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. 2019 Sep 16;8:e44474. doi: 10.7554/eLife.44474

Figure 1. The LYK5-CERK1-PBL27 receptor complex and SLAH3 are required for chitin-induced stomatal closure.

(A–C) Stomatal aperture measurements in mutants of all members of the LysM-RLK family (A), pbl27 (B), slac1 and slah3 (C). Mature leaf discs were soaked in opening buffer (10 mM MES, 50 mM KCl, pH 6.15) and kept under light (100 μmol.m-2 s −1) for 2 hr. Stomatal apertures were measured 2 hr after treatment with 1 mg/ml chitin. Values are mean ± SD (n > 60; two-way ANOVA). Different letters indicate significantly different values at p<0.05. These experiments were repeated three times with similar results.

Figure 1—source data 1. Source data for stomatal measurements shown in Figure 1 and Figure 1—figure supplements 1 and 2, and for Figure 4.
elife-44474-fig1-data1.xlsx (335.9KB, xlsx)
DOI: 10.7554/eLife.44474.005

Figure 1.

Figure 1—figure supplement 1. Functional complementation and guard cell expression of the LYK5-CERK1-PBL27 receptor complex.

Figure 1—figure supplement 1.

Stomatal aperture measurements in transgenic cerk1/pCERK1::CERK1-eGFP, lyk5/pLYK5::LYK5-eGFP and pbl27-1/pPBL27::PBL27-eGFP. Mature leaf discs of six independent transgenic T1 lines were soaked in opening buffer (10 mM MES, 50 mM KCl, pH 6.15) and kept under light (100 μmol.m-2 s −1) for 2 hr. Stomatal aperture was measured 2 hr after treatment with 1 mg/ml chitin. Values are mean ± SD (n > 50; two-way ANOVA). Different letters indicate significantly different values at p<0.05. These experiments were repeated three times with similar results.
Figure 1—figure supplement 2. Current ejection of chitosan induces stomatal closure in Arabidopsis.

Figure 1—figure supplement 2.

Chitosan was current ejected into the cell wall of guard cells in intact leaves via a microelectrode, by application of 1 nA positive current for 1 min. The tip of electrode was filled with 0.3 mg/ml chitosan or 10 mM MES/BTP (pH6, control), and rest with 300 mM KCl. Data are presented as average ± SE. Col-0 control, n = 8 stomata; Col-0 chitosan, n = 8 stomata; slah3-1 control, n = 3 stomata; slah3-1 chitosan, n = 8 stomata. The statistical significance indicates with different characters (one-way ANOVA, Tukey’s test, p<0.05).