To elucidate the characteristics and biological functions of the Rho gene family in Procambarus clarkii，members of this gene family were identified at the genome-wide level.Quantitative real-time PCR （qRT-PCR） was employed to analyze the expression profiles of these genes across various tissues，and multiple transcriptomic datasets were utilized to investigate their responses to viral infection，low-temperature stress，and hypoxic stress.A total of 4 Rho gene family members （PcRhoA，PcCdc42，PcRac，and PcRhoBTB） were identified from the P.clarkii genome，corresponding to the Rho，Cdc42，Rac，and RhoBTB subfamilies，respectively.These Rho family members exhibited distinct tissue-specific expression patterns in the muscle，hepatopancreas，gill，stomach，antennal gland，epidermis，eyestalk，heart，and intestine.Among them，PcRhoA and PcCdc42 were highly expressed in both the heart and intestine，PcRac displayed high expression in the epidermis，while PcRhoBTB was predominantly expressed in the gill.Following a 7-day challenge with white spot syndrome virus （WSSV），the expression levels of PcRhoA and PcRhoBTB in the hepatopancreas were significantly up-regulated，with PcRhoA showing an extremely significant increase.After 31 days of low-temperature treatment （1 ℃），the expression levels of PcRhoA，PcCdc42，and PcRhoBTB in muscle tissue were significantly downregulated.Under hypoxic conditions ［dissolved oxygen （DO） = （2.5±0.2） mg/L］ for 3 hours，PcRac and PcRhoBTB were significantly downregulated in the hepatopancreas.Stress-response analyses demonstrated that PcRhoA and PcRhoBTB responded significantly to both biotic and abiotic stressors，suggesting their potential involvement in comprehensive anti-stress mechanisms.