• Microbiology and Biotechnology Letters
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• v.46 no.2
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• pp.154-161
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• 2018

Currently, urban greening projects and research are attracting attention as a way to mitigate urban heat island phenomenon. In this study, nitrogen fixative bacteria were isolated and their effects on plant growth were confirmed. First, enrichment was performed in a nitrogen-free medium to isolate the nitrogen-fixing bacteria, and the colony showing high growth in a medium with limited nitrogen source was isolated and purified. Separated bacterial isolates were reduced by more than 90% acetylene by ARA and indirectly confirmed the activity of nitrogenase by ethylene production. Cedecea sp. MK7 and Enterobacter sp. Y8 with confirmed reproducibility were selected as nitrogen fixative bacteria. Nitrogen fixing bacteria were applied to the growth of perennial rye grass, and it was found that the dry weight increased to 34.80 mg (186.60%) compared with the control with 18.65 mg dry weight. After plant growth, microbial community analysis of soil applied by bacteria showed similarity to the control group. Therefore, in this study, it is expected that the efficiency will be increased if plant growth is promoted by using nitrogen fixing bacteria in urban greenery system.

• Microbiology and Biotechnology Letters
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• v.34 no.1
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• pp.78-82
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• 2006

Since there could be more and rather various diazotrophs in rhizosphere of wild crops than those in rhizosphere of cultivars, some wild gramineous crops grown in Korea were collected for isolating nitrogen-fixing bacteria. Six diazotrophs were purified from their roots using nitrogen-free media. The isolated bacteria were partially identified as 4 genera by 16S rDNA sequence analysis: Stenotrophomonas sp., Bosea sp., Klebsiella sp., and Azorhizobium sp. By PCR amplification and sequence analysis, DNA fragments extracted from all isolates turned out to have an individual nifH homologous gene. Five isolates (KNUC163, KNUC165, KNUC169, KNUC170, and KNUC171) showed auxin activity and four isolates (KNUC163, KNUC166, KNUC170, and KNUC171) produced siderophores. Especially,3 strains of S. maltophilia showed both auxin and siderophore activities. In conclusion, the isolated nitrogen-fixing bacteria might have capabilities for plant growth promotion.

• The Korean Journal of Ecology
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• v.26 no.2
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• pp.71-74
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• 2003

Effects of nitrogen addition on the growth of Indigofera pseudo-tinctoria (Leguminosae) in the waste landfill site was investigated. Nitrogen fertilization in the nitrogen poor soils of waste landfill may influence the growth of nitrogen fixing plants beneficially or detrimentally. When I. pseudo-tinctoria was fertilized with three different levels of nitrogen, the coverage of plants treated with 46 g N/$m^2$ and 460 g N/$m^2$ was significantly less than that of plants treated with 23 g N/$m^2$. The growth rates of plant height treated with 46 g N/$m^2$ and 460 g N/$m^2$ were significantly less than those of plants treated with 23 g N/$m^2$. The growth rates of plant diameter treated with 46 g N/$m^2$ and 460 g N/$m^2$ were significantly less than those of plants treated with 23 g N/$m^2$. Dry weights of whole plants in control sites were higher than those of all the others nitrogen treatment sites. Nodule numbers were higher in control plants than those of plants in all the other nitrogen treatment sites. It is suggested that nitrogen fertilizer addition over 23 g N/$m^2$ affect the growth of some nitrogen fixing plants, such as I. pseudo-tinctoria, negatively.

• Journal of Plant Biology
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• v.29 no.2
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• pp.109-115
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• 1986

Actinorhizal plants being capable of fixing atmospheric nitrogen in symbiotic association with Frankia were surveyed for their root nodules, and the root nodules were collected for further studies. Three species of Alnus and Elaeagnus (A. hirsuta, a. firma, A. japonica; E. glabra, e. umbellata, E. macrophylla) and one species of Myrica (M. rubura) were confirmed to bear the root nodules of typical external shape. Morphological studies using light microscope and scanning electron microscope revealed the presence of vesicle clusters in the cortex region of the root. Two tentative endophyte colonies were isolated from the root nodule of A. hirsuta.

• Journal of Plant Biology
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• v.32 no.3
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• pp.137-144
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• 1989

In order to clarify the effect of the associative nitrogen fixing bacteria on the growth of corn(Zea mays L. Suwon19), we inoculated corn seedlings with Azospirillum amazonense Y1, a micro-aerobic and acid-tolerant nitrogen fixer, and analyzed the growth and dry matter production and changes of nitrogen and phosphorus quantity of the plant during the growing period. The inoculation of associative N-fixing bacgteria increased the growth of height, leaf area, dry weight and total nitrogen quantity of the plant by 15.4%, 65.4%, 33.7% and 38.0%, respectively, on the day of 56th after sowing. Especially the inoculation of A. amazonense Y1 showed two to three-fold accumulation of phosphorus in each organ of plant. The associative bacteria accelerated the growth of the under ground parts more than those of the upper ground parts of corn plant, and caused decrease in T/R ratios.

• ALGAE
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• v.19 no.2
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• pp.145-148
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• 2004

As a result of the 2-year monthly monitoring of the phytoplankton community at 3 stations in Mankyeong Estuary, Korea, we learned that cyan bacterial species of the genus Anabaena occurred at most sampling points with huge salinity differences (0.1-32.5 psu). We isolated several clones of Anabaena spp. from the monitoring stations, and screen out two euryhaline and nitrogen-fixing Anabaena clones, CB-MAL21 and CB-MAL22. The two clones were grown under various environmental gradients such as temperature (20, 30, 35 and 40$^{\circ}C$), salinity (0, 2, 5, 15 and 30psu), and $PO_4^{3-}$-P concentration (0, 1.6, 8.0, 40 and 200 ${\mu}M$M). Growth of CB-MAL21 and CB-MAL22 was measured by daily monitoring of chlorophyll fluorescence from each experimental culture for more than three serial transfers. Both the two experimental clones did not grow at 0psu. Maximal growth rates of the two clones were markedly reduced at lower $PO_4^{3-}$-P concentrations showing negligible growth at 0 and 1.6 ${\mu}M$M. However, growth of CB-MAL21 was not affected by low $NO_3^--$ concentration in culture media, showing the nitrogen-fixing ability. Maximum biomass yields of the two clones decreased dramatically at 35 and 40$^{\circ}C$. Optimal growth conditions for the two experimental clones were determined to be 20-30$^{\circ}C$, 40 ${\mu}M$M $PO_4^{3-}$-P, and wide salinity range from 5.0 to over 30psu. Best growth of CB-MAL21 was shown at (20$^{\circ}C$-15psu), which is less saline and cooler condition than those (i.e., 30$^{\circ}C$-30psu) for the best growth of CB-MAL22. The euryhaline and nitrogen-fixing CB-MAL21 strain thus can be a candidate laboratory culture for the future cyan bacterial marine biotechnology in temperate coastal waters.

• Journal of Microbiology and Biotechnology
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• v.20 no.8
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• pp.1251-1258
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• 2010

During the nitrogen-fixing process, ammonia ($NH_3$) is incorporated into glutamate to yield glutamine and is generally not secreted. However, in this study, $NH_3$-excreting strains of nitrogen-fixing Paenibacillus were isolated from soil. The ammonium production by the Paenibacillus strains was assayed in different experiments (dry biomass, wet biomass, cell-free extract, and cell-free extract adsorbed on nano $TiO_2$ particles) inside an innovative bioreactor containing capsules of $N_2$ and $H_2$. In addition, the effects of different $N_2$ and $H_2$ treatments on the formation of $NH_3$ were assayed. The results showed that the dry biomass of the strains produced the most $NH_3$. The dry biomass of the Paenibacillus strain E produced the most $NH_3$ at 1.50, 0.34, and 0.27 ${\mu}M$ $NH_3$/mg biomass/h in the presence of $N_2$ + $H_2$, $N_2$, and $H_2$, respectively, indicating that a combined effluent of $N_2$ and $H_2$ was vital for $NH_3$ production. Notwithstanding, a cell-free extract (CFE) adsorbed on nano $TiO_2$ particles produced the most $NH_3$ and preserved the enzyme activities for a longer period of time, where the $NH_3$ production was 2.45 ${\mu}M$/mg CFE/h over 17 h. Therefore, the present study provides a new, simple, and inexpensive method of $NH_3$ production.

• 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.

• Korean Journal of Microbiology
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• v.25 no.2
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• pp.129-136
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• 1987

Nitrogen fixing activity associated with 40 varieties of rice was assayed at heading stage by an in situ acetylene reduction method. The in situ acetylene reduction activity and population of nitrogen fixing bacteria obtained on nitrogen-free malate medium for Azospirillum spp. enrichment showed positive correlation. Six Azospirillum spp. with high nitrogenase activity were isolated from the rice roots, from which five spp. were identified as A. lipoferum and one was A. brasilense. The physiological characteristics of the six Azospirillum isolates, that is, carbon source utilization, biotin requirement, antibiotic resistance, indole acetic acid excretion, plasmid profile and protein patterns were compared.

• Proceedings of the Plant Resources Society of Korea Conference
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• 2008.04a
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• pp.5-13
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• 2008

The aquatic fern Azolla spp. is of value as a bio-fertilizer for wetland paddy, and also can be used as an ideal feed for cattle, pigs and poultry. It is popular and cultivated widely in other countries like China, Vietnam, and the Philippines, but has yet to be taken up in Korea, in a big way. It fixes nitrogen as high as 3-5kg N per day, because it contains nitrogen fixing blue-green algae, Anabaena azollae. Azolla's ability to create a light-proof mat that suppresses other weeds has been used for centuries in rice production. Azolla spp. has also the capacity to take up the heavy metals ($75{\sim}100%$) and may be used as a bioaccumulator. Moreover, Azolla meal as an unconventional feed resource has a potential as a feedstuff for livestock.