Achara Nuntagij

Characterization of root-nodule bacteria isolated from the medicinal legume Indigofera tinctoria

Fourteen root-nodule bacteria isolated from the medicinal legume Indigofera tinctoria were characterized for their phenotypic features including growth curves, utilization of carbon and nitrogen sources, antibiotic resistance, vitamin requirement and growth under different conditions. The partial sequences of the 16S rDNA of these strains were obtained and BLASTN analysis revealed that the microsymbionts of I. tinctoria were related to members of five distinct genera: Rhizobium, Sinorhizobium, Bradyrhizobium, Cupriavidus and Pseudoalteromonas. The partial nifH gene of Pseudoalteromonas-like strain DASA 57075 had 96% similarity with nifH genes of members of Bradyrhizobium. The partial nodC gene of Pseudoalteromonas-like strain DASA 57075 showed 88% similarity with the nodC gene of several rhizobia including Sinorhizobium, Bradyrhizobium and Mesorhizobium. We propose a bacterium that is related to Pseudoalteromonas from the gamma-class of Proteobacteria as a new legume symbiont. This is also the first report that the same species of legume can be nodulated by bacteria from up to five different genera in three distinct classes.

Heavy metal tolerant Metalliresistens boonkerdii gen. nov., sp. nov., a new genus in the family Bradyrhizobiaceae isolated from soil in Thailand

Bacterial strains from inoculated soybean field soil in Thailand were directly isolated using Bradyrhizobium japonicum selective medium (BJSM), on the basis of Zn(2+) and Co(2+) resistance of B. japonicum and B. elkanii. The isolates were classified into symbiotic and non-symbiotic groups by inoculation assays and Southern hybridization of nod and nif genes. In this study, a nearly full-length 16S rRNA gene sequence showed that the non-symbiotic isolates were more closely related to members of Rhodopseudomonas and to a number of uncultured bacterial clones than to members of Bradyrhizobium. Therefore, a polyphasic study was performed to determine the taxonomic positions of four representatives of the non-symbiotic isolates. Multilocus phylogenetic analysis of individual genes and a combination of the 16S rRNA and three housekeeping genes (atpD, recA and glnII) supported the placement of the non-symbiotic isolates in a different genus. The ability of heavy metal resistance in conjunction with phenotypic analyses, including cellular fatty acid content and biochemical characteristics, showed that the non-symbiotic isolates were differentiated from the other related genera in the family Bradyrhizobiaceae. Therefore, the non-symbiotic isolates represented a novel genus and species, for which the name Metalliresistens boonkerdii gen. nov., sp. nov. is proposed. The type strain is NS23 (= NBRC 106595(T)=BCC 40155(T)).