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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.