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. 2018 Jul 11;9:1004. doi: 10.3389/fpls.2018.01004

FIGURE 3.

FIGURE 3

SlLYK1 is required for chitin responses. (A,B) Relative transcript levels of SlLYK1, SlLYK12, and SlLYK13 after chitooctaose (CO8) treatment in leaves (A) and roots (B). RNA was extracted from 2-week-old wild type leaves and roots 30 min after CO8 treatment. Transcript levels were detected by qRT-PCR. CO8-induced immune responses were analyzed in leaves (C–E) and roots (F,G) 4 weeks after infiltration with VIGS-SlLYK1, -12, -13, and VIGS-GUS. (C) CO8-induced reactive oxygen species (ROS) accumulation. ROS was measured using a chemiluminescence assay. Signals were recorded for 30 min and ROS were quantified as the total amount of light emitted (RLU). Data are expressed as means ± SD (n = 8). (D,F) CO8-induced SlWRKY53 (Solyc08g008280) gene expression in leaves (D) and roots (F). RNA was extracted 30 min after CO8 treatment, and gene expression was detected by qRT-PCR. Asterisks indicate significant differences from the VIGS-GUS control (Student’s t-test: ∗P ≤ 0.05, ∗∗P ≤ 0.01). (E,G) CO8-induced MAP kinase phosphorylation in leaves (E) and roots (G). After CO8 treatment, MAP kinase phosphorylation was detected by immunoblot using the α-P42/P44 MAPK antibody and α-cFBPase (CYTOSOLIC FRUCTOSE-1,6-BISPHOSPHATASE) as a loading control. The experiment was repeated twice with similar results.