In order to investigate the response of male and female Nile tilapia (Oreochromis niloticus) to continuous high temperature habitat, the morphological characteristics of female and male fish were continuously monitored in the 36℃ experimental group and the 28℃ control group, and the brain, back muscles and gill tissues of female and male fish treated for 70 days were analyzed by TUNEL staining method, and back muscles were selected for transcriptome sequencing. The results showed:The growth rate of the high-temperature treatment group was significantly slower than that of the control group, and the male fish grew faster than the female fish; TUNEL staining showed that the signal of apoptosis was the strongest in back muscle tissues, but weaker in other tissues; The results showed after high temperature treatment, 3 405 DEGs were identified in females and 4 645 DEGs were identified in males, with more up-regulated genes than down-regulated genes. Through the KEGG pathway cluster analysis, female fish were mainly involved in the cell cycle, purine metabolism and DNA replication and other pathways, and male fish were significantly enriched in the pathways of adrenaline signaling, myocardial contraction and tight connection in cardiomyocytes. The research results provide basic information for further exploring the adaptation mechanism of different sex Nile tilapia to high temperature environment.