PurposeThe purpose of this paper is to examine the effect of quality-oriented human resource practices (QHRP) on organizational ambidexterity. Furthermore, the mediating role of ambidextrous culture in the relationship between QHRP and organizational ambidexterity was assessed.Design/methodology/approachDrawing on data from 350 green agro-food companies with two respondents in each company, structural equation models were used.FindingsThis paper has been drawn up to provide some responses to the needs of the companies to be ambidextrous while applying QHRP. The findings show that there is a positive effect of QHRP on organizational ambidexterity. In addition, ambidextrous culture mediated the relationship between QHRP and organizational ambidexterity.Practical implicationsThis research reveals key managerial aspects for QHRP implementation that facilitate firms to be more ambidextrous, and thus more efficient and innovative.Originality/valueThe authors illustrate the connection between quality-oriented human resource practices (QHRPs) and organizational ambidexterity under the dynamic capabilities theory. The findings contribute to the empirical evidence on the antecedents of organizational ambidexterity, and suggest that these specific QHRPs influence an organization's baseline beliefs and values and support the development of ambidextrous capabilities by means of an ambidextrous culture.