Lakes are potential sources of nitrous oxide (N₂O). The N₂O emission pattern of lakes also has special and regional characteristics due to the differences in natural environment as well as human activities. In order to explore the spatial and temporal characteristics of N₂O flux from Lake Chaohu, this study observed N₂O emissions from different regions (east, middle, and west) of the lake from March 2018 to December 2019 using a dark floating chamber. The results showed that the annual average N₂O flux from Lake Chaohu was (25.14 ± 55.01) μg/(m²·h), which behaved as a “net source” of N₂O and had a obvious spatial and temporal distribution pattern. In terms of temporal variation, the seasonal trend showed an “M” pattern, with a minimum value in July ((-12.97±16.32) μg/(m²·h)), a peak in June and August, and a maximum value in August ((68.25±78.05) μg/(m²·h)). The extreme values all happened over the summer. Maximum N₂O emission flux ((43.32±79.96) μg/(m²·h)) occurred in the west sub-lake zone and the minimum ((10.37±19.31) μg/(m²·h)) occurred in the east sub-lake zone, showing an overall trend of gradually decreasing from the shore to the center of the lake. However, only the west sub-lake zone showed a similar pattern from different lake areas, the central sub-lake zone had a maximum value of 4 km from the lake shore and the east sub-lake zone had a minimum value of 4 km from the lake shore, which were mainly influenced by exogenous input. The N₂O emission from Lake Chaohu was significantly negatively correlated with pH, but the relationships between environmental factors and N₂O emission varied depending on the season and region of the lake. The flux of N₂O from lakes is not controlled by a single or fixed environmental factor, but is a combination of multiple environmental factors.