• Journal of Korean Society of Forest Science
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• v.85 no.1
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• pp.96-106
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• 1996

Red alder(Alnus rubra Bong.) seedlings inoculated with Frankia only or both Frankia and spores of Alpova diplophloeus(Zeller & Dodge) Trappe & Smith were grown in a greenhouse for ten weeks. The ten-week-old seedlings were fertilized with six nitrogen(N) and phosphorus(P) fertility regimes (no fertilization, 10mM $NH_4NO_3$, 50mM $NH_4NO_3$, 5mM $KH_2PO_4$, 10mM $NH_4NO_3+5mM$ $KH_2PO_4$, and 50mM $NH_4NO_3+5mM$ $KH_2PO_4$) three times a week for ten weeks. The higher N-fertilization significantly increased mycorrhiza formation by greenhouse contaminant mycorrhizal fungi, but decreased N-fixation and P concentration in nodule tissues. P-fertilization significantly increased nodule and shoot dry weight, and P concentration in plant tissues. When N was highly fertilized, however, the P-fertilization effect disappeared in nodule P concentration but doubled in leaf P concentration. A. diplophloeus inoculation significantly increased diameter growth and $CO_2$ exchange rate, but decreased leaf dry weight. Our results suggest that the higher N- or P-fertilization affect nitrogenase activity and mycorrhizal development but the effects are changed by their interactions.

• Journal of Microbiology and Biotechnology
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• v.25 no.8
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• pp.1339-1348
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• 2015

Until now, considerable effort has been made to engineer novel nitrogen-fixing organisms through the transfer of nif genes from various diazotrophs to non-nitrogen fixers; however, regulatory coupling of the heterologous nif genes with the regulatory system of the new host is still not well understood. In this work, a 49 kb nitrogen fixation island from P. stutzeri A1501 was transferred into E. coli using a novel and efficient transformation strategy, and a series of recombinant nitrogen-fixing E. coli strains were obtained. We found that the nitrogenase activity of the recombinant E. coli strain EN-01, similar to the parent strain P. stutzeri A1501, was dependent on external ammonia concentration, oxygen tension, and temperature. We further found that there existed a regulatory coupling between the E. coli general nitrogen regulatory system and the heterologous P. stutzeri nif island in the recombinant E. coli strain. We also provided evidence that the E. coli general nitrogen regulator GlnG protein was involved in the activation of the nif-specific regulator NifA via a direct interaction with the NifA promoter. To the best of our knowledge, this work plays a groundbreaking role in increasing understanding of the regulatory coupling of the heterologous nitrogen fixation system with the regulatory system of the recipient host. Furthermore, it will shed light on the structure and functional integrity of the nif island and will be useful for the construction of novel and more robust nitrogen-fixing organisms through biosynthetic engineering.

• Applied Biological Chemistry
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• v.40 no.5
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• pp.438-441
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• 1997

The relatedness of naturalized Bradyhizobium japonitum populations with soil physico-chemical characteristics as affected by paddy rice-upland soybean rotation cropping with conventional and none fertilization in Chilgog clay loam soils were determined as follows. The populations of B. japonicum in soils were increased from about $10^1$ in continuous paddy upto $10^1cells/g.soil$ only in one-year rotation of upland use with soybean cropping. Compared to the densities in plots of conventional fertilization, those in none fertilization were high ranging from 1.9 to 10 fold in 2-year upland use rotation and both in 3-year upland use rotation and 4-year upland use, respectively. The populations were positively correlated with soil organic matter $contents(r=0.83^*),\;Ca/K(r=0.74^*),\;and(Ca+Mg)/K(r=0.72^*)$ and were negatively correlated with soil $hardness(r=-0.73^*)$. And the soil populations increased by paddy-upland rotation resulted in superior symbiotic potentials to those in continuous paddy use in terms of nodule mass, nitrogenase activity, and soy-bean shoot dry weight.

• 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.26 no.3
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• pp.197-203
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• 1993

Greenhouse experiments were conducted to avaluate strain competition, nodulation, patterns of nodule occupancy and population changes of Bradyrhizobium sp. strain HCR-46 $str^{r}cep^{r}$ and CB756 $str^{r}rif^{r}$ in the rhizosphere of peanut(Arachis hypogaea L.) under different root temperatures. Inoculated with two strains using seed coating with peat slurry under different root temperatures, population of each strain in the rhizosphere increased with plant growth and multiplication rate of inoculum in the unit weight of root were showed the highest from 10 to 15days after sowing. The multiplication rate of inoculum in the rhizosphere was $28^{\circ}C$>$34^{\circ}C$>$22^{\circ}C$. The density of HCR-46 $str^{r}cep^{r}$ was more increased than that of CB756 $str^{r}rif^{r}$ under $22^{\circ}C$ and $28^{\circ}C$. While the density of two strains showed no difference under $34^{\circ}C$. Inoculated with HCR-46 $str^{r}cep^{r}$ and CB756 $str^{r}rif^{r}$, respectively at 22, 28 and $34^{\circ}C$, nodulation of each strain was dominated in its inoculation portion. Inoculated with the mixture of HCR-46 $str^{r}cep^{r}$ and CB756 $str^{r}rif^{r}$, occupancy rate of HCR-46 $str^{r}cep^{r}$ was dominated over that of CB756 $str^{r}rif^{r}$ at $22^{\circ}C$ and $28^{\circ}C$, but that was similar between them at $34^{\circ}C$. Dry mass, nodulation, nitrogen content per plant and nitrogenase activity showed higher at $28^{\circ}C$ than at $32^{\circ}C$ and $22^{\circ}C$, while those were higher in HCR-46 $str^{r}cep^{r}$ and mixing HCR-46 $str^{r}cep^{r}$ with CB756 $str^{r}rif^{r}$ than in CB756 $str^{r}rif^{r}$.