Extended Data Fig. 6: The contribution of NLS and cTP to SlSAD8-mediated fruit ripening.
a, Immunoblot analysis was performed to check the subcellular localization of SlSAD8, SlSAD8ΔcTP, and SlSAD8ΔNLS when expressed in SlSAD8-KO (#5) fruits. Toc75 and histone H3 proteins were used as plastid and nuclear markers, respectively. Tot., Total; P., plastids; N., nuclei. Experiments were repeated at three independent times with similar results. b-f, Compared with transient expression of GFP, transient expression of SlSAD8, but not SlSAD8ΔNLS, resulted in a delayed ripening initiation and impaired ethylene biosynthesis. b, Representative ripening phenotypes. Bars, 1 cm. c. Days from anthesis to breaker stage. d, Ethylene production at the Br stage. e, Relative transcript levels of SlACS2. f, Relative transcript levels of SlACS4. g-i, Transient expression of SlSAD8, but not SlSAD8ΔcTP, resulted in delayed color change (g), elevated chlorophyll accumulation (h), and reduced carotenoid levels (i) at the Br + 3 stage. Bars in g, 1 cm. In c, data are shown in a violin plot (n = 50), and differences were assessed using two-tailed t-test (***, P < 0.001; NS, no significant). In d-f, h, and i, data are shown as mean ± s.d. of three biological replicates, and differences were assessed using two-tailed t-test (***, P < 0.001; **, P < 0.01; *, P < 0.05; NS, no significant).
