The DENSE AND ERECT PANICLE1-GRAIN NUMBER ASSOCIATED module enhances rice yield by repressing CYTOKININ OXIDASE 2 expression

The phytohormone cytokinin (CK) positively regulates the activity of the inflorescence meristem (IM). Cytokinin oxidase 2/Grain number 1a (OsCKX2/Gn1a)-mediated degradation of CK in rice (Oryza sativa L.) negatively regulates panicle grain number, whereas DENSE AND ERECT PANICLE 1 (DEP1) positively regulates grain number per panicle (GNP). However, the detailed regulatory mechanism between DEP1 and OsCKX2 remains elusive. Here, we report the GRAS (GIBBERELLIN ACID INSENSITIVE, REPRESSOR OF GA1, and SCARECROW) transcription factor GRAIN NUMBER ASSOCIATED (GNA), previously thought to be involved in the Brassinosteroids (BRs) signaling pathway, directly inhibits OsCKX2 expression in the IM through a DEP1-GNA regulatory module. Overexpressing GNA leads to increased CK levels and consequently higher branch number, GNP, and yield. Both DEP1 and dep1 enhance the inhibitory effect of GNA on OsCKX2 expression through interacting with GNA. GNA promotes the translocation of DEP1 to the nucleus, while the gain-of-function mutant dep1 translocates into the nucleus in the absence of GNA. Our findings provide insight into the regulatory mechanism underlying OsCKX2 and a strategy to improve rice yield.