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. 2020 May 21;32(8):2621–2638. doi: 10.1105/tpc.19.00971

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Figure 8. — Cooperative Regulation of Camalexin Biosynthesis by CPK5/CPK6 and MPK3/MPK6.

(A) to (D) Coexpression of CPK5^CA and MKK5^DD has additive effects on the induction of camalexin biosynthesis and the activation of camalexin biosynthetic genes. Col-0 and homozygous lines of Est:CPK5^CA-HA and Dex:FLAG-MKK5^DD were pairwise crossed to generate Est:CPK5^CA-HA^+/−, Dex:FLAG-MKK5^DD+/−, and Est:CPK5^CA-HA^+/−/Dex:FLAG-MKK5^DD+/− F1 plants. Camalexin produced from 2-week-old seedlings was measured after treatment with 10 μM Est for 24 h and 5 μM Dex for 16 h (A). The transcript levels of CYP71A13 (B) and PAD3 (C) after treatment with Est and Dex were analyzed by RT-qPCR. The expression of CPK5^CA-HA and FLAG-MKK5^DD and the activation of MPK3/MPK6 by FLAG-MKK5^DD were analyzed by immunoblotting (IB), and total protein loading was assessed by Ponceau S staining (PS [D]). FW, fresh weight.

(E) CPK5/CPK6 and MPK3 contribute additively to B. cinerea-induced camalexin production. Camalexin produced from 2-week-old Col-0 and mutant seedlings indicated was measured at 24 h postinoculation with B. cinerea spores.

In (A), (B), (C), and (E), error bars indicate sd (n = 3 biological repeats), small circles represent individual data points, and different letters above the columns indicate significant differences (P < 0.05), as determined by one-way ANOVA.