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Journal of Bacteriology, March 2000, p. 1748-1753, Vol. 182, No. 6
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.

Glutathione Is Involved in Environmental Stress Responses in Rhizobium tropici, Including Acid Tolerance

Pablo M. Riccillo,1,2 Cecilia I. Muglia,1 Frans J. de Bruijn,2 Andrew J. Roe,3 Ian R. Booth,3 and O. Mario Aguilar1,*

Instituto de Bioquímica y Biologia Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, La Plata, Argentina1; MSU-DOE Plant Research Laboratory and Department of Microbiology, Michigan State University, East Lansing, MI 488242; and Department of Molecular and Cell Biology, Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, United Kingdom3

Received 20 September 1999/Accepted 20 December 1999

The isolation of rhizobial strains which exhibit an intrinsic tolerance to acidic conditions has been reported and has facilitated studies on the basic mechanisms underlying acid tolerance. Rhizobium tropici strain CIAT899 displays a high intrinsic tolerance to acidity and therefore was used in this work to study the molecular basis of bacterial responses to acid conditions and other environmental stresses. We generated a collection of R. tropici CIAT899 mutants affected in acid tolerance using Tn5-luxAB mutagenesis, and one mutant strain (CIAT899-13T2), which fails to grow under acid conditions, was characterized in detail. Strain CIAT899-13T2 was found to contain a single Tn5-luxAB insertion in a gene showing a high degree of similarity with the Escherichia coli gshB gene, encoding the enzyme glutathione synthetase. Intracellular potassium pools and intracellular pH levels were found to be lower in the mutant than in the parent. The glutathione-deficient mutant was shown to be sensitive to weak organic acids, osmotic and oxidative stresses, and the presence of methylglyoxal. Glutathione restores responses to these stresses almost to wild-type levels. Our data show that in R. tropici the production of glutathione is essential for growth in extreme environmental conditions. The mutant strain CIAT899-13T2 induced effective nodules; however, it was found to be outcompeted by the wild-type strain in coinoculation experiments.

* Corresponding author. Mailing address: Instituto de Bioquímica y Biologia Molecular, Facultad de Ciencias Exactas, Universidad Nacional de La Plata, Calles 47 y 115 (1900), La Plata, Argentina. Phone: 54 221 4250497, ext. 61. Fax: 54 221 4226947. E-mail: aguilar{at}nahuel.biol.unlp.edu.ar.

Journal of Bacteriology, March 2000, p. 1748-1753, Vol. 182, No. 6
0021-9193/00/$04.00+0
Copyright © 2000, American Society for Microbiology. All rights reserved.


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