ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR330/029/SRR33001429/SRR33001429_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR330/032/SRR33001432/SRR33001432_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR330/030/SRR33001430/SRR33001430_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR330/031/SRR33001431/SRR33001431_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR330/029/SRR33001429/SRR33001429_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR330/032/SRR33001432/SRR33001432_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR330/031/SRR33001431/SRR33001431_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR330/030/SRR33001430/SRR33001430_1.fastq.gzprimaryOK200GenomicsGuangdong Ocean Universityhttps://www.ebi.ac.uk/ena/browser/view/PRJNA1246497Salt stress significantly impairs rice productivity by disrupting ion homeostasis and generating oxidative damage that undermines protein synthesis. In rice, the translation elongation factor eEF1 plays a critical role in the accurate, GTP-dependent delivery of aminoacyl-tRNAs to the ribosome, a process that becomes compromised under stress conditions. Here, we report the design and comprehensive characterization of a nucleic acid aptamer (S2-A) that binds rice eEF1 with nanomolar affinity. Using iterative SELEX from both a fully randomized (N40) and a stem-enriched (Stem2) library, we enriched aptamers that converge on a conserved stem-bulge architecture. Binding analyses via EMSA revealed an apparent dissociation constant of 5-10 nM for S2-A, while structural predictions using RNAstructure and AlphaFold-based modeling, together with MDockPP docking, indicated that S2-A targets the GTP-binding domain of eEF1. Site-directed mutagenesis and fluorescence polarization assays identified Ile585, Lys621, and Arg625 as critical for the aptamer-eEF1 interaction, with the K621A mutation causing the most pronounced loss of binding. Functionally, rice seedlings transfected with the S2-A aptamer under 150 mM NaCl stress exhibited improved growth, enhanced chlorophyll content, reduced lipid peroxidation, and a coordinated upregulation of key salt stress-responsive genes (OsSOS1, OsHKT1, OsDREB2A). These findings demonstrate that aptamer-mediated stabilization of eEF1 preserves translational efficiency and contributes to enhanced salt tolerance in rice, offering a novel strategy for crop improvement under adverse environmental conditions.ENAfalseAptamer mediated modulation of eEF1 enhances salt stress tolerance in rice2025-07-042025-05-30PRJNA1246497