• Korean Journal of Soil Science and Fertilizer
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• v.32 no.1
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• pp.62-67
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• 1999

As a basic experiment to enhance the symbiotic utility of atmospheric nitrogen on soybean, host affinities and serological distribution of Bradyrhizobium japonicum indigenous to five Korean upland soils were measured. Based on nodulation, the symbiotic affinities between indigenous B, japonicum and eight soybean cultivars were remarkably different among soil inocula. On the whole, the averaged affinities of B. japonicum populations to soybeans were favorable in order of Goseong > Milyang > Suweon soils, but those in Iksan and Namjeju soils were not, especially for Danweonkong, Jangkyeongkong, and Eunhakong soybean cultivars. Regression analyses between nodules mass and shoot dry weight of soybean yielded model with $R^2=0.51$ for Goseong, $R^2=0.45$ for Milyang, $R^2=0.38$ for Iksan, $R^2=0.28$ for Namjeju, and $R^2=0.24$ for Suweon soils. B. japonicum from sampled soils were serologically fell into more than seven serogroups such as YCK 117(34.1%), YCK 141(6.5%), YCK 321(6.5%), YCK 445(4.7%), YCK 338(2.9%), YCK 150(1.2%), YCK 258(0.6%). The dominant serogroup YCK 117 was distributed 51.9 for Namjeju, 45.8 for Goseong, 41.7 for Iksan. 34.2 for Sueveon, and 11.1% for Milyang soils.

• Korean Journal of Microbiology
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• v.37 no.2
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• pp.114-119
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• 2001

Many isolates from different forest soil layers did not show appreciable growth on full strength of the conventional nutrient broth (NB medium) but grow on its 100-fold dilution (DNB medium). These isolates were divided into four types according to organic nutrient concentration in the growth medium from $1^{-1}\;to\;10^{-4}$dilution of normal NB medium. Oligotrophic bacteria were type II and type IV which grew in $10^{-4}$ dilution of NB (1 mg C/l) medium. Sixty strains were isolated for obligate oligotrophic bacteria. Chemotaxonomic and phylogenetic characteristics of eleven isolates of acetylene-reducing (nitrogen-fixing) oligotrophic bacteria from forest soil were investigated. They showed similar characteristics: the cellular fatty acid mainly consisted of straight-chain unsaturated $C_{18:1}$ (60-84% of total fatty acids). Ubiquinone Q-10 and a high guanine plus-cytosine content(61-64 mol%) were found. Eleven isolates of nitrogen-fixing oligotrophic bacteria were found to be closely related by full 16S rDNA sequence simility and many common taxonomic traits. Analysis of full 16S rDNA sequences of eleven isolates indicated that they were more closely related to Bradyrhizobium (similarity values: 98.1-98.8%), Agromonas, Nitrobacter, and Afipia.

• Korean Journal of Soil Science and Fertilizer
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• v.23 no.1
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• pp.60-66
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• 1990

Soybean [Glycin max (L.)] cv. Jangbaekkong was inoculated with 5 cultivated- and 5 uncultivated upland soils, in Yeongnam area, as soil inoculum and NifTAL peat inoculum as standard for soil inoculum potentials by Bradyrhizobium japonicum. 120 Bradyrhizobium japonicum isolates out of the soil populations were scored of three colony morphologies, designed "Dry", "Wet", and "Dry/Wet", and symbiotic effectiveness between "Dry" and "Wet" was compared. The results obtained were summarized as follows: 1. Indigenous populations of B. japonicum were above $10^4cells/g$. soil at the cultivated upland soils but were a few at the uncultivated upland soils except a colluvivum, orchard previously, in Yeongnam area. 2. Inoculum potentials of the cultivated upland soils were higher than the NifTAL inoculum and generally, nodule mass compensated nodule number for symbiotic effectiveness of soil populations. 3. Colony morphologies of soil populations showed the different proportions of "Dry" and "Wet" so that "Dry" types were dominant at the cultivated upland soils while "Wet" types at the uncultivated upland soils. 4. "Dry" colony morphology significantly exhibited higher symbiotic effectiveness than "Wet" types in nodule fresh weight, shoot dry weight, and shoot dry weight/nodule fresh weight. Therefore, as long as soil inoculum potentials, the growth of soybean at the cultivated upland soils could presumedly be affected by soil populations of Bradyrhizobium japonicum of "Dry" colony morphology.

• Korean Journal of Soil Science and Fertilizer
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• v.25 no.4
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• pp.387-393
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• 1992

The fate of inoculum strain of Bradyrhizobium japonicum was studied by using genetically marked strain. RJB6 $str^rnal^rneo^r$. A spontaneous mutant of B. japonicum isolated from nodules was made to have antibiotic resistance against streptomycin and nalidixic acid. In order to make genetically marked strain, neomycine resistant gene(Tn5) was introduced into this spontaneous mutant by conjugation with E. coli containing pSUP2021. The southern hybridization was carried out to confirm the plasmid insertion. Hybridization of chromosome DNA using pSUP2021(Tn5) as a probe showed that Tn5 was located on the 4.9kb fragment of chromosome. Soybean seeds were planted into a soil previously cultivated with soybean and inoculated with different cell densities of marked strain. Fourty days after planting, the inoculation effects on nodule number, nodule fresh weight, plant height and nitrogen content in the plot inoculated with heavy cell suspension was a little better than those in the plot with low inoculation. The recovery percentage of the marked strains was about 12% in the plot inoculated with heavy density cell suspension, while 5% in the plot inoculated with low cell suspension.

• Applied Biological Chemistry
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• v.30 no.2
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• pp.153-162
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• 1987

Soybean rhizobia were isolated from 101 soybean (Glycine max.) cultivar which had been grown for the breeding experiment in Korea. Seven strains of the fast-growing soybean rhizobia and nine strains of the slow-growing soybean rhizobia were selected on the basis of their growth rate in AMA medium and their high ability of nodulation. The slow-growing soybean rhizobia were identified as Bradyrhizobium japonicum in the acetylene-reducing activity, microbial characteristics, and biochemical characteristics whereas the fast-growing soybean rhizobia were very similar to Rhizobium fredii.

• Korean Journal of Soil Science and Fertilizer
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• v.29 no.2
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• pp.190-198
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• 1996

The Neomycin resistant gene in transposon 5 (Tn5) was introduced into a mutant of Bradyrhizobium japonicum RJB 6 $str^rnal^r$ by conjugation. This marked strain was used in coinoculation of soybean with Pseudomonas fluorescens(Ps-3) and Bacillus subtilis(BCAC-4) and in antibiotic treatment of soybean for studying rate of nodule formation and growth promoting effect on soybean plant. When the marked strain RJB 6 $str^rnal^rneo^r$ with Ps-3, BCAC-4, and Ps-3 plus BCAC-4 was coinoculated into two soils, the recovery rate of the marked strain was 8.5, 4.3 and 2.9 percent in soybean cultivated soil, and 10.3. 6.0 and 5.0 percent in soybean uncultivated soil. The best growth promoting effect of coinoculation on soybean plant was found with the marked strain plus Ps-3. When the marked strain was inoculated into soybean with antibiotic treatment, the rate of nodule formation in soybean cultivated soil was between 12.5 and 25.4 percent, while that in soybean uncultivated soil was between 23.7 and 43.2 percent. The highest rate of nodule formation with the marked strain was found in streptomycin 1000 ppm treatment plot.

• Korean Journal of Soil Science and Fertilizer
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• v.20 no.3
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• pp.275-283
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• 1987

In order to find out the effectiveness of nitrogen fixation in rhizobia-legume symbiotic relationship, ecological and physiological characteristics of indigenous rhizobia distributed in Korean soils, that is, dissimilatory nitrate reduction patterns of indigenous soybean rhizobia isolated from four different soils, and differences in one-and two-dimensional polyacrylamide gel electrophoretic pattern of proteins among the each subgroups of Bradyrhizobium japonicum and Rhizobium fredii, were investigated. The results were summarized as follows: 1. The indigenous soybean rhizobia isolated from four different soils could be classified into 4 groups depending on growth rate and dissimilatory nitrate reduction pattern, that is, $S_1$ (slow-grower; Bradyrhizobium japonicum and nitrate denitrifier), $S_2$ (slow-grower; Bradyrhizobium japonicum and nitrate respirer), $F_1$ (fast-grower; Rhizobium fredii and denitrifier), and $F_2$ (fast-grower; Rhizobium fredii and nitrate respirer). 2. The population ratio of fast- to slow-growing R. japonicum was 39% to 61%, and the ratio of denitrifier to nitrate respirer was 31% to 69% and 89% to 11% in fast and slow-grower, respectively. Some differences were observed between fast- and slow-growing R. japonicum but no significant difference was observed between denitrifier and nitrate respirer within same growth type by one and two dimensional SDS-polyacrylamide gel electrophoretic patterns.

• Current Research on Agriculture and Life Sciences
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• v.11
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• pp.121-132
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• 1993

This study was carried out to research the effect of the chemotaxis of Bradyrhizobium japonicum KCTC 2422 and its mutant toward soybean root exudate and to elucidate the effect of the lectin of host specificity (Host Recognition) in soybean-Bradyrhizobium symbiosis. The results obtained were as follows: The homogeneities of the purified lectins from soybean and pea seed was ascertained chromatographically and electrophoretically. Gel electrophoresis of soybean lectin in the presence of sodium dodecyl sulfate appeared a single protein band, whereas pea lectin appeared two protein bands. Soybean lectin from 2 cultivars formed immunoprecipitin arcs at same position with anti-soybean lectin rabbit IgG, but pea lectin did not form immunoprecipitin lines with anti-soybean lectin rabbit IgG. Chemotactic responses of KCTC 2422, LPN-100 and LCR-101 toward proline in capillary assays were 3.1, 1.3 and 1.0-fold above background, respectively. The chemotactic responses of KCTC 2422, LPN-100, and LCR-101 toward Paldal crude root exudate in capillary assays were 3.5, 1.4 and 1.4-fold above background, respectively. The present work shows that B. japonicum and its mutants are capable of very different responses toward root exudate fraction. The chemotactic responses of KCTC 2422 was most with neutral fraction, least with anionic fraction and intermediate with cationic fraction. The nitrogenase activity of soybean nodule was shown in 15days after inoculation with LCR-101. However, we couldn't find out the nodules when soybean was inoculated with LPN-100. From these result we can suppose that the chemotaxis of Bradyrhizobium plays inportant the role of forming the nodule (host recognition) in the soybean-B. japonicum symbiosis.

• Korean Journal of Soil Science and Fertilizer
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• v.29 no.2
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• pp.165-172
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• 1996

Symbiotic effectiveness and significance of Bradyrhizobium japonicum strains in five serogroups which were consisted of three corresponding serotype strains, respectively, indigenous to Korean soils were evaluated in terms of utility of strain serogroup for symbiotic improvement on soybean plant. The nodulation by rhizobia of each serogroup on four soybean cultivars(Danweonkong, Kwangkyo, Pangsakong, and Eunhakong) was good in order of USDA 123 > YCK 150 > YCK 117 > YCK 141 > USDA 110 serogroup members. Shoot dry weight of soybean was relatively high with USDA 110 serogroup members as well as with YCK 141 serogroup members, whereas the effectiveness of USDA 123 serogroup members was the lowest among the serogroups examined. In particular, Pangsakong among soybeans inoculated with five-serogroup members was positively outstanding on nodulation and shoot dry weight of the plant. Overall, symbiotic parameters of serogroup members associated with soybean plant such as nodule number, nodule mass, $N_2$ase activity, and shoot dry weight showed significantly different responses at level of 1% probability among both rhizobial serogroups and soybean cultivars, respectively. The rate in symbiotic similarity of the members of each serogroup from F-test ($$P{\leq_-}0.05$$) was 100% for nodule No., 90% for $N_2$ase activity. and 80% for soybean shoot dry weight. Taken together, the results indicated that the serological grouping of B. japonicum could be strongly useful for improving the symbiotic effectiveness hetween soybean and Rhizobium.

• Current Research on Agriculture and Life Sciences
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• v.15
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• pp.25-32
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• 1997

This study was carried out to research the biological characteristics among rhizobia and soybean seed and root lectins, and to obtain a basic imformation of host specificity in biological nitrogen symbiosis system. The results obtained were as follows: Purified seed lectin from soybean varieties of paldal, backwoon and hwangkeum formed immunoprecipitin lines with standard soybean seed lectin and the root lectins from soybean seedlings immunoelectrophoretically. Soybean seed and root lectins interacted with Rhizobium japonicum and Bradyrhizobium japonicum, but didn't interacted with Rhizobium. viceae, whereas pea lectin conjugated with R. viceae, but didn't bind with R. japonicum and B. japonicum. Lipopolysaccharides of B. japonicum and R. viceae were fractionated into LPS I and LPS II on the sephadox G-50. Lipopolysaccharides from B. japonicum showed rhe binding acitivity both with soybean seed lectin and root lectin, but those from R. viceae didn't show it with soybean seed and root lectins.