OsBIR3 maintains the homeostasis of OsBRI1, OsREM4.1, and Brd2 protein levels in brassinosteroid pathways in rice

Brassinosteroids (BRs) are a crucial class of plant hormones and regulate many important agronomic traits in crops. In Arabidopsis (Arabidopsis thaliana), BIR3 interacts with the BR receptor BRI1 and coreceptor BAK1 to negatively regulate BR signalling. In contrast, OsBIR3 interacts with OsBRI1 and OsBAK1 to positively regulate BR signalling in rice (Oryza sativa). However, our understanding of OsBIR3 remains incomplete. In this study, we isolated a reduced upper branch (rub1) mutant of rice, exhibiting a significant reduction in grain number. The causal gene for the mutant phenotype was LOC_Os04g41030 (OsBIR3). OsBIR3 interacts with both OsBRI1 and the remorin protein OsREM4.1, but interactions of the mutated Osbir3 with both OsBRI1 and OsREM4.1 were decreased. Furthermore, BL interferes with the interaction of OsBIR3 with OsBRI1, but promotes the interaction of OsBIR3 with OsREM4.1. Overexpression of OsREM4.1 and OsBRI1 individually in the rub1 mutant caused an exacerbation of the mutant phenotype. Additionally, OsBIR3 interacts with Brd2, involving BR biosynthesis in the early stage, and the interaction of the mutated brd2 with wild-type OsBIR3 was increased. Besides, BL promotes the interaction between OsBIR3 and Brd2. Collectively, the data indicate that OsBIR3 plays a key role in maintaining the homeostasis of OsBRI1, OsREM4.1, and Brd2 at their respective protein levels. This work provides insight into the roles of OsBIR3 in BR signalling and biosynthesis pathways of rice.