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. 2021 Mar 29;12:1952. doi: 10.1038/s41467-021-21694-3

Fig. 8. ALMT9 but not ALMT9F243C/Y245C restores the GABA sensitivity of almt9-2.

Fig. 8

a Stomatal aperture measurement of A. thaliana WT, almt9-2 and complementation lines. Epidermal strips were pre-incubated in stomatal measurement buffer for 1 h under dark, followed by a 1.5 h dark-to-light transition, as indicated above graphs by black (dark) or white (light) bars, ±2 mM GABA; n  = 189 (control) and n = 195 (GABA) for WT-like 2 (segregated from almt9-2)21, n = 197 (control) and n = 153 (GABA) for almt9-2, n = 213 (control) and n = 178 (GABA) for almt9-2 complement with 35S::ALMT9 #1 (almt9-2/ALMT9 #1), n = 219 (control) and n = 127 (GABA) for almt9-2/ALMT9 #2, n = 195 (control) and n = 115 (GABA) for almt9-2 complemented with 35S::ALMT9 with double mutation F243C/Y245C (ALMT9F243C/Y245C) targeting the putative GABA interaction residues18,36,39 (almt9-2/F243C/Y245C #1), n = 221 (control) and n = 109 (GABA) for almt9-2/F243C/Y245C #2 with control treatment. b–g Leaf feeding assay of almt9-2 and complementation lines. Stomatal conductance of detached leaves from 5–6-week-old Arabidopsis almt9-2, almt9-2/ALMT9 #2 and almt9-2/F243C/Y245C #1 plants was recorded using a LI-COR LI-6400XT in response to dark (shaded region) and 200  µmol m−2 s−1 light (white region), fed with artificial xylem sap solutions ± 4 mM GABA (b, d, f). The iWUE of almt9-2 (c), almt9-2/ALMT9 #2 (e) and almt9-2/F243C/Y245C #1 (g) detached leaves was calculated based on the ratio of photosynthetic rate (Supplementary Fig. 17b, e, h) versus stomatal conductance (b, d, f); n  = 14 (control) and n = 13 (GABA) for almt9-2 (b, c); n  = 15 (control and GABA) for almt9-2/ALMT9 #2 (d, e); n  = 13 (control) and n = 12 (GABA) for almt9-2/F243C/Y245C #1 (f, g). All data are plotted with box and whiskers plots: whiskers plot represents minimum and maximum values, and box plot represents second quartile, median and third quartile (a, c, e, g), or data are represented as mean ± s.e.m (b, d, f); statistical difference was determined by two-sided Student’s t test; *P < 0.05, **P < 0.01, ****P < 0.0001 (a–g).