Integrated single-nucleus and spatial transcriptomics captures transitional states in soybean nodule maturation

Legumes form symbiosis with rhizobium leading to the development of nitrogen-fixing nodules. By integrating single-nucleus and spatial transcriptomics, we established a cell atlas of soybean nodules and roots. In central infected zones of nodules, we found that uninfected cells specialize into functionally distinct subgroups during nodule development, and revealed a transitional subtype of infected cells with enriched nodulation-related genes. Overall, our results provide a single-cell perspective for understanding rhizobium–legume symbiosis. The integration of single-nucleus and spatial transcriptomics is used to profile dynamic changes during soybean nodule maturation. The approach identifies a rare cell type that specifically expresses numerous genes required for symbiotic nitrogen fixation.