To investigate the effects of high-temperature stress on the chlorophyll fluorescence induction kinetics of peony and to determine indicators for the rapid screening of varieties responding to high temperatures, three four-year-old peony variety, ‘Fengdanbai’, ‘Huhong’, and ‘Yinhongqiaodui’, were selected as materials. The photosynthetic curves (Pn-PAR) and fast chlorophyll fluorescence curves (OJIP curves) of peony leaves were measured at different times under high-temperature stress conditions (40 °C), the changes in the photosynthetic characteristics of different peony varieties under high-temperature stress were analyzed, and the heat tolerance of peony was evaluated. The results showed that ‘Huhong’ grew well within 16 days, while all of the other varieties showed obvious wilting at 6–9 days. High temperatures damaged the structure and function of the photosystem of peony leaves, indicating that the maximum net photosynthetic rate (Pnmax), apparent quantum efficiency (AQE), maximum photochemical efficiency (Fv/Fm), and photosynthetic performance index (PIABS) all tended to decrease under high-temperature stress, while the rate of closing the PS II reaction center (Mo) and the absorption per reaction center (ABS/RC), the capture (TRo/RC), and the dissipation (Dio/RC) of light energy per reaction center showed an overall increasing trend. The ability to cope with high-temperature stress differed among varieties, and the heat tolerance was determined to be in the descending order of ‘Fengdanbai’ < ‘Yinhongqiaodui’ < ‘Huhong’. The correlation analysis among the parameters and the analysis of the morphological change patterns in peony leaves concluded that PIABS, Dio/RC, and Mo could be used as indicators of peony tolerance to high-temperature stress. The results of the study can provide a basis for the screening of heat-tolerant peony species and peony heat defense in the Jiangnan area.