The development of rural tourism plays an important role in promoting rural culture. By integrating 3833 household questionnaires from the 2020 China Rural Revitalization Survey (CRRS) database with remote sensing data, we constructed an evaluation system to measure the level of rural culture. Then, we analyzed the impacts of rural tourism on rural culture from macro and micro perspectives. Our research results show the following: (1) Villages with developed rural tourism show a 85.9% increase in rural culture compared to those without tourism; (2) mechanism tests show that rural tourism promotes the rural culture by improving households’ risk-sharing behavior, human resources, and self-identification, leading to increases of 3.4%, 55% and 10.9%, respectively; (3) with micro-level (fieldwork survey) and macro-level analysis (remote sensing), we analyzed the various impacts of rural tourism on rural culture under different income levels, demographic structures, geographical locations and topographical conditions. The results show that at the micro level, the promotion effect of rural tourism on rural culture increases by 2.214% and 1.679% with the increase in per capita income and the proportion of women, respectively. For geographical location, macro-level data suggest that rural tourism in the east of China increases the rural culture by 3.416%. Moreover, in plain areas, both micro- and macro-level analysis indicated that rural tourism promotes rural culture by 2.323% and 4.607%, respectively. This is the first time rural culture has been evaluated on a large scale with two cross-validated approaches.