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bioRxiv · 10.64898/2026.09.12.750751

The plant specific Histone Lysine Demethylase MtPKDM9B mediates the root nodule symbiosis by controlling H3K27me3 levels and expression of symbiotic genes

Ferrari, M.·Yacullo, M.·Traubenik, S.·Ibarra, E.·Blanco, F. A.·Reynoso, M.·Zanetti, M. E.
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Abstract

Under nitrogen limiting conditions, legume plants interact with nitrogen fixing bacteria known as rhizobia, resulting in the formation of a new organ, the nodule. This process is accompanied by dramatic changes in gene expression, which operate at different levels. A previous study revealed that histone methylation is differentially modulated during nodulation. However, the histone methyl transferases and demethylases involved in this modulation have not been characterized. In this study we report the identification of the Medicago truncatula putative histone lysine demethylase MtPKDM9B, which is subject to alternative splicing (AS), and the differential modulation of AS variants at translational level during nodule symbiosis. Knockdown of MtPKDM9B impaired infection by rhizobia, nodule development, bacterial viability and the expression of the leghemoglobin coding gene MtLHB1. MtPKDM9B is the putative ortholog of Arabidopsis EARLY FLOWERING 6 (ELF6/AtPKDM9B) gene involved in the removal of the repressive mark H2K27me3. A combination of ChIP-seq and RNA-seq experiments revealed that MtPKDM9B is required for demethylation of H3K27me3 in regions nearby or contained within gene bodies of symbiotic genes and the upregulation of the cognate mRNAs in response to rhizobia, including those encoding the putative ubiquitin ligase MtPUB2, the MYB transcription factor MtMYB040 and the auxin conjugating enzyme MtGH3 (Gretchen Hagen 3). Our findings illustrate how AS and translational regulation of this plant specific histone lysine demethylase contributes to the removal of the repressive mark H3K27me3, promoting transcriptional activation of symbiotic genes required for the formation of functional nitrogen fixing nodules.

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Ferrari, M., Yacullo, M., Traubenik, S., Ibarra, E., Blanco, F. A., Reynoso, M., Zanetti, M. E.. 2026-09-17. The plant specific Histone Lysine Demethylase MtPKDM9B mediates the root nodule symbiosis by controlling H3K27me3 levels and expression of symbiotic genes. https://doi.org/10.64898/2026.09.12.750751

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