This study investigates how quality-oriented human resource practices (QHRP) influence organizational ambidexterity. It also examines whether ambidextrous culture functions as a mediating mechanism linking QHRP to organizational ambidexterity. Data were obtained from 350 green agro-food firms, each represented by two respondents. Structural equation modeling techniques were employed to analyze the relationships among the variables. The study responds to the growing need for organizations to achieve ambidexterity while implementing QHRP. Results indicate that QHRP significantly enhances organizational ambidexterity. Furthermore, an ambidextrous culture serves as a mediating factor in the relationship between QHRP and organizational ambidexterity. This study establishes a link between quality-oriented human resource practices (QHRPs) and organizational ambidexterity through the lens of dynamic capabilities theory. It extends empirical research on the antecedents of ambidexterity by demonstrating that QHRPs shape organizational values and shared assumptions, thereby enabling ambidextrous capabilities by fostering an ambidextrous culture. Findings highlight important managerial considerations for implementing QHRP to strengthen organizational ambidexterity, ultimately improving both operational efficiency and innovation capacity.