Fig. 3: AHL genes are essential for 2,4-D-induced and BBM-induced SE.

a The effect of ahl loss-of-function on the capacity to induce somatic embryos on IZEs by 2,4–D. Dots indicate the values of three biological replicates per plant line with 50 IZEs per replicate, bar indicates the mean, and error bars the s.e.m. For the ahl15/+ pAHL15:AHL15-GUS combination IZEs were harvested from ahl15/+ pAHL15:AHL15-GUS plants and scored and genotyped after culture. Only scores of ahl15/+ pAHL15:AHL15-GUS explants were used. The letters indicate statistically different values, as determined by a one-way ANOVA with Tukey’s post hoc test. The p-values are provided in the source data file. b RT-qPCR analysis of the fold change in expression of AHL15, AHL19 and AHL20 in IZEs cultured for 7 days on medium with 5 μM 2,4-D relative to medium without 2,4-D. c, d pAHL15:AHL15-GUS IZEs cultured for 7 days in the absence (c) or presence (d) of 5 μM 2,4-D and histochemically stained for GUS activity. Similar results were obtained from two independent experiments. Size bar indicates 1 mm. e Predominant phenotype of wild-type (upper panel), ahl15 ahl19 amiRAHL20 (middle panel), or ahl15/+ pAHL15:AHL15-GUS (lower panel) IZEs cultured for 2 weeks on 2,4-D medium. Numbers indicate the frequency with which similar phenotypes were observed in two independent experiments. Size bar indicates 1 mm. f–i ChIP-seq BBM binding profiles for AHL15 (f), AHL19 (g), AHL20 (h), and AHL29 (i). The binding profiles from the pBBM:BBM-YFP (upper profile) and p35S:BBM-GFP (lower profile) ChIP-seq experiments are shown. The x-axis shows the position of DNA binding relative to the location of the selected gene (TAIR 10 annotation), the y-axis shows the ChIP-seq score (fold enrichment of the BBM-GFP/YFP ChIP to the control ChIP), and the arrow brackets around the gene name indicate the direction of gene transcription. j RT-qPCR analysis of the fold change in expression of AHL genes in DEX + CHX treated p35S:BBM-GR seedlings relative to that in DEX + CHX treated Col-0 wild-type seedlings. b, j Dots indicate the values of three or four biological replicates, bar indicates the mean, and error bars the s.e.m. The p-values were determined by a two-sided Student’s t-test.