Land degradation issues and declining fertility are driving the need for agroecological practices. This research analysed the determinants of acceptance and actual use of five main agroecological practices (contour farming techniques, organic fertiliser, crop association, improved seeds and integrated crop management practices) by farmers in Mali. The extended Unified Theory of Acceptance and Use of Technology (UTAUT) was used to develop the conceptual model. Data were collected from 505 randomly selected farming households in the cotton and cereal production zones in Mali. Partial Least Square–Structural Equation Modelling (PLS-SEM) was used to estimate technology acceptance and use. The findings revealed that behavioural intention is significantly and positively influenced by the expected performance and social influence. The expected effort is a key influential factor of the behavioural intention to adopt organic fertiliser. Experience has a mediating effect on the relationship between social influence and behavioural intention to adopt improved seeds adapted to the agroecological conditions. The actual use behaviour is directly and positively affected by the behavioural intention, facilitating conditions and expected net benefit. These findings align with the UTAUT model, have useful implications for both farmers and decision-makers and offer directions for technical approaches to agroecological practices’ development.