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The yield potential of large-panicle rice is often limited by grain-filling barriers caused by the development of inferior spikelets (IS). Photoassimilates, which are the main source of rice grain filling, mainly enter the caryopsis through the dorsal vascular bundle. The distribution of assimilates between superior spikelets (SS) and IS is influenced by auxin-mediated apical dominance; however, the mechanism involved is still unclear. In this study, the effect of auxin signaling on the grain filling of SS and IS was investigated in two large-panicle japonica rice varieties, W1844 and CJ03. Compared to SS, IS displayed delayed initiation of filling and a significantly lower grain weight. Furthermore, the endosperm development in IS remained stagnant at the coenocytic stage. The development of the dorsal vascular bundle in the IS was also slow, and poor sucrose-unloading was observed during the initial grain filling stage. However, the endosperm development in IS immediately started after the improvement of dorsal vascular bundle development. GUS activity staining further revealed that indole-3-acetic (IAA) was localized in the dorsal vascular bundle and surrounding areas, suggesting that the low IAA content observed in the IS during the initial grain filling stage may have delayed the development of the dorsal vascular bundle. Therefore, these results demonstrate that IAA may control sugar transport and unloading by regulating dorsal vascular bundle development, consequently affecting endosperm development in IS., (© 2024. The Author(s).)