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A histidine kinase sensor protein gene is necessary for induction of low pH tolerance in Sinorhizobium sp. strain BL3

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Abstract

The aim of this investigation was to identify and isolate genes involved in acid tolerance from Sinorhizobium sp. strain BL3. It was hypothesized that acid tolerance of strain BL3 could be enhanced by high level expression of certain genes involved in acid tolerance, following insertion of these genes in a multiple copy plasmid. A cosmid clone library of BL3 was introduced into BL3, and the transconjugant colonies were selected at low pH. A single cosmid containing genes for acid tolerance was isolated from 40 different colonies. By transposon–insertion mutagenesis, subcloning and DNA sequencing, a gene involved in acid tolerance, actX, was identified in a 4.4-kb fragment of this cosmid. The actX mutant of BL3 showed increased acid sensitivity and was complemented by the 4.4-kb subcloned fragment. Phaseolus lathyroides seedlings inoculated with recombinant strains containing multiple copies of actX showed increased symbiotic performance at low pH. By constructing an actX::gus fusion, it was shown that actX was induced at low pH. actX encodes a putative histidine kinase sensor protein of a two-component regulatory system. The method of gene identification used in this study for isolation of actX may be applied for the isolation of other genes involved in tolerance to adverse environmental factors.

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Acknowledgements

This work was supported by the Royal Golden Jubilee Ph.D. Program of the Thailand Research Fund, Suranaree University of Technology, Nakhon Ratchasima, Thailand, and University of Hawaii, USA. We thank Jonathan Awaya for helpful suggestions and technical help throughout the project.

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Authors and Affiliations

  1. School of Biotechnology, Suranaree University of Technology, Nakhon Ratchasima, 30000, Thailand

    Panlada Tittabutr, Waraporn Payakapong, Neung Teaumroong & Nantakorn Boonkerd

  2. Department of Tropical Plant and Soil Sciences, University of Hawaii at Manoa, Honolulu, HI, 96822, USA

    Paul W. Singleton

  3. Department of Molecular Biosciences and Bioengineering, University of Hawaii at Manoa, Honolulu, HI, 96822, USA

    Dulal Borthakur

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  1. Panlada Tittabutr
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  2. Waraporn Payakapong
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  3. Neung Teaumroong
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  4. Nantakorn Boonkerd
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  5. Paul W. Singleton
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  6. Dulal Borthakur
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Correspondence to Dulal Borthakur.

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Tittabutr, P., Payakapong, W., Teaumroong, N. et al. A histidine kinase sensor protein gene is necessary for induction of low pH tolerance in Sinorhizobium sp. strain BL3. Antonie Van Leeuwenhoek 89, 125–134 (2006). https://doi.org/10.1007/s10482-005-9015-0

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