Phylogeny based on 16S rDNA andnifHsequences ofRalstonia taiwanensisstrains isolated from nitrogen-fixing nodules ofMimosa pudica, in India

Abstract

Bacterial symbionts present in the indeterminate-type nitrogen (N)-fixing nodules of Mimosa pudica grown in North and South India showed maximum similarity to Ralstonia taiwanensis on the basis of carbon-source utilization patterns and 16S rDNA sequence. Isolates from the nodules of M. pudica from North India and South India showed identical ARDRA (Amplified Ribosomal DNA Restriction Analysis) patterns with Sau3AI and RsaI, but AluI revealed dimorphy between the North Indian and South Indian isolates. Alignment of 16S rDNA sequences revealed similarity of North Indian isolates with an R. taiwanensis strain isolated from M. pudica in Taiwan, whereas South Indian isolates showed closer relatedness with the isolates from Mimosa diplotricha. Alignment of nifH sequences from both North Indian and South Indian isolates with that of the related isolates revealed their closer affinity to α-rhizobia, suggesting that nif genes in the β-rhizobia might have been acquired from α-rhizobia via lateral transfer during co-occupancy of nodules by α-rhizobia and progenitors of R. taiwanensis, members of the β-subclass of Proteobacteria. Immunological cross-reaction of the bacteroid preparation of M. pudica nodules showed strong a positive signal with anti-dinitrogenase reductase antibody, whereas a weak positive cross-reaction was observed with free-living R. taiwanensis grown microaerobically in minimal medium with and without NH4Cl. In spite of the expression of dinitrogenase reductase under free-living conditions, acetylene reduction was not observed under N-free conditions even after prolonged incubation.Key words: symbiotic nitrogen fixation, Mimosa pudica, rhizobia, phylogeny, 16S rDNA, nifH, Ralstonia taiwanensis.