Extended Data Fig. 8: SlNTRC is indispensable for the normal growth of tomato plants.
From: Redox modification of m6A demethylase SlALKBH2 in tomato regulates fruit ripening
a, Genotyping of slntrc-21 and slntrc-29 mutants generated by CRISPR/Cas9-mediated gene editing. The single guide RNAs (sgRNAs) containing different target sequences (T1 and T2) were designed to specifically target the fourth and fifth exons of SlNTRC. The red letters indicate the protospacer adjacent motif (PAM). b, Schematic illustration for the predicted truncated SlNTRC proteins in the slntrc-21 and slntrc-29 mutants. c, Changes in the transcript levels of SlNTRC in the leaves of slntrc-21 and slntrc-29 mutants, as determined by RT-qPCR analysis. Actin gene was used as an internal control. d, The growth phenotypes of slntrc mutant lines. Scale bar, 1 cm. e, Changes in redox state of endogenous SlALKBH2 proteins in the slntrc mutant lines. Total proteins extracted from 35-day-old leaves were subjected to SDS-PAGE with (+) or without (-) DTT (50 mM) and analyzed using a polyclonal anti-SlALKBH2 antibody. Actin was used as a loading control. f, Quantification of SlALKBH2 homodimer versus monomer shown in e (-DTT). g, Quantification of SlALKBH2 protein levels shown in e (+DTT). h, Genotyping of SlNTRC heterozygous mutants generated by CRISPR/Cas9-mediated gene editing. i, Changes in the transcript level of SlNTRC in fruits of slntrc-H1 and slntrc-H3, as determined by RT-qPCR analysis. j, The OE-NTRC-2 and OE-NTRC-6 overexpression lines displayed no obvious change in vegetative growth compared to the WT. Scale bar, 1 cm. k, Changes in transcript levels of ripening-related genes in fruits of OE-NTRC-2 and OE-NTRC-6 at 42 day post-anthesis, as determined by RT-qPCR analysis. For c, f, g, i and k, data are means ± s.d. (n = 3 biological replicates). Statistical significance was assessed through one-way ANOVA analysis, with lowercase letters indicating significant differences (P < 0.05, Tukey’s test).
