• Journal of Plant Biotechnology
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• v.1 no.1
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• pp.31-38
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• 1999

An artificial symbiosis was established between diazotropic Azomonas insignis and strawberry (Fragaria x ananassa). The partnership was created by in vitro techniques through callus induction and organogenesis. The basis of this partnerships is the bacterial dependence on the plants metabolic activity, using maltose in the medium as a carbon and energy source which can be utilized by the plant cells only. The presence of bacteria in the intercellular spaces of the callus tissues and regenerated plants was proven by microscopic techniques. Nitrogenase activity could also be detected in the plant tissues. For successful and high frequency introduction of bacteria to the plant tissues, biolistic gun method was used. On the basis of the DNA transfer method, Azotobacter vinelandii bacteria were delivered directly into strawberry tissues by the particle bombardment. This was the first use of living bacteria as microprojectils for bombardment of plant tissues. The treatment was successful, the presence of bacteria in the developing callus tissue and regenerated plants were detected by light and electron microscopy.

• Journal of Plant Biology
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• v.30 no.4
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• pp.257-266
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• 1987

Investigation on the ability of nitrogen usage by N2-fixing Nostoc muscorum and cultured tobacco cells when they were associately cultured on nitrogen-free media was carried out. Also, effect of polyamines on the associated cultured condition was carried out. In addition, morphological changes of N. muscorum and cultured tobacco cells in associate culture were observed to detect the possibility of induction of nitrogen fixing ability on cultued plant cells. The activity of nitrogenase increased markedly when N. muscorum was grown exclusively on nitrogen-free media. When N. muscorum was cultured associately with cultured tobacco cells on nitrogen-free media containing polyamines, high activity was detected in 10-4 M spermine treated group. Investigation on the change of polyamine amounts showed two times increase in spermidine and eight times increase in spermine on a associate culture. These effects of associated culture were shown through morphological change such as dense loclization of N. muscorum around the cultured tobacco cells as well as inside the cells. These results indicate the viability of N. muscorum in cultured tobacco cells and possible induction of nitrogen fixation ability by symbiosis.

• Journal of Life Science
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• v.22 no.8
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• pp.1009-1017
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• 2012

The orf282 gene of Rhodobacter sphaeroides is located between the ccoNOQP operon encoding $cbb_3$ terminal oxidase and the fnrL gene encoding an anaerobic activator, FnrL. Its function remains unknown. In an attempt to reveal the function of the orf282 gene, we disrupted the gene by deleting a portion of the orf282 gene and constructed an orf282-knockout mutant. Two FnrL binding sites were found to be located upstream of orf282, and it was demonstrated that orf282 is positively regulated by FnrL. The orf282 gene is not involved in the regulation of spectral complex formation. The $cbb_3$ oxidase activity detected in the orf282 mutant was comparable to that in the wild-type sample, indicating that the orf282 gene is not involved in the regulation of the ccoNOQP operon and the biosynthesis of the cbb3 cytochrome c oxidase. The elevated promoter activity of the nifH and nifA genes, which are the structural genes of nitrogenase and its regulator, respectively, in the orf282 mutant, suggests that the orf282 gene product acts as a negative effector for nifH and nifA expression.

• Applied Biological Chemistry
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• v.40 no.2
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• pp.134-138
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• 1997

The objective of this study was to examine the effect of phosphorus deficiency on nitrogen fixation and photosynthesis of nitrogen fixing soybean plant under $CO_2$ enrichment condition. The soybean plants(Glycine max [L.] Merr.) inoculated with Bradyrhizobium japonicum MN 110 were grown with P-stressed(0.05 mM-P) and control(1 mM-P) treatment under control$(400\;{\mu}l/L\;CO_2)$ and enrichment$(800\;{\mu}l/L\;CO_2)$ enviromental condition in the phytotron equipped with high density lamp$(1000\;{\mu}Em^{-2}S^{-1})$ and $28/22^{\circ}C$ temperature cycle for 35 days after transplanting(DAT). At 35 DAT, phosphorus deficiency decreased total dry mass by 64% in $CO_2$ enrichment condition, and 51% in control $CO_2$ condition. Total leaf area was reduced significantly by phosphorus deficiency in control and enriched $CO_2$ condition but specific leaf weight was increased by P deficiency. Phosphorus deficiency significantly reduced photosynthetic rate(carbon exchange rate) and internal $CO_2$ concentration in leaf in both $CO_2$ treatments, but the degree of stress was more severe under $CO_2$ enrichment condition than under control $CO_2$ environmental condition. In phosphorus sufficient plants, $CO_2$ enrichment increased nodule fresh weight and total nitrogenase activity(acetylene reduction) of nodule by 30% and 41% respectively, but specific nitrogenase activity of nodule and nodule fresh weight was not affected by $CO_2$ enrichment in phosphorus deficient plant at 35 DAT. Total nitrogen concentrations in stem, root and nodule tissue were significantly higher in phosphorus sufficient plant grown under $CO_2$ enrichment, but nitrogen concentration in leaf was reduced by 30% under $CO_2$ enrichment. These results indicate that increasing $CO_2$ concentration does not affect plant growth under phosphorus deficient condition and phosphorus stress might inhibit carbohydrate utilization in whole plant and that $CO_2$ enrichment could not increase nodule formation and functioning under phosphorus deficient conditions and phosphorus has more important roles in nodule growth and functioning under $CO_2$ enrichment environments than under ambient condition.

• The Korean Journal of Ecology
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• v.16 no.2
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• pp.159-168
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• 1993

The seasonal changes of symbiotic nitrogen-fixation activity and environmental factors of autumn olive plant (Elaeagnus umbellata Thunb.), which is an important constituent species of temperate vegetation and a non-leguminous root nodule plant interacting with Frankia sp., were quantitatively analyzed inthe natural vegetations during hegrowing period. The acetylene redution April and showed two peaks of 133 and $145{\mu}M\;C_2H_4\;{\cdot}\;gfw^{-1}\;{\cdot}\;hr^{-1}$ in early June and mid September. The nitrogenase activity decreased to 10~30% during hot dry summer frommid June to the end of August, and disappeared during the dormant period of winter. The optimum rhizoshere. diurnal change showed the maximum activity in the mid-day and the minimum in the mid-night. The average contents of total nitrogen in each organ changed in the ranges of 42.5~40.1, 40.2~36.3, 30.3~28.6 and 18.4~16.2mgN $gdw^{-1}$ for nodule, leaf, root, and stem, respectively. The soil conditions of rhizosphere were weak acidic, ad seasonal variations of $NO_3^--N,\;NH_4^+-N,\;PO_4^{3-}-P$ and water contents were in the ranges of 48.3~79.5ppm,5.1~13.9ppm, 4.4~$9.9{\mu}M$ and 14.5~39.4%, respectively.

• Microbiology and Biotechnology Letters
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• v.13 no.1
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• pp.79-85
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• 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 Microbiology and Biotechnology
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• v.25 no.7
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• pp.1119-1128
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• 2015

1-Aminocyclopropane-1-carboxylate (ACC) deaminase, which is encoded by some bacteria, can reduce the amount of ethylene, a root elongation inhibitor, and stimulate the growth of plants under various environmental stresses. The presence of ACC deaminase activity and the regulation of ACC in several rhizospheric bacteria have been reported. The nitrogen-fixing Pseudomonas stutzeri A1501 is capable of endophytic association with rice plants and promotes the growth of rice. However, the functional identification of ACC deaminase has not been performed. In this study, the proposed effect of ACC deaminase in P. stutzeri A1501 was investigated. Genome mining showed that P. stutzeri A1501 carries a single gene encoding ACC deaminase, designated acdS. The acdS mutant was devoid of ACC deaminase activity and was less resistant to NaCl and NiCl₂ compared with the wild-type. Furthermore, inactivation of acdS greatly impaired its nitrogenase activity under salt stress conditions. It was also observed that mutation of the acdS gene led to loss of the ability to promote the growth of rice under salt or heavy metal stress. Taken together, this study illustrates the essential role of ACC deaminase, not only in enhancing the salt or heavy metal tolerance of bacteria but also in improving the growth of plants, and provides a theoretical basis for studying the interaction between plant growth-promoting rhizobacteria and plants.

• Korean Journal of Microbiology
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• v.24 no.3
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• pp.211-220
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• 1986

Nitrogen fixation (nif) genes of Rhizobium leguminosarum were transferred to nif Klebsiella pneumoniae and E. coli by conjugation after partial heat induction of $RP_4$ :: Mu cts in Rhizobium $R^+$ transconjugant, and the hybrid plasmids in the transconjugant strains were isolated and characterized. In order to transfer the nif genes from Rhizobium, the hybrid plasmid $RP_4$ :: Mu cts was transferred by conjugation from E. coil to the symbiotic nitrogen fixer, R. leguminosarum. After stabillity test, the $RP_4$ :: Mu cts in Rhixobium $R^+$ transconjugant was subjected to partial heat induction by culturing it statically at $38^{\circ}C$ for 16 hours, and then conjugated with the nif defective mutant strains of K. pneumoniae or nif mutant strains of E. coli having whole nif gene plasmid. Recombinant strains of K. pneumoniae, which could grow in a N-free medium and exhibit the nitrogenase activity were selected. However, in the case of E. coli, they could grow well in a NA medium containing antibiotices, but hardly frow in a N-free medium. The hybrid plasmids in these transconjugal strains were isolated by gel electrophoresis and compared their molecular sizes.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.43 no.1
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• pp.23-27
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• 1998

Inhibition of nodule formation and nitrogen fixation by soil nitrogen, primarily nitrate, is well known in legume plants. The present study was undertaken to evaluate the effect of ${NO_3}^-$ on the nodulation, nitrogenase activity, and growth of supernodulating soybean mutant and its wild type. A greenhouse study was conducted to compare two of supernodulating mutants, 'SS2-2' and 'nts 382', with the normal nodulating cultivar 'Sinpaldalkong 2' when grown in a 1-l styroform cup filled with sand, and fertilized with five levels of ${NO_3}^-$ (0, 2, 4, 8, and 12 mM). During the growth period, each plant was supplied two or three times a week with 50 mL of nutrient solution. Supernodulating soybean mutants, SS2-2 and nts 382, showed more nodules and nodule mass, and greater $C_2\;H_2$ activity than the wild type, Sinpaldalkong 2, regardless of the level of exogeneous nitrogen supply. On the other hand, total dry weight of SS2-2 mutant, which was smaller than Sinpaldalkong 2, did not respond to the various ${NO_3}^-$-N levels. This suggested that supernodulating SS2-2 mutant could maintain fairly high total dry weight at the low ${NO_3}^-$-N level, even in the absence of exogeneous ${NO_3}^-$-N in the nutrient solution. From the reduced top growth and high nitrogen fixing ability of supernodulating mutants, it was surmised that supernodulating mutant could potentially protect agricultural environments from pollution through the reduction in nitrogen fertilization as well as maintain fairly high yield with increasing planting density.

• 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.