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. 2020 Jun 30;8(1):nwaa149. doi: 10.1093/nsr/nwaa149

Figure 2.

Figure 2.

LRX3/4/5-RALF22/23-FER module negatively regulates JA signaling pathway. (A) qRT-PCR analysis of the transcript levels of PDF1.2, PDF1.3, PR1 and PR5 in wild type, lrx345 and fer-4 seedlings. ACTIN8 was used as an internal control. (B) qRT-PCR analysis of the transcript levels of PDF1.2, PDF1.3, PR1 and PR5 in wild type and two independent RALF22 overexpressing lines (#5 and #6). ACTIN8 was used as an internal control. (C) Jasmonic acid (JA) and salicylic acid (SA) contents in wild type, lrx345 and fer-4 seedlings. (D) JA and SA contents in transgenic plants overexpressing RALF22. (E) Immunoblotting analysis of JAZ1 protein in wild type, lrx345 and fer-4 seedlings expressing JAZ1-YFP. (F) qRT-PCR analysis of the JAZ1 transcript levels in the transgenic plants. (G, H) Transgenic plants expressing JAZ1-GFP (G) and JAZ9-GFP (H) were treated with mature RALF22 (mRALF22) for 0, 3, 6, 9, 12 and 24 h. Immunoblottings were performed using anti-GFP antibody. (I) Ten-day-old seedlings were treated with mRALF22 (1 μM) for 0, 6 and 12 h. Immunoblotting assays were performed using anti-GFP and anti-Actin antibodies. The band intensity in (E), (G) and (H) was qualified using ImageJ software. Values in (A–D) are means of three biological replicates ± SD, *P < 0.05 and **P < 0.01 (Student's t-test).