Supplemental Figure 1. Profiles of JA and 12-OPDA Levels Post Drought Stress

Supplemental Figure 1. Profiles of JA and 12-OPDA levels post drought stress

Levels of (A) 12-OPDA and (B) JA 12 and 36 hours post drought stress.Means ±SD of three independent biological replicates with three technical replicates for each are shown.

Supplemental Figure2. Drought stress signaling uncouples conversion of 12-OPDA to JA

(A) Western blot analyses of transgenic lines expressing HPL-GFP fusion constructs for the two extraplastidial (HPL2-2 and HPL3-TP-2) and one plastidial HPL (HPL3-2) enzyme, using anit-GFP antibody. Lower panel is the Coomassie blue staining of large Rubiscosubunit displaying equal protein loading.(B-E) Levels of HPL-derived metabolites, 3-hexenal and 3-hexenol; (E-F) AOS derived metabolites, 12-OPDA and JA; and (F) ABA in untreated control (while bars), 90 min after wounding (black bars) and drought stressed (grey bars) WT plants and transgenic lines overexpressing HPL enzymes. Means ±SD of three independent biological replicates with three technical replicates for each are shown. Letters above bars indicate significant differences between stress treatments and the corresponding control in various ecotypes (p<0.05).

Supplemental Figure 3.Plants overexpressing extra plastidial HLP are more drought resistance than the WT control

(A) Images of WT and HPL overexpressing plants survived after drought treatment. Plants were grown in individual pots with equal amount of soil. Water was withheld from 3-week-old plants for 10 days followed by 7 days of watering. (B) Percent survival of WT and transgenic plants overexpressing plastidial (HPL3-2) and extraplastidial (HPL2-2 and HPL3-TP-2) HPLs. The results represent the means ±SE from four independent experiments with 10-12 plants of each genotype per experiment. (C) Measurements of stomatal aperture in epidermal peels of WT and HPL overexpressing plants. Representative stomatal images are shown. Data represent the mean of at least 90 measurements ±SE. Asterisk denotes significant differences in stomata aperture between different transgenic lines as compared with the WT control (p<0.05).

Supplemental figure 4. External application of 12-OPDA promotes stomatal closure

Measurements of stomatal aperture in epidermal peels of Col-0 after treatment with various concentrations of (A) 2-hexenal and 3-hexenyl acetate; (B) 12-OPDA alone or together with either ABA or 3-hexenal applied. Data represent the means of minimum 90 measurements ±SE.

Supplemental Table 1. List of primer sequences