• Microbiology and Biotechnology Letters
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• v.38 no.3
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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.

• Microbiology and Biotechnology Letters
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• v.37 no.3
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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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• v.34 no.3
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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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• v.31 no.1
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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.

• 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.

• Korean Journal of Microbiology
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• v.37 no.2
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• pp.114-119
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• 2001

Many isolates from different forest soil layers did not show appreciable growth on full strength of the conventional nutrient broth (NB medium) but grow on its 100-fold dilution (DNB medium). These isolates were divided into four types according to organic nutrient concentration in the growth medium from $1^{-1}\;to\;10^{-4}$dilution of normal NB medium. Oligotrophic bacteria were type II and type IV which grew in $10^{-4}$ dilution of NB (1 mg C/l) medium. Sixty strains were isolated for obligate oligotrophic bacteria. Chemotaxonomic and phylogenetic characteristics of eleven isolates of acetylene-reducing (nitrogen-fixing) oligotrophic bacteria from forest soil were investigated. They showed similar characteristics: the cellular fatty acid mainly consisted of straight-chain unsaturated $C_{18:1}$ (60-84% of total fatty acids). Ubiquinone Q-10 and a high guanine plus-cytosine content(61-64 mol%) were found. Eleven isolates of nitrogen-fixing oligotrophic bacteria were found to be closely related by full 16S rDNA sequence simility and many common taxonomic traits. Analysis of full 16S rDNA sequences of eleven isolates indicated that they were more closely related to Bradyrhizobium (similarity values: 98.1-98.8%), Agromonas, Nitrobacter, and Afipia.

• Environmental Engineering Research
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• v.12 no.4
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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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• 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.

• Microbiology and Biotechnology Letters
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• v.13 no.3
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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.