Fig. 3: Mutation of FvALKBH10B leads to a delay in strawberry fruit ripening.
From: The FvABF3-FvALKBH10B-FvSEP3 cascade regulates fruit ripening in strawberry
a Schematic diagram of sgRNAs with two target sequences (T1 and T2) designed to specifically target the exons of FvALKBH10B. Red letters, protospacer adjacent motif (PAM); red vertical line, target sequence location; red horizontal line, Alkb domain location. b Genotyping of mutations mediated by CRISPR/Cas9 gene editing in the Fvalkbh10b-1, Fvalkbh10b-2, Fvalkbh10b-3 mutant lines. Sequences of the isolated mutant alleles are aligned to the wild type (WT). The inserted/deleted nucleotides are colored with red to indicate the editing sites. Red arrowheads indicate the sites of frameshift in the translated mutant proteins. Red asterisks indicate protein translation termination site. c LC-MS/MS assay revealing the total m6A level in fruits of WT and Fvalkbh10b mutants at 25 days post anthesis (DPA). d, e Phenotypes of strawberry flowers (d) and fruits (e) from WT and Fvalkbh10b mutants. Scale bar, 0.5 cm. f–i Changes in anthocyanin contents (f) and sugar contents, including sucrose (g), glucose (h), and fructose (i) in the mutant lines at 27 and 33 DPA. j LC-MS/MS assay revealing the changes in total m6A level in fruits of overexpression lines (FvALKBH10B-OE) at 28 DPA. k Relative expression of FvALKBH10B in control and overexpression lines as determined by RT-qPCR. ACTIN was used as an internal control. l Phenotypes of fruit ripening in overexpression lines. EV empty vector. Scale bar, 0.5 cm. m Changes in anthocyanin contents in overexpression lines at 27 and 33 DPA. For c, f–k, and m, values are means ± SEM (n = 3 biological replicates). Data are analyzed using one-way ANOVA with Tukey’s HSD test (P < 0.05). Different lowercase letters represent significant differences. Source data are provided as a Source Data file.
