Interaction of MdDof2.4 with MdPAT10proIn-868 Increases the Expression of MdPAT10 and the Accumulation of PC. (IMAGE)
Caption
Interaction of MdDof2.4 with MdPAT10proIn-868 increases the expression of MdPAT10 and the accumulation of PC. (A) The Y1H assay shows the binding of MdDof2.4 to the insertion-containing promoter of MdPAT10 (MdPAT10proIn-868). (B) EMSA demonstrates the binding of MdDof2.4 to the 5′-AAAAG-3′ motif in MdPATA10proIn-868. The hot probe is a biotin-labeled fragment containing the 5′-AAAAG-3′ motif; while the cold probe is a nonlabeled competitive probe, prepared at concentrations 150- and 250-fold, respectively, of the hot probe; the mutant motif is 5’-CCCCC-3′. (C) Diagrams of the effector and reporter constructs. (D) Interaction between MdDof2.4 and MdPAT10proIn-868, as determined by dual luciferase reporter assays in Nicotiana benthamiana leaves. Data are presented as the mean ± SE (n = 9). Statistical analyses were performed using the one-way ANOVA, with statistically significant differences indicated by different lowercase letters. (E–F) qRT-PCR analysis of MdDof2.4 (E) and MdPAT10 (F) genes in WT, transiently overexpressed, or silenced MdDof2.4 fruits. 35S: MdDof2.4 or pTRV2-MdDof2.4 were transiently transformed in ‘Gan Hong’ fruit at 100 DAF. (G) Determination of the PC content in WT, transiently overexpressed, or silenced MdDof2.4 fruits. Error bars indicate SD (n = 20). Statistical analyses were performed using the Student’s t-test, with statistically significant differences indicated by *P < 0.05, **P < 0.01, or ***P < 0.001 in (E–G). N.s. denotes no significant difference. (H) A proposed working model of the regulation of PC accumulation by the MdDof2.4-MdPAT10 module in apples.
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Horticulture Research
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