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Figure 1—figure supplement 3. The biochemistry, physiology and stomatal features of single and double mutants, and testing of PAP action via pathogen signaling.(A) Stomata density, epidermal cell density, stomatal index and total stomatal opening area per leaf area from CryoSEM images of abaxial surface of leaves of six week-old plants. Stomatal index is calculated according to the equation: Stomatal index = [(stomatal density) / (stomatal density + epidermal cell density)]*100. Calculations were performed from five different fields of view per leaf per genotype, n > 30. The experiments were performed twice with similar results. Significant differences at p<0.05 (*) or no significant difference (n.s.) are shown for abi1-1 sal1-8 to abi1-1 and ost1-2 sal1-8 to ost1-2, respectively. (B) Effect of ABA on the stomatal aperture of Col-0 vs ColLer hybrid of leaf peels from five- to six-week old plants after 2 hr of 50 µM ABA. (C) Effect of ABA on the stomatal aperture of ost1 (Col-0) treated with either mock control buffer or 100 µM ABA for 30 min. Results are mean ± SE of 15 stomata per treatment. (D) Stomatal closure, as assayed by leaf temperature using infrared thermography, in well-watered and drought-stressed WT, ost1 and ost1sal1-6 (all in Col-0 background). Values are mean ± SE of five individual plants per genotype. Significant differences are shown for p<0.05 (*). (E) Effect of the sal1-8 mutation on total leaf ABA content. The ABA content in sal1-8 and ost1-2 sal1-8 were marginally higher but not statistically different to ColLer or ost1-2 (ANOVA, p=0.09). Results are means and standard error of 3 × 100 mg pooled two-week old seedlings. (F) Stomatal closure in leaf peels of the chloroplastic calcium sensor mutant, cas1, after treatment with 100 µM ABA, 100 µM PAP or control measuring buffer (Means ± SEM of >40 stomata per genotype). Significant differences at t = 30 min (a, b; p<0.05) are shown.DOI: http://dx.doi.org/10.7554/eLife.23361.005 Image published in: Pornsiriwong W et al. (2017) © 2017, Pornsiriwong et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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