Summary In mammalians and yeast, the splicing factor U2AF65/Mud2p functions in precursor messenger RNA (pre‐mRNA) processing. Arabidopsis AtU2AF65b encodes a putative U2AF65 but its specific functions in plants are unknown. This paper examines the function of AtU2AF65b as a negative regulator of flowering time in Arabidopsis. We investigated the expression and function of AtU2AF65b in abscisic acid (ABA)‐regulated flowering as well as the transcript abundance and pre‐mRNA splicing of flowering‐related genes in the knock‐out mutants of AtU2AF65b. The atu2af65b mutants show early‐flowering phenotype under both long‐day and short‐day conditions. The transcript accumulation of the flowering repressor gene FLOWERING LOCUS C (FLC) is reduced in the shoot apex of atu2af65b, due to both increased intron retention and reduced transcription activation. Reduced transcription of FLC results, at least partially, from the abnormal splicing and reduced transcript abundance of ABSCISIC ACID‐INSENSITIVE 5 (ABI5), which encodes an activator of FLC in ABA‐regulated flowering signaling. Additionally, the expression of AtU2AF65b is promoted by ABA. Transition to flowering and splicing of FLC and ABI5 in the atu2af65b mutants are compromised during ABA‐induced flowering. ABA‐responsive AtU2AF65b functions in the pre‐mRNA splicing of FLC and ABI5 in shoot apex, whereby AtU2AF65b is involved in ABA‐mediated flowering transition in Arabidopsis.

Pre-mRNA splicing is an important step for gene expression regulation. Yeast Bud13p (bud-site selection protein 13) regulates the budding pattern and pre-mRNA splicing in yeast cells; however, no Bud13p homologs have been identified in plants. Here, we isolated two mutants that carry T-DNA insertions at the At1g31870 locus and shows early embryo lethality and seed abortion. At1g31870 encodes an Arabidopsis homolog of yeast Bud13p, AtBUD13. Although AtBUD13 homologs are widely distributed in eukaryotic organisms, phylogenetic analysis revealed that their protein domain organization is more complex in multicellular species. AtBUD13 is expressed throughout plant development including embryogenesis and AtBUD13 proteins is localized in the nucleus in Arabidopsis. RNA-seq analysis revealed that AtBUD13 mutation predominantly results in the intron retention, especially for shorter introns (≤100 bases). Within this group of genes, we identified 52 genes involved in embryogenesis, out of which 22 are involved in nucleic acid metabolism. Our results demonstrate that AtBUD13 plays critical roles in early embryo development by effecting pre-mRNA splicing.

Abscisic acid (ABA) is an important phytohormone regulating plant growth, development and stress responses. A multitude of key factors implicated in ABA signaling have been identified; however, the regulation network of these factors needs for further information. AtS40.4, a plant-specific DUF584 domain-containing protein, was identified previously as a senescence regulator in Arabidopsis. In this study, our finding showed that AtS40.4 was negatively involved in ABA signaling during seed germination and early seedling growth. AtS40.4 was highly expressed in seeds and seedlings, and the expression level was promoted by ABA. AtS40.4 was localized both in the nucleus and the cytoplasm. Moreover, the subcellular localization pattern of AtS40.4 was affected by ABA. The knockdown mutants of AtS40.4 exhibited an increased sensitivity to ABA, whereas the overexpression of AtS40.4 decreased the ABA response during seed germination and seedling growth of Arabidopsis. Furthermore, AtS40.4 was involved in ABRE-dependent ABA signaling and influenced the expression levels of ABA INSENTIVE (ABI)1-5 and SnRK2.6. Further genetic evidence demonstrated that AtS40.4 functioned upstream of ABI4. These findings support the notion that AtS40.4 is a novel negative regulator of the ABA response network during seed germination and early seedling growth.

During the opening and closing of stomata, guard cells undergo rapid and reversible changes in their volume and shape, which affects the adhesion of the plasma membrane (PM) to the cell wall (CW). The dynamics of actin filaments in guard cells are involved in stomatal movement by regulating structural changes and intracellular signaling. However, it is unclear whether actin dynamics regulate the adhesion of the PM to the CW. In this study, we investigated the relationship between actin dynamics and PM-CW adhesion by the hyperosmotic-induced plasmolysis of tobacco guard cells. We found that actin filaments in guard cells were depolymerized during mannitol-induced plasmolysis. The inhibition of actin dynamics by treatment with latrunculin B or jasplakinolide and the disruption of the adhesion between the PM and the CW by treatment with RGDS peptide (Arg-Gly-Asp-Ser) enhanced guard cell plasmolysis. However, treatment with latrunculin B alleviated the RGDS peptide-induced plasmolysis and endocytosis. Our results reveal that the actin depolymerization is involved in the regulation of the PW-CW adhesion during hyperosmotic-induced plasmolysis in tobacco guard cells.

Sorbitol dehydrogenase (SDH) catalyses the reversible oxidation of sorbitol, xylitol and ribitol to their corresponding ketoses. In this study, we investigated the expression and role of Arabidopsis SDH in salt and osmotic stress tolerance, and abscisic acid (ABA) response. The expression patterns of SDH were investigated using transgenic Arabidopsis plants expressing beta-glucuronidase (GUS) under control of the promoter with the first intron of SDH. qRT-PCR and histochemical assay of GUS activity were used to study SDH expression regulation by ABA, salt and osmotic stress. SDH-overexpression lines of Arabidopsis were used to investigate the role of SDH in salt and osmotic stress, and ABA response. Arabidopsis SDH was predominantly expressed in source organs such as green cotyledons, fully expanded leaves and sepals, especially in vascular tissues of theses organs. SDH expression was inhibited by NaCl and mannitol treatments. Seed germination and post-germination growth of SDH-overexpressing lines exhibited decreased sensitivity to salt and osmotic stress compared to WT plants. The transcript of SDH was induced by ABA. Overexpression of SDH decreased sensitivity to ABA during seed germination and post-germination growth. Expression of AAO3 increased but ABI5 and MYB2 decreased in SDH-overexpressing lines after ABA treatment. This study demonstrates that expression of SDH is regulated by ABA, salt and osmotic stress. SDH functions in plant tolerance to salt and osmotic stress, and ABA response via specific regulating gene expression of ABA synthesis and signalling in Arabidopsis.