Fig. 6

Target genes of MaSPL4 identified via combined genome-wide DAP-seq and RNA-seq analyses. a Distribution of the MaSPL4-binding peaks and the putative DNA-binding motif of MaSPL4 identified by DAP-seq in the promoter region of the target genes. b KEGG categorization of MaSPL4-binding genes. Enriched KEGG pathways of MaSPL4 compared with all Musa acuminata genes are shown. A false discovery rate (FDR) cut-off was set on the basis of a P < 0.05. c Venn diagram displaying the numbers of overlapping genes among MaSPL4 target genes (orange circle) revealed by DAP-seq and differentially expressed genes (DEGs, green and purple circles) identified by RNA-seq in OE-MaSPL4 samples. DEGs refer to those differentially expressed after 14 days of cold treatment when fruit showed obvious chilling injury. d Enriched KEGG pathways and e GO terms of 68 OE-MaSPL4 induced targets. f Expression analysis of selected MaSPL4 target genes related to lipid metabolism and REDOX, respectively. The RPS2 gene was used as an internal reference. g Changes in POD activity in OE-MaSPL4 and control (empty vector) banana peel. Error bars indicate the SD of three biological replicates (n = 3) and asterisks indicate significant differences between control and transient transgenic samples at each time point, according to two-tailed unpaired Student’s t-test (*, P < 0.05; **, P < 0.01, ns, not significant). h DLR assay showing the activation of selected lipid metabolism and REDOX-related genes by MaSPL4. The LUC/REN ratio of the empty vector plus promoter-reporter was set as 1. Values are presented as means ± SD (n = 6). The asterisks (**) indicate significant differences at 0.01 level according to two-tailed unpaired Student’s t-test. i Binding peaks of MaSPL4 in the promoter of MIR528, MIR397 and MIR156e identified by DAP-seq. Both miRNA precursor and upstream 1.5 kb regions are shown. The arrows indicate the direction of MIRNA gene transcription and the solid orange lines indicate the positions of MaSPL4 binding motif