Fig. 4: Functional analysis of PYL4 phosphorylated by CIPK1 in vivo.
a Germinating seeds were grown on 1/2 Murashige-Skoog (MS) medium containing 3 μM ABA and imaged after 5 d. Scale bar, 1.4 cm. The experiments was repeated thrice with similar results. b Images of 14-day-old seedlings growing on 1/2 MS with or without 20 μM ABA. Protein levels of transgenic materials used in Fig. 4. The PYL4 protein was detected using Anti-FLAG antibodies. Scale bar, 1.4 cm. Three independent replications of the experiment yielded similar results. c Images of PYL4 transgenic plants growing normally for 40–50 d in the greenhouse. Scale bar, 8.0 cm. Three independent replicate experiments obtained similar results. (d) Phenotype of PYL4/12458, PYL4S129A/12458, and PYL4S129D/12458 under drought conditions. Scale bar, 2.0 cm. The experiment was repeated three times with similar results. e water loss measure in detached leaves of PYL4 transgenic plants. The experiment was repeated three times with independent treatments. Data are means ± SD of 3 replicates. Asterisks indicate significance compared to the wild type. ***P < 0.001, Student’s t-test (two-sided). f Leaf temperature of PYL4 transgenic plants. Leaf temperature was measured on nine leaves in triplicates. Scale bar, 2.0 cm. Box plots show the 25% quantile, median (line), and 75% quantile. The whiskers plots (Tukey method) represent minimum and maximum values. Statistical analysis was performed by one-way ANOVA analysis followed by Dunnett’s multiple comparison test. Asterisks indicate significance compared to the wild type (**P < 0.01).
