• Microbiology and Biotechnology Letters
• /
• v.13 no.1
• /
• pp.79-85
• /
• 1985

Rhizobium japonicum wild type strains isolated from local soybean variety Jangback root nodules with higher nitrogenase activity than R. japonicum 3I1110 or 61A76 was mutangenized by N-methyl-N'-nitro-N-nitrosoguanidine and UV-irradiation, and screened by effectiveness assay with soybean. One mutant strain JB65 nodulated the roots earlier than the wild type and also expressed higher acetylene-reducing activity in the presence and absence of fixed nitrogen. The selected mutant was compared with SM35 strain and showed greater nodulation and symbiotic nitrogen fixing activity with local soybean variety Jangback than SM35 strain.

• Journal of The Korean Society of Grassland and Forage Science
• /
• v.11 no.3
• /
• pp.145-152
• /
• 1991

This experiment was carried out to investigate the effects of liming, manure application, and inoculation with Rhizobium on the early growth, nodulation, seedling establishment, winter survival, and forage yield and quality of alfalfa(Medicago sativa L.) at the Experimental Livestock Farm, College of Agriculture, Seoul National University, Suweon in 1985 and 1986. The results obtained are as follows: 1. Seedling vigor and nodule formation of alfalfa at early stage were the best in the treatments of liminginoculation( L1) and liming-manure-inoculation(LM1). 2. Seedling establishment was not significantly affected by the treatments of liming, manure application, nodulation and the combination of these. 3. The LI and LMI treatments showed the highest winter survival of 94.3 and 83.5%, respectively, but the lowest winter su.rvival of 55.9 and 58.5% was found in no treatment(None) and inoculation( I ), respectively. 4. No differences in crude protein(CP1, acid detergent fiber(ADF) and neutral detergent fiber(NDF) contents were found among the treatments. 5. There were significant differences in the total dry matter yield of alfalfa among the treatments. The total dry matter yields of 10,618, 11,830, 11,855 and 13,456 kg/ha from the treatments of L, LM, LI and LMI, respectively, were significantly higher than those of None, M and I treatments. 6. Based on the results of the experiment, it appears that the dry matter yield of alfalfa could be enhanced by the treatments of liming with Rhizobium inoculation and also liming, manure application and Rhizobium inoculation as a practical method.

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

• Journal of Microbiology and Biotechnology
• /
• v.12 no.3
• /
• pp.522-525
• /
• 2002

A very rapid and efficient separation technique for cellular rhizobial cyclosophoraoses was developed based on fractional precipitation and partition chromatography. Cyclosophoraoses are known to function in the osmotic regulation and root nodule formation of legumes during the nitrogen fixation process. Cyclosophoraoses are produced as unbranched cyclic (1longrightarrow12)-${\beta}$-D-glucans in Agrobacterium or Rhizobium species. Recent research has shown that cyclosophoraoses can form inclusion complexation with various unstable or insoluble guest chemicals, thereby implying great potential for industrial application. Typical separation of pure cellular cyclosophoraoses has been so far carried out by several time-consuming steps, including size exclusion, anion exchange, and desalting liquid chromatographies, with a relatively poor recovery. However, the proposed method demonstrated that the successive application of fractional ethanol precipitation and one step of silica gel-based flash column chromatography was enough to simultaneously purify neutral or anionic forms of cyclosophoraoses. This novel technique is very rapid and provides a high recovery.

• Journal of Microbiology and Biotechnology
• /
• v.20 no.2
• /
• pp.346-349
• /
• 2010

Coenzyme $Q_{10}$ ($CoQ_{10}$) production from Rhizobium radiobacter T6102 was monitored under various oxygen supply conditions by controlling the agitation speeds, aeration rates, and dissolved oxygen levels. As the results, the $CoQ_{10}$ production was enhanced by limited oxygen supply. To investigate whether the $CoQ_{10}$ production is associated with its physiological functions of electron carrier and antioxidant, the effects of sodium azide and hydrogen peroxide on the $CoQ_{10}$ production were studied, showing that the $CoQ_{10}$ contents were slightly enhanced with increasing sodium azide (up to 0.4 mM) and hydrogen peroxide (up to $10\;{\mu}M$) concentrations. These results suggest the plausible mechanism where the limited electron transfer stimulating the environments of limited oxygen supply and oxidative stress could accumulate the $CoQ_{10}$, providing the relationship between the $CoQ_{10}$ physiological functions and its regulation system.

• Journal of Microbiology and Biotechnology
• /
• v.21 no.11
• /
• pp.1101-1108
• /
• 2011

The rise in global energy demand has prompted researches on developing strategies for transforming coal into a cleaner fuel. This requires isolation of microbes with the capability to degrade complex coal into simpler substrates to support methanogenesis in the coal beds. In this study, aerobic bacteria were isolated from an Indian coal bed that can solubilize and utilize coal as the sole source of carbon. The six bacterial isolates capable of growing on coal agar medium were identified on the basis of their 16S rRNA gene sequences, which clustered into two groups; Group I isolates belonged to the genus Rhizobium, whereas Group II isolates were identified as Chelatococcus species. Out of the 4 methods of whole genome fingerprinting (ERIC-PCR, REP-PCR, BOX-PCR, and RAPD), REP-PCR showed maximum differentiation among strains within each group. Only Chelatococcus strains showed the ability to solubilize and utilize coal as the sole source of carbon. On the basis of 16S rRNA gene sequence and the ability to utilize different carbon sources, the Chelatococcus strains showed maximum similarity to C. daeguensis. This is the first report showing occurrence of Rhizobium and Chelatococcus strains in an Indian coal bed, and the ability of Chelatococcus isolates to solubilize and utilize coal as a sole source of carbon for their growth.

• Korean Journal of Soil Science and Fertilizer
• /
• v.11 no.1
• /
• pp.1-7
• /
• 1978

Laboratory and pot experiment were carried out to find the symbiotic nitrogen fixing activities and the yield of dry matter on the several selected wild soybean plant. 1. Very high acetylene reducing(nitrogen fixing)activities were observed in isolated strains of Rhinchosis volubilies Lour. (924 nmole), Desmodium oldhami Oliver (844 nmole) and Glycine soja s. et z. (271 nmole/test tube/hr) while Rhizobium japonicum was only 0.6 nmole/test tube/hr. 2. On the serological reaction, eight isolated Rhizobium strains were sufficiently different to distinguish from the Rhizobium japonicum. 3. On the pot experiment with Glycine soja s. et z., A. Number of total nodules were higher in NPK+rice straw+Ca+Mo than in NoPK. However, little differences were observed in mumber of effective nodules. B. Yield of dry matter was increased with increasing of nitrogen content in plants. C. Estimated amount of fixed nitrogen in plant by an individual nodule was higher in NoPK than in NPK+rice straw+Ca+Mo.

• Applied Biological Chemistry
• /
• v.38 no.4
• /
• pp.313-319
• /
• 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 CROP SCIENCE
• /
• v.34 no.4
• /
• pp.400-407
• /
• 1989

This experiment was carried out to study the effects of nitrogen concentration of cultural solution, Rhizobium inoculation, and planting density on the growth and yield of soybean cultivars, Hwanggeumkong, Jangbaegkong, Paldalkong, Clark, and non-nodulation isoline of Clark. Rhizobium inoculation increased the stem length, particularly in Hwanggeumkong, Jangbaegkong, and decreased it significantly in non-nodulation Clark. Stem length was increased by the increase in nitrogen fertilization by the 195ppm level and decreased by the increase in plant population density. Rhizobium inoculation also increased the shoot dry weight, but significantly decreased it in non-nodulation Clark. As nitrogen concentration in the cultural solution increased the shoot dry weight decreased in Jangbaegkong and paldalkong. However, the shoot dry weight was decreased by the increase in plant population density. Rhizobium inoculation and the increase in nitrogen concentration of cultural solution increased the ratio of shoot dry weight to root weight. The Rhizobium inoculation and the increase in nitrogen concentration of cultural solution increased the grain yield per pot in Hwanggeumkong and paldolkong, While non-nodulating Clark showed significant decrease in grain yield. Grain yield per pot was also increased by the increase of plant population density. Grain yield was significantly correlated with shoot dry weight, nodule number, and nitrogen content of the soybean plants. The correlation between nitrogen contents of the soybean plants and stem length, shoot dry weight, and nodulation was significant. The allantoin-N content in stem was also significantly correlated with nodulation.

• Proceedings of the Korean Society of Crop Science Conference
• /
• 1980.05a
• /
• pp.13-14
• /
• 1980