Identification and functional characterization of Rhizobium leguminosarum bv. viciae genetic systems involved in nickel homeostasis.

A collection of Rhizobium leguminosarum bv. viciae strains isolated from ultramafic and contaminated soils in Italy and Germany, respectively, was analyzed for resistance to nickel and cobalt ions. These assays led to the identification of strain UPM1137, which is able to grow at high concentrations of nickel and cobalt. In order to identify genetic systems involved in the homeostasis to these metals, a random mutagenesis was carried out in UPM1137 by inserting a Tn5-derivative minitransposon. As a result 4313 transconjugants were obtained, being 39 of them (0.90%) unable to grow at 1.5 mM NiCl2. The identification of the transposon insertion site in these mutants showed that the disrupted genes encode proteins belonging to different functional categories, where the secreted and membrane proteins were the most numerous. The analysis of heavy metal resistance and phenotypes in symbiotic and free –living cells will define the contribution of these genes to metal homeostasis.