Fig. 1: Nuclear localization and all three domains of RCD1 are essential for its function.
a Schematic representation of RCD1 domain structure containing a bipartite NLS, WWE, PARP-like and RST domains. Intrinsically disordered regions are marked as IDR1-4. b Early flowering phenotype of rcd1 is not reverted by expression of RCD1ΔWWE-HA (ΔW), RCD1ΔPARP-like-HA (ΔP), RCD1ΔRST-HA (ΔR), and RCD1S/TIDR2A-HA in rcd1 background. Picture shows 5-week-old plants of representative lines under standard growth conditions. c Wild-type MV sensitivity is not restored in lines expressing RCD1ΔWWE-HA (ΔW), RCD1ΔPARP-like-HA (ΔP), and RCD1ΔRST-HA (ΔR) constructs. PSII inhibition (Fv/Fm) by MV was measured in indicated lines using 0.5 μM MV. For each experiment, leaf discs from three individual rosettes were used. The experiment was performed three times with similar results. Mean ± SD are shown. * – P value < 0.05 with Tukey corrected post hoc test at the selected time point between rcd1: RCD1ΔPARP-like-HA and rcd1: RCD1-HA lines; *** – P value < 0.001 with Tukey corrected post hoc test at the selected time point between rcd1: RCD1ΔWWE-HA and rcd1: RCD1-HA lines. Source data and full statistics are presented in Supplementary Data 1. d Leaf shape phenotype of rcd1 can be complemented by re-introduction of wild-type RCD1-HA, but not by RCD1 with mutated NLS (RCD1nls-HA) into the mutant background. The photo shows 3-week-old plant rosettes of two independent lines (A and B) for each construct under standard growth conditions. e RCD1 requires its NLS to complement the rcd1-specific MV tolerance. PSII inhibition (Fv/Fm) by methyl viologen (MV) was measured in indicated lines using 1 μM MV. For each experiment, leaf discs from three individual rosettes were used. The experiment was performed three times with similar results. Mean ± SD are shown. *** – P value < 0.001 with Tukey corrected post hoc test; n.s. – nonsignificant difference. Source data and statistics are presented in Supplementary Data 1.
