• 한국미생물·생명공학회지
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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.

• 한국작물학회:학술대회논문집
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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.

• 한국미생물·생명공학회지
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• 제13권3호
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• pp.251-255
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• 1985

국내 논의 흙으로부터 벼과식물에 관련된 질소 고정 미생물을 238주 분리하여 비교적 활성이 높은 4주의 질소고정활성 미생물을 선별하여 이중 3주는 K. pneumoniae, 1주는 E. agglomerans로 동정하였다. NFB3, NFB264, NFB278의 3주는 1.7-84Mdal. 의 plasmid들을 가지고 있었으나 이들 plasmid에 질소고정유전자 단편을 함유하고 있지는 않았다. 선별미생물의 chromosomal DNA상에 존지하는 질소고정 유전자는 K. pneumoniae M5al의 질소고정유전자와 homology를 나타냈으며, 이들의 질소고정유전자는 K. pneumoniae M5al의 질소고정유전자와 제한효소 절단부위가 상이하였다.

• 한국토양비료학회지
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• 제20권2호
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• pp.185-192
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• 1987

담수상태(湛水狀態)의 논토양(土壤)에서 광합성미생물(光合成微生物) 및 타양성미생물(他養性微生物)에 의한 질소고정력(窒素固定力)을 알기 위하여 우리나라 전국(全國) 수도(水稻) 삼요소시비량시험지(三要素施肥量試驗地) 16개소(個所)의 무질소구(無窒素區) 표토(表土)를 채취(採取)하여 초자실(硝子室)에서 항온(恒溫)하면서 토양통(土壤統), 농업기후대(農業氣候帶), 논토양유형(土壤類型), 토성(土性) 및 배수정도(排水程度)에 따른 질소고정량(窒素固定量) 차이조사(差異調査)를 한 결과(結果)를 요약(要約)하면 다음과 같다. 1. 담수항온기간중(湛水恒溫期間中) 광합성질소고정미생물(光合成窒素固定微生物)에 의한 Acetylene 환원력(還元力)은 항온(恒溫) 7일째, 그리고 타양성질소고정미생물(他養性窒素固定微生物)에 의한 Acetylene 환원력(還元力)은 항온(恒溫) 35일째 가장 높았다. 2. 타양성미생물(他養性微生物)에 의한 질소고정력(窒素固定力)이 가장 높은 토양통(土壤統)은 장계(長溪), 옥천(沃泉) 및 화동통(華東統)이었으며 질소고정력(窒素固定力)이 낮은 토양(土壤)은 부용(芙蓉) 및 대정통(大靜統)이었다. 광합성미생물(光合成微生物)에 의한 질소고정력(窒素固定力)이 가장 높은 토양(土壤)은 대정(大靜) 및 지산통(芝山統)이였으며 낮은 토양(土壤)은 장계(長溪), 함창통(咸昌統)이였다. 3. 항온(恒溫) 105 일간(日間) Acetylene 환원법(還元法)에 의한 생물질소고정량(生物窒素固定量)은 광합성미생물(光合成微生物)에 의하여 3.0mg, 그리고 타양성미생물(他養性微生物)에 의하여 4.9mg/100g이었다. 4. 농업기후대별(農業氣候帶別) 생물질소고정량(生物窒素固定量)은 광합성미생물(光合性微生物)의 경우 기온(氣溫)이 온난(溫暖)한 남부해안지역(南部海岸地域)에서 높고 타양성미생물(他養性微生物)은 산간지(山間地)와 충청내륙지역(忠淸內陸地域)에서 높았다. 5. 답토양유형별(沓土壤類型別) 질소고정량(窒素固定量)은 광합성미생물(光合成微生物)의 경우 보통답(普通沓) 토양(土壤)에서 높고 타양성미생물(他養性微生物)의 경우는 조립질(粗粒質)의 사질답(砂質畓) 토양(土壤)에서 높았다. 6. 토양(土壤)의 특성별(特性別) 질소고정력(窒素固定力)을 볼 때 광합성미생물(光合成微生物)은 배수(排水) 약간불량(若干不良)한 식질(埴質) 혹(或)은 식양질토양(埴壤質土壤)에서 높고 타양성미생물(他養性微生物)은 배수양호(排水良好)한 사질토양(砂質土壤)에서 높았다.

• 한국미생물·생명공학회지
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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.

• 한국미생물·생명공학회지
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• 제31권1호
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• pp.18-24
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• 2003

벼과식물과 관련된 질소고정균을 분리하기 위하여 한반도 남부지방의 19지역에서 쌀, 밀, 귀리, 보리, 호밀, 옥수수 등을 채집하여 이들의 뿌리로부터 free living bacteria와 endophyte를 분류하였다. Colony의 형태, 크기, 색깔 등과 16S rDNA 염기서열을 근거로 63종으로 분류하였다. nifH 유전자 염기서열 결정과 nitrogenase 활성 측정을 통하여 분리균주 모두 질소고정 능력이 있음을 확인하였다. 그리고 식물에서 분리한 내생균의 특징 중 하나인 cellulase활성이 대부분의 분리 균주에서 확인되었다.

• 한국토양비료학회지
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• 제16권1호
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• pp.77-83
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• 1983

논토양(土壌)의 생물적(生物的) 질소고정활성(窒素固定活性) 및 수도생육(水稻生育)에 미치는 볏짚시용(施用) (특(特)히 시용방법(施用方法); 표면시용(表面施用)과 혼층시용(混層施用))효과(効果)를 밝히기 위하여 황천(荒川) 충적토양(沖積土壌)과 회목(栃木) 화산회토양(火山灰土壌)을 충분(充墳)한 라이시메타 시험(試験)을 실시(実施)하여 다음과 같은 결과(結果)를 얻었다. 1. 볏짚시용(施用)은 혼층시용(混層施用), 표면시용(表面施用) 어느 경우에도 수도(水稻) 생육초기(生育初期) 1~2개월(個月) 사이에 작토(作土) 표층부(表層部) (0~1cm)의 질소고정활성(窒素固定活性), 특(特)히 광합성미생물(光合成微生物)에 의한 질소고정활성(窒素固定活性)을 현저(顕著)하게 증대(増大)시켰다. 2. 볏짚 시용방법(施用方法)에 따라서는 혼층시용(混層施用)보다도 표면시용(表面施用)하므로 더욱 현저(顕著)한 질소고정활성(窒素固定活性)의 증대(増大)를 보였으며, 표면시용(表面施用)은 수도(水稻)의 생육(生育) 정도(程度)에도 양호(良好)한 결과(結果)를 가져왔다. 3. 유수형성기(幼穗形成期)에 작토(作土)에 잔존(殘存)하는 볏짚, 수도근(水稻根), 토양(土壌)을 각각(各各) 채취(採取)하여 질소고정활성(窒素固定活性)을 비교(比較)한 결과(結果), 볏짚시용(施用)에 의한 표층부(表層部)에서의 질소고정활성(窒素固定活性)의 증대(増大)는 시용(施用)되어진 볏짚 및 그 근방(近傍)에서 발현(発現)된 것으로 판명(判明)되었다. 4. 제초제(除草剤)의 산포(散布)(단(但) MO입제(粒剤))는 제초제(除草剤) 무산포구(無散布区)보다 오히려 표층부(表層部) 볏짚의 질소고정활성(窒素固定活性)의 증대(増大)를 가져왔다.

• 한국토양비료학회지
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• 제23권1호
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• pp.67-72
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• 1990

수종(數種)의 비두과식물(非荳科植物)의 뿌리로 부터 질소고정균(窒素固定菌)을 분리(分離)하여 분리(分離)된 균(菌)에 대(對)해서 숙주식물(宿主植物)의 접종실험(接種實驗)을 하여 유효균(有效菌)의 선발(選拔)을 한 결과(結果) 1. 9개(個)의 식물(植物)에 대(對)해서는 시금치, 배추, 오이, 갓 및 가지에서 비교적(比較的) 높은 아세칠렌 환원능(還元能)을 가진 균주(菌株)를 얻었고 2. 그 중(中)에서 아세칠렌 환원능(還元能)이 큰 4개(個)의 숙주식물(宿主植物)중 시금치, 배추, 오이에서 양호(良好)한 생육(生育)을 하는 균주(菌株)를 찾았다. 특(特)히 시금치에서 접종효과(接種效果)가 현저(顯著)하였다. 3. 이상(以上)의 결과(結果)로 부터 비두과식물(非荳科植物)의 뿌리에도 약한 협생(協生)을 하는 질소고정균(窒素固定菌)이 생육(生育)되고 있어 뿌리의 생장(生長) 및 분화(分化)를 촉진(促進)시켜 지상부(地上部)의 생육(生育)을 왕성하게 하는 것으로 판단(判斷)된다.