Figure 7.

MdNAC7 interacts with MdWRKY31 to alleviate the transcriptional inhibition effect of MdWRKY31 on MdXTH2. (a) Yeast two‐hybrid (Y2H) assay to determine the interacting of MdNAC7 and MdWRKY31. pGBD‐MdNAC7/pGAD and pGBD/pGAD‐MdWRKY31 were negative controls. Different co‐transformed AH109 yeast cells were dropped on a culture medium (SD/−Trp/−Leu/−Ade/‐His). (b) In vitro GST pull‐down between MdWRKY31 and MdNAC7. The purified MdWRKY31‐His protein was incubated with MdNAC7‐GST and GST control. Proteins were eluted from the magnetic beads and immunoblotted with anti‐His antibody and anti‐GST antibody. (c) MdWRKY31 interacted with MdERF72 in a LUC assay using tobacco leaf cells. MdNAC7‐nLUC and MdWRKY31‐cLUC interacted in the nucleus of tobacco leaf cells. (d) The interact ability was expressed as a ratio of LUC to REN. Data values represent three independent experiments with similar results, and SD in all the points is <5%. (e) MdWRKY31‐MdNAC7 interaction alleviates the transcriptional repression of MdXTH2 by MdWRKY31. (f) Quantitative statistics of fluorescence intensity. Data are averages ± SD from three independent experiments.