Isolation of salt-tolerant mutants of Mesorhizobium ciceri strain Rch125 and identification of genes involved in salt sensitivity

Mesorhizobium ciceri Rch125 is a salt-sensitive strain isolated from root nodules of chickpea (Cicer arietinum L.). The aim of this work was to investigate the genes responsible for the sensitivity to salinity. Twelve Rch125 salt-tolerant mutants were isolated after random Tn5 mutagenesis and selected using a medium containing 300 mM NaCl, where growth of the wild-type is totally inhibited. In addition to this NaCl tolerance, the mutants also displayed higher tolerance to LiCl, CaCl and sucrose. Genes that were disrupted in the salt-tolerant mutants were in one of three functional categories: membrane transporters, outer membrane proteins, and genes of unknown function. Genetic complementation experiments demonstrated that the genes identified were involved in the salt sensitivity of the Rch125 strain. In most cases, disruption of the salt-sensitivity genes did not negatively affect the free-living or the symbiotic capabilities of Rch125 under non-saline conditions.This work was supported by grants AECI 173/03/P and AECI A/6935/06 from Agencia Española de Cooperación Internacional to JS and BB and grants BIO2005-08089-C02-01 (Ministerio de Educación y Ciencia, Spain) and BIO2008-02447 (Ministerio de Ciencia e Innovación) to JS.Peer Reviewe