Identifying ecosystem service bundles and their long-term evolutionary characteristics is essential for the overall enhancement of regional ecosystem services, as well as the division and management of functional areas, providing a basis for decision-making in formulating ecological and environmental protection policies, as well as regional development planning. Based on land use, remote sensing, and meteorological data obtained from Anhui Province, this study assessed six important ecosystem service functions, including food production (FP), water yield (WY), carbon sequestration (CS), soil conservation (SC), habitat quality (HQ), and landscape aesthetics (LA), at the township scale in 2000, 2010, and 2020. On this basis, the k-means clustering method was used to identify ecosystem service bundles, analyze the spatio-temporal evolution trajectory of service bundles, and explore the driving factors of the spatio-temporal evolution of ecosystem service bundles using GeoDetector 2015 The results indicate the following: (1) At the spatial level, diverse ecosystem services demonstrate pronounced spatial differentiation. The distribution pattern of HQ, carbon fixation, and SC services is generally lower in the north and higher in the south, with areas of high value predominantly located in the western Dabie Mountains and the mountains of Southern Anhui. Conversely, FP services exhibit the reverse pattern, and WY services display a gradual increase from north to south, while cultural services are more dispersed, with areas of high value primarily located in the western Dabie Mountains, the Yangtze River Basin, and other locations. On the temporal scale, WY, SC, and FP services mainly exhibit an increasing trend, marked by a significant increase, whereas other services tend to present a decreasing trend. (2) Anhui Province can be categorized into four distinct types of service bundles: the grain production bundle (GPB), mountain ecological conservation bundle (MECB), urban living bundle (ULB), and core protection bundle (CPB). Ecosystem service bundles exhibit clear spatial differentiation, and identical service bundles demonstrate substantial clustering in space. Between 2000 and 2020, ecosystem service bundles displayed a marked spatio-temporal evolution, with the prevalence of GPBs diminishing, whereas the share of ULBs progressively increased, and the number of MECBs and CPBs remained largely stable. (3) In the spatio-temporal evolution process, the average annual precipitation, the proportion of forest land, and slope constitute the principal natural factors influencing the spatio-temporal evolution of ecosystem service bundles, while the proportion of construction land represents the primary socio-economic factor, with natural factors exerting a more significant influence than socio-economic factors.