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. 2022 Jul 21;34(10):4088–4104. doi: 10.1093/plcell/koac209

Figure 7.

Figure 7

Phosphorylation of CBP60g regulates CBP60g activity and resistance to V. dahliae. A and B, The cpk4 cpk11, cpk5 cpk6, and cpk4 cpk5 cpk6 cpk11 mutants are compromised in resistance to V. dahliae. Disease symptoms (A) and index analyses (B) of Arabidopsis WT, cpk4 cpk11, cpk5 cpk6, or cpk4 cpk5 cpk6 cpk11 mutant plants infected with V. dahliae. The plants were photographed and subjected to disease index analyses. Disease indexes were evaluated with three replicates generated from 24 plants for each inoculum. Error bars indicate SD of three biological replicates. Student’s t test was carried out to determine the significance of difference. Different letters indicate significant differences. C, TCH3 enhances CBP60g-activated ProICS1:LUC reporter activity. ProICS1:LUC reporter was transfected alone, or together with CBP60g and TCH3, as indicated. Pro35S:RLuc was co-transfected as internal control. Error bars indicate SD of three biological repeats. Student’s t test was carried out to determine the significance of difference. Different letters indicate significant differences. D, CPK5 enhances the DNA-binding activity of CBP60g. MBP-CBP60g, MBP-CBP60gSSAA, or MBP-CBP60gSSDD were incubated with [γ-32P]ATP-labeled 60-bp double-stranded DNA probe within the ICS1 promoter, and subjected to EMSA. Unlabeled probe was used as competitors (150×) for binding. MBP-CBP60g was preincubated with or without His-CPK5 or His-CPK5Km as indicated for 30 min at room temperature as indicated before EMSA. The experiments were repeated 3 times with similar results. E, Ser450 and Ser495 are required for transcription activator activity of CBP60g. ProICS1:LUC was transfected alone or together with CBP60g, CBP60gS450A, CBP60gS450D, CBP60gSSAA, or CBP60gSSDD as indicated. LUC reporter activity was determined 16 h post-transfection. Error bars indicate SD of three biological repeats. Student’s t test was carried out to determine the significance of difference. Different letters indicate significant differences between CBP60g, CBP60gS450A, CBP60gS450D, CBP60gSSAA, or CBP60gSSDD. F and G, Ser450 and Ser495 are required for resistance to V. dahliae. Disease symptoms (F) and index analyses (G) of Arabidopsis WT, cbp60g-1 mutant, cbp60g-1/CBP60g-FLAG, and cbp60g-1/CBP60gSSAA-FLAG transgenic plants infected with V. dahliae. The plants were photographed and subjected to disease index analyses. Disease indexes were evaluated with three replicates generated from 24 plants for each inoculum. Error bars indicate SD of three biological replicates. Student’s t test was carried out to determine the significance of difference. Different letters indicate significant differences.