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. 2018 Feb 22;19(7):1624–1638. doi: 10.1111/mpp.12635

Figure 8.

Figure 8

Function of GhMKK4–GhMPK20 is essential for Fusarium oxysporum‐induced GhWRKY40 expression. (A) GhMPK20 interacts with GhWRKY40 according to the yeast two‐hybrid (Y2H) system assay results. The indicated BD and AD fusion constructs were co‐transformed into the Y2H Gold yeast strain and grown on the SD media DDO and QDO/X. (B) Co‐immunoprecipitation (Co‐IP) analysis was performed using MYC antibody in cotton protoplasts. GhMPK20‐MYC and GhWRKY40‐FLAG fusion constructs were co‐transformed into cotton protoplasts. Western blot analysis shows protein expression levels. (C) GhWRKY40 expression levels in GhWRKY40‐silenced cotton plants. The data are presented as the mean ± standard error (SE) of three independent experiments (n = 9). The letters above the columns represent significant differences (P < 0.05) based on Tukey's honestly significant difference (HSD) test. (D) GhWRKY40 expression levels in GhMPK20‐silenced cotton plants with or without F. oxysporum infection. The error bars indicate the mean ± SE of three independent experiments (n = 9). Asterisks above the lines indicate significant differences (*P < 0.05, **P < 0.01) based on Tukey's HSD test. (E) Representative phenotypes of GhWRKY40‐silenced cotton after infection with F. oxysporum for 5 days. DAB, diaminobenzidine. (F) GhMPK20 and GhWRKY40 expression levels in GhMKK4‐silenced cotton plants with or without F. oxysporum infection. The error bars indicate the mean ± SE of three independent experiments (n = 9). Asterisks above the lines indicate significant differences (*P < 0.05, **P < 0.01) based on Tukey's HSD test. SD, synthetically defined medium; SD medium DDO, SD medium without Leu and Trp; SD medium QDO/X, SD medium with X‐a‐gal and without Ade, His, Leu, Trp.