To investigate the associations between dietary patterns and biological ageing, identify the most recommended dietary pattern for ageing and explore the potential mediating role of gut microbiota in less-developed ethnic minority regions (LEMRs). This prospective cohort study included 8288 participants aged 30–79 years from the China Multi-Ethnic Cohort study. Anthropometric measurements and clinical biomarkers were utilised to construct biological age based on Klemera and Doubal’s method (KDM-BA) and KDM-BA acceleration (KDM-AA). Dietary information was obtained through the baseline FFQ. Six dietary patterns were constructed: plant-based diet index, healthful plant-based diet index, unhealthful plant-based diet index, healthy diet score, Dietary Approaches to Stop Hypertension (DASH), and alternative Mediterranean diets. Follow-up adjusted for baseline analysis assessed the associations between dietary patterns and KDM-AA. Additionally, quantile G-computation identified significant beneficial and harmful food groups. In the subsample of 764 participants, we used causal mediation model to explore the mediating role of gut microbiota in these associations. The results showed that all dietary patterns were associated with KDM-AA, with DASH exhibiting the strongest negative association (β = −0·91, 95 % CI (–1·19, −0·63)). The component analyses revealed that beneficial food groups primarily included tea and soy products, whereas harmful groups mainly comprised salt and processed vegetables. In mediation analysis, the Synergistetes and Pyramidobacter possibly mediated the negative associations between plant-based diets and KDM-AA (5·61–9·19 %). Overall, healthy dietary patterns, especially DASH, are negatively associated with biological ageing in LEMRs, indicating that Synergistetes and Pyramidobacter may be potential mediators. Developing appropriate strategies may promote healthy ageing in LEMRs.