• Title/Summary/Keyword: 16s rRNA 유전자

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Bacterial Stripe of Proso Millet Caused by Acidovorax avenae subsp. avenae in Korea (Acidovorax avenae subsp. avenae에 의한 기장 세균성줄무늬병)

  • Yoon, Young-Nam;Jung, Ji-Hun;Lee, Yeong-Hoon;Kim, Hyun-Joo;Bae, Soon-Do;Choi, Byeong-Ryeol;Nam, Min-Hee;Lee, Young-Kee
    • Research in Plant Disease
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    • v.18 no.3
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    • pp.236-239
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    • 2012
  • In July, 2009, proso millet (Panicum miliaceum), which showing the bacterial brown stripes on leaf sheaths, was collected in Miryang in Korea. Symptoms were systemic brown necrotic stripe lesions on the leaf sheaths and stems, and these symptoms were found in the entire field. The causal agent isolated from symptomatic plants was identified as an Acidovorax avenae subsp. avenae, based on its biochemical and physiological characteristics and also confirmed by the Biolog data and 16S rRNA gene sequence analysis. Also it caused hypersensitive response (HR) when it was inoculated onto the tobacco and tomato. It caused similar symptoms when inoculated onto proso millet. This is the first report of A. avenae subsp. avenae, the causal agent of bacterial brown stripe of the proso millet in Korea.

Characterization of Microbial Nitrate Uptake by Bacillus sp. PCE3 (Bacillus sp. PCE3 균주에 의한 질산이온 흡수 특성)

  • Yun, Yeong-Bae;Park, Soo-Jin;Han, Min-Woo;Kim, Young-Kee
    • Journal of Applied Biological Chemistry
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    • v.56 no.4
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    • pp.241-244
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    • 2013
  • Nitrate is one of the major nutrients in plants, and nitrate fertilizer often overused for the high yields of crops. Nitrate deposit in soil became one of the major reasons causing salt stress. Specially, salt stress is a serious problem in the soils of plastic film or glass houses. In this study, six microorganisms have been isolated from the wet soils near the disposals of livestock farms and their nitrate uptake activities were investigated. These bacteria were able to remove nitrate as high as 1,000-3,000 ppm (10-50 mM). The strain PCE3 showed the highest nitrate uptake activity and it removed more than 3,700 ppm. In order to identify these bacteria, genes of 16S rRNA were sequenced and analyzed. Phylogenetic trees were constructed with the neighbor-joining methods. Among these bacteria, strain PCE3 was identified as Bacillus species. When the growth and nitrate uptake activities were measured, both were maximal at $37^{\circ}C$ and optimal pH was pH 7-9. Bacillus sp. PCE3 removed nitrate up to 40-60 mM (2,500-3,700 ppm) depending on the nitrate concentration in media. Therefore, Bacillus sp. PCE3 can be a good candidate for the microbial remediation of nitrate-deposited soils in glass and plastic film houses.

Isolation and characterization of cellulolytic bacteria, Bacillus sp. EFL1, EFL2, and EFP3 from the mixed forest (혼효림으로부터 셀룰로오스분해 박테리아 분리 및 효소학적 특성규명)

  • Park, Hwa Rang;Oh, Ki-Cheol;Kim, Bong-Gyu
    • Journal of Applied Biological Chemistry
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    • v.61 no.1
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    • pp.59-67
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    • 2018
  • This study was conducted to isolate the cellulolytic bacteria able to grow on LB- Carboxymethyl cellulose (CMC) agar trypan blue medium from the mixed forest and Larix leptolepis stands. Three bacterial strains with high activity against both CMC and xylan were isolated. Both API kit test and 16S rRNA gene sequence analysis revealed that the three different isolates belong to the gene Bacillus. Therefore, the isolates named as Bacillus sp. EFL1, Bacillus sp. EFL2, and Bacillus sp. EFP3. The optimum growth temperature of Bacillus sp. EFL1, EFL2, and EFP3 were $37^{\circ}C$. The optimum temperature for CMCase and xylanase from Bacillus sp. EFL1 were $50^{\circ}C$. The optimum pH of Bacillus sp. EFL1 xylanase was pH 5.0 but the optimum pH of CMCase from Bacillus sp. EFL1 was pH 6.0. The optimum temperature of CMCase and xylanase from Bacillus sp. EFL2 was $60^{\circ}C$, respectively. The optimum pH of CMCase of Bacillus sp. EFL2 was 5.0, whereas xylanase showed high activity at pH 3.0-9.0. The optimum temperature for CMCase and xylanase of Bacillus sp. EFP3 was $50^{\circ}C$. The optimum pH for CMCase and xylanse was 5.0 and 4.0, respectively. CMCases from Bacillus sp. EFL1, EFL2, and EFP3 were thermally unstable. Although xylanase from Bacillus sp. EFL1 and EFP3 showed to be thermally unstable, xylanase from Bacillus sp. EFL2 showed to be thermally stable. Therefore, Bacillus sp. EFL2 has great potential for animal feed, biofuels, and food industry applications.

Isolation and Characterization of Soil Bacteria Degrading a Fungicide Defenoconazole (살균제 디페노코나졸 분해 세균 분리 및 특성 분석)

  • Ahn, Jae-Hyung;Ro, Yu-Mi;Lee, Gwan-Hyeong;Park, InCheol;Kim, Wan-Gyu;Han, Byeong-Hak;You, Jaehong
    • The Korean Journal of Pesticide Science
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    • v.20 no.4
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    • pp.349-354
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    • 2016
  • Triazole fungicides occupy an important portion in the global fungicide market and are relatively persistent in soil compared to the other fungicides, suggesting possible adverse effects of the fungicides on human health and environment. In this study, we tried to isolate microorganisms from orchard soils, which can decompose the triazole fungicides, tebuconazole, fluquinconazole, and difenoconazole. Only difenoconazole was completely degraded in the enrichment culture, from which several difenoconazole-degrading bacteria were isolated. They showed the same rep-PCR pattern thus only one strain, C8-2, was further studied. The strain was identified as Sphingomonas sp. C8-2 based on its 16S rRNA gene sequence and decomposed 100 mg/L of difenoconazole in a minimum medium to an unknown metabolite with a molecular weight of 296 within 24 hours. The inhibition effect of the metabolite against representative soil microorganisms significantly decreased compared to that of difenoconazole thus the bacterial strain is expected to be used for the detoxification of difenoconazole in soil and crop.

Optimization of γ-Aminobutyric Acid Production by Enterococcus faecium JK29 Isolated from a Traditional Fermented Foods (전통발효식품 유래 Enterococcus faecium JK29에 의한 γ-aminobutyric acid의 생산 최적화)

  • Lim, Hee Seon;Cha, In-Tae;Lee, Hyunjin;Seo, Myung-Ji
    • Microbiology and Biotechnology Letters
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    • v.44 no.1
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    • pp.26-33
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    • 2016
  • Dominant lactic acid bacteria (LAB) strains were isolated from traditional fermented foods to obtain rare ${\gamma}$-aminobutyric acid (GABA)-producing LAB. Out of 147 isolates, 23 strains that could produce GABA with 1% (w/v) L-monosodium glutamate (MSG) were first isolated. After further screening of these rare GABA-producing LAB by analysis of the glutamate decarboxylase and 16S rRNA gene sequences, Enterococcus faecium JK29 was isolated, and 1.56 mM of GABA was produced after 48 h cultivation in basic de Man, Rogosa, and Sharpe (MRS) medium. To enhance GABA production by E. faecium JK29, the culture conditions were optimized. When E. faecium JK29 was cultivated in optimized MRS medium containing 0.5% (w/v) sucrose and 2% (w/v) yeast extract with 0.5% (w/v) MSG, GABA production reached 14.86 mM after 48 h cultivation at initial conditions of pH 7.5 and $30^{\circ}C$.

Isolation and characterization of marine bacteria with alginate degrading activity (알긴산 분해능을 갖는 Pseudoalteromonas 및 Vibrio 속 해양세균들의 분리 및 특성분석)

  • Yoon, Young-Jun;Kim, Jung-Wan
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.364-373
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    • 2015
  • As an effort to utilize alginate, 103 bacterial isolates that were positive for the alginate lyase activity were isolated from various clams and seawater samples collected in Incheon coastal area. Among them, 3 strains (M1-2-1, M6-1, and C8-15) were finally selected for further analysis based on their activities at higher levels than others. These isolates were all Gram-negative and rod shaped halophilic bacteria with motility. According to their physiological and biochemical properties as well as DNA sequence of their 16S rRNA genes, M1-2-1 and M6-1 were identified as a member of genus Pseudoalteromonas and C8-15 belonged to genus Vibrio. They exhibited the alginate degrading activity at the maximal level when they were cultured in APY broth for 6-8 h at $25^{\circ}C$. Both their growth and the enzyme activity were greatly enhanced when NaCl was added to the growth medium. The crude alginate lyases from the supernatants of the bacterial cultures showed the highest activity at $45^{\circ}C$ and pH 7.0-8.0. M1-2-1 and M6-1 produced 2.723 and 1.976 g/L of reducing sugar from alginate, respectively, suggesting that they have potential for commercial application.

Isolation and characterization analysis of the halophilic archaea isolated from solar saltern, Gomso (곰소 염전에서 분리한 호염성 고세균의 특성 분석)

  • Koh, Hyeon-Woo;Kim, So-Jeong;Rhee, Sung-Keun;Park, Soo-Je
    • Korean Journal of Microbiology
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    • v.51 no.4
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    • pp.427-434
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    • 2015
  • Most of halophilic archaea are found in the various hypersaline environments including solar saltern, salt lake with very high salt concentration. The present study is about isolation and characterization of halphilic archaea from Gomso solar saltern known as a representative high salt environment in Korea. In order to isolate the halophilic archaea, we prepared and used high salt medium. Finally, total 7 strains obtained were tentatively identified based on comparative similarity analysis for 16S rRNA gene sequence and physiological traits. All halophilic archaea belonged to Haloruburm, Halogeometriucm, Halobacterium, and Haloarcula genera. These isolates were all Gram-staining negative, and growth was not observed using nitrate as an alternative electron acceptor under anaerobic conditions. In addition, all isolates required about 12-30% (w/v, NaCl) salt. This case study might provide basic information on microbial isolation technologies and related research in halophilic microorganisms from domestic halophilic environments, and contribute to obtaining useful indigenous halophilic archaea in a variety of extreme environmental conditions.

Microbial Diversity in Three-Stage Methane Production Process Using Food Waste (음식물 쓰레기를 이용한 3단계 메탄생산 공정의 미생물 다양성)

  • Nam, Ji-Hyun;Kim, Si-Wouk;Lee, Dong-Hun
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.125-133
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    • 2012
  • Anaerobic digestion is an alternative method to digest food wastes and to produce methane that can be used as a renewable energy source. We investigated bacterial and archaeal community structures in a three-stage methane production process using food wastes with concomitant wastewater treatment. The three-stage methane process is composed of semianaerobic hydrolysis/acidogenic, anaerobic acidogenic, and strictly anaerobic methane production steps in which food wastes are converted methane and carbon dioxide. The microbial diversity was determined by the nucleotide sequences of 16S rRNA gene library and quantitative real-time PCR. The major eubacterial population of the three-stage methane process was belonging to VFA-oxidizing bacteria. The archaeal community consisted mainly of two species of hydrogenotrophic methanogen (Methanoculleus). Family Picrophilaceae (Order Thermoplasmatales) was also observed as a minor population. The predominance of hydrogenotrophic methanogen suggests that the main degradation pathway of this process is different from the classical methane production systems that have the pathway based on acetogenesis. The domination of hydrogenotrophic methanogen (Methanoculleus) may be caused by mesophilic digestion, neutral pH, high concentration of ammonia, short HRT, and interaction with VFA-oxidizing bacteria (Tepidanaerobacter etc.).

Probiotic Properties and Immunomodulator Evaluation of the Potential Feed Additive Pediococcus acidilactici SRCM102607 (잠재적 사료첨가제로서 Pediococcus acidilactici SRCM102607의 생균제 특성 및 면역활성 효과)

  • Shin, Su-Jin;Ha, Gwangsu;Jeong, Su-Ji;Ryu, Myeong Seon;Kim, Jinwon;Yang, Hee-Jong;Kwak, Mi-Sun;Sung, Moon-Hee;Jeong, Do-Youn
    • Journal of Life Science
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    • v.30 no.10
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    • pp.896-904
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    • 2020
  • The purpose of this study was to investigate the probiotic characteristics and immune activities of selected lactic acid bacterial (LAB) strains as feed additives in livestock. 301 LAB strains isolated from traditional fermented foods were first assessed for their antibacterial activity potential. Of the 301 isolates, five showed antibacterial activity against five livestock pathogens (Esherichia coli KCCM11234, Listeria monocytogens KCTC3710, Salmonella Typhimurium KCTC1926, Staphylococcus aureus KCCM11593, and Shigella flexneri KCTC2517). The probiotic characteristics of the five selected strains were also investigated by antioxidative activity, hemolysis, bile salt hydrolase, acid resistance and bile tolerance. The SRCM102607 strain was found to have superior probiotic properties and was selected for further experimentation. 16S rRNA gene sequencing showed that SRCM102607 is Pediococcus acidilactici, which was labeled as P. acidilactici SRCM102607 (KCCM 12246P). The survival characteristics of P. acidilactici SRCM102607 in artificial gastrointestinal conditions were assessed under exposed acidic (pH 2.0) and bile (0.5% and 1.0%) conditions. P. acidilactici SRCM102607 was also confirmed to have resistance to various antibiotics, including amikacin, gentamicin, vancomycin, and etc. The TNF-α production by P. acidilactici SRCM102607 was 171.86±4.00 ng/ml. These results show that P. acidilactici RCM102607 has excellent potential for use as a probiotic livestock feed additive.

Characterization of Agarase from an Isolated Marine Bacterium, Simiduia sp. SH-1 (해양성 Simiduia sp. SH-1 균주의 분리 및 한천분해효소의 특성조사)

  • Lee, Sol-Ji;Oh, Soo-Jeong;Lee, Dong-Geun;Lee, Sang-Hyeon
    • Journal of Life Science
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    • v.25 no.11
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    • pp.1273-1279
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    • 2015
  • Agarase from a novel agar-degrading bacterium isolated from seawater in Namhae at Gyeongsangnamdo province of Korea was characterized. The SH-1 strain was selected from thousands of colonies on Marine agar 2216 media. Almost full 16S rRNA gene sequence of the agarolytic SH-1 strain showed 99% similarity with that of bacteria of Simiduia genus and named as Simiduia sp. SH-1. Agarase production was growth related, and activity was declined from stationary phase. Secreted agarase was prepared from culture media and characterized. It showed maximum activity of 698.6 units/L at pH 7.0 and 30℃ in 20 mM Tris-HCl buffer. Agarase activity decreased as the temperature increased from an optimum of 30℃, with 90% and 75% activity at 40℃ and 50℃, respectively. Agarase was not heat resistant. Slightly lower agarase activity was observed at pH 6.0 than at pH 7.0, without statistical difference, and 80% and 75% activity were observed at pH 5.0 and 8.0, respectively. Neoagarotetraose and neoagarobiose were the main final products of agarose, indicating that it is β-agarase. Simiduia sp. SH-1 and its β-agarase would be useful for the industrial production of neoagarotetraose and neoagarobiose, which have a whitening effect on skin, delaying starch degradation, and inhibiting bacterial growth.