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Figure 2. Exogenous PAP interacts with ABA signaling and acts in stomatal closure in both Arabidopsis and barley.(A) Stomatal aperture, calculated using measurements of pore width and length, in leaf peels of wild type (ColLer) plants treated with either 100 µM PAP or 100 µM ABA over a period of 1 hr. Values are means, expressed as a percentage compared to t = 0 min, of at least 20 stomata ± SEM. Rates of closure were compared by modelling the closure between 10–25 min (log-transformed data), significant difference groups (p<0.05) are denoted by #, *. Final level of closure was also considered by ANOVA across the final 30 min; significant difference (p<0.05) denoted a, b, c. (B) Stomatal aperture in leaf epidermal peels of three-week old barley plants in measuring buffer (Control) for 10 min before adding 100 µM ABA or 100 µM PAP for another 50 min. Values are means ± SEM (n = 17–20 stomata of four plants). Significant difference (p<0.05) is denoted a, b. (C) Stomatal aperture as in (A) but treated with either 100 µM PAP or 1 mM ATP alone or in combination, in measuring buffer. Values are means of at least eight stomata ± SEM. The control treatment for (A), (B) and (C) was 1 hr of measuring buffer. (D) Thermography of 35-day old wild type leaves petiole fed with 250 µL of different combinations of 20 µM ABA, 100 mM LiCl, 1 mM PAP and 10 mM ATP in infiltration buffer or buffer alone (Control). Mean and SEM of leaf temperature from three leaves from three plants per genotype are shown. Leaves in solution were returned to growth chamber and temperature measured at indicated timepoints. Significant differences to control are shown (*p<0.05; **p<0.01). Also see Figure 2—figure supplement 1.DOI: http://dx.doi.org/10.7554/eLife.23361.006Figure 2—figure supplement 1. Exogenous PAP feeding to plant leaves via epidermal leaf peels or petiole feeding.Petiole feeding of PAP for 1 hr results in accumulation of PAP in leaves. Levels were significantly enhanced by co-application with LiCl, an inhibitor of the PAP catabolic enzyme SAL1, or with ATP, which outcompetes PAP for transport into plastids where PAP is degraded. ATP also allows PAP to be localized to its sites of action, the nucleus/cytoplasm. Results averaged from three individual plants ± SEM. a, b and c represent significant differences (p<0.05).DOI: http://dx.doi.org/10.7554/eLife.23361.007 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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