Fig. 5: Influence of PT^H-regulated FUL2 expression on the development of fruit with pointed tips.

a Relative transcript levels of FUL2 in the distal end of fruit from PT^H-overexpressing lines, PT^H mutants, and the wild-type control (TS-9, PT^H allele). The fruit were harvested at 14 DPA. Error bars indicate mean ± SE. n = 3 biological replicates. Statistically significant differences were determined using a one-way ANOVA with Tukey’s post hoc test. Different letters indicate statistically significant differences (P < 0.05). b Schematic diagram of the FUL2 promoter region. The red line indicates the probe used in e. c PT^H binding of the FUL2 promoter in yeast. Positive control: p53-AbAi + pGAD-Rec2-53. Negative control: pAbAi-FUL2 + pGADT7. The transformants were grown on a selective medium (SD/-Ura-Leu) containing 40 or 50 ng mL⁻¹ aureobasidin A (AbA). d Co-expression of the PT^H/R and the FUL2 promoter: β-glucuronidase (GUS) reporter gene in N. benthamiana leaves. Co-transformation of FUL2_pro:GUS and the empty vector pHELLSGATE8 was used as a control. e PT protein binding the FUL2 promoter in an electrophoretic mobility shift assay (EMSA). A 50-bp double-stranded fragment labeled or unlabeled with 5′ 6-FAM was used as a DNA probe or DNA competitor. The FAM-labeled probe with the substitutions of four conserved bases was used as a mutant probe. The + and − symbols indicate the presence and absence, respectively, of the DNA probe or the protein. Arrows indicate the protein–DNA complex and unbound labeled DNA probe. Three independent experiments were performed. f, g Binding affinity of PT^H (f) and PT^R (g) for the FUL2 promoter fragment was measured using an open surface plasmon resonance (SPR) assay. h Schematic illustration of the two sgRNA target sites in FUL2. Red lines indicate the binding sites of sgRNAs. i Genomic DNA sequences of FUL2 in CR-ful2-1, CR-ful2-3, and the wild-type (TS-9). j Phenotype of fruit produced by the CR-ful2 and CR-pt^H single mutants, the CR-ful2 CR-pt^H double mutant, and the pertinent wild-type (TS-9, PT^H allele). All of the mutants were derived from the same wild-type background, TS-9. Source data underlying Fig. 5a, e are provided as a Source Data file.