Nodulation competitiveness of Tn5-induced mutants of Rhizobium fredii USDA208 that are altered in motility and extracellular polysaccharide production

The role of motility and extracellular polysaccharide production in nodulation competitiveness of Rhizobium fredii was examined. Transposon Tn5 was used to mutagenize strain USDA208, and mutants with reduced motility on semisolid agar medium were isolated. One such mutant, 208M3, migrated to only one-seventh the distance of the parental strain. Solid medium amended with triphenyltetrazolium chloride was used to identify mutants altered in extracellular polysaccharide production. Type 1 colonies, typified by mutant 208T13, were heavily mucoid, while type 2 colonies, represented by mutant 208T3, were dry and nonmucoid. Compared with strain USDA208, these mutants produced 4- to 5-fold more extracellular polysaccharide and 20% as much extracellular polysaccharide, respectively. Marker exchange of 208T3 genomic DNA containing Tn5 into strain USDA208 resulted in a mutant, 208K1, that produced extracellular polysaccharide levels similar to mutant 208T3. Mutants 208M3, 208T3, and 208T13 contained single Tn5 insertions. All formed pink nodules on 'Peking' soybean that were structurally indistinguishable and contained proteins with similar profiles. Rates of nodulation were similar in the mutants and the parental strain. Mutants 208M3 and 208T13 were as competitive against an isolate of Bradyrhizobium japonicum serogroup 123 as was strain USDA208. In contrast, mutants 208T3 and 208K1 were competitively superior. Key words: nodulation competition, motility, extracellular polysaccharide, Rhizobium.