• Microbiology and Biotechnology Letters
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• 제38권3호
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• pp.302-307
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• 2010

Nitrogen is an element need to grow plants growth. Plants take up nitrogen in the form of nitrate or ammonium. Most of plants absorb nitrogen source as fertilizers. But from 50 to 70% of fertilizers applied were washed away. This study was conducted to isolate free-living nitrogen fixing bacteria from reed and to examine its beneficial traits for developing sustainable biofertilizers. Enriched consortium obtained from a reed in Ansan was developed for the fixing of nitrogen. Nitrogen fixing bacteria isolated from an enriched culture in Congo Red Medium was analyzed by 16s rDNA sequencing. AKC-10 was isolated and shown to have excellent nitrogen fixing ability. The optimum conditions of nitrogen fixing ability were $25^{\circ}C$ ($237.50{\pm}39.65\;nmole{\cdot}mg-protein^{-1}{\cdot}h^{-1}$ and pH 7 ($168.335{\pm}12.84$ nmole/hr mg-protein). It was identified as Microbacterium hominis [(AKC-10 (similarity : 99%)]. This strain was had to IAA (indole-3-acetic acid) productivity and ACC(1-aminocyclopropane-1-carboxylic acid) deaminase activity. Therefore, Microbacterium hominis AKC-10 stimulated plant development in the soil, enhancing the efficiency of remediation.

• Proceedings of the Korean Society of Crop Science Conference
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.190-190
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• 2017

Chemical fertilizers have been used to increase crop production and contributed to escaping food shortages. However, excessive use of chemical fertilizers over a long period caused many problems such as environmental pollution and the hampered production potential of the land. Thus, it is necessary to develop eco-friendly bio-fertilizers that can replace the use of chemical fertilizers. Here, we tested the effect of some nitrogen-fixing microorganims on the plant growth promotion. Seventy free-living nitrogen fixing microorganisms were isolated from rhizosphere of crop cultivation fields, streamside soils and sludge in Ansung, Korea. Of them, three strains (NF2-4-1, Yeast; EMM409, Mesorhizobium; Gsoil662, Burkholderia) were selected to be most efficient in the capacity of N-fixing nitrogen based on colony forming cell assay in N-free media. To investigate the ability to promote plant growth, these strains were inoculated into the soil and cabbage were grown for 4 weeks in the grown chamber. Fresh weight, dry weight, and leaf area were measured from 4-week-old plants. Phenotypic analysis revealed that the growth of the plants inoculated with NF2-4-1 and EMM409 strains were significantly promoted compared to the mock-treated control plants, while Gsoil662-inoculated plants did not show statically significant promotion. These results indicate that these nitrogen-fixing microorganims can be used to develop plant growth promoting bio-fertilizers. Further analysis on nitrogen fixing level in soil by these strains will be tested.

• Journal of Microbiology and Biotechnology
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• 제34권3호
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• pp.570-579
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• 2024

Root-nodule nitrogen-fixing bacteria are known for being specific to particular legumes. This study isolated the endophytic root-nodule bacteria from the nodules of legumes and examined them to determine whether they could be used to promote the formation of nodules in other legumes. Forty-six isolates were collected from five leguminous plants and screened for housekeeping (16S rRNA), nitrogen fixation (nifH), and nodulation (nodC) genes. Based on the 16S rRNA gene sequencing and phylogenetic analysis, the bacterial isolates WC15, WC16, WC24, and GM5 were identified as Rhizobium, Sphingomonas, Methylobacterium, and Bradyrhizobium, respectively. The four isolates were found to have the nifH gene, and the study confirmed that one isolate (GM5) had both the nifH and nodC genes. The Salkowski method was used to measure the isolated bacteria for their capacity to produce phytohormone indole acetic acid (IAA). Additional experiments were performed to examine the effect of the isolated bacteria on root morphology and nodulation. Among the four tested isolates, both WC24 and GM5 induced nodulation in Glycine max. The gene expression studies revealed that GM5 had a higher expression of the nifH gene. The existence and expression of the nitrogen-fixing genes implied that the tested strain had the ability to fix the atmospheric nitrogen. These findings demonstrated that a nitrogen-fixing bacterium, Methylobacterium (WC24), isolated from a Trifolium repens, induced the formation of root nodules in non-host leguminous plants (Glycine max). This suggested the potential application of these rhizobia as biofertilizer. Further studies are required to verify the N₂-fixing efficiency of the isolates.

• Microbiology and Biotechnology Letters
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• 제13권3호
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• pp.251-255
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• 1985

Nitrogen-fixing bacteria were isolated from the rice rhizosphere of various paddy fields in our country. The screening of 235 isolates for nitrogen-fixing ability resulted in the isolation of Enterobacter agglomerans NFB264 and three Klebsiella pneumoniae NFB 3, NFB 320. Plasmids of various molecular weight from 1.7 to more than 84 Mal. were detected by agarose gel electrophoresis in three out of four isolates. But, these plasmids had not any nitrogen-fixing genes. Hybridization experiments using Klebsiella pneumoniae M5al nitrogen-fixing genes, nif Q-K and nif DH, as probes revealed the presence of homologous sequences in the chromosomal DNA of all isolates. However the restriction patterns of nif genes of the isolates by various restriction endonucleases were different to those of Klebsiella pneumoniae M5al.

• Korean Journal of Soil Science and Fertilizer
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• 제20권2호
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• pp.185-192
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• 1987

A green house experiment was conducted to find out the acetylene reducing and $N_2$-fixing activity from photosynthetic and aerobic heterotrophic nitrogen fixing microorganisms in submerged paddy soil under different agricultural locality, soil series, soil texture, soil type, and drainage condition in which samples taken from without nitrogen treatment plot of NPK trials on 16 sites of the farmer's field. The results obtained were summarized as follows: 1. The highest acetylene reducing activity was observed at 7 days after incubation in the light condition (photo synthetic microbes+heterotrophic bacteria) while it was observed at 35 days incubation in the dark condition (heterotrophic bacteria). 2. Among the soil series, photosynthetic nitrogen fixing activity was pronounced more in Jangae, Ogcheon and Hwadong series while lower was obtained in Buyong and Daejeong series. Aerobic heterotrophic nitrogen fixing activity was high in Buyong and Daejong series. 3. Estimated amount of $N_2$-fixation from acetylene reducing activity was equivalented to 3.0 mg in light condition and 4.9 mg/100g/105 days in dark condition. 4. Among the agricultural locality, photosynthetic nitrogen fixing activity was high in rather warm southern part while heterotrophic nitrogen fixing activity was predominated more in mountainous area and Chungcheong continental. 5. Photosynthetic nitrogen fixing activity was predominated in high productive soil while aerobic heterotrophic nitrogen fixing activity was pronounced more in crose coarse sandy soil. 6. The soils properties of high photosynthetic nitrogen fixing activity were constituted of poorly or imperfectly drained clay or clay loam soil while heterotrophic nitrogen fixing activity was pronounced more in well to moderately well drained sandy or sandy loam soil.

• Microbiology and Biotechnology Letters
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• 제37권3호
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• pp.189-196
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• 2009

Bacteria were isolated from roots of plants belonging to family Solanaceae and Gramineae, inhabited in Dokdo island. Fifty six endospore-forming bacteria grown on tryptic soy broth (TSB) agar medium and 23 nitrogen-fixing bacteria (NFB) grown on nitrogen free agar medium were isolated, respectively. The isolates were partially identified by analyzing the 16S rDNA and categorized into phylogenetic groups. The 16S rDNA sequences of each identified isolates were compared with sequences of each type strains to analyze phylogenetic relationship by phylogenetic tree. As a result, endospore-forming bacteria and nitrogen-fixing bacteria were classified into 4 and 6 lineage groups, respectively. Among these isolated, 18 were presumed to be novel species candidates based on the similarity (lower than 98%) analysis of the l6S rDNA sequences.

• Journal of Plant Biotechnology
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• 제49권4호
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• pp.300-306
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• 2022

The mitogen-activated protein kinase (MAPK) signaling cascade is essential for a wide range of cellular responses in plants, including defense responses, responses to abiotic stress, hormone signaling, and developmental processes. Recent investigations have shown that the stress, ethylene, and MAPK signaling pathways negatively affect the formation of nitrogen-fixing nodules by directly modulating the symbiotic signaling components. However, the molecular mechanisms underlying the defense responses mediated by MAPK signaling in the organogenesis of nitrogen-fixing nodules remain unclear. In the present study, I demonstrate that the Medicago truncatula mitogen-activated protein kinase kinase 5 (MtMKK5)-Medicago truncatula mitogen-activated protein kinase 3/6 (MtMPK3/6) signaling module, expressed specifically in the symbiotic nodules, promotes defense signaling, but not ethylene signaling pathways, thereby inhibiting nodule development in M. truncatula. U0126 treatment resulted in increased cell division in the nodule meristem zone due to the inhibition of MAPK signaling. The phosphorylated TEY motif in the activation domain of MtMPK3/6 was the target domain associated with specific interactions with MtMKK5. I have confirmed the physical interactions between M. truncatula nodule inception (MtNIN) and MtMPK3/6. In the presence of high expression levels of the defense-related genes FRK1 and WRKY29, MtMKK5a overexpression significantly enhanced the defense responses of Arabidopsis against Pseudomonas syringae pv. tomato DC3000 (Pst DC3000). Overall, my data show that the negative regulation of symbiotic nitrogen-fixing nodule organogenesis by defense signaling pathways is mediated by the MtMKK5-MtMPK3/6 module.

• Microbiology and Biotechnology Letters
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• 제31권1호
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• pp.18-24
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• 2003

For researching nitrogen-fixing bacteria associated with gramineous crops, we collected growing roots of rices, wheats, oats, barleys, ryes, and maizes at 19 sites of southern Korean peninsula. Endophytes and free living bacteria were isolated from those crop roots. Sixty-three isolates were classified on the basis of different morphology, size, color, host of colony, and the 16S rDNAs sequence. The analyses of PCR amplification for nifH gene and nitrogenase activity assay, revealed that all isolates contained nitrogen-fixing abilities. In addition, most of them have cellulase activity which is one of the common features of endophytic bacteria from plant.

• Korean Journal of Soil Science and Fertilizer
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• 제16권1호
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• pp.77-83
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• 1983

Effects of rice straw application (particularly application method; surface placement and deep placement) on the biological nitrogen fixing activity of paddy fields and on the growth of rice plant have been investigated by the lysimeter experiment with Aragawa alluvial soil and Tochigi ando soil. 1. Nitrogen fixing activity of plow layer (0~1cm) was increased by the rice straw application, notably by the phototrophs, both deep placement and surface placement, between 1 and 2 months of initial growth stage of rice plant. 2. Surface placement method stimulated the nitrogen fixing activity more significantly them the deep placement method, and also showed good effects on the growth of rice plant. 3. The increased of effect on the nitrogen fixing activity of surface placed area was found to be originated from the applied rice straw and its neigh bouring area. 4. Nitrogen fixing activity of surface placed rice straw was rather promoted by the application of herbicides (2, 4, 6-arichlorophenyl ether) than the non-applicated plot.

• Korean Journal of Soil Science and Fertilizer
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• 제23권1호
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• pp.67-72
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• 1990

A number of nitrogen-fixing microorganisms were isolated from roots of non-legumious crops and evaluated for their nitrogen-fixing ability on their host crops. The results are summarized as follows: 1. Among nitrogen-fixing isolates obtained from 9 different crops, several isolates showed relatively higher level of acetylene reduction activity on Spinach, Chinese cabbage, Cucumber, mustard and Egg plant. 2. When the isolates were inoculated into 4 host crops, a number of isolates were found to grow well on Spinach, Chinese cabbage and Cucumber, but particularly well on the former. 3. Results of this study suggest that nitrogen-fixing a symbiotic microorganisms are also present and able to grow well on roots of non-legumious crops, and stimulate plant growth by promoting growth and differentiation of roots.