[Construction of high-effective symbiotic bacteria: evolutionary models and genetic approaches].

Using the example of N2-fixing legume-rhizobial symbiosis, we demonstrated that the origin and evolution of bacteria symbiotic for plants involve the following: 1) the formation of novel sym gene systems based on reorganizations of the bacterial genomes and on the gene transfer from the distant organisms; 2) the loss of genes encoding for functions that are required for autonomous performance but interfere with symbiotic functions (negative regulators of symbiosis). Therefore, the construction of effective rhizobia strains should involve improvement of sym genes activities (for instance, nif, fix, and dct genes, encoding for nitrogenase synthesis or for the energy supply of N2 fixation), as well as the inactivation of negative regulators of symbiosis identified in our lab (eff genes encoding for the transport of sugars, and the production of polysaccharides, and storage compounds, as well as for oxidative-reductive processes).

[LPS mutants of Sinorhizobium meliloti and their nodulation competitiveness]. / LPS-mutanty Sinorhizobium meliloti i ikh noduliatsionnaia konkurentosposobnost'.

Four Tn5-transposon LPS mutants of Sinorhizobium meliloti (Tb9, Tb29, Ts22 and Ts32) have been studied. Each of four mutants has been established to contain a single insertion of Tn5-transposon in its genome. All mutations are located on a chromosome. Nodulation competitiveness (NC) of mutants towards the parent strain of S. meliloti CXM1-188 was investigated by resistant method using coinoculation of mutant and parent strain in the ratio 1:1. It was shown that NC was only 19-31% and 8-10%, for two strains Tb29 and Ts22, respectively which had lost the capability to synthesize higher molecular weight form of lipopolysaccharide (LPS1). Nodulation competitiveness of two other strains (Tb9 and Ts32) which retained the capability to synthesize LPS1 although in modified form varied from 49 to 62% and did not differ from NC of strain CXM1-188. The investigation of nodule formation rate has shown that four LPS-mutants did not differ from the parent strain by the number of root nodules. However the appearance of nodules induced by the mutant Tb29 was registered 7 days later than the nodules formed by other LPS-mutants and CXM1-188 strain. Obtained data concerning a single Tn5-insertion in genome of each of four S. meliloti LPS-mutants testify to the fact that both the disturbance of lipopolysaccharide synthesis and change of nodulation competitiveness in mutants Tb29 and Ts22 are results of a single mutation.