In order to examine the role of glutaminase in Rhidium etli, we isolated and characterized a R. etli glutaminase mutant (LM16). This mutant was selected for its impaired ability to grow on glutamine as nitrogen and carbon source while retaining the ability to grow on other nitrogen and carbon sources. The mutant showed very low levels of glutaminase activity under various growth conditions in comparison with the wild-type strain. With glutamine as the only nitrogen and carbon source, LM16 showed poor growth, with a very high content of glutamine, low glutamate content, and reduced ammonium excretion and 14C02 evolution from [U-l4C]glutarnine compared to the wild-type strain. This indicates that the main role of R. etli glutaminase is in the use of glutamine as carbon source. R. etli glutaminase also plays a role in maintaining the balance between glutamate and glutamine, as shown by the accumulation of glutamine and the low glutamate content of the mutant under different growth conditions. These results also indicate that glutaminase participates in a glutamine cycle in which it degrades glutamine which is then resynthesized by glutamine synthetase. The higher glutamine and lower glutamate content found in bacteroids of LM16 in comparison with bacteroids of the wild-type strain indicate that glutamine degradation by glutaminase plays an important role during the symbiosis between R. etli and Phaseolus vulgaris.