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Open Access Methodology article

Tools for genetic manipulation of the plant growth-promoting bacterium Azospirillum amazonense

Fernando H Sant'Anna1, Dieime S Andrade1, Débora B Trentini1, Shana S Weber1 and Irene S Schrank2*

Author affiliations

1 Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul (Av. Bento Gonçalves, 9500, Campus do Vale), Porto Alegre, RS, Brazil

2 Departamento de Biologia Molecular e Biotecnologia - Centro de Biotecnologia, Universidade Federal do Rio Grande do Sul (Av. Bento Gonçalves, 9500, Campus do Vale), RS, Brazil

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Citation and License

BMC Microbiology 2011, 11:107  doi:10.1186/1471-2180-11-107

Published: 16 May 2011

Abstract

Background

Azospirillum amazonense has potential to be used as agricultural inoculant since it promotes plant growth without causing pollution, unlike industrial fertilizers. Owing to this fact, the study of this species has gained interest. However, a detailed understanding of its genetics and physiology is limited by the absence of appropriate genetic tools for the study of this species.

Results

Conjugation and electrotransformation methods were established utilizing vectors with broad host-replication origins (pVS1 and pBBR1). Two genes of interest - glnK and glnB, encoding PII regulatory proteins - were isolated. Furthermore, glnK-specific A. amazonense mutants were generated utilizing the pK19MOBSACB vector system. Finally, a promoter analysis protocol based on fluorescent protein expression was optimized to aid genetic regulation studies on this bacterium.

Conclusion

In this work, genetic tools that can support the study of A. amazonense were described. These methods could provide a better understanding of the genetic mechanisms of this species that underlie its plant growth promotion.