Evening complex component ELF3 interacts with LUX proteins to repress soybean root nodulation.

Formation of root nodules is a unique hallmark of the symbiotic interaction between legume host plants and rhizobia and is governed by a complex regulatory framework that balances the appropriate orchestration of rhizobial infection and subsequent nodule organogenesis. In contrast to prominent model species such as Medicago truncatula and Lotus japonicus, research on symbiotic signal transduction in the staple-crop soybean Glycine max remains relatively insufficient. Here, we identified a soybean mutant with ~25% additional root nodules over wild-type, designated as increased number of nodules 1 (inn1). Through map-based cloning, INN1 encodes the EARLY FLOWERING 3a (ELF3a) protein component of the soybean Evening Complex, together with LUX1 and LUX2. INN1 is co-expressed with LUX1 and LUX2 in roots, and knockout of INN1 or knockdown of LUX1 and LUX2 enhances root nodulation. The function of INN1 in negatively regulating nodulation is genetically and biochemically dependent upon LUXs, as the INN1-LUX complex binds to the promoter of the downstream pro-nodulation target ENOD40, repressing its expression. ELF3a/INN1's repression of root-nodule formation extends beyond its established roles in diverse above-ground developmental and physiological processes and offers a theoretical basis for enhancing the biological-nitrogen fixation capacity of soybean.