FIGURE 4.

AtSMXL7 degradation in N. benthamiana is NbD14‐dependent. (a) Diagram of the ratiometric AtSMXL7 reporter expressed in pRATIO3212. mScarlet‐I is a fluorescent reporter protein translationally fused to the C‐terminus of AtSMXL7, *F2A is a modified “self‐cleaving” peptide, and Venus is a yellow fluorescent protein used for normalization. Diagram adapted from Khosla, Rodriguez Furlan, et al. (2020). (b) Background‐corrected AtSMXL7‐mScarlet‐I to Venus fluorescence in N. benthamiana wt and Nbd14a,b leaf discs after 16‐h treatment with solvent control (0.02% acetone [v/v]), or 10 μM rac‐GR24, GR24^5DS, or GR24 ^ent‐5DS. (c) Background‐corrected AtSMXL7‐mScarlet‐I to Venus fluorescence in Nbd14a,b leaf discs after 16‐h treatment with solvent control (0.02% acetone [v/v]), or 10‐μM rac‐GR24. pRATIO3212‐AtSMXL7 was coexpressed with pGWB415 empty vector (e.v.), AtD14, or AtD14^S97A. Box plots show median with 25th and 75th percentiles, and whiskers represent minimum and maximum values (n = 3–6 leaf discs). *p < .05, two‐way ANOVA with (b) Dunnetts multiple comparisons test or (c) Bonferronis multiple comparisons test, comparing GR24 treatments with control for each genotype