Genomic and Molecular Interaction Analysis of NodD1 in a Novel Bradyrhizobium yuanmingense sp. B64 Isolate for Nodulation and Symbiosis of Legume Plants.

Aura Aslan; Rumella Simarmata; Dwi Santosa; Tiwit Widowati; Sylvia Lekatompessy; Ananda Merrisa; Misbakhul Bait; Rendi Palar

doi:10.1007/s10528-026-11403-4

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Genomic and Molecular Interaction Analysis of NodD1 in a Novel Bradyrhizobium yuanmingense sp. B64 Isolate for Nodulation and Symbiosis of Legume Plants.

Aura Aslan·Rumella Simarmata·Dwi Santosa·Tiwit Widowati·Sylvia Lekatompessy·Ananda Merrisa·Misbakhul Bait·Rendi Palar

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Abstract

Rhizobial bacteria are known for their ability to fix nitrogen for leguminous plants and their essential function for sustainable agriculture. This study characterizes the taxonomic status and functional potential of the Bradyrhizobium B64 isolate using integrated genomic and molecular approaches. The whole genome of the B64 isolate was sequenced via Illumina paired-end technology. Species delimitation was performed using average nucleotide identity (ANI) and digital DNA-DNA Hybridization (dDDH). The NodD1 protein structure was modeled using AlphaFold3 and validated by Ramachandran plot analysis. Molecular docking was then conducted to evaluate interactions between NodD1 and four signaling flavonoids: Apigenin, Daidzein, Genistein, and Naringenin. Genomic analysis revealed a maximum ANI of 94.4% and dDDH values between 51.4 and 62.4%. Since these values fall below the standard prokaryotic thresholds (ANI < 95%; dDDH < 70%), the B64 isolate is identified as a novel species. Physiological assays confirmed nitrogen fixation (1.97 ppm), IAA production (3.67 ppm), and phosphate solubilization (26.10 ppm). Structural validation showed 100% of NodD1 residues in allowed regions, ensuring high model reliability. Docking simulations demonstrated strong binding affinities across all flavonoids, with binding free energies ranging from - 8.8 to - 9.0 kcal/mol. Daidzein exhibited the highest thermodynamic stability (- 9.0 kcal/mol), whereas apigenin showed the most extensive residue interaction network. The B64 isolate is a novel Bradyrhizobium species with a high symbiotic capacity. The stable NodD1-flavonoid interactions provide a molecular basis for efficient nodulation, positioning B64 as a promising candidate for developing lipo-chitooligosaccharide (LCO)-based biofertilizers.

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Bradyrhizobium Symbiosis Bacterial Proteins Fabaceae Molecular Docking Simulation Plant Root Nodulation Nitrogen Fixation Genome, Bacterial Isoflavones Flavanones Biofertilizer Flavonoids Molecular docking NodD1 Novel species Journal Article

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Aura Aslan, Rumella Simarmata, Dwi Santosa, Tiwit Widowati, Sylvia Lekatompessy, Ananda Merrisa, Misbakhul Bait, Rendi Palar. 2026-06-08. Genomic and Molecular Interaction Analysis of NodD1 in a Novel Bradyrhizobium yuanmingense sp. B64 Isolate for Nodulation and Symbiosis of Legume Plants.. https://doi.org/10.1007/s10528-026-11403-4

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