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Applied and Environmental Microbiology, April 2007, p. 2635-2643, Vol. 73, No. 8
0099-2240/07/$08.00+0     doi:10.1128/AEM.01823-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Evidence of Horizontal Transfer of Symbiotic Genes from a Bradyrhizobium japonicum Inoculant Strain to Indigenous Diazotrophs Sinorhizobium (Ensifer) fredii and Bradyrhizobium elkanii in a Brazilian Savannah Soil{triangledown}

Fernando Gomes Barcellos, Pâmela Menna,{dagger} Jesiane Stefânia da Silva Batista,{dagger} and Mariangela Hungria*

Embrapa Soja, Cx. Postal 231, 86001-970, Londrina, PR, Brazil; Departamento Microbiologia, Universidade Estadual de Londrina, Londrina, PR, Brazil; and Conselho Nacional de Desenvolvimento Científico e Tecnológico, Brasilia, DF, Brazil

Received 2 August 2006/ Accepted 4 February 2007

The importance of horizontal gene transfer (HGT) in the evolution and speciation of bacteria has been emphasized; however, most studies have focused on genes clustered in pathogenesis and very few on symbiosis islands. Both soybean (Glycine max [L.] Merrill) and compatible Bradyrhizobium japonicum and Bradyrhizobium elkanii strains are exotic to Brazil and have been massively introduced in the country since the early 1960s, occupying today about 45% of the cropped land. For the past 10 years, our group has obtained several isolates showing high diversity in morphological, physiological, genetic, and symbiotic properties in relation to the putative parental inoculant strains. In this study, parental strains and putative natural variants isolated from field-grown soybean nodules were genetically characterized in relation to conserved genes (by repetitive extragenic palindromic PCR using REP and BOX A1R primers, PCR-restriction fragment length polymorphism, and sequencing of the 16SrRNA genes), nodulation, and N2-fixation genes (PCR-RFLP and sequencing of nodY-nodA, nodC, and nifH genes). Both genetic variability due to adaptation to the stressful environmental conditions of the Brazilian Cerrados and HGT events were confirmed. One strain (S 127) was identified as an indigenous B. elkanii strain that acquired a nodC gene from the inoculant B. japonicum. Another one (CPAC 402) was identified as an indigenous Sinorhizobium (Ensifer) fredii strain that received the whole symbiotic island from the B. japonicum inoculant strain and maintained an extra copy of the original nifH gene. The results highlight the strategies that bacteria may commonly use to obtain ecological advantages, such as the acquisition of genes to establish effective symbioses with an exotic host legume.

* Corresponding author. Mailing address: Embrapa Soja, Cx. Postal 231, 86001-970 Londrina, Paraná, Brazil. Phone: 55 43 33716206. Fax: 55 43 33717100. E-mail: hungria{at}cnpso.embrapa.br

{triangledown} Published ahead of print on 16 February 2007.

{dagger} P.M. and J.S.D.S.B. contributed equally to this work.

Applied and Environmental Microbiology, April 2007, p. 2635-2643, Vol. 73, No. 8
0099-2240/07/$08.00+0     doi:10.1128/AEM.01823-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.



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