Applied Microbiology and Biotechnology

, Volume 93, Issue 5, pp 2035–2049 | Cite as

Polyphasic approach for the characterization of rhizobial symbionts effective in fixing N2 with common bean (Phaseolus vulgaris L.)

  • Juscélio Donizete Cardoso
  • Mariangela Hungria
  • Diva S. Andrade
Applied genetics and molecular biotechnology
First Online:
Received:
Revised:
Accepted:

Abstract

Common bean (Phaseolus vulgaris L.) is a legume that has been reported as highly promiscuous in nodulating with a variety of rhizobial strains, often with low effectiveness in fixing nitrogen. The aim of this work was to assess the symbiotic efficiency of rhizobial strains isolated from common bean seeds, nodules of Arachis hypogaea, Mucuna pruriens, and soils from various Brazilian agroecosystems, followed by the characterization of elite strains identified in the first screening. Forty-five elite strains were analyzed for symbiotic properties (nodulation, plant-growth, and nitrogen-fixation parameters) under greenhouse conditions in pots containing non-sterile soil, and variation in symbiotic performance was observed. Elite strains were also characterized in relation to morpho-physiological properties, genetic profiles of rep-polymerase chain reaction (PCR; BOX), and restriction fragment length polymorphism (RFLP)-PCR of the 16S rRNA. Sequence analyses of the 16S rRNA were obtained for 17 strains representative of the main groups resulting from all previous analyses. One of the most effective strains, IPR-Pv 2604, was clustered with Rhizobium tropici, whereas strain IPR-Pv 583, showing lower effectiveness in fixing N2, was clustered with Herbaspirillum lusitanum. Surprisingly, effective strains were clustered with unusual symbiotic genera/species, including Leifsonia xyli, Stenotrophomonas maltophilia, Burkholderia, and Enterobacter. Some strains recognized in this study were outstanding in their nitrogen-fixing capacity and therefore, show high biotechnological potential for use in commercial inoculants.

Keywords

Biological nitrogen fixation BOX-PCR RFLP-PCR 16S rRNA Phaseolus vulgaris Principal component analysis (PCA) 
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Notes

Acknowledgments

The first author thanks Capes (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior) for the PhD scholarship. The authors are grateful to Walter M. Kranz for help with the soil sampling. Authors also thank Maria Elisabeth Vasconcellos (IAPAR) for help with the multivariate analyses and L.F.A. Fregonesi to organize the map, to Mauro S Parra (IAPAR) for helping with the greenhouse experiment, and Dr. Allan R. J. Eaglesham for suggestions on the manuscript. The research was partially supported by CNPq (Conselho Nacional de Desenvolvimento Científico e Tecnológico)/MCT/MAPA (577933/2008) and CNPq-Repensa (562008/2010-1). M. Hungria also acknowledges a research fellowship from CNPq (300547/2010-2).

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Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Juscélio Donizete Cardoso
    • 1
    • 2
    • 3
  • Mariangela Hungria
    • 2
  • Diva S. Andrade
    • 3
    • 4
  1. 1.Department of MicrobiologyUniversidade Estadual de LondrinaLondrinaBrazil
  2. 2.Embrapa SojaLondrinaBrazil
  3. 3.Instituto Agronômico do ParanáLondrinaBrazil
  4. 4.IAPARLondrinaBrazil

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