The indica transcription factor gene OsERF65 confers rice drought tolerance via upregulation of late embryogenesis abundant genes

• The expressions of OsERF65 were induced by drought stress. • Overexpression of OsERF65 from indica rice improved drought tolerance. • OsERF65 binds to the promoters and promotes the transcription of LEA genes. • Five haplotypes of OsERF65 were identified and OsERF65 Hap1 has stronger trans-activation activity than that of OsERF65 Hap2 . Drought stress severely impacts crop productivity. As a subclass of plant transcription factors, APETALA2/Ethylene Responsive Factors (AP2/ERFs) play crucial roles in plant development and abiotic stress responses. In rice, the functions of many members of the ERF subfamily of AP2/ERFs in drought resistance remain unclear. Here, we characterized the function of OsERF65 , an ERF transcription factor gene from indica rice cultivar ‘Zhenshan97B’(ZS97B), in drought tolerance. Overexpression of OsERF65 ZS97B significantly enhanced the drought tolerance of transgenic rice. In contrast, its knockout reduced rice drought tolerance. Transcriptome sequencing analysis revealed that late embryogenesis abundant (LEA) genes were significantly up-regulated in OsERF65- overexpressing rice plants. Further assays verified that OsERF65 binds to the promoters and activates the transcription of LEA genes. Yeast two-hybrid analysis showed that OsERF65 could interact with OsMYB48 and other transcription factors. Additionally, five haplotypes of OsERF65 were identified in rice germplasm, with Hap1 dominant in indica rice and Hap2 dominant in japonica rice. The OsERF65 Hap1 encoded protein exhibited stronger transcriptional activation activity than OsERF65 Hap2 . Collectively, our findings reveal an indica rice-derived ERF transcription factor that positively regulates drought tolerance via upregulation of LEA genes.