• Microbiology and Biotechnology Letters
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• v.32 no.2
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• pp.190-194
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• 2004

The gene product of dctD (DctD) activates transcription from the dctA promoter regulatory region by the $\sigma^{54}$ -holoenzyme form ofRNA polymerase ($E\sigma^{54}$ ) in Rhizobium meliloti and R. leguminosarum. The Escherichia coli integration host factor (IHF) stimulated DctD-mediated activation from the dctA promoter regulatory region of R. leguminosarum but not R. meliloti. In the absence of UAS, IHF inhibited DctD-mediated activation from both of these promoter regulatory regions. IHF also inhibited activation from R. leguminosarum dctA by nitrogen regulatory protein C (NtrC), another activator of $E\sigma^{54}$ but not by one which lacks a specific binding site in this promoter regulatory region. IHF, however, stimulated NtrC-mediated activation from the R. meliloti dctA promoter. Upon removal of the UAS, IHF inhibited NtrC-mediated transcription activation from the R. meliloti dctA promoter regulatory region. These data suggest that IHF likely faciliates productive contacts between the activators NtrC or DctD and $E\sigma^{54}$ to stimulate activation from dctA promoter.

• Applied Biological Chemistry
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• v.38 no.4
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• pp.313-319
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• 1995

The involvement of the cell-wall degrading enzymes in Rhizobium has long been an unsolved question about the infection process in the formation of root nodule. To assess the contribution of the cellulase to the nodulation of rhzobia, here we report the production of cellulase from R. meliloti TAL1372 which degrade carboxymethylcellulose (CMC) model substrate with CMC-plate method. We constructed a genomic library by cloning Sau3A-digested genomic DNA from R. meliloti TAL1372 into the BamHI site of the cosmid vector pLAFR3. Out of more than one thousand transductants of E. coli, one clone (pRC8-71) had CM-cellulase activity and contained pLAFR3 cosmid with 30 kb insert of R. meliloti DNA The product of CM-cellulase gene was analyzed by native PAGE. About 45 kD protein was considered to be a product of the gene. Tn5 mutagenesis reveals that the structural gene located in a ca. 3 kb KpnI fragment. The cellulase-minus mutants of R. meliloti TAL1372 were obtained by Tn5 mutagenesis of pRC8-71 and marker exchange techniques. Analyses of the nodulation ability of these Tn5 mutants showed that the CM-cellulase gene of R. meliloti TAL1372 may be involved in early nodulation development on alfalfa (Medicago satiua).

• Korean Journal of Microbiology
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• v.26 no.4
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• pp.312-317
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• 1988

The genes for ribosomal RNA in Rhizobium meliloti and Bradyrhizobium japonicum were analyzed by southern hybridization of BamHI, EcoRI, HindIII digested chromosomal DNA with purified 5' $^{32}P$-labeled 16S and 23S rRNA. The big differences in the hybridization pattern of both rhizobia were found. The comparative results were discussed in relation to the copy number and conservativity of restriction sites in the rRNA genes of both rhizobia.

• Korean Journal of Soil Science and Fertilizer
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• v.20 no.4
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• pp.369-374
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• 1987

This study was conducted to evaluate the degree of acid tolerance of indigenous Rhizobium in natural soils, and to select the superior Rhizobia for legume forage on hilly land. 153 Rhizobial strains isolated from two host genera, Medicago and Trifolium in the pastures of Yeongnam area were screened on acidic agar plates. The results obtained are summarized as follows: 1. The degrees of acid tolerance of R. meliloti and R. trifolii were ranged from 4.3 to pH 5.5 and from 4.0 to pH 5.5 respectively. 2. Among acid tolerant Rhizobia isolates, two strains of R. meliloti at pH 4.3 and three strains of R. trifolii at pH 4.0 were shown to have the activities of nodulation and nitrogenase. 3. R. trifolii strains were shown to survive about 80 to 99% at the pH range 4.8 to 5.0, while R. meliloti were about 35 to 47% at that pH range. Therefore, more acid tolerant R. meliloti should be developed for legume forage in Korean hilly land.

• Microbiology and Biotechnology Letters
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• v.21 no.2
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• pp.95-100
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• 1993

Effect of dual inoculation with vesicular-arbuscular(VA) mycorrhizal fungi, Glomus mosseae, and Rhizobium meliloti R455 on growth, nodulation, and nitrogenase activity of alfalfa (Medicago sativa L. cv. Vernal) were examined in pot experiment. After 63 days growth, shoot length, number of leaf, and leaf size of alfalfa were increased as as result of dual inoculation with Glomus mosseae and Rhizobium meliloti. Total dry weight of alfalfa plant was increased 1.4 times compared to single inoculation with Rhizobium meliloti. Nodule number and mean fresh weight of nodule per plant were also increased due to the mycorrhizal infection.

• Applied Biological Chemistry
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• v.40 no.6
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• pp.551-555
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• 1997

This experiment was carried out to elucidate the biological activity and absorption characteristics of flavonoids in Rhizobium and Bradyrhizobium and to obtain basic information on host specific nodulation by flavonoids in rhizobium-legume symbiosis. The purpose of the present study was to explore the biological activity and the flavonoid absorption indicates that host-specificity is induced by flavonoids in symbiotic nitrogen fixation. Biological activity increased by daidzein and genistein treatment on B. japonicum KCTC 1539 whereas decreased by luteolin treatment but increased by luteolin treatment on R. meliloti whereas decreased by daidzein and genistein treatment. Daidzein and genistein are absorbed by B. japonicum, KCTC 1539 at higher rate than other flavonoids. Especially, luteolin was absorbed at a least rate. Luteolin are absorbed by R. meliloti KCTC 2353 at higher rate than other flavonoids. Especially, daidzein and genistein was absorbed at a least rate.

• Korean Journal of Soil Science and Fertilizer
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• v.25 no.3
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• pp.275-280
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• 1992

Inoculation responses of alfalfa[Medicago sativa(L.) Vernal] to Rhizobium meliloti with nitrogen fertilization were characterized by soil populations and plant yields from 1989 to 1991 at cultivated upland soil, which contained $3.1{\times}10$cells of R. meliloti/g.soil. The results obtained accordingly to fertilizing with 8Kg N/10a and differently inoculating the host such as initial and annual inoculation methods were as follows : 1. Soil populations of R. meliloti were increased more at no nitrogen fertilized condition compared to nitrogen fertilized condition to 2nd year experiment from 1st year that came up $1.6{\times}10^3$cells/g.soil, but at 3rd year the trend was reversed. Between inoculation methods, on the whole, the annual inoculation caused more populations, which were $2.0{\times}10^4$cells/g.soil in maximum number. And the populations declined after winter but recovered passing through summer season. 2. Alfalfa yields were mainly influenced by rhizobial populations rather than by fertilizing nitrogen showing a significant correlation(Y=0.36+0.287X, $r^2=0.58^{**}$) with the former. The increased extents of yields obtained by inoculation at no nitrogen and nitrogen fertilized conditions, respectively, were 66 and 10% in 1st : 13 and 20% in 2nd : 19 and 13% in 3rd year experiment with the initial inoculation, and were 66 and 10% in 1st ; 30 and 20% in 2nd : 35 and 36% in 3rd year experiment with the annual inoculation. 3. The results demonstrated the importance of inoculating, if possible, annual inoculating alfalfa to get much yields even at cultivated upland soil.

• Korean Journal of Soil Science and Fertilizer
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• v.22 no.1
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• pp.72-77
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• 1989

A study was conducted to obtain acid tolerant and superior Rhizobium meliloti strain for alfalfa on hilly acid soils. With host plant, eight isolates of R, meliloti selected in the vicinity of Milyang were evaluated for their ability to establish symbiotic effectiveness in acidified tube culture medium and vermiculite pot with different urea levels. Among isolates "YA03" was characterized for the ability to manifest to acid tolerance in three different soils of which pH were 5.0, 6.0, and 7.5. The results obtained were summarized as follows: 1. Of eight isolates "YA03" and "YA09" performed nodule in the tube medium of pH5.0, and in the symbiotic effectiveness YA03 was superior to others. 2. Alfalfa growth and $N_2-fixing$ activity by the inoculation of "YA03" isolate were better than others at the level of urea 1.25mM as well as nonurea. 3. Application of urea with inoculation of "YA03" islate to alfalfa was one of the effective factors for symbiotic effectiveness. 4. In infertile soil of pH5.0 inoculation of R. meliloti "YA03" to alfalfa caused the increase of shoot dry matter of 320% as compared to the control, and the total amount of yield was 131% as much yield in moderate fertile soil of pH7.5. Finally R. meliloti "YA03" isolate was selected as an acid tolerant strain.

• Journal of Microbiology and Biotechnology
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• v.16 no.5
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• pp.791-794
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• 2006

Cyclosophoraose (cyclic $\beta-(1,2)-glucan$, Cys) isolated from Rhizobium meliloti, a soil microorganism, was used as a solubility enhancer for flavonoids. The complexes of the cyclic oligosaccharide with flavonoids were confirmed through $^1H$ nuclear magnetic resonance (NMR) spectroscopic analysis. Flavonoids solubilized by Cys were quantitatively analyzed through high-performance liquid chromatography (HPLC). Among the flavonoids tested, the solubility of naringenin was greatly enhanced by Cys, compared with other compounds. The solubility of naringenin was enhanced about 7.1-fold by adding 10 mM Cys, compared with a control. $^1H$ NMR spectroscopic analysis indicated that the H-6 and H-8 protons, which are located on the A ring of naringenin, were greatly shifted upfield upon the complexation with Cys. This result suggested that Cys showed a regioselective interaction with the naringenin molecule upon the complexation, resulting in the solubility enhancement of naringenin.

• Applied Biological Chemistry
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• v.28 no.3
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• pp.149-155
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• 1985

590 strains of Rhizobia were isolated from root nodules of the legumes collected at 223 sites in Korea. According to their host specificities they were classified into R. japonicum(218 strains), R. phaseoli(101 strains), R. trifolii(97 strains), R. meliloti(4 strains), R. leguminosarium(1 strain), Rhizobium species(101 strains), and unidentified species(159 strains). 3 potent strains R-138, R-168, and R-214 of R. japonicum have been selected based on the infectivity to soybean cultivar and effeciency of nitrogen fixation. It was observed that the fast-growing strains of R. japonicum contained 1 to 4 plasmids of M.W. of 35-300 Md. However, plasmids were hardly detected for the slow-growing strains.