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Rhizobium, Bradyrhizobium and Agrobacterium strains isolated from cultivated legumes

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Abstract

The present study was conducted to isolate and characterize rhizobial strains from root nodules of cultivated legumes, i.e. chickpea, mungbean, pea and siratro. Preliminary characterization of these isolates was done on the basis of plant infectivity test, acetylene reduction assay, C-source utilization, phosphate solubilization, phytohormones and polysaccharide production. The plant infectivity test and acetylene reduction assay showed effective root nodule formation by all the isolates on their respective hosts, except for chickpea isolate Ca-18 that failed to infect its original host. All strains showed homology to a typical Rhizobium strain on the basis of growth pattern, C-source utilization and polysaccharide production. The strain Ca-18 was characterized by its phosphate solubilization and indole acetic acid (IAA) production. The genetic relationship of the six rhizobial strains was carried out by random amplified polymorphic DNA (RAPD) including a reference strain of Bradyrhizobium japonicum TAL-102. Analysis conducted with 60 primers discriminated between the strains of Rhizobium and Bradyrhizobium in two different clusters. One of the primers, OPB-5, yielded a unique RAPD pattern for the six strains and well discriminated the non-nodulating chickpea isolate Ca-18 from all the other nodulating rhizobial strains. Isolate Ca-18 showed the least homology of 15% and 18% with Rhizobium and Bradyrhizobium, respectively, and was probably not a (Brady)rhizobium strain. Partial 16S rRNA gene sequence analysis for MN-S, TAL-102 and Ca-18 strains showed 97% homology between MN-S and TAL-102 strains, supporting the view that they were strains of B. japonicum species. The non-infective isolate Ca-18 was 67% different from the other two strains and probably was an Agrobacterium strain.

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Acknowledgements

The research work was funded partly by the IDB BIRCEN project and FAO/IAEA TC project no. PAK/5/037. We are thankful to Ms. Fathia Mubeen and Mr. Mazhar Hussain for their help in DNA sequencing work, as well as to Dr. Farooq Latif for his help with HPLC to detect phytohormones.

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  1. National Institute for Biotechnology and Genetic Engineering (NIBGE), Jhang Road, P.O. Box 577, Faisalabad 3800, Pakistan

    Sohail Hameed, Sumera Yasmin, Kauser A. Malik, Yusuf Zafar & Fauzia Y. Hafeez

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  1. Sohail Hameed
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  2. Sumera Yasmin
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  3. Kauser A. Malik
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  4. Yusuf Zafar
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  5. Fauzia Y. Hafeez
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Correspondence to Fauzia Y. Hafeez.

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Hameed, S., Yasmin, S., Malik, K.A. et al. Rhizobium, Bradyrhizobium and Agrobacterium strains isolated from cultivated legumes. Biol Fertil Soils 39, 179–185 (2004). https://doi.org/10.1007/s00374-003-0697-z

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  • DOI: https://doi.org/10.1007/s00374-003-0697-z

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