Dajiuhu Peatland in Shennongjia forestry region is a rare subalpine Sphagnum wetland in central China. It is regarded as an ideal site to study climate change and ecological effects in the middle reach of the Yangtze River. Based on the records of geochronology, elements and subfossil chironomids of a peat core collected from Dajiuhu Peatland, combined with regional paleoclimate data, this paper aims to explore past changes in surface wetness and biotic response in Dajiuhu Peatland. Our results showed that three major stages were identified based on paleohydrology and chironomid assemblages in Dajiuhu Peatland during the past 400 years. Before the 1820s, especially between the 1720s and the 1820s, a cold and wet climate favored carbon accumulation in the peatland, and a high surface moisture promoted the growth of chironomids. Correspondently, limnic chironomid taxa were abundant and diverse during this period. Between the 1820s and the 1940s, a drop in summer precipitation, accompanying with evident regional climate warming, led to obvious drought and low surface humidity in the peatland, and hence rare aquatic midges and extremely low density of subfossil chironomid head capsules were found in sediments. Chironomid biomass and semi-terrestrial taxa then rebounded after the 1940s (especially after the 1970s), while the abundance of limnic taxa remarkably declined. This suggested that peatland surface wetness was still lower than that in the period before the 1820s, despite of a relatively increase in comparison with the previous stage (1820s-1940s). Warm and humid climate likely accelerated carbon decomposition in this period. This study explored the response of chironomid assemblages to surface wetness in peatland in the context of climate change, which not only launched a new field for the application of subfossil chironomids as a biotic proxy in climate change studies, but also provided new clues for tracking comprehensive and accurate information on peatland development and regional environmental change.