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Transfer RNA-dependent asparagine biosynthesis in Gluconacetobacter diazotrophicus and its influence on biological nitrogen fixation

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Abstract

Background and aims

Gluconacetobacter diazotrophicus is a nitrogen-fixing endophytic bacterium isolated from sugarcane, rice, elephant grass, sweet potato, coffee, and pineapple. These plants have high level of asparagine, which promotes microbial growth and inhibits nitrogenase activity. The regulation of intracellular concentrations of this amino acid is essential for growth and biological nitrogen fixation (BNF) in this diazotroph; however its asparagine metabolic pathway has not yet been clearly established.

Methods

The work reported here is the first to demonstrate the use of an alternative route for asparaginyl-tRNA (Asn-tRNA) and asparagine formation in an endophytic nitrogen-fixing bacterium by using in silico and in vitro analysis.

Results

The indirect route involves transamidation of incorrectly charged tRNA via GatCAB transamidase. Nitrogenase activity was completely inhibited by 20 mM Asn in LGI-P medium, which in contrast promotes protein synthesis and microbial growth.

Conclusions

The analysis carried out in this work shows that intracellular levels of asparagine regulate the expression of nitrogenase nifD gene (GDI0437), suggesting that the presence of an alternative route to produce asparagine might give the G. diazotrophicus a tighter control over cell growth and BNF, and may be of importance in the regulation of the endophytic plant-microbe interaction.

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Acknowledgments

The authors thank Genome Sequencing facilities core Johanna Döbereiner and Embrapa Agrobiologia staff for their help and support in laboratory analyses and the team of Dieter Söll Lab (Yale University) for support and discussion. The research was partly supported by Fundação Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro (FAPERJ) and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq).

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Authors and Affiliations

  1. Instituto de Bioquímica Médica, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brazil

    Sylvia M. Alquéres, Alexander M. Cardoso, Jordano Brito-Moreira & Orlando B. Martins

  2. Instituto Nacional de Metrologia, Qualidade e Tecnologia, Diretoria de Programa, DIPRO, Av. Nossa Senhora das Graças, 50-prédio 6, LaBio, Xerém, 25250-020, Duque de Caxias, RJ, Brazil

    Alexander M. Cardoso

  3. Laboratório de Genética e Bioquímica, Embrapa Agrobiologia, Seropédica, Brazil

    Jose I. Baldani

Authors
  1. Sylvia M. Alquéres
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  2. Alexander M. Cardoso
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  3. Jordano Brito-Moreira
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  4. Jose I. Baldani
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  5. Orlando B. Martins
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Corresponding author

Correspondence to Alexander M. Cardoso.

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Responsible Editor: Euan K. James.

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Alquéres, S.M., Cardoso, A.M., Brito-Moreira, J. et al. Transfer RNA-dependent asparagine biosynthesis in Gluconacetobacter diazotrophicus and its influence on biological nitrogen fixation. Plant Soil 356, 209–216 (2012). https://doi.org/10.1007/s11104-011-0952-2

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  • DOI: https://doi.org/10.1007/s11104-011-0952-2

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