Figure 8.

Binding of MdWRKY79 Transcriptional Factor to the W-Box Motif in MdNLR16 Promoter.

(A) Binding sites in the MdNLR16 promoter; numbers indicate positions of the various cis-acting elements upstream of the MdNLR16 coding sequence with the first nucleotide of its start codon designated as 0; yellow indicates the W-box site.

(B) Expression patterns of 22 WRKY genes in CK, A4 and A10 lines based on RNA-seq data, with the three downregulated WRKY genes outlined in red (RPKM > 1; adjusted P < 0.01 for the RPKM ratios of A4/CK and A10/CK).

(C) Y1H assay on binding of MdWRKY79 protein to the promoter region of MdNLR16. N1 to N4 represent different regions of the MdNLR16 promoter shown in (A). Yellow line shows the location of the W-box site. Blue triangle represents the range of yeast concentrations (10⁰ to 10⁻³). pGADT7-p53+pAbAi-p53 was used as a positive control and pGADT7+pAbAi-p53 as a negative control.

(D) ChIP-PCR confirming the binding of WRKY79 to the NLR16 promoter. Actin was used as an internal control. W1 to W4 represent the different primers shown in Supplemental Data Set 5. Data are mean ± sd of three biological replicates. **P < 0.01 between MdWRKY79-GFP and GFP alone using Student’s t test.