• Title/Summary/Keyword: DNB세균

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Study on the Improvement of Soil for High Efficient and Sustainable Agriculture-II Changes of Population of Soil Microorganisms in the Fertilized Soil with Organic Materials (지속적 농업을 위한 고성능토양의 개발 연구-II 유기질비료 시용에 따른 토양미생물상의 변화)

  • Yoo, Sung-Joon;Chang, Ki-Woon;Whang, Kyung-Sook
    • Applied Biological Chemistry
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    • v.41 no.6
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    • pp.457-464
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    • 1998
  • For investigating the effect of amount of applied fertilizer with pig and chicken manures in the field, the number of soil bacteria were counted on the full strength conventional nutrient broth (NB) medium and its 100-fold dilution (DNB) medium. From the control soil samples without organic amendments, the number of bacteria on DNB medium was 5 to 10 times higher than that on NB medium. However, population density on NB medium was higher than on DNB medium from the treated soil with 60 and 120 Mg/㏊ for 3 years. Most of isolates from DNB plates did not grow on the NB plates. There were only grown on the 100-fold dilution medium, so it was called as DNB organisms. The DNB organisms were occupied as dominant group over 60% fo isolates in control soil. However, their occupation rates were rapidly decreased in the treatment soil with pig and chicken manures above 60 and 120 Mg per ha. These DNB organisms (oligotrophs) were significantly low population desities in the treatment soil with organic materials.

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Phylogenetic Analysis of Oligotrophic Bacteria Found in Potable Groundwater (음용 지하수중에 분포하는 저영양세균의 계통학적 해석)

  • ;Tomoyoshi Hashimoto
    • Korean Journal of Microbiology
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    • v.38 no.4
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    • pp.293-298
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    • 2002
  • In order to investigate the ecological aspect of bacteria on groundwater, water samples were collected from various regions. Total of 318 strains were isolated from diluted nutrient broth (DNB) agar medium, and investigated their growth pattern on nutrient broth (NB) medium. As a result, all the isolated strains were divided into two groups, NB and DNB organisms. Growth of DNB organisms were suppressed in full strength NB medium but not in DNB medium, which were called oligotrophic bacteria in this study. Proportion of DNB organisms occurred in the frequency of 50-98% in potable groundwaters (CW, CJ, DPG, CJG1), however, it was 23,46% in polluted site (TJ, NPG1). One hundred and two strains were identified as oligotrophic bacteria and their phylogenetic characteristics were determined by using 16S rDNA sequencing. Based on the phylogenetic analysis, they were found to fall into three major phylogenetic groups: belonging to the Proteobacteria $\alpha$-(49 strains), $\beta$-(50 strains), $\gamma$ -(3 strains) subdivisions. The phylogenetic analysis suggested that microbial diversity of potable groundwater is more complex than that obtained in the past investigation.

The Detection and a Quantitative Evaluation of Viable but Non-Culturable Soil Bacteria Using a Modified Direct Viable Count Method (변형된 DVC법을 이용한 난배양성 토양세균의 검출 및 정량적 평가)

  • 황경숙;양희찬;염곡효
    • Korean Journal of Microbiology
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    • v.39 no.3
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    • pp.181-186
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    • 2003
  • This study was performed to analyze quantitatively the number of living bacteria in forest soil samples collected from Mt. Keryong using improved direct viable count (DVC) and plate count (PC) methods. The number of living bacteria by DVC comprised 18~44% of the total direct count (TDC), whereas the number of living bacteria by PC was less than 1% of TDC. These results showed that viable but non-culturable (VBNC) bacteria existed in the soil with high percentages. Besides, DVC was proved to make it possible to make a quantitative detection of the VBNC bacteria. On the other hand, upon measuring the value from the conventional nutrient broth (NB) and $10^{-2}$ folded diluted nutrient broth (DNB), the values from the DNB showed 5 to 10 times higher than those from the conventional NB medium. These results indicate that oligotrophic bacterial groups, which could multiply in the low nutrient broth, abundantly exist in the soil ecosystem. It would also be possible to apply this kind of method to other substrate to make a quantitative detection of soil bacterial groups.

Effect of Organic Amendments on Rhizosphere Microflora of Tomato Plant (유기질 비료 시용이 토마토 근권 미생물 상에 미치는 영향)

  • Yoo, Sung-Joon;Whang, Kyung-Sook;Kim, Sun-Ik;Chang, Ki-Woon
    • Korean Journal of Soil Science and Fertilizer
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    • v.29 no.3
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    • pp.297-302
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    • 1996
  • The effect of application rate of pig or chicken manure on the seasonal change of tomato(Minicarol) rhizosphere microflora was investigated by field experiment were surface soil(16cm) mixed with manures 1 weeks before transplanting. 1. Rhizobacteria population of control plot decreased 5 weeks after transplanting(WAT) than 1 WAT and 1.5~2 times higher colony counting was found in DNB(Diluted nutrient broth medium) than NB(Nutrient broth medium) at 15 WAT. 2. Rhizobacteria population at 1 WAT inereased in NB medium with the application rate of both manures but decreased in DNB with chicken manure. Colony counting in NB of 60 and 120 mg/ha treatment was 100 times higher than that of the control. However, rhizobacteria in DNB at 15 WAT(harvest stage) was much higher than that of NB. 3. Application rate did not affect fungi population 1 WAT in pig manure but decresed in chicken manure. At 15 WAT fungi population was 2 times higher than 1 WAT with chieken manure and highest in 30 mg/ha but with pig manure decreased with rate than the control. 4. Actinomycetes population at 1WAT was not different among pig manure rates and decreased with chicken manure than control. At 15 WAT population increased in all manure rates. especially in 10 mg/ha pig manure and 30 mg/ha chicken manure 4 times that 1 WAT.

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The Observation and a Quantitative Evaluation of Viable but Non-Culturable Bacteria in Potable Groundwater Using Epifluorescence Microscopy (형광현미경을 이용한 음용 지하수내 배양불능 세균의 관찰 및 정량적 평가)

  • ;Takashi Someya
    • Korean Journal of Microbiology
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    • v.38 no.3
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    • pp.180-185
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    • 2002
  • The direct viable count (DVC) and plate count (PC) methods was used to measure the number of bacteria in potable groundwater samples collected from bottled water from the market, mineral water, and edible groundwater near the urban areas and the stock farming congested areas. As a result, the number of living bacteria by DVC was comprised 30~80% of the total direct count (TDC), whereas the number of living bacteria by PC was around l~30% of DVC. Such results show that viable but non-culturable (VBNC) bacteria exist in the potable groundwater with high percentages. On the other hand, upon measuring the value from the conventional nutrient broth (NB), $10^-2$ fold diluted nutrient broth (DNB), and R2A broth, the values from the DNB and R2A showed 2~50 times higher than the conventional NB medium. These results indicate that oligotrophic bacterial groups which can multiply in the low nutrient broth abundantly exist in the oligotrophic environment like potable groundwater.

Taxonomic Characteristics of Nitrogen-Fixing Oligotrophic Bacteria from Forest Soil (산림토양으로부터 분리한 저영양성-질소고정세균의 분류학적 특성)

  • 황경숙
    • 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.

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Phylogenetic Characteristics of viable but Nonculturable Bacterial Populations in a Pine Mushroom (Tricholoma matsutake) Forest Soil (송이 자생군락 토양 내 난배양성 세균군집의 계통학적 특성)

  • Kim, Yun-Ji;Whang, Kyung-Sook
    • Korean Journal of Microbiology
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    • v.43 no.3
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    • pp.201-209
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    • 2007
  • The CFDA (6-carboxyfluorescein diacetate) direct viable count method and plate count (PC) method using conventional nutrient broth (NB) medium and $10^{-2}$ diluted NB (DNB) medium were applied to samples collected from Mt. Yongdoo In Andong, in an effect to determine the number of living bacteria pine mushroom forest soil. The number of living bacteria determined via plate count in NB medium comprised $5{\sim}8%$ of the CFDA direct viable count, and the bacteria in the DNB medium comprised $40{\sim}47%$. This result indicated that viable but nonculturable (VBNC) bacteria existed in the pine mushroom forest soil at a high percentage. The phylogenetic characteristics of the VBNC bacterial populations in the samples of pine mushroom (Tricholoma matsutake) forest soil were analyzed via the direct extraction of DNA and 16S rDNA-ARDRA. The 115 clones from pine mushroom forest soil were clustered into 31 different RFLP phylotypes by ARDRA. Based on the 16S rDNA sequences, the 31 ARDRA clusters were classified into 6 phylogenetic groups: ${\alpha}-,\;{\beta}-,\;{\gamma}-Proteobacteria$, Acidobacteria, Actinobacteria and Firmicutes. Among these bacterial populations, approximately 85% were classified as members of phylum Acidobacteria. The Acidobacteria phylum was shown to exist abundantly in the pine mushroom forest soil.

Isolation and Phylogenetic Characterization of Chitinase Producing Oligotrophic Bacteria (Chitinase생산 저영양세균의 분리 및 계통분류학적 특성)

  • Kim Soo-Jin;Kim Min-Young;Koo Bon-Sung;Yoon San-Hong;Yeo Yun-Soo;Park In-Cheol;Kim Yoon-Ji;Lee Jong-Wha;Whang Kyung-Sook
    • Korean Journal of Microbiology
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    • v.41 no.4
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    • pp.293-299
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    • 2005
  • Many isolates from soil of Korean ginseng rhizosphere did not show remarkable growth on full strength of the conventional nutrient broth (NB medium) but grew on its 100-fold dilution (DNB medium). Six hundred-forty strains were isolated as oligotrophic bacteria. In the course of screening for new bioactive compounds from oligotrophic bacteria from soil, 8 strains which had appeared to form of clear zone on a medium containing colloidal chitin as a sole carbon source were selected for further studies. Strain CR42 hydrolyzed a fluorogenic analogue of chitin, 4-methylumbelliferyl-D-glucosaminide (MUF-NAG) . Mo st of the culture supernatant of these isolates hydrolyzed 4-methylumbelliferyl-D-N,N'-diacetylchitobioside (MUF-diNAG). The isolates were heterogeneous and categorized to gamma- and beta-proteobacteria, Bacillaceae, Actinobactepia, and Bacteroides by 16S rRNA analysis. Two strains, WR164 and CR18, had a 16S rRNA sequence of $95-96\%$ identical to uncultured bacteria. It was observed that CR2 and CR75 could inhibit the growth of Colletotrichum gloeosporioides with hyphal extention-inhibition assay on PDA plate supplemented with $1\%$ colloidal chitin.