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Denitrification activity of Bradyrhizobium sp. isolated from Argentine soybean cultivated soils

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Abstract

Two hundred and fifty strains, all of them representatives of native Bradyrhizobium sp., isolated from soils cultivated with soybean have been characterized by their denitrification activity. In addition, the denitrification potential of those soils was also measured by evaluating the most-probable-number (MPN) of denitrifying bacteria and the denitrification enzyme assay (DEA). Of the 250 isolates tested, 73 were scored as probable denitrifiers by a preliminary screening method. Only 41 were considered denitrifiers because they produced gas bubbles in Durham tubes, cultures reached an absorbance of more than 0.1 and NO3− and NO2− were not present. Ten of these 41 were selected to confirm denitrification and to study denitrification genes. According to N2O production and cell protein concentration with NO3−, the isolates could be differentiated in three categories of denitrifiers. The presence of the napA, nirK, norC and nosZ genes was detected by production of a diagnostic PCR product using specific primers. RFLP from the 16S-23S rDNA spacer region (IGS) revealed that denitrifiers strains could be characterized as Bradyrhizobium japonicum and strains which were non-respiratory denitrifiers as B. elkanii.

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Acknowledgements

The authors are grateful to Universidad Nacional del Sur for financial support of the research work, through PGI No. 24/A112 and to CONICET for providing a fellowship to Leticia A. Fernández. We would like to thank Eulogio Bedmar from Zaidín for provided the primers to the detection of denitrifying genes and for helpful discussion and advice. We would also like to thank Alejandro Pidello from Universidad Nacional de Rosario for providing laboratory facilities to perform the chemical analysis; Viviana Echenique and Marina Díaz from CERZOS CONICET for providing the necessary facilities to do the molecular analysis and Carolina Fernández for editing the figures.

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Authors and Affiliations

  1. Laboratorio de Microbiología Agrícola, Departamento de Agronomía, Universidad Nacional del Sur, Altos del Palihue s/n, 8000, Bahia Blanca, Provincia de Buenos Aires, Argentina

    Leticia Andrea Fernández, Marcelo Antonio Sagardoy & Marisa Anahí Gómez

  2. Laboratorio de Química Biológica, Facultad de Ciencias Veterinarias, Universidad Nacional de Rosario, B. O. Lagos y Ruta 33, 2170, Casilda, Provincia de Santa Fe, Argentina

    Elda Beatriz Perotti

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  1. Leticia Andrea Fernández
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  2. Elda Beatriz Perotti
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  3. Marcelo Antonio Sagardoy
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  4. Marisa Anahí Gómez
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Correspondence to Leticia Andrea Fernández.

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Fernández, L.A., Perotti, E.B., Sagardoy, M.A. et al. Denitrification activity of Bradyrhizobium sp. isolated from Argentine soybean cultivated soils. World J Microbiol Biotechnol 24, 2577–2585 (2008). https://doi.org/10.1007/s11274-008-9828-x

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  • DOI: https://doi.org/10.1007/s11274-008-9828-x

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