Abstract: Salt stress is a major abiotic stress factor limiting rice growth and development. Elucidating salt stress-responsive molecular mechanisms is vital for clarifying the plant salt tolerance regulatory network and breeding salt-tolerant rice cultivars. WRKY transcription factors play key regulatory roles in plant stress responses. However, the specific regulatory network of OsWRKY50 in rice salt stress responses remains unclear. OsWRKY50 overexpression lines (W50 OE) and wild-type Nipponbare (Nip) were used as materials. RNA-Seq was performed to analyze transcriptional differences under different treatments. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted to identify potential regulatory pathways. Selected differentially expressed genes were further validated by RT-qPCR. The results showed that the W50 OE lines exhibited stronger salt stress tolerance and a higher survival rate. Transcriptome analysis revealed that a total of 2250 differentially expressed genes (DEGs) were identified in the NaCl_Nip vs CK_Nip comparison, while 3463 DEGs were identified in the NaCl_W50 OE vs CK_W50 OE comparison. RT-qPCR results were largely consistent with the RNASeq data. Functional enrichment analysis showed that these DEGs were mainly involved in MAPK signaling pathways, and phenylpropanoid biosynthesis. Promoter W-box element screening revealed that some DEGs may serve as potential target genes of OsWRKY50. OsWRKY50 regulates the expression of genes related to ion homeostasis, MAPK signaling and secondary metabolism at the transcriptional level, and participates in the salt stress response. The study provides a basis for further elucidating its molecular regulatory mechanisms.