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.

Low-temperature stress during the jointing stage severely disrupts the coordination of the source–flow–sink system in winter wheat. To elucidate the underlying mechanism, three wheat cultivars with different winter habits (Zhenmai 12, Jimai 22, and Shannong 38) were selected and subjected to six temperature levels (−6 °C to 8 °C) and three stress durations (2–6 days). The effects of vascular bundle traits on the transport of photosynthetic products, dry matter distribution, and yield formation were analyzed. The results showed that Zhenmai 12 and Jimai 22 completely ceased photosynthesis under 0 °C and −3 °C, respectively. The leaf vascular bundle area continuously decreased with increasing low-temperature stress, while the proportion of xylem and phloem initially increased by approximately 15% and 10%, respectively, before rapidly decreasing to 65% of the control value. In the stem, the three vascular bundle parameters initially increased by 20%, 25%, and 20%, respectively, before quickly decreasing to 50%. Changes in the vascular bundle structure weakened the transport capacity of assimilates, with dry matter in leaves and stems decreasing by 15–20% and 10%, respectively, while the root dry matter increased by 20–30%. Correlation analysis revealed highly significant relationships (p < 0.001) between vascular bundle parameters and yield components. Principal component and cluster analyses indicate that the area of leaf and stem vascular bundles, maximum net photosynthetic rate, and water use efficiency may be key indicators in explaining the variation in yield. Radar plots further validated this finding, showing that Zhenmai 12 and Jimai 22 are more sensitive to changes in the maximum net photosynthetic rate, while Shannong 38 exhibits a greater sensitivity to changes in water use efficiency. Based on existing research on photosynthetic pathways and dry matter distribution, this study innovatively investigates the potential relationship between material transport and yield formation under low-temperature stress during the jointing stage from the perspective of anatomical structure and functional coupling. The findings provide new insights into understanding the structural impact of low-temperature stress on crop yield formation and offer theoretical support for identifying the structural basis of limited material transport under stress and for developing disaster diagnostic models driven by structural parameters.