Encouraging sustainability in science education demands a collaborative approach, involving the integration of diverse knowledge systems, including indigenous and formal scientific knowledge. Nevertheless, educators’ perceptions of these knowledge systems are crucial factors to consider. This study investigated the perceptions of pre-service science teachers about the role of indigenous knowledge in promoting sustainability science education. The study was conducted at the Department of Integrated Science Education University of Education Winneba, Ghana and predominantly adopted a qualitative approach. A sample size of 54 Level 300 undergraduate pre-service science teachers were purposively chosen. These pre-service teachers have engaged with the science curriculum and syllabus through on-campus peer teaching and are prepared to take their internships in the country’s various second-cycle institutions. Emphasis is laid on the teaching of science to promote education for sustainable development in these institutions. Data were collected through an online questionnaire comprising open- and closed-ended items. The closed-ended items were quantified and analysed for clearer inference whereas thematic analysis was employed for qualitative data. The findings of the study established pre-service teachers’ affirmation of the potential of indigenous knowledge in providing education for sustainability if integrated into the teaching of science. The study recommended targeted interventions including the integration of indigenous knowledge into pre-service science teachers’ curriculum, training programmes, workshops, courses and campaigns aimed at increasing awareness, fostering positive attitudes, and promoting understanding and appreciation of indigenous knowledge and its role in sustainability.