LuxR family transcription factors can inhibit or stimulate the expression of various functional genes to maintain the stability of cell function. [Objective] To study the role of LuxR family transcription factors in Aeromonas hydrophila. [Methods] In this study, we have knocked out the AHA_1581 gene by homologous recombination technology to understand their role in virulence and drug resistance. [Results] The deletion of AHA_1581 gene not only enhanced the extracellular protease enzyme activity, the tolerance of A. hydrophila to survive under low temperature, different antibiotic stress, but also reduced the biofilm formation capability. Further the quantitative proteomic and bioinformatics analysis showed that the deletion of AHA_1581 gene 59 proteins were down-regulated and 142 proteins were up-regulated, which affected several important biological processes and virulence pathways in A. hydrophila. [Conclusion] Overall, the outcome of this study on the role of transcription factor AHA_1581 in the biological processes and virulence of A. hydrophila may pay the way for controlling their infections.