Fig. 2: Impact of hydropriming on ROS levels in ‘67/3’ seeds.

a Experimental design showing the selected timepoints for molecular analyses. Seeds were collected during priming (HP72 protocol) at 24, 48, and 72 h, as well as at the end of dry-back (DB, 2 h). Germination of unprimed (UP) and primed (HP) seeds was monitored by collecting samples throughout imbibition at 2, 4, 8, 16, 24, 48, and 72 h. The last timepoint, corresponding to the phenological stage of radicle protrusion (RD), was anticipated at 4 days for the primed seeds, compared to unprimed (8 days). ROS levels were measured during hydropriming b and subsequent germination c at the selected timepoints using the DCFH-DA fluorescent dye. Values are expressed as mean ± SD of three independent replications with 20 seeds for each replication. Asterisks indicate statistically significant differences determined using Student’s t-test (P < 0.05). DS dry seed, R.F.U. relative fluorescence unit, ROS reactive oxygen species, DCFH-DA dye 2′,7′-dichlorofluorescein diacetate