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3D genome reorganization and symbiotic nodulation

Three-dimensional genome reorganization enables cytokinin activation of symbiotic nodulation

Legumes establish symbiotic interactions with soil-dwelling rhizobial bacteria, leading to the formation of nitrogen-fixing root nodules. This process requires extensive transcriptional reprogramming, which is associated with dynamic changes in DNA methylation and histone modifications. However, the role of three-dimensional (3D) genome architecture in symbiosis remains largely unexplored. In a new collaborative study published in Plant Communications between the teams of Florian Frugier (SILEG) and Moussa Benhamed (ChromD) at IPS2, we revealed using High-throughput Chromosome Conformation Capture (Hi-C), that the 3D chromatin landscape undergoes major reorganizations in nitrogen-fixing symbiotic nodules compared with non-symbiotic roots and non-nitrogen-fixing nodules. These changes involve alterations in A / B compartmentalization and the establishment of enhancer-promoter loops linked to the symbiotic program. Strikingly, we identified a long-range chromatin loop bridging a 15 kb distal enhancer to the proximal promoter of the NODULE INCEPTION (NIN) gene, a master regulator of nodulation. This enhancer region contains multiple putative cytokinin (CK) response elements, and the CK-dependent induction of NIN in roots is associated with this enhancer-promoter interaction. Moreover, we demonstrated that the CK signaling transcription factor RESPONSE REGULATOR B3 (RRB3) binds specifically to this distal enhancer region and participates in forming enhancer-promoter looping. Altogether, these results uncovered a critical role for 3D chromatin reorganization in regulating gene expression during the legume-rhizobium symbiosis, highlighting a regulatory mechanism through which hormonal signaling shapes genome architecture and modulates NIN activation via long-range chromatin looping.

The distal enhancer region located upstream of the NODULE INCEPTION (NIN) gene becomes accessible in response to cytokinins ATAC-Seq signals in untreated and 1h cytokinin-treated roots. The position of the Cytokinin Element (CE) chromatin loop is indicated at the top.
The distal enhancer region located upstream of the NODULE INCEPTION (NIN) gene becomes accessible in response to cytokinins ATAC-Seq signals in untreated and 1h cytokinin-treated roots. The position of the Cytokinin Element (CE) chromatin loop is indicated at the top.

23/07/2026