GmNIGT2a/2b-mediated transcriptional cascade buffers high-nitrogen-triggered nodule senescence in soybean.

Soil inorganic nitrogen (N) availability is a critical determinant of symbiotic nitrogen fixation efficiency, making it essential for legumes to respond appropriately and effectively to N fluctuations. Here, we identify a pair of high N response factors GmNIGT2a/2b (NITRATE-INDUCIBLE GARP-TYPE TRANSCRIPTIONAL REPRESSOR). GmNIGT2a/2b are activated by GmNLP4a/b (NIN-like proteins) under high N. The nodules of gmnigt2a/2b double mutants exhibit exacerbated reduction in nitrogenase activity and accelerated senescence in response to high N. Integration of RNA-seq and DAP-seq analyses reveals that GmNIGT2a/2b negatively regulate a suite of core N-induced genes, including NAC, WRKY, and bZIP transcription factors as well as trehalose metabolism genes. GmNIGT2b binds to the promoters of SNAP3 and NAC039 and represses their expression, thereby delaying nodule senescence. Our results suggest that GmNIGT2a/2b-mediated transcriptional regulation prevents excessive nodule senescence in response to high N, highlighting the complexity of transcriptional reprogramming for environmental adaptation in nodules.