Abstract: 【Objective】This study aimed to establish a hypoxia stress model for Pelteobagrus vachelli， clarify the response mechanism of related genes in the gill tissue to hypoxia stress， providing data support for further exploration of its gill tissue hypoxia adaptive regulation and theoretical basis for screening highly adaptive populations of Pelteobagrus vachelli.【Method】One-year-old Pelteobagrus vachelli were used as research subjects， and three treatment groups were set control group （C: dissolved oxygen content of 6.50 ± 0.10 mg/L）， hypoxia group （H: dissolved oxygen content of 0.75 ± 0.10 mg/L， 6 h）， and recovery group （R: dissolved oxygen content of 6.50 ± 0.10 mg/L， 6 h）. Gill tissue samples were collected to extract RNA， and differentially expressed genes （DEGs） were screened by transcriptome sequencing. GO functional annotation analysis and KEGG signaling pathway enrichment analysis were performed， and real-time fluorescence quantitative PCR was used for verification.【Result】The results showed that raw data of 57.72 Gb were detected through sequencing nine samples from Group C， Group H， and Group R. According to the reference genome， a total of 22116 genes were annotated and 1824 new genes were found. For the Group C vs Group H， 554 DEGs were detected， in which 270 were up-regulated and 284 were down-regulated； for the Group C vs Group R， 42 DEGs were detected， in which 12 were up-regulated and 30 were down-regulated； for the Group H vs Group R， 993 DEGs were detected， in which 470 were up-regulated and 523 were down-regulated. DEGs were mainly annotated to GO functional items such as cellular process， single-organism process， metabolic process， binding， catalytic activity， nucleic acid binding transcription factor activity， signal transducer activity， cell part， membrane， and cell. KEGG signaling pathway enrichment analysis found that DEGs were mainly enriched in signaling pathways such as breast cancer， pathways in cancer， basal cell carcinoma， cytokine-cytokine receptor interaction， chemical carcinogenesis-DNA adducts， and metabolism of xenobiotics by cytochrome P450. The real-time fluorescence quantitative PCR detection results showed that the expression trends of partial DEGs between groups were overall consistent with the transcriptome sequencing results.【Conclusion】Hypoxia stress inhibits the cell proliferation and signaling pathways of Pelteobagrus vachelli to some extent， lea-ding to inhibition of the tricarboxylic acid cycle and anaerobic respiration activation， and gill tissue up-regulates expression of glycometabolism gene to promote glucose breakdown to produce energy. Activation of the glycolytic pathway leads to pyruvate accumulation， meanwhile， pyruvate kinase expression is inhibited at the transcriptional level， accompanied by a partial activation of gluconeogenesis pathway. Changes in heat shock protein gene expression may counteract hypoxic stress by inhibiting the autophagy pathway to reduce energy consumption.