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Rewiring systemic nodulation control to engineer heat-tolerant nitrogen fixation in Medicago truncatula and chickpea

Tiwari, M.; Nagalla, S.; Swain, R. K.; Kumar, S.; Infante, V.; Maeda, J.; Yadav, M.; Mishra, D.; Ane, J.-M.

2026-10-02 plant biology
10.64898/2026.10.01.755958 bioRxiv
Show abstract

Root nodule symbiosis allows legumes to meet their nitrogen requirements. However, the causes of symbiosis failure at high temperatures remain unknown. We demonstrated that elevated temperatures affect both the host and the microbial partner in Medicago truncatula and chickpea. Rhizobial viability and nod gene expression persisted under heat stress, but plant transcriptome analysis revealed a strong downregulation of the nodule organogenesis machinery. Nodule organogenesis was restored by constitutive overexpression of NIN-LBD16-NFYA1-NFYB16, miR2111, and CLE5 knockdown but failed to restore nitrogen fixation. In contrast, the genetic manipulation of the competence pathway involving CEP peptides significantly improved both nodulation and nitrogen fixation. Constitutive loss of the autoregulation of nodulation (AON) pathway in the SUNN alleles also rescued root nodulation and nitrogen fixation. Therefore, heat stress perturbs the balance between AON and the competence pathway, and genetic manipulation of these pathways is a promising approach to engineer thermotolerant nitrogen fixation in crop legumes.

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