The phytohormone abscisic acid (ABA) regulates plant growth, the developmental process, and abiotic stresses. ABA signaling is induced in response to mediate plant acclimation to environmental challenges, including high salinity and drought. The ABA-binding receptors (RCAR/PYR1/PYL), composing of 14 members, are the core components of the ABA-signaling pathway. Here, we observed that the three subfamilies within the RCARs showed different expression patterns at the basal and exogenous ABA levels. Subsequently, we generated transgenic plants overexpressing subfamily III, RCAR11–RCAR14, respectively. The transgenic plants showed increased ABA sensitivity in seed germination and post-germination seedling establishment and root length. Further studies revealed that the overexpressing subfamily III transgenic plants enhanced drought resistance, increased water-use efficiency, and accelerated stress-responsive gene expression compared with the wild-type plants. These findings confirm that the subfamily III plays a key role in ABA-mediated developmental processes and, more importantly, is involved in drought tolerance in the ABA-dependent pathway.

Abscisic acid (ABA) signaling plays important roles in plant growth, development and adaptation to various stresses. RCAR1/PYL9 has been known as a cytoplasm and nuclear ABA receptor in Arabidopsis. To obtain further insight into the regulatory mechanism of RCAR1/PYL9, a yeast two-hybrid approach was performed to screen for RCAR1/PYL9-interacting proteins and an R2R3-type MYB transcription factor, AtMYB44, was identified. The interaction between RCAR1/PYL9 and AtMYB44 was further confirmed by glutathione S-transferase (GST) pull-down and bimolecular fluorescence complementation (BiFC) assays. Gene expression analysis showed that AtMYB44 negatively regulated the expression of ABA-responsive gene RAB18, in contrast to the opposite role reported for RCAR1/PYL9. Competitive GST pull-down assay and analysis of phosphatase activity demonstrated that AtMYB44 and ABI1 competed for binding to RCAR1/PYL9 and thereby reduced the inhibitory effect of RCAR1/PYL9 on ABI1 phosphatase activity in the presence of ABA in vitro. Furthermore, transient activation assay in protoplasts revealed AtMYB44 probably also decreased RCAR1/PYL9-mediated inhibition of ABI1 activity in vivo. Taken together, our work provides a reasonable molecular mechanism of AtMYB44 in ABA signaling.

Drought is a major limiting factor for plant growth and crop productivity. Many Calcineurin B-like interacting protein kinases (CIPKs) play crucial roles in plant adaptation to environmental stresses. It is particularly essential to find the phosphorylation targets of CIPKs and to study the underlying molecular mechanisms. In this study, we demonstrate that CIPK11 acts as a novel component to modulate drought stress in plants. The overexpression of CIPK11 (CIPK11OE) in Arabidopsis resulted in the decreased tolerance of plant to drought stress. When compared to wild type plants, CIPK11OE plants exhibited higher leaf water loss and higher content of reactive oxygen species (ROS) after drought treatment. Additionally, a yeast two hybrid screening assay by using CIPK11 as a bait captures Di19-3, a Cys2/His2-type zinc-finger transcription factor that is involved in drought stress, as a new interactor of CIPK11. Biochemical analysis revealed that CIPK11 interacted with Di19-3 in vivo and it was capable of phosphorylating Di19-3 in vitro. Genetic studies revealed that the function of CIPK11 in regulating drought stress was dependent on Di19-3. The transcripts of stress responsive genes, such as RAB18, RD29A, RD29B, and DREB2A were down-regulated in the CIPK11OE plants. Whereas overexpression of CIPK11 in di19-3 mutant background, expression levels of those marker genes were not significantly altered. Taken together, our results demonstrate that CIPK11 partly mediates the drought stress response by regulating the transcription factor Di19-3.

Because they are sessile organisms, plants need rapid and finely tuned signaling pathways to adapt to adverse environments, including salt stress. In this study, we identified a gene named Arabidopsis thaliana stress-induced BTB protein 1 (AtSIBP1), which encodes a nucleus protein with a BTB domain in its C-terminal side and is induced by salt and other stresses. The expression of the β-glucuronidase (GUS) gene driven by the AtSIBP1 promoter was found to be significantly induced in the presence of NaCl. The sibp1 mutant that lost AtSIBP1 function was found to be highly sensitive to salt stress and more vulnerable to salt stress than the wild type WT, while the overexpression of AtSIBP1 transgenic plants exhibited more tolerance to salt stress. According to the DAB staining, the sibp1 mutant accumulated more reactive oxygen species (ROS) than the WT and AtSIBP1 overexpression plants after salt stress. In addition, the expression levels of stress-induced marker genes in AtSIBP1 overexpression plants were markedly higher than those in the WT and sibp1 mutant plants. Therefore, our results demonstrate that AtSIBP1 was a positive regulator in salinity responses in Arabidopsis.