# Isolation, biochemical characterization, and greenhouse authentication of chickpea (Cicer arietinum L.) rhizobia collected from some major chickpea growing areas of Woldia, North Wollo, Ethiopia

**Authors:** Kelemu Yimenu Gebre, Abebe Girma Demissie, Aderajew Adgo Tesema, Habtamu Zegeye Belay, Mekashaw Worku Akalye, Agere Bereket Friew, Fisseha Getachew Baye, Habtie Bassie Felatie, Mezgebu Abunie Yohannes, Marye Alemu Eshetu, Wondye Ayalew Shiferaw

PMC · DOI: 10.1371/journal.pone.0330169 · PLOS One · 2025-08-14

## TL;DR

This study identifies effective native Rhizobium bacteria from chickpea fields in Ethiopia that can boost plant growth and reduce fertilizer use.

## Contribution

The study isolates and validates native Rhizobium strains from Woldia, Ethiopia, showing their potential as bio-inoculants for chickpea.

## Key findings

- Five Rhizobium isolates improved chickpea germination and seedling growth in greenhouse trials.
- Isolate WUSFDG-23 showed the highest symbiotic effectiveness and biomass accumulation.
- Native Rhizobium strains can serve as sustainable bio-inoculants for chickpea cultivation.

## Abstract

Chickpea (Cicer arietinum L.) is a vital legume crop worldwide, valued for its high nutritional content and significant contribution to food security and soil fertility through biological nitrogen fixation. Despite its importance, chickpea yields remain suboptimal in many regions, including Ethiopia, primarily due to constraints such as poor soil fertility and inadequate use of effective rhizobia inoculants. This study aimed to isolate and characterize native Rhizobium strains from chickpea root nodules collected from fields in the Woldia region and to assess their potential to promote plant growth. A total of 41 bacterial isolates were obtained, of which 12 were presumptively identified as Rhizobium based on growth characteristics on Congo red and bromothymol blue media. These isolates were further characterized morphologically and biochemically. Five biochemically promising isolates were selected for evaluation in a controlled 45-day greenhouse experiment under sterile conditions. Inoculation with these isolates significantly enhanced seed germination and early seedling growth compared to uninoculated controls. The symbiotic effectiveness of the isolates ranged from 74.3% to 121.9%, with isolates WUSFDG-23, WUSFMC-31, and WUSFMC-23 demonstrating high effectiveness, isolate WUSFDG-23 markedly increased nodulation and biomass accumulation. This study highlights the potential of native Rhizobium isolates from Woldia chickpea fields, especially WUSFDG-23, as effective bio-inoculants to promote sustainable chickpea production and reduce dependence on chemical fertilizers.

## Full-text entities

- **Genes:** Catalase [NCBI Gene 101513499], Urease [NCBI Gene 101514697]
- **Diseases:** fungal (MESH:D009181), infection (MESH:D007239)
- **Chemicals:** CaCl2 2H2O (-), Congo Red (MESH:D003224), Copper sulphate pentahydrate (MESH:D019327), HCl (MESH:D006851), Safranin (MESH:C009195), Calcium chloride dihydrate (MESH:D002122), H2O2 (MESH:D006861), iron chloride (MESH:C024555), gibberellins (MESH:D005875), carbon (MESH:D002244), nucleotides (MESH:D009711), Boric acid (MESH:C032688), calcium hypochlorite (MESH:C004488), H2S (MESH:D006862), polythene (MESH:D020959), isoamyl alcohol (MESH:C029683), potassium hydroxide (MESH:C029943), N (MESH:D009584), Indole (MESH:C030374), IAA (MESH:C030737), Carbohydrate (MESH:D002241), para-dimethyl amino benzaldehyde (MESH:C510241), starch (MESH:D013213), NaCl (MESH:D012965), ketoconazole (MESH:D007654), citrate (MESH:D019343), urea (MESH:D014508), ferrous sulfate heptahydrate (MESH:C020748), sugar (MESH:D000073893), dextrose (MESH:D005947), sucrose (MESH:D013395), Dipotassium phosphate (MESH:C013216), aluminum (MESH:D000535), potassium iodide (MESH:D011193), reactive oxygen species (MESH:D017382), phenol red (MESH:D010637), cytokinin (MESH:D003583), water (MESH:D014867), ethanol (MESH:D000431), Mannitol (MESH:D008353), amino acids (MESH:D000596), sodium hypochlorite (MESH:D012973), tryptophan (MESH:D014364), oil (MESH:D009821), NaOH (MESH:D012972), lactose (MESH:D007785), ammonium (MESH:D064751), auxin (MESH:D007210), EDTA (MESH:D004492), agar (MESH:D000362), Magnesium sulphate heptahydrate (MESH:D008278), Fe (MESH:D007501), Ammonia (MESH:D000641), Iodine (MESH:D007455), glycerol (MESH:D005990), Zinc sulphate heptahydrate (MESH:D019287), flavonoids (MESH:D005419), CaCO3 (MESH:D002119), Manganese sulphate monohydrate (MESH:C039798), Bromothymol Blue (MESH:D001979)
- **Species:** Bradyrhizobium (genus) [taxon 374], Vicia faba (broad bean, species) [taxon 3906], Ovis aries (domestic sheep, species) [taxon 9940], Rhizobium (genus) [taxon 379], Cicer arietinum (chickpea, species) [taxon 3827], Sorghum bicolor (broomcorn, species) [taxon 4558], Bos taurus (bovine, species) [taxon 9913], Saccharomyces cerevisiae (baker's yeast, species) [taxon 4932], Capra hircus (domestic goat, species) [taxon 9925], Lens culinaris (lentil, species) [taxon 3864], Equus asinus (African ass, species) [taxon 9793], Agrobacterium (genus) [taxon 357]

## Full text

_Full body text omitted from this summary view._ Fetch the complete paper as Markdown: https://tomesphere.com/paper/PMC12352659/full.md

## Figures

3 figures with captions in the complete paper: https://tomesphere.com/paper/PMC12352659/full.md

## References

60 references — full list in the complete paper: https://tomesphere.com/paper/PMC12352659/full.md

---
Source: https://tomesphere.com/paper/PMC12352659