In eukaryotes, Histone acetylation levels directly regulate downstream gene expression. As a plant-specific Histone deacetylase (HDAC), HD2D is involved in plant development and abiotic stress. However, the response of HD2D to drought stress and its interaction proteins are still unclear. In this study, HD2D gene expression patterns were analyzed, revealing that the HD2D gene was highly expressed in roots and rosette leaves, but poorly expressed in the other tissues examined. The HD2D gene expression was induced by D-mannitol. The study investigated the responses of the wild-type plant (WT), HD2D-overexpression lines, and hd2d mutants to drought stress, further demonstrating that HD2D-overexpressing lines showed Abscisic acid (ABA) hypersensitivity and drought tolerance, and these phenotypes were not present in hd2d mutants. RNA-seq analysis revealed the transcriptome changes by HD2D under drought stress and showed that HD2D responded to drought stress through the ABA signaling pathway. In addition, the study demonstrated that CASEIN KINASE II (CKA4) directly interacted with HD2D. The phosphorylation of Ser residues of HD2D by CKA4 enhanced HD2D enzymic activity. Furthermore, the phosphorylation of HD2D was shown to contribute to lateral root development and ABA sensing in Arabidopsis, but, these phenotypes could not be reproduced by the overexpression of Ser-phosphorylated mutant of HD2D. Collectively, this study suggested that the HD2D responded to drought stress by regulating the ABA signaling pathway and the expression of drought stress-related genes. The regulation mechanism of HD2D mediated by CKII phosphorylation provides new insights for its response to ABA and lateral root development in Arabidopsis.