• 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.

• The Korean Journal of Ecology
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• v.10 no.4
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• pp.175-182
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• 1987

Soybeans(Glycine max Merr. cv. Kwanggyo), inoculated with Rhizobium japonicum 110 and then sand-cultured with nitrate gradients (0, 1, 3, 10 and 30mM KNO3). were studied on the growth analysis, nitrogen fixation and nitrogen economy during the growing period. The maxium values of total leaf area, biomass and nitrogen quantity were increased 139%, 122% and 161%, respectively with higher concentration of nitrate treatment. Nodulation showed significant linear correlation with leaf area growth for each treatment of nitrate concentration increased. The more nitrate concentration increased, the more distribution ratios of dry matter and nitrogen to nodule decreased, and the more T/R ratios, CGR and N content increased. On the other hand, F/C ratios and RGR showed little changes. The amounts of nitrogen fixation of soybean alloted to 0, 1, 3, 10 and 30mM nitrate treatments were 100, 46, 14, 0.1 and 0.004% for the total nitrogen assimilation, respectively. The nitrogen utility of soybean plant was smaller than that of other plants and ranged from 23 to 30 at varying nitrate gradients.

• Journal of Plant Biology
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• v.30 no.3
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• pp.205-213
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• 1987

Soybean, inoculated with effective Rhizobium japonicum 110, were grown by sand culture with nutrient solution containing either of 0, 1, 3, 10 or 30mM NO3-/l, and analyzed growth characteristics, NR activity, N2-fixation activity, and changes of ureide contents during the growing period. The amount of nodule formation decreased abruptly by nitrate treatment, the maximum nodule dry weight was 1.59, 1.05, 0.78, 0.09 and 0.008 g plant-1, respectively for each treatment on the 98th day. Specfic activity of N2-fixation showed the maximum rates of 140, 101, 37, 5 and 2.2 nM dw.mg-1.hr-1, respectively for each treatment in the earlier growth period. The maximum acetylene reduction activity on the 98th day after sowing was 81.5, 35.3, 14.3, 0.1 and 0.0045 $\mu$M C2H4 plant-1.hr-1, respectively for 0, 1, 3, 10 and 30 mM of NO3- gradients. Nitrate reduction activity increased along with nitrate gradients, and decreased abruptly with age. Relative abundance of ureides in plant organs was high in reproductive growth, and showed the maximum value in fully symbiotic dependent plant. Relative abundance of ureides in stem is a useful indication for the evaluation of nitrogen fixation in nodules of symbiotic plant.

• Korean Journal of Soil Science and Fertilizer
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• v.24 no.1
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• pp.61-68
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• 1991

Thirty Bradyrhizobium japonicum isolates (10 strains per each soil) from 1 uncultivated [Sangnam(Soil 1), Milyang]- and 2 cultivated [Dong(Soil 2)and Chinbuk(Soil 3), Changweon] upland soils in Yeongnam area were evaluated on their symbiotic effectiveness to soybean [Glycin max (L.)] cv. Korean Jangbaekkong and American Clark and examined on their serological diversity. The results obtained were summarized as follows : 1. On symbiotic effectiveness of B. japonicum with plant genotypes, isolates showed a relatively high value of nodule mass in Jangbaekkong cv. and of shoot dry weight and total nitrogen in Clark cv. demonstrating the order of Soil 1> Soil 2> Soil 3 isolates. 2. Among 30 B. japonicum isolates, YCK 141 showed the best effectiveness on mean nitrogen fixation of two cultivars. 3. Thirty indigenous B. japonicum showed 6 types of serological diversities in the immunoblot analysis which were present in various proportions at Soil 2(5) and Soil 3(5) except Soil 1 where all isolates fell into the YCK 117 serogroup. And their distribution order was serotype YCK 117( 12 strains) > USDA 1l0(5strains), USDA 123(5 strains) > YCK 150(4 strains) > YCK 141(3 strains) > YCK 226(1 strain). 4. Especially, 10 isolates from Soil 1, an uncultivated orchard, showed a very homologous pattern in not only effectiveness but serological distribution. It seemed to indicate that the isolates were typically affected by numerous physical and environmental factors of the soil.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.54 no.3
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• pp.299-306
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• 2009

In nitrogen-limited conditions, rhizobia lead to formation of nitrogen-fixing nodules on the roots of leguminous plants. The process of nodulation is autoregulated by pre-existing nodules in the same root system. The altered profile of sap proteins by inoculation with B. japonicum may indicate presence of a signal responsible for autoregulation transferred through stem. The 20 kDa protein enhanced by innoculation significantly decreased in intensity from 2.5 to 7 days after inoculation (DAI). However 6 kDa protein did increase during such a transition period. Western blot analysis showed that both 20 kDa and 6 kDa were cross-reacted with the SKTI antiserum. This suggests that SKTI may be involved in soybean nodulation by specific induction and degradation in stem sap during early stage of nodulation. RNAi technique and Agrobacterium rhizogenes-mediated transformation were applied to investigate the function of SKTI in nodulation. We have found that the number of rhizobium-induced nodule was much less in SKTIi-silenced hairy roots than the non-silenced. Indeed the quantitative RT-PCR showed that the expression level of SKTI gene was reduced over 40% in the transgenic hairy roots compared to the non-transgenic. It appears that the observed early induction of SKTI and degradation into small peptide in a specific time manner may be involved in autoregulation of nodulation in soybean and the specific mechanism of such regulation remains to be investigated.

• Korean Journal of Microbiology
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• v.30 no.4
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• pp.246-251
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• 1992

Molccular cloning of nod genes from Bradvrhizobium sp. SNU001, a nitrogen-fixing symbiont isolated from thc root nodules of soybean (Clycine trim) . was carried out. nod genes were found to be located on thc genome of the symbiont by gcnomic hybridization with 4.5 kb EcoRI/HndIII fragment (nod DABC) of Rhizohium meliloti as probe. Genomic library of this symbiont was constructed using h phage EMBL3-BanlHI vector. from which five nod positive clones were sclectcd by primary and secondary screening methods. The partial restriction map of inserted genomic DNA of h CNS-l(c1one 2) was constructed. and 3.9 kh Bun7HI fragment. which showed strong hybridization signal to the probe, was subcloned into pBS KS(+) plasmid vector. Partial restriction inap ot' a selected subclone (pBjCNS-I) was constructed and nod DABC was found to be located on the 1.8 kb KpnI/Sacl fragment of this subclone.

• Korean Journal of Soil Science and Fertilizer
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• v.25 no.2
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• pp.181-188
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• 1992

This study was carried out to evaluate the fate of inoculant Bradyrhizobium japonicum and the inoculation effect on soybean in complex soil environment. To moniter Rhizobium strains from the root, streptomycine and streptomycine and nalidixic acid resistant marker strains were prepared by spontaneous mutagenesis. The characteristics and properties of antibiotic marked strains were not altered by the mutagenesis. The comparison of properties of wild type and antibiotic resistant Bradyrhizobium strains are summarized as follow : 1) The strains of USDA110K-$STR^r$, USDA110N-$STR^r$ and R318-$STR^r$ showed weak tolerance to pH 9.0. The utilization of carbon sources by fast growing group was different from that of slow growing group. The marked strains of R214-$STR^rNAL^r$, USDA110K-$STR^r$ and USDA110N-$STR^r$ was doubtful in utilization of sorbitol and R138-$STR^rNAL^r$ was doubtful in utilization of xylose as a carbon source. 2) By examining the agglutination reaction of serogroups, the strains used were identified as different ones. There were no differences between wild type and marked strains in agglutination titer values. 3) The plasmid size of fast group was slightly greater than that of slow group. However, there was no differences in plasmid size between the wild type and antibiotic resistant strains. This result indicates that the antibiotic resistance was not encoded in plasmid. 4) The recovery of the inoculated strains was up to 12.5 % in soybean cultivated soil and was up to 25 % in soybean uncultivated soil. 5) When the wild type or marked strains were inoculated. there was no significant effect on soybean plant, whereas the inoculation effect was pronounced in soybean uncultivated soil. The inoculation effect seemed to be more pronounced in wild type strains than antibiotic resistant strains. however, the difference was not significant.

• Applied Biological Chemistry
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• v.17 no.1
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• pp.31-41
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• 1974

This study was conducted to determine the effects of soybean cultivation on yield and other characters in a newly reclaimed soil. Four factois - (1) plant density (2) compost application (3) innoculation of nodule bacteria (Rhizobium japonicum) combined with lime application (4) NPK application-were examined in a 24 factorial experiments arranged in randomized complete block design with 4 replicates. Kangio variety was used and the results obtained are summarized as follows; (1) Dense plant population, Application of Compost and application of fertilizer (NPK) gave the yields 1.7, 1.4 and 2.1 times the check respectively. The Combinations of, dense $population{\times}compost$, dense $compost{\times}fertilizer$ (NPK), $population{\times}compost{\times}fertilizer$ (NPK), and dense $population{\times}compost{\times}nodule$ bacteria with $lime{\times}fertilizer$ (NPK) increased the yields by 2.0, 3.0, 2.6 and 5.4 times than the check respectively. But little effect on yield was noted in the treatment inoculated with nodule bacteria with lime. (2) In the case of higher yield, the increased weight of 100 seeds was found. (3) A markedly increased PH was observed in all of the plots under study after the completion of experiment. (4) The compost treatment and the ferttilizer (NPK) treatment greatly increased the organic matter, total nitrogen and the available phosphorus in the soil. (5) The variation in the grain yield appeared to have a close correlation with the content of available $P_2O_5$ in the soil.

• Korean Journal of Soil Science and Fertilizer
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• v.19 no.2
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• pp.157-165
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• 1986

This experiment was conducted to find out the some physiological characteristics and nitrogen fixation activities under free-living conditions of indigenous rhizobia isolated from soybean-cultivated (30 fields) and un-cultivated (30 fields) soil. The results were summerized as follows: 1. In free-living condition, only 12.8% and 6.4% of the indigenous rhizobia isolated from soybean cultivated (133 strains) and un-cultivated (125 strains) soils were nitrogenase positive as more than 4 n mole $C_2H_4$ per tube per hour by acetylene reduction assay. 2. The acid-producing rhizobia on litmus milk test was observed to be 20% of the total 160 strains isolated from soybean cultivated soil but about 34% of 166 strains isolated from un-cultivated soil. And the serum zone positive strains were higher in the soybean un-cultivated soil than cultivated soil. 3. The population ratio of fast-to slow-growing indigenous rhizobia based on growth pattern of AMA medium was 35.6% to 64.4% of the total 346 strains. 4. The population of indigenous Rhizobium japonicum counted by MPN method was ranged from $9.2{\times}10^2$ cells per gram of soil in soybean un-cultivated soil to $2.3{\times}10^4$ cells per gram of soil soybean cultivated soil. The number of indigenous R. japonicum in 0-10cm depth of surface layer was higher than low layer.

• Korean Journal of Soil Science and Fertilizer
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• v.24 no.1
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• pp.69-76
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• 1991

A pot experiment was conducted to find out the effects of lime and phosphate application on the changes of number of soil microorganisms, indigenous Rhizobium japonicum, nodule formation, and ureide-and amide-N in leaf and stem exudate of soybean plant under uncultivated hillside red earth in very low pH value, organic matter, available phosphate, and cation exchange capacity. The results obtained were summarized as follows : 1. The plant height, stem length, root dry weight and nodule weight were significantly increased with the application of lime and phosphate application than that of control plot. 2. The concentration of amide-N in soybean plant at the 45 days after sowing was obtained as high in order of control>lime> lime+phosphate while the concentration was obtained in order of Iime+phosphate>lime> control at flowering stage 3. However, concentration of ureide-N in the soybean leaf at the 45 days after sowing was obtained as high in order of control>lime>lime+hosphate while reversed concentration was obtained in stem. 4. The number of soil microorgan isms were increased with increase of pH value, available phosphate and soil exchangeable cation. 5. Significantly negative high correlation were obtained with the concentration of Al, Fe in soil and the concentration of amide-and ureide-N in soybean plant at flowering stage while positive correlation was obtained with plant growth and the concentration of ureide-N.