Fig. 2: COI1-independent function of cis-OPDA. | Nature Communications

Fig. 2: COI1-independent function of cis-OPDA.

From: Downstream metabolites of (+)-cis-12-oxo-phytodienoic acid function as noncanonical bioactive jasmonates in Arabidopsis thaliana

Fig. 2

A–H Gene expression analysis by RT-qPCR in 10-day-old wild-type (WT, Col-0; white bars) and coi1-1 mutant (gray bars) seedlings treated with 30 μM ( − )-JA or cis-OPDA for 30 min, or mock-treated. Gene expression levels were normalized to UBQ10. Data are presented as mean ± SD, with n = 4 biologically independent samples per condition. Statistical significance was determined using one-way ANOVA followed by Tukey-Kramer post hoc test (two-sided). Different letters represent a significant difference at P  <  0.05. The experiment was repeated independently three times with similar results. I Evaluation of the affinity between cis-OPDA and COI1-JAZs. Pull-down assay of GST-AtCOI1 with Fl-AtJAZPs in the presence of JA-Ile (1 µM) or cis-OPDA (1/30 µM). Fl-AtJAZ13 was used as a negative control because JAZ13 has no canonical JAZ degron sequence, which is necessary for JA-Ile perception. GST-AtCOI1 bound to Fl-AtJAZPs was pulled down with anti-fluorescein antibody and Protein G magnetic beads and analyzed by immunoblotting (anti-GST-HRP conjugate for detection of GST-AtCOI1). Arrowheads indicate GST-AtCOI1 (95 kDa). Experiments were repeated three times with similar results (shown in Supplementary Figs. 2–3).

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