Thirteen Gram-negative, aerobic, motile with polar flagella, rod-shaped bacteria were isolated from root nodules of Centrolobium paraense Tul. grown in soils from the Amazon region of Brazil. Growth of strains was observed at temperature range 20-36 °C (optimal 28 °C), pH ranges 5-11 (optimal 6.0-7.0), and 0.1-0.5%NaCl (optimal 0.1-0.3%). Analysis of 16S rRNA gene placed the strains into two groups within Bradyrhizobium. Closest neighbouring species (98.8%) for group I was B. neotropicale while for group II were 12 species with more than 99% of similarity. Multi-locus sequence analysis (MLSA) with dnaK, glnII, recA, and rpoB confirmed B. neotropicale BR 10247 as the closest type strain for the group I and B. elkanii USDA 76 and B. pachyrhizi PAC 48 for group II. Average Nucleotide Identity (ANI) differentiated group I from the B. neotropicale BR 10247 (79.6%) and group II from B. elkanii USDA 76 and B. pachyrhizi PAC 48 (88.1% and 87.9%, respectively). Fatty acid profiles [majority C and Summed feature 8 (18:1ω6c/18:1ω7c) for both groups], DNA G + C content, and carbon compound utilization supported the placement of the novel strains in the genus Bradyrhizobium. Gene nodC and nifH of the new strains have in general low similarity with other Bradyrhizobium species. Both groups nodulated plants from the tribes Crotalarieae, Dalbergiae, Genisteae, and Phaseoleae. Based on the presented data, two novel species which the names Bradyrhizobium centrolobii and Bradyrhizobium macuxiense are proposed, with BR 10245 (=HAMBI 3597) and BR 10303 (=HAMBI 3602) as the respective-type strains.
This study describes two Bradyrhizobium strains, UFLA03-164 T and UFLA03-153, which share more than 99% sequence similarity of the 16S rRNA with the type strains of 15 species in this genus. The concatenation of three housekeeping genes (recA, gyrB, and dnaK) indicated that both strains formed a single clade separate from known Bradyrhizobium species. B. viridifuturi, represented by SEMIA 690 T , is the closest neighboring species (96.2%). Low (< 92%) average nucleotide identity (ANI) was observed between strain UFLA03-164 T and any of the closest species on the phylogenetic trees based on concatenated housekeeping genes. The DNA G+C content of UFLA03-164 T is 63.25%. Phenotypic characteristics were determined for both UFLA strains. Based on the data, the two strains represent a new species for which the name Bradyrhizobium uaiense is proposed, with UFLA03-164 T (= LMG 31509 T ) as type strain.
Identification and selection of nitrogen-fixing bacterial strains for inoculation into native leguminous tree species can assist in the recovery of degraded areas. Additionally, native strains from these areas are genetic resources adapted to these conditions and are thus suitable for selection. The aim of this study was to symbiotically and genetically characterize 18 bacterial strains from the Rhizobium and Bradyrhizobium genera isolated from Machaerium nyctitans, Platypodium elegans, and Ormosia arborea grown in a nursery in an iron mining area. Three experiments were conducted under axenic conditions in a greenhouse. The nodulation capacity of the strains was evaluated by the number (NN) and dry matter (NDM) of nodules. Symbiotic efficiency was evaluated based on the following parameters: SPAD index (SPAD), shoot dry matter (SDM), root dry matter (RDM), and total dry matter (TDM) of the plants, relative efficiency (RE), shoot nitrogen content (SNC), and total nitrogen content in the plant (TNC). The atpD and gyrB housekeeping genes and the nifH gene were sequenced for phylogenetic analysis, and the nodC and nodD symbiotic genes of the strains were amplified. Out of the 18 strains, 16 were authenticated by nodulation capacity in the species of origin. The SPAD variable allowed for the detection of differences between treatments before the SDM. Additionally, the SPAD index showed correlation with TNC, and the strain Bradyrhizobium sp., UFLA01-839, which may represent a new species, was outstanding in Machaerium nyctitans. The nifH, nodD, and nodC genes were detected in UFLA01-839.
In some leguminous plants, associations with nitrogen-fixing microorganisms allow their nutrition with nitrogen (N) from the atmosphere. This process is known as Biological Nitrogen Fixation (BNF), where through nitrogenase enzymes, N2 is converted to an available form. This process can replace in part, or in total, nitrogen fertilizers. Cowpea bean is a legume species that is recognized for its high capacity to carry out BNF. In the last decades, studies have encouraged small farmers from north and northeast Brazil to use inoculants with rhizobia species since the results of researches have demonstrated that inoculation is an interesting strategy to improve cowpea production. Considering the specific function of molybdenum (Mo) in the N assimilation, different doses of Mo were tested in this study in order to find doses that could improve and enhance BNF. Therefore, this study aimed to compare nitrogen fertilization and BNF in the N assimilation by plants with different Mo doses. Inoculation was performed with the strains UFLA 03-84 and INPA 03-11B. Doses of Mo were applied in seeds and each pot contained five seeds. Thirty-five days after germination, the plants were analyzed for shoot dry matter and fresh matter, N contents and accumulation, as well as the Soil-Plant Analysis Development (SPAD) Index and nodulation in inoculated plants. The different doses of Mo and also the nodulation treatments did not show significant differences in the contents of N. Plants with N fertilization had significant higher shoot dry matter and root dry matter production, in addition to higher N foliar contents and N accumulation. Therefore, BNF was not as efficient as nitrogen fertilization in the evaluated experimental conditions using cowpea beans.
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