Global warming and its relevant effects have aroused increasing human concerns in recent decades. These anomalies are likely influencing vegetation dynamics and ecosystem stability. This paper aims to dissect extreme temperature variations in both space and time, and related regional responses to global warming in the Huai River Basin. Using the daily maximum and minimum air temperature at 153 stations across the Huai River Basin, China, covering the period of 1961–2014, trends and relations amongst extreme air temperature indices were analyzed, and regional responses of the Huai River Basin to global warming process were also investigated. The results indicated that: (1) widespread increasing trends can be observed in maximum and minimum air temperature, with the largest increasing magnitude of 0.4 °C per decade and 1.3 °C per decade, respectively. The minimum air temperature regimes have a larger increasing magnitude than the maximum air temperatures. (2) There is an increasing trend in the extreme maximum temperature indices, and the increasing rates of TN90p and TR20 are greater than those of the other extreme maximum temperature indices. However, the extreme minimum temperature indices, except for consecutive icy days (CID), show significant decreasing trends. The growing season length (GSL) would increase due to the joint action of (i) an increase in maximum temperature indices, and (ii) a decrease in minimum temperatures indices. Although increases in the GSL would be beneficial for increasing the growing time of crops, a decreased extreme minimum temperature and increased extreme maximum temperature may reduce the winter wheat yield. (3) Extreme low temperature indices show a larger changing magnitude than the extreme high temperature indices. Temporally, a larger changing magnitude can be identified for temperature indices during night-time than during daytime. Moreover, a larger changing magnitude and higher significance of trends of extreme temperature indices can be observed in the eastern Huai River Basin than in the western Huai River Basin. Particularly, we detect an even higher changing rate of extreme temperature indices in the southeastern Huai River Basin. These findings can be well explained by urban heat island effects resulting from urban aggregation across the Yangtze Delta. (4) The correlation between the extreme low temperature indices is slightly higher than that of the extreme high temperature indices. The changing trends of the most extreme temperature indices are closely related to latitude and altitude. Trends in the main extreme temperature indices of the Huai River Basin are essentially similar to those in other regions around the world, implying consistent spatial patterns of extreme temperature indices across the globe.