• Journal of Microbiology
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• v.44 no.3
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• pp.363-368
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• 2006

In this study, native Byadyrhizobium strains were isolated from the host plant, Glycine max, harvested from fields in Madhya Pradesh, India, and were typed by Iytic rhizobiophages. Eight indigenous (Soy2, ASR011, ASR031, ASR032, MSR091, ISR050, ISR076 and ISR078) and two exotic strains (USDA123 and CB1809), all of which evidenced a distinct reaction with six phages, were employed in this study. The symbiotic interaction of these strains was studied initially using soybean cultivar JS335 in a sand culture in a controlled environment, and the efficiency was assessed based on the nodule number, nodule dry weight, plant dry weight, nitrogenase activity, and total accumulation of N per plant. Symbiotic effectiveness was found to be highest with the native phage-sensitive isolate ASR011, whereas it was at a minimum with the phage-resistant isolates, ISR050 and ISR078. Additionally, the effectiveness of these strains was evaluated using six soybean cultivars belonging to different maturity groups; namely, Brags, Lee, Pusa20, PK416, JS33S and NRC37. Analysis of variance data evidenced significant differences due to both symbionts, for the majority of the tested parameters. The CB1809, USDA123, and ASR011 strains evidenced relatively superior symbiotic effectiveness with soybean cultivars Brags, Lee and JS335. Strain ISR078 evidenced no significant responses with any of the cultivars. The ASR031 strain performed moderately well with all tested cultivars. The symbiotic response of all the strains was quite poor with cultivar PK416. Our studies showed that a significant relationship existed between the phage sensitivity and symbiotic efficiency of the bacterial strains with the host-cultivars.

• Korean Journal of Microbiology
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• v.26 no.4
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• pp.305-311
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• 1988

The spectinomycin resistant strain of slow growing R. japonicum R-168 was selected to be participated in conjugation with E. coli WA803/pGS9. Tn5 was introduced from suicide vector pGS9 into R. japonicum R-168 $spr^{r}$ chromosome at the frequency of $1.0\times 10^{-5}-5.0\times 10^{-7}$ and the transconjugante were selected on the yeast extract-mannitol plate containing kanamycin ($50{\mu}$g/ml) and spectinomycin ($100{\mu}$g/ml) after 8-9 days incubation. All transconjugants we tested were found to contain Tn 5 DNA on their genome, which was confirmed by Southern hybridization experiments. R. japonicum RNa75, which had been selected through plant test, was found to be defective in symbiotic nitrogen fixing ability and the production of leghemoglobin in soybean nodules formed by the inoculation of this mutant. In addition, this mutant strain hardly developed nitrogenase activity asymbiotically in contrast with the wild type strain, indicating that some nitrogen fixing gene might be blocked in this strain and the production of leghemoglobin could be decreased by the interference in nitrogen fixing genes.

• Korean Journal of Microbiology
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• v.32 no.2
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• pp.97-101
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• 1994

Fifteen strains of the nitrogen fixer Axospirillum were isolated from the rhizosphere of rice collected from Kyonggi-do and Chungcheongnam in Korea. They had strong acetylene-reducing activity of 400 of 900 nmol $C_2H_4$ per hour vial had a similar morphology in succinate-malate medium: vibrioid cells having a diameter of 1.0 ${\mu}m$ and a monopolar single flagellum in liquid media. According to their physiological and morphological characteristics, they were divided into two distinct groups, group I and group II. Group I strain were, unlike group II, distinguished by their ability to use glucose as a sole carbon source in nitrogen-free medium, requirement for biotin, and formation of wider, longer, and S-shaped cells in semisolid nitrogen-free malate medium. On the basis of their characteristics, strains belonging to group I were identified as Azospirillum lipoferum, while those belonging to group II were identified as Azospirillum brasilense.

• Korean Journal of Soil Science and Fertilizer
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• v.17 no.4
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• pp.415-427
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• 1984

This experiment was carried out to investigate the distribution of associative nitrogen fixers, Azospirillum spp. and their nitrogenase activities measured by ARA in the rhizosphers of rice, soybean, and weed grown in the rice paddy field at ear formation stage of rice. Nitrogenase activities produced by Azospirillum spp. enriched from histophere ranged from 16 to 53 n mole/tube/hr.. High nitrogen fixing activities more than 30 n mole/tube/hr. were observed in the histophere of the Echinochloa crus-galli L., Finbristylis miliacea L., and Monochoria vaginalis var.. Nitrogen fixing activities of Azospirillum spp. obtained from single colonies which originated from the rhizoplane of rice (pot-kwang var.), Finbris tylis miliacea L., Monochoria vaginaliz var., Glycine max L. were higher over 100 n mole/tube/hr. than those histophere. Genus of Azopsirillum isoltated from roots of the Graminease (Oriza sativa L., $C_3$-plant, Echinochloa crus=galli L., $C_4$-plant, Cyperus difforuis L.. $C_4$-plant), and Aeschynomene indica L. (Leguminosae, $C_3$-plant) was identified as A. brasilense. However, both strains, A. lipoferum and A. brasilense ($nir^-$ or $nir^+$ strain) were isolated from other plant roots, Both $nir^-$ and $nir^+$ strains of A. brasilense were associated with the same host plant.

• Korean Journal of Soil Science and Fertilizer
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• v.20 no.1
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• pp.43-53
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• 1987

In order to improve effectiveness of rhizobia- legume symbiotic nitrogen fixation, ecological and physiological characteristics of indigenous rhizobia distributed in Korea, that is, symbiotic effectiveness of indigenous soybean rhizobia, nitrate reductase activities of the soybean bacteroid from five different soils, and differences of host-infection abilities among the soybean cultivars under population densities of the same indigenous soybean rhizobia, were investigated. The results were summarized as follows: 1. The number of indigenous soybean rhizobia was ranged from $9.2{\times}10^2$ cells per gram of soil in calcareous soil II to $42.4{\times}10^3$ cells per gram of soil in calcareous soil I in Danyang. 2. The symbiotic effectiveness of indigenous soybean rhizobia from five different soils was high in the case of soybean continuously cultivated, and calcareous soil I that population densities of indigenous soybean rhizobia were observed highly. 3. Inverse relationship was observed between total nitrogenase activity (TNA) and nitrate reductase activity (NRA) from the soybean bacteroids ($r=-0.502^*$), but the correlation between nitrate reductase and specific nitrogenase activities (SNA) could be devided into two groups. It was classified into group I which is high in SNA and low in NRA, and group II which is low in SNA and high in NRA. 4. The infection ability of the indigenous soybean rhizobia in the same soil conditions showed the reciprocal difference among each soybean cultivars. In Kwangkyo and Jangyeup, the symbiotic effectiveness appeared by infection of indigenous soybean rhizobia was higher than it of the other soybean cultivars.

• Korean Journal of Soil Science and Fertilizer
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• v.21 no.3
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• pp.307-315
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• 1988

This experiment was conducted to find out the effects of fertilizer-N and co-inoculation of the Rhizobium japonicum and the Azospirillum lipoferum on nodulation, $N_2$-fixation, and growth of soybean under in situ conditions. The results obtained were summarized as follows: 1. The yield of soybean dry matter was significantly greater in the R. japonicum alone, and the mixed inoculation of R. japonicum and A. lipoferum than those of un-inoculation. But inoculum applied by different strains did not significant effect on plant growth. The effects of nitrogen applied on soybean dry matter were higher in the ammonium sulfate than potassium nitrate, and decreased with increasing rates of two forms of nitrogen applied regardless of nitrogen source. 2. Acetylene redution activity was more increased in a single inoculation of R. japonicum than those of the mixed inoculation of the R. japonicum and the A. lipoferum, in cases of Danyeup cultivar, regardless of the form of combined nitrogen used. 3. Nodule mass and total nitrogenase activity per plant showed the positively significant effect in the interrelationship between dry matter of soybean and some factors related to nitrogen fixation efficiency. 4. The highest symbiotic effect in Danyeup cultivar was obtained when a single R. japonicum 84 Dy-1 strain was inoculated and fertilized with 18 mM potassium nitrate.

• Current Research on Agriculture and Life Sciences
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• v.4
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• pp.27-35
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• 1986

This experiment was carried out to elucidate the factor of nitrogenase formation and to establish the nitrogen fixation system in mixed culture of cultured cells and rhizobia through tissue culture technique using three soybean varieties, Hwangkeum, Namcheon and D 68-0099 as host plants. The results obtained were as follows; The callus was induced in embryo and radicle, but not in hypocotyl. The most favorable callus induction was caused by the individual application of 2,4-D and NAA at the concentration of 2mg/1 and 4mg/1, respectively, but in case of treating both 2,4-D and kinetin, that was done at the concentration of 0.2mg(2,4-D)/0.05mg(kinetin)per liter. The growth of cultured cell was good at the concentration of 2.0mg(2,4-D)/1 and 0.2mg(2,4-D)/0.05mg(kinetin)per liter. When cultured cells were inoculated with R. japonicum 019 and 011, their growthes were considerably inhibited. The addition of single amino acid inhibited the growth of cultured cells. Hwangkeum was inhibited considerably by methionine and leucine. The inhibition of growth by single amino acid can be abolished by the addition of certain amino acids. The differentiation of adventitious root was good at the concentration of 2.0mg 2,4-D and 0.2mg 2,4-D/0.05mg kinetin per liter. Of three host plants tested with 25 R. japonicum strains, Hwangkeum had affinity for 10 strains, Namcheon for 7 strains and D68-0099 for none. The nitrogen fixing abilities of Hwangkeum and Namcheon caused by cultured cell-Rhizobium association were high in strain 019, 007, and in 007 mixed with 119, respectively.

• Korean Journal of Soil Science and Fertilizer
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• v.37 no.1
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• pp.46-53
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• 2004

In this study, we compared the levels of methylotrophic bacterial community diversity in the leaf, stem, grain, root and rhizosphere soil sainples of four rice cultivars collected from three regions of Korea. Thirty five pigmented and five non-pigmented isolates showing characteristic growth on methanul were obtained. When phylotypes were defined by performing numerical analysis of 42 characteristics, four distinct clusters were formed. While two clusters, I and IV diverged on the basis of nitrate and nitrite reduction, other two clusters, comprising only pink pigmented colonies, diverged on the basis of cellulase activity. Out of the two reference strains used in the analysis, Methyhbacterium extorquens AM1 diverged from all the clusters and M. fujisawaense KACC 10744 grouped under cluster III. All the isolates were positive for urease, oxidase, catalase and pectinase activity and negative for indole production, MR and VP test, $H_2S$ production, starch, and casein hydrolysis. No clusters were found to possess thermotolerant isolates, as no growth of the isolates was observed at $45^{\circ}C$. Two strains in cluster I were found to possess gelatin hydrolysis and methane utilizing properties respectively. Most of the isolates in all the four clusters utilized monosaccliarides, disaccharide and polyols as carbon source. Six isolates showed considerable nitrogenase activity ranging from 86.2 to $809.9nmol\;C_2H_4\;h^{-1}\;mg^{-1}$ protein.

• Korean Journal of Soil Science and Fertilizer
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• v.23 no.3
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• pp.220-226
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• 1990

This experiment was carried out to study the effects of Bradyrhizobium moculation and fertilizer N application on symbiotc $N_2$ fixation of peanut (Arachis hypogaea L. cv. Younghotangkong) at newly reclaimed soil. Treatments consisting of B. sp. (Vigna) HCR-3 and HCR-46 with control and five levels of nitrogen (0, 4, 8, 16, 32kg/10a) were arranged with split design of four repetitions. The results obtained were as follows. The number of viable Bradyrhizobium in rhizosphere was decreased to 20 days after sowing and thereafter it was remarkably increased, until full ripe stage of pods, decreased as fertilizern rates increased. Nodulation and nitrogenase activity were reduced with increasing levels of fertilizer, whereas top dry matter and nitrogen accumulation where increased. Seed yield of Bradyhizobium inoculation under the fertilizer N zero level was similar to that N : 8kg/10a under uninoculation. It suggested that Bradyrhizobium inoculation could replace the effect of nitrogen application.

• 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.