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High-latitude permafrost peatlands are sensitive to global warming, and understanding their paleohydrological evolution is critical for assessing future hydrological and environmental changes. This paper presents a diatom record from a peat deposit covering the past 3300 years to reconstruct the moisture and acidity conditions in the Tuqiang permafrost peatland in the northern Greater Khingan Mountains of northeast China. Our findings are integrated with previously published records of plant macrofossils, palynology, and geochemistry from the same profile. From 3300 to 2800 cal yr BP, planktonic and benthic diatoms were equally abundant, indicating relatively high moisture conditions, while the low abundance of acidophilous diatoms reflects relatively high pH during this interval. Pollen data show dominance by sedges with input from the surrounding Pinus forest, while LOI data and lithology suggest that the site was a non-peat accumulating wetland. During the period 2800–1100 cal yr BP, the dominance of planktonic diatoms indicates a wetter local environment, concurrent with the dominance of peat-forming sedges (Eriophorum), indicating the development of a peatland. Between 1100 and 500 cal yr BP, a short-lived increase in acidophilous diatoms (Eunotia) marks a decrease in pH. Since 500 cal yr BP, the dominance of benthic diatoms reflects a drier environment, while a vegetation shift from an Eriophorum -dominated rich fen to a Sphagnum- and Ericaceae-dominated poor fen. An increase in Eunotia diatoms prior to the Sphagnum peak may indicate that diatoms are more sensitive to local acidified habitats than moss. Peatland development in the northern Greater Khingan Mountains was likely induced by the onset of a cooler and wetter climate starting at approximately 2800 cal yr BP, a scenario supported by regional paleoclimate records. Since 700 cal yr BP, the peatland has been affected by both anthropogenic activities and climatic changes, as a drying climate and intensified human activities around the study area reduced diatom species diversity and peat accumulation. Under the background of future global warming and enhanced human activities, the decreasing species diversity and carbon accumulation rate of peatlands in high-latitude permafrost regions need more attention. • Diatoms can indicate local moisture and acidity in permafrost peatlands. • Diatoms are likely more sensitive to microhabitats than bryophytes. • TQ peatland formed after 2800 cal yr BP caused by the cooler and wetter climate. • Rich-fen shifted to poor-fen which co-influenced by deforestation and dry climate at LIA. [ABSTRACT FROM AUTHOR]