Rhizobium tibeticum sp. nov., a symbiotic bacterium isolated from Trigonella archiducis-nicolai (Sirj.) Vassilcz

Abstract

Isolated from root nodules of Trigonella archiducis-nicolai (Sirj.) Vassilcz. grown in Tibet, China, cells of the bacterial strains CCBAU 85039(T) and CCBAU 85027 were Gram-negative, aerobic, motile, non-spore-forming rods that formed colonies that were semi-translucent and opalescent on yeast extract-mannitol agar. In numerical taxonomy, SDS-PAGE analysis of whole-cell proteins and DNA-DNA hybridization, the two strains were very similar and were different from reference strains of defined Rhizobium species. In the phylogeny based on 16S rRNA gene sequences, they were most similar to Rhizobium etli CFN 42(T) (98.2 % similarity) and R. leguminosarum USDA 2370(T) (97.6 %). Sequence analyses of the housekeeping genes recA, atpD and glnII and the 16S-23S rRNA intergenic spacer, phenotypic characteristics and cellular fatty acid profiles strongly suggested that these two strains represented a novel species within Rhizobium. Cross-nodulation tests and sequencing of nifH and nodA genes showed that these two strains were symbiotic bacteria that nodulated Trigonella archiducis-nicolai, Medicago lupulina, Medicago sativa, Melilotus officinalis, Phaseolus vulgaris and Trigonella foenum-graecum. Based on the results, the novel species Rhizobium tibeticum sp. nov. is described to accommodate the two strains. The type strain is CCBAU 85039(T) (=LMG 24453(T) =CGMCC 1.7071(T)). The DNA G+C content of this strain is 59.7 mol% (T(m)).