Fig. 5: ANAC102 controls petal abscission.
From: Petal abscission is promoted by jasmonic acid-induced autophagy at Arabidopsis petal bases
a Identification of evolutionally conserved G-box motifs at the ANAC102 promoter. Top, IGV browser view of MYC2 binding peaks by ChIP-seq. Middle, Bit score comparison of 5′ regions of the promoter between nine Brassicaceae species by Gene Slider. Bottom, diagram of the ANAC102 promoter with and without mutations in the G-box motifs (shown as asterisks). PCR amplicons for ChIP–qPCR are shown in e. b Web logos of three evolutionarily conserved G-box motifs. Reporter gene expression of intact (c) and G-box mutants of the ANAC102 (ANAC102Δ) (d) promoter. Scale bar = 100 µm. e Association of MYC2 with the ANAC102pro:GUS and ANAC102Δpro:GUS transgenes based on ChIP–qPCR. Values are means ± SE (n = 4 independent experiments). Asterisks indicate statistically significant differences of MYC2 binding between the wild-type and mutated (Δ) ANAC102 promoter based on two-tailed Student’s t test. f Relative ANAC102 expression in WT and JA-related mutants, as determined by RT–qPCR. Values are means ± SE (n = 3 independent experiments). Different letters indicate statistically significant differences based on one-way ANOVA (p = 1.9 ×10−6) and post-hoc Tukey’s HSD test (p < 0.05: See the Data Source file for all combinations of the exact p-values). g GUS reporter gene expression of the ANAC102 promoter in the dad1 mutant. Scale bars = 100 µm. h Side view of wild-type, snac septuple, dad1 and dad1 InMYB1pro:ANAC102 inflorescences. Opened flowers with petals are indicated by asterisks. Scale bar = 1 cm. i Quantification of abscission timing, shown as individual data points (left) and violin plots (right) for each genotype. Black dots and vertical lines indicate mean and SD, respectively. n = 20. Different letters indicate statistically significant differences based on one-way ANOVA test (p = 1.1 × 10−16) and post-hoc Tukey’s HSD test (p < 0.05: See the Data Source file for all combinations of the exact p-values). j DAB staining of WT, snac, dad1 and dad1 InMYB1pro:ANAC102 petals from position +3 flowers. Scale bar = 100 µm. k Quantification of DAB-stained area (µm2), shown as individual data points (left) and violin plots (right) for each genotype. Black dots and vertical lines indicate mean and SD, respectively (WT: n = 37; snac septuple: n = 45; dad1: n = 30; dad1 InMYB1pro:ANAC102: n = 38). Different letters indicate statistically significant differences based on one-way ANOVA test (p = 9.9 × 10−11) and post-hoc Tukey’s HSD test (p < 0.05: See the Data Source file for all combinations of the exact p-values). l Trypan blue staining of WT, snac, dad1 and dad1 InMYB1pro:ANAC102 petals of position +3 flowers. Scale bar = 100 µm. m SEM images of petal base in snac and dad1 InMYB1pro:ANAC102 plants from position +3 flowers. Top, Higher magnification of the green square in Fig. 1i. Bottom, Higher magnification of position d cells. Scale bar = 5 µm. EM images of position d cells (n) and neighboring cells (o) from position +3 petals in snac and dad1 InMYB1pro:ANAC102 plants. Scale bar = 1 µm.
