Signal intensity is shown in rainbow spectrum. Scale bar, 100 μm. d, DMBQ does not induce apoplastic ROS production. External ROS production was measured against DMSO (control), DMBQ or flg22 (positive control), using a chemiluminescence probe (L-012). Data are mean ± s.d. from 48 seedlings. The experiment was repeated twice, with similar results obtained. e, DMBQ induces [Ca2+]cyt elevation in Arabidopsis seedlings in a dose-dependent manner, following Michaelis–Menten kinetics. f, Maximum concentration of Ca2+ influx at any given DMBQ concentration using data from e. g, Lineweaver–Burk plot of f. Km was calculated from x-axis intercept. Pretreatment with protein kinase inhibitor or chemicals that interfere with Ca2+ influx reduced DICE in a dose-dependent manner. h–j, WTAEQ Arabidopsis seedlings were pretreated with K252a (n = 6 seedlings) (h), LaCl3 (n = 4 seedlings) (i) or EDTA (n = 6 seedlings) (j) for 30 min before addition of 5 μM DMBQ concentration, as indicated. Data are mean ± s.d. Error bars are indicated every 30 s for clarity. For a–c, e–g, experiments were repeated 3 times with similar results obtained.
