Optimizing plant spatial competition can change phytohormone content and promote tillering, thereby improving wheat yield.

As an important type of interplant competition, line-spacing shrinkage and row-spacing expansion (LSRE) can increase the number of tillers and improve resource utilization efficiency in wheat. Wheat tillering is closely related to various phytohormones. However, it is unclear whether LSRE regulates phytohormones and their relationship to tillering and wheat yield. This study evaluated tillering characteristics, phytohormone content in tiller nodes at the pre-winter stage, and grain yield factors for the winter wheat variety Malan1. We used a two-factor randomized block trial design with two sowing spacings of 15 cm (15RS, conventional treatment) and 7.5 cm (7.5RS, LSRE treatment) at the same density and three sowing-date groups (SD1, SD2, and SD3). LSRE significantly promoted wheat tillering and biomass at the pre-winter stage (average increases of 14.5% and 20.9% in the three sowing-date groups, respectively) and shortened the accumulated temperature required for a single tiller. Changes in the levels of phytohormones, including decreased gibberellin and indole acetic acid and increased zeatin riboside and strigolactones, were determined by high-performance liquid chromatography and were shown to be responsible for the tillering process under LSRE treatment in winter wheat. LSRE treatment can improve crop yield by increasing the number of spikes per unit area and grain weight. Our results clarified the changes in tillering and phytohormones content of winter wheat under LSRE treatment and their correlation with grain yield. This study also provides insights into the physiological mechanisms of alleviating inter-plant competition to improve crop yield.