Sugar regulators integrate the plant’s sugar status into endogenous signaling pathways to coordinate developmental decisions. Three major conserved regulators TARGOT OF RAPAMYCIN (TOR), SNF1-RELATED PROTEIN KINASE1 (SnRK1), and HEXOKINASE1 (HXK1), have been extensively studied for their role in plant growth. But despite the fact that sugars are pivotal as energy for beneficial plant-associated microbes, such as rhizobia during nodulation, the role of TOR, SnRK1, and HXK1 in these interactions remains largely unknown. In the proposed project, we will advance the state-of-the art by studying these sugar regulators during soybean nodulation. First, we will perform a large-scale exploratory analysis to study where, when and which of the regulators is expressed during nodulation by combining physiological assays with pharmacological treatments. Changes in nodulation and expression will be linked with nutrient levels by means of local and real-time quantification of primary metabolites. Next, the mode-of-action of one regulator will be elucidated in more detail by cutting-edge single-cell multi-omics approaches, i.e. single nuclei transcriptomics and tissue/cell-type specific (phospho)interactomics, allowing the integration of the cellular complexity of a nodule. Finally, we will study the effect of five candidates on nodulation and metabolites. Our findings will benefit the international interest in soybean nodulation by providing novel insights in how sugars regulate nodulation.