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. 2022 Jan 11;12:706633. doi: 10.3389/fpls.2021.706633

FIGURE 3.

FIGURE 3

pxc3 mutants have reduced sensitivity to TDIF peptide treatment. (A) Longitudinal confocal sections of wild type and mutant hypocotyls (7 DAG seedlings) treated or not with the TDIF (5 μM) peptide for 4 days as indicated. The double-sided arrow indicates the stele width. Scale bars represent 20 μm. (B) Quantification of the stele width (n ≥ 15). This experiment was done three times. In two out of three replicates, a significant interaction between treatment and genotype was found for pxc3 mutants compared to wild type. Numbers indicate fold change in the mean stele width relative to the untreated sample for each genotype. A two-way ANOVA followed by a Dunnett’s test was performed and the asterisks indicate a significant genotype and treatment interaction compared to wild type (***P < 0.001). (C) WOX4 and WOX14 transcript levels after 8 h TDIF treatment of wild-type and mutant seedlings. Fold change relative to wild type (mock) is shown. Numbers indicate fold change in the mean value (line) relative to the untreated sample for each genotype (n = 5 independent replicates). A one-way ANOVA followed by a Tukey’s test was performed on Log2-transformed peptide/mock gene expression ratio, and the asterisks indicate a significant genotype and treatment interaction compared to wild type (*P < 0.05, ** P < 0.01, and ***P < 0.001, and ns indicates that no significant difference was found). (D) Longitudinal confocal sections of wild type and mutant hypocotyls (7 DAG seedlings) treated or not with the TDIF (5 μM) peptide for 4 days as indicated. The double-sided arrow indicates the stele width. Scale bars represent 20 μm. (E) Quantification of the stele width (n ≥ 15). This experiment was done two times. Numbers indicate fold change in the mean stele width relative to the untreated sample for each genotype. A two-way ANOVA followed by a Dunnett’s test was performed and the asterisks indicate a significant genotype and treatment interaction (*P < 0.05, **P < 0.01, and ****P < 0.0001, and ns indicates that no significant difference was found). (F) WOX4 and WOX14 transcript levels after 8 h TDIF treatment of wild-type and mutant seedlings. Fold change relative to wild type (mock) is shown. Numbers indicate fold change in the mean value (line) relative to the untreated sample for each genotype (n = 3 independent replicates). A one-way ANOVA followed by a Tukey’s test was performed on Log2-transformed peptide/mock gene expression ratio, and the asterisks indicate a significant genotype and treatment interaction (*P < 0.05, *** P < 0.001, and ***P < 0.001, and ns indicates that no significant difference was found).