This study aimed to investigate the impact of low-temperature (LT) stress during the jointing stage on three most representative wheat varieties in the Huang-Huai-Hai region: “Shannong 38” (a robust winter wheat variety), “Jimai 22” (a semi-winter wheat variety), and “Zhenmai 12” (a weak winter wheat variety). The objective was to explain the sensitivity and change thresholds of various physiological and yield indicators of three winter wheat varieties to low temperatures during the jointing stage and to construct an index for the grading of LT disasters during the jointing stage using principal component analysis. Controlled environment experiments were conducted using an artificial climate chamber during the jointing stage of winter wheat. Five daily minimum temperature treatments were applied, namely (T1, −6 °C); (T2, −3 °C); (T3, 0 °C); (T4, 3 °C); (T5, 6 °C); and control (CK, 8 °C). The duration of treatments was divided into three levels: (D1: 2 days), (D2: 4 days), and (D3: 6 days). It was found that the photosynthetic parameters and chlorophyll content showed a decreasing trend with the increase in the degree of LT stress. The activities of protective enzymes and endogenous hormones increased during the early stages of LT stress or at relatively high temperatures. However, they decreased significantly with an increase in LT stress. Among the varieties, “Zhenmai 12” exhibited a yield reduction rate exceeding 10% under 3 °C LT stress for more than 4 days and a yield reduction rate exceeding 20% under 0 °C LT stress for more than 6 days. “Jimai 22” showed a yield reduction rate exceeding 10% under 3 °C LT stress lasting more than 2 days, and a yield reduction rate exceeding 20% under −3 °C LT stress lasting more than 4 days. “Shannong 38” experienced a yield reduction rate exceeding 10% under 0 °C LT stress lasting more than 4 days and a yield reduction rate exceeding 20% under −6 °C LT stress lasting more than 6 days. Principal component analysis (PCA) conducted on all trait indicators of the three winter wheat varieties revealed that “Zhenmai 12” experienced mild LT stress at 6 °C for 2 days, moderate LT stress at 0 °C for 6 days, and severe LT stress at −3 °C for 6 days. “Jimai 22” experienced mild LT stress under 6 °C for 6 days, moderate LT stress under 0 °C for 4 days, and severe LT stress under −6 °C for 2 days. “Shannong 38” experienced mild LT stress under 3 °C for 4 days, moderate LT stress under 0 °C for 4 days, and severe LT stress under −6 °C for 6 days.
