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Rice (Oryza sativa L.) is a main food crop in China and is crucial for the maintenance of national food security. The growth of rice seedling roots is regulated by a variety of genes and is closely related to abscisic acid (ABA) metabolism and ABA signaling pathways. In this study, we found that OsABT could increase the length of rice root tip meristem cells and upregulate root development-related genes, thereby alleviating ABA's inhibitory effects on rice root growth and seed germination. The overexpression of OsABT reduced the ABA content by downregulating ABA synthesis genes (OsNCED3 and OsNCED5) and upregulating the ABA catabolic gene (OsABA8ox2). In addition, OsABT interacted with OsPYL4, OsPYL10, and OsABIL2 via the ABA signal transduction pathway. By inhibiting the expression of positive regulatory genes (OsPYL9 and Rab16a) and increasing the expression of a negative regulatory gene (OsABIL1), OsABT negatively regulates the ABA signal transduction pathway. Transcriptome analysis revealed that OsABT inhibited the activity of Gene Ontology entries in response to ABA. Thus, OsABT increased the length of the rice root meristem, reduced the accumulation of ABA in the roots, and negatively regulated the ABA signal transduction pathway by interacting with key proteins in this pathway, ultimately relieving the inhibitory effect of ABA on rice root length and seed germination. [ABSTRACT FROM AUTHOR]