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Identification of salt stress inducible genes that control cell envelope related functions in Azospirillum brasilense Sp7

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Abstract

Plant growth promoting rhizobacteria such as Azospirillum brasilense are agronomically important as they are frequently used for crop inoculation. But adverse factors such as increasing soil salinity limit their survival, multiplication and phytostimulatory effect. In order to understand the role of the genes involved in the adaptation of A. brasilense Sp7 to salt stress, a mutant library (6,800 mutants) was constructed after random integration of a mini-Transposon Tn5 derivative containing a promoterless gusA and oriV. The library was screened for salt stress inducible Gus activity on minimal malate agar medium containing NaCl and 5-bromo-4-chloro-3-indolyl-β-d-glucuronide. Salt stress responsiveness of the promoters was estimated by quantifying GusA activity in the presence and absence of NaCl stress using p-nitrophenyl-β-d-glucuronide as a substrate. In 11 mutants showing high levels of gusA expression in the presence of salt-stress, the partial nucleotide sequence of the DNA region flanking the site of Tn5 insertion was determined and analysed using the NCBI-BLAST programs. Similarity searches revealed that 10 out of the 11 genes sequenced showed notable similarity with genes involved in functions related to modulation in the composition of exopolysaccharides, capsular polysaccharides, lipopolysaccharides, peptidoglycan and lipid bilayer of the cell envelope. Induction of cell envelope related genes in response to salt stress and salt sensitive phenotype of several mutants in A. brasilense indicate a prominent role of cell envelope in salt-stress adaptation.

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Acknowledgments

We acknowledge the financial support to TN from International KU-Leuven Scholarship and ASM-UNESCO travel grant. We thank Veerle Keijers for technical assistance; Jan Michiels and Anne Van Dommelen, for their valuable suggestions.

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Authors and Affiliations

  1. Bacterial Genetics Laboratory, School of Biotechnology, Faculty of Science, Banaras Hindu University, Varanasi, 221005, India

    Thirunavukkarasu Nagarajan & Anil Kumar Tripathi

  2. Centre for Microbial and Plant Genetics, K.U. Leuven, Kasteelpark Arenberg 20, 3001, Leuven, Belgium

    Jos Vanderleyden

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  1. Thirunavukkarasu Nagarajan
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  2. Jos Vanderleyden
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  3. Anil Kumar Tripathi
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Correspondence to Anil Kumar Tripathi.

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Communicated by G. Klug.

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Nagarajan, T., Vanderleyden, J. & Tripathi, A.K. Identification of salt stress inducible genes that control cell envelope related functions in Azospirillum brasilense Sp7. Mol Genet Genomics 278, 43–51 (2007). https://doi.org/10.1007/s00438-007-0224-2

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  • DOI: https://doi.org/10.1007/s00438-007-0224-2

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