• 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.

• 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 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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• 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.

• 한국미생물·생명공학회지
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• 제46권2호
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• pp.154-161
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• 2018

현재 도시의 도시열섬현상을 완화시키는 방안으로 도시 녹화사업 및 연구가 주목 받고 있다. 이 연구에서는 질소고정균을 분리하고, 식물 성장에 미치는 영향을 확인했다. 먼저 질소고정 균을 분리하기 위해, 질소원이 없는 배지에서 enrichment를 실시했고, 질소원이 제한된 배지에서 높은 성장을 보인 colony를 분리하여 순수분리 했다. 순수 분리된 균은 ARA를 통해 acetylene이 90% 이상 감소되고, ethylene 생성을 통해 nitrogenase의 활성을 간접적으로 확인했다. 재현성이 확인된 Cedecea sp. MK7과 Enterobacter sp. Y8을 선별했다. 선별된 질소고정 균을 perennial rye grass의 성장에 적용한 결과 건조중량이 18.65 mg인 대조군에 비해 34.80 mg (186.60%)으로 증가한 것을 확인했다. 식물 성장 후, 질소고정 균이 접종된 토양의 미생물 군집 분석은 대조군과 유사했다. 따라서 본 연구에서는 도시녹화 시스템에 질소고정 균을 이용하여 식물 성장을 촉진한다면 그 효율이 증대될 것이다.

• 한국미생물·생명공학회지
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• 제34권1호
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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.

• 미생물학회지
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• 제37권2호
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• pp.114-119
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• 2001

산림토양의 각 토양층으로부터 분리된 세균 중에는 통상 농도의 NB배지에서는 중식이 현저히 저해되고 희석한 DNB 배지에서만 중식이 가능한 세균이 다수 포 되어 있었으며, 이들 세균은 NB배지를 $10^{-1}$~$10^{-4}$배로 희석한 배지에서의 증식양상에 따라 4가지형으로 구분되었다. 본 연구에서는 저영양세균의 기준에 따라 $10^{-4}$NB(1 mg C/liter)배지에서 양호하게 증식하는 Type II와 IV세균을 저영양세균으로 분류하였고, 60개의 Type IV(편성저영양세균; obligate oligotrophic bacteria)균주를 순수분리 하였다. 이들 저영양성 세균중 질소고정능을 갖는 11균주에 대하여 화학분류 및 계통분류학적 특성을 검토한 결과 모든 균주는 주요 균체지방산으로 $C_{18:1}$)을, 퀴논종은 Q-10을 함유하였으며, G+C함량은 61-64 mol%범위를 나타내었다. 16S rDNA염기서열을 결정한 결과 각 균주는 Proteobacteria $\alpha$-subdivision의 BANA domain (Bradyrhizobium, Agromonas, Nitrobacter 및 Afipia)에 속하였고 Bradyrhizobium japonicum 및 Bradyrhizobium elkanii와 98% 이상의 높은 상동성을 나타내었다. 나타내었다.

• Environmental Engineering Research
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• 제12권4호
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• pp.136-147
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• 2007

In an effort to identify possible microbes for seeking bioagents for remediation of herbicide-contaminated soils, seven species of phototrophic nonsulfur bacteria (Rhodobacter capsulatus and sphaeroides, Rhodospirillum rubrum, Rhodopseudomonas acidophila, blastica and viridis, Rhodomicrobium vannielii) were grown in the presence of the herbicide, butachlor, and bacterial growth rates and nitrogen fixation were measured with different carbon sources. Under general conditions, all species showed 17-53% reductions in growth rate following butachlor treatment. Under nitrogen-fixing conditions, Rb. capsulatus and Rs. rubrum showed 1-4% increases in the growth rates and 2-10% increases in nitrogen-fixing abilities, while the other 5 species showed decreases of 17-47% and 17-85%, respectively. The finding that Rp. acidophila, Rp. blastica, Rp. viridis and Rm. vannielii showed stronger inhibitions of nitrogenase activity seems to indicate that species in genera Rhodobacter and Rhodospirillum are less influenced by butachlor than those in Rhodopseudomonas and Rhodomicrobium in terms of nitrogen-fixing ability. Overall, nitrogenase activity was closely correlated with both growth rate and glutamine synthetase activity (representing nitrogen metabolism). When the carbon sources were compared, pyruvate (three carbons) was best for all species in terms of growth rate and nitrogen fixation, with malate (four carbons) showing intermediate values and ribose(five carbons) showing the lowest; these trends did not change in response to butachlor treatment. We verified that each of the 7 species had a plasmid ($12.2{\sim}23.5\;Kb$). We found that all 7 species could use butachlor as a sole carbon source and 3 species were controlled by plasmid-born genes, but it is doubtful whether plasmid-born genes were responsible to nitrogen fixation.

• Journal of Plant Biology
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• 제32권3호
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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.

• 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.