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. 2019 Feb 5;10:67. doi: 10.3389/fpls.2019.00067

FIGURE 5.

FIGURE 5

BnABI3 regulates mucilage secretion by targeting MUM1 and GALT5 expression. (A–C) Relative expression of MUM1, PMEI6, and GALT5 in BnABI3GR transgenic plant siliques treated with 20 μM DEX or mock for 0, 3, or 6 h. The expression of the corresponding genes in mock-treated plants was set to 1.0. Values are given as mean ± SD, n = 3. ∗p < 0.05 by Mann–Whitney test. (D–F) Relative expression level of MUM1, PMEI6, and GALT5 in BnABI3GR transgenic plant siliques treated with 20 μM DEX, 100 μM CHX, 20 μM DEX plus 100 μM CHX, or mock. The gene expression in mock-treated plants was set at 1.0. Values are given as mean ± SD, n = 3. ∗p < 0.05 by Kruskal–Wallis test. (G) Schematic diagram indicating the locations of ABI3 binding sites (D1 to D3) in the GALT5 and MUM1 genes promoters. (H,I) Transient dual-luciferase reporter assay. The pGreenII-0800 luciferase (LUC) construct containing the GATL5 and MUM1 promoters and the p62-SK construct with the BnABI3 coding region and truncated BnABI3 without DNA binding domain were transiently co-transformed into Col-0 protoplasts. Firefly LUC and Renilla luciferase (REN) activities were measured after culturing the protoplasts under low-light conditions for 16 h. Values are given as mean ± SD, n = 4. (J) ChIP-qPCR analysis of the ability of BnABI3 to bind to the promoters of GATL5 and MUM1. An anti-HA monoclonal antibody was used for DNA immunoprecipitation from 2-week-old 35S::BnABI3-HA transgenic plants. Gray bars indicate the enrichment fold changes normalized to ACT2. Values are given as mean ± SD, n = 3. ∗p < 0.05 by Kruskal–Wallis test.