# Genomic and Phenotypic Bases of Salt Tolerance in Sinorhizobium meliloti : Candidate Traits for Bioinoculant Development Addressing Saline Soils

**Authors:** Agnese Bellabarba, Camilla Fagorzi, Giovanni Bacci, Francesca Decorosi, Alice Checcucci, Gaio Cesare Pacini, Abdelkader Bekki, Amina El Hadj Mimoune, Khalid Azim, Majida Hafidi, Alessio Mengoni, Francesco Pini, Carlo Viti

PMC · DOI: 10.1111/1751-7915.70304 · 2026-01-29

## TL;DR

This study identifies genetic and metabolic traits in Sinorhizobium meliloti that help the bacteria survive in salty soils, which could improve legume growth in such environments.

## Contribution

The study identifies candidate genes and metabolic traits associated with salt tolerance in Sinorhizobium meliloti using genome-wide and phenotypic analyses.

## Key findings

- Genome-wide association analysis identified loci linked to salt tolerance, including genes involved in cell wall organization and quorum sensing.
- Salt-tolerant strains showed enhanced carbon source usage, indicating broader metabolic adaptability.
- Phenotype Microarray confirmed a connection between salt tolerance and carbohydrate metabolism.

## Abstract

Soil salinity poses a major challenge to the legume‐rhizobia symbiosis development, thereby affecting sustainable agriculture. Selecting NaCl‐tolerant strains and enhancing the native strains' fitness under salt stress are essential steps for the restoration of marginal areas. In this work, 49 
Sinorhizobium meliloti
 strains, the rhizobial species forming symbiotic nitrogen‐fixing associations with alfalfa—including 21 de novo‐sequenced field isolates—were subjected to a thorough in vitro screening for salt tolerance at progressively higher NaCl concentrations. Field isolates showed genome‐based geographical clustering but contrasting salt tolerance abilities. Indeed, genome‐wide association (GWA) analysis on the strains' whole‐genome sequencing data indicated several loci associated with the variability in salt tolerance. Candidate genes were involved in various processes including cell wall organisation, LPS biosynthesis, quorum sensing, and carbohydrate transport and metabolism. The relationship with carbohydrate metabolism was further confirmed by Phenotype Microarray analysis which indicated salt‐tolerant strains having enhanced capacity in carbon source usage. These findings reveal synergistic pathways underlying salt tolerance and suggest candidate traits (e.g., quorum sensing, carbohydrate synthesis and modification) for developing bioinoculants to enhance legume performance in saline soils.

Genome‐wide association analysis on 49 
Sinorhizobium meliloti
 strains (21 de novo sequenced) reveals extensive phenotypic heterogeneity in salt tolerance, closely associated with quorum sensing, cell surface stability and adaptation. Salt‐tolerant strains exhibit a wider metabolic capacity, especially in carbon sources exploitation, underscoring their potential for adaptability in harsh conditions.

## Linked entities

- **Chemicals:** NaCl (PubChem CID 5234)
- **Species:** Sinorhizobium meliloti (taxon 382)

## Full-text entities

- **Diseases:** Salt Tolerance (MESH:D013651)
- **Chemicals:** CaCl2 (MESH:D002122), NH4Cl (MESH:D000643), D-tagatose (MESH:C030192), K (MESH:D011188), C (MESH:D002244), glycogen (MESH:D006003), spermidine (MESH:D013095), D-mannitol (MESH:D008353), amine (MESH:D000588), FeCl3 (MESH:C024555), Sucrose (MESH:D013395), sorbitol (MESH:D013012), Carbohydrate (MESH:D002241), UDP-galactose (MESH:D014531), Adonitol (MESH:D012255), tetrazolium (MESH:D013778), Glycerol (MESH:D005990), chloride (MESH:D002712), Acetyl-CoA (MESH:D000105), D-mannose (MESH:D008358), thiamine-HCl (MESH:C000712172), agar (MESH:D000362), L-fucose (MESH:D005643), aromatic hydrocarbons (MESH:D006841), D-arabitol (MESH:C014999), NaCl (MESH:D012965), trehalose (MESH:D014199), LPS (MESH:D008070), Maltose (MESH:D008320), proline (MESH:D011392), H (MESH:D006859), polysaccharide (MESH:D011134), glycerol-3-phosphate (MESH:C029620), UDP-glucose (MESH:D014532), aromatic amino acid (MESH:D024322), N-Acetyl-D-Glucosamine (MESH:D000117), nitrogen (MESH:D009584), L-Arabinose (MESH:D001089), Fatty acid (MESH:D005227), glutamate (MESH:D018698), lipid (MESH:D008055), amino acids (MESH:D000596), amide (MESH:D000577), Turanose (MESH:C029454), glycine betaine (MESH:D001622), sugar alcohols (MESH:D013402), Salt (MESH:D012492), mycothiol (MESH:C089265), sugar (MESH:D000073893), Melibionic acid (MESH:C017485), CoCl2 (MESH:C018021), N-acetylmuramic acid (MESH:C031651), Na+ (MESH:D012964), MgSO4 (MESH:D008278), fructose (MESH:D005632), Cl- (MESH:D002713), L-rhamnose (MESH:D012210), D-cellobiose (-), CTAB (MESH:D000077286), Maltotriose (MESH:C008317)
- **Species:** Sinorhizobium meliloti BO21CC (strain) [taxon 928723], Escherichia coli (E. coli, species) [taxon 562], Sinorhizobium medicae (species) [taxon 110321], Saccharomyces cerevisiae (baker's yeast, species) [taxon 4932], Sinorhizobium meliloti (species) [taxon 382], Cajanus cajan (pigeon pea, species) [taxon 3821], Sinorhizobium meliloti RU11/001 (strain) [taxon 1401243], Sinorhizobium sp. (species) [taxon 42445], Mycolicibacterium smegmatis (species) [taxon 1772], Rhizobium sp. (species) [taxon 391], Medicago sativa (alfalfa, species) [taxon 3879], Bacteria Latreille et al. 1825 (Bacteria stick insect, genus) [taxon 629395], Sinorhizobium meliloti 2011 (strain) [taxon 1286640], Ensifer (genus) [taxon 106591]
- **Mutations:** A0641M
- **Cell lines:** SO14 — Homo sapiens (Human), Chronic myelogenous leukemia, BCR-ABL1 positive, Cancer cell line (CVCL_6905), MO26 — Homo sapiens (Human), Lung small cell carcinoma, Cancer cell line (CVCL_0C22), RU11/001 — Homo sapiens (Human), Cutaneous melanoma, Cancer cell line (CVCL_H242), RMO17 — Gnathopogon caerulescens (Willow shiner), Spontaneously immortalized cell line (CVCL_Y563), MO56 — Mus musculus (Mouse), Factor-dependent cell line (CVCL_C4TM), Rm41 — Homo sapiens (Human), Retinitis pigmentosa, Induced pluripotent stem cell (CVCL_WI46), BO21CC — Homo sapiens (Human), Transformed cell line (CVCL_E492), SM15 — Homo sapiens (Human), Lung small cell carcinoma, Cancer cell line (CVCL_7004), SM15(1 — Homo sapiens (Human), AIDS-related immunoblastic lymphoma, Cancer cell line (CVCL_IU19), MO28 — Oryctolagus cuniculus (Rabbit), Transformed cell line (CVCL_6E94), MO53 — Homo sapiens (Human), Galactosemia, Finite cell line (CVCL_2778)

## Figures

5 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12855168/full.md

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Source: https://tomesphere.com/paper/PMC12855168