• Title/Summary/Keyword: 16s rRNA sequence

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Isolation, characterization, and phylogenetic position of a new sulfur-oxidizing bacterium

  • Chang, So Youn;Yoon, Joon Sik;Park, Yong Ha;Yang, Song Suk;Yoon, Seong Myeong;Lee, In Hwa;Kim, Si Wouk
    • Journal of Microbiology
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    • v.35 no.3
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    • pp.165-171
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    • 1997
  • A sulfer-oxidizing bacterium was isolated from mine wastewater and characterized. The isolate was gra-negative, rod (0.2 * 1.2-1.5.mu.m), nonmotiloe, catalase positive, and oxidase prositive. The opotimal pH and temperature for growth were 7.0 and 30.deg.C. respectively. The optimum thiosulfate concentration was 70 mM and the maximum growth rate was 0.081 hr. The major ubiquinone contained in the isolate was Q-8. The cellular fatty acid composition was $C_{16 : 0}$, $C_{18 : 1}$, $C_{17cyc}$,and $C_{19cyc}$ as nonpolar fatty acids, and 3-OH C10 : 0 and 3-OH $C_{12 : 0}$ as hydroxylated fatty acids. The isolate was a facultative chemolithoautotroph which can grow autotrophically on sodium thiosulfate and sodium sulfide and which can grow heterotrophically on yeast extract. It can also grow mixotrophically on sodium thiosulfate and yeast extract. Comparison of the 16S rRNA gene sequence of the isolate with that of Thiobacillus species and Paracoccus thiocyanatus revealed that it is closely related to T. caldus which belongs to the .betha.-subclass of the class Proteobacteria. However, the isolated could not grow at extremely low pH (pH 1-3.5). On the basis of the phenotypic, chemotaxonomic and phylogenetic data, the isolate was tentatively named Thiobacillus sp. strain C.ain C.

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Isolation and Optimal Culture Conditions of Prodigiosin-like Pigment Produced by Zooshikella sp. JE-34 (Zooshikella sp. JE-34가 생산하는 Prodigiosin계 색소의 분리 및 최적 배양조건)

  • Kim, Ju-Sang;Kim, Man-Chul;Lee, Kyeong-Jun;Heo, Moon-Soo
    • Microbiology and Biotechnology Letters
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    • v.37 no.3
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    • pp.219-225
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    • 2009
  • A bacterial strain, JE-34, producing a high concentration of red pigment was isolated from a sediment in East China Sea. It was identified as Zooshikella sp. JE-34 based on the 16S rRNA gene sequence analysis. The red pigment was purified by solvent extraction and HPLC was identified as prodigiosin-like compound. Nutritional and cultural conditions were optimized for the production of prodigiosin-like pigment in the flask level. Optimal culture conditions were at initial medium pH $6.0{\sim}7.0$, $30^{\circ}C$ and 4 days incubation. For carbon and, nitrogen sources were soluble starch and malt extract.

Selection and immunomodulatory evaluation of lactic acid bacteria suitable for use as canine probiotics (개 생균제 사용에 적합한 유산균주의 선발 및 면역활성 평가)

  • Park, Su-Min;Park, Ho-Eun;Lee, Wan-Kyu
    • Korean Journal of Veterinary Research
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    • v.55 no.2
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    • pp.81-88
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    • 2015
  • This study was conducted to isolate lactic acid bacteria (LAB) from dog intestine and identify potential probiotic strains for canine use. One hundred and one LAB were isolated from feces of 20 healthy dogs. Acid, bile, and heat resistance along with adherence to Caco-2 cells and antimicrobial activity against pathogens were examined. To analyze immunomodulative effects, the production of nitric oxide (NO), TNF-${\alpha}$, and IL-$1{\beta}$ was measured using RAW 264.7 macrophages. Additionally, RAW BLUE cells were used to evaluate nuclear factor-${\kappa}B$ (NF-${\kappa}B$) generation. Ultimately, three strains were selected as canine probiotics and identified as Lactobacillus reuteri L10, Enterococcus faecium S33, and Bifidobacterium longum B3 by 16S rRNA sequence analysis. The L10 and S33 strains showed tolerance to pH 2.5 for 2 h, 1.0% Oxgall for 2 h, and $60^{\circ}C$ for 5 min. These strains also had strong antimicrobial activity against Escherichia coli KCTC 1682, Salmonella Enteritidis KCCM 12021, Staphylococcus aureus KCTC 1621, and Listeria monocytogenes KCTC 3569. All three strains exerted better immunomodulatory effects than Lactobacillus rhamnosus GG (LGG), a well-known commercial immunomodulatory strain, based on NO, NF-${\kappa}B$, IL-$1{\beta}$, and TNF-${\alpha}$ production. These results suggested that the three selected strains could serve as canine probiotics.

Development of Molecular Markers for the authentication of Zanthoxyli Pericarpium by the analysis of rDNA-ITS DNA barcode regions (rDNA-ITS DNA 바코드 부위 분석을 통한 산초(山椒) 기원종 감별용 유전자 마커 개발)

  • Kim, Wook Jin;Ji, Yunui;Lee, Young Mi;Kang, Young Min;Choi, Goya;Moon, Byeong Cheol
    • The Korea Journal of Herbology
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    • v.30 no.3
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    • pp.41-47
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    • 2015
  • Objectives : Due to the morphological similarity of the pericarp and description of multi-species in National Pharmacopoeia of Korea and China, the Zanthoxylum Pericarpium is difficult to authenticate adulterant in species levels. Therefore, we introduced the sequence analysis of DNA barcode and identification of single nucleotide polymorphism(SNP) to establish a reliable tool for the distinction of Zanthoxylum Pericarpium from its adulterants. Methods : To analyze DNA barcode region, genomic DNA was extracted from twenty-four specimens of authentic Zanthoxylum species and inauthentic adulterant and the individual internal transcribed spacer regions (rDNA-ITS and ITS2) of nuclear ribosomal RNA gene were amplified using ITS1, ITS2-S2F, and ITS4 primer. For identification of species-specific sequences, a comparative analysis was performed using entire DNA barcode sequences. Results : In comparison of four Zanthoxylum ITS2 sequences, we identified 16, 4, 6, and 4 distinct species-specific nucleotides enough to distinguish Z. schinifolium, Z. bungeanum, Z. piperitum, and Z. simulans, respectively. The sequence differences were available genetic marker to discriminate four species. Futhermore, phylogenetic relationship revealed a clear classification between different Zanthoxylum species showing 4 different clusters. These results indicated that comparative analysis of ITS2 DNA barcode was an useful genetic marker to authenticate Zanthoxylum Pericarpium in species levels. Conclusions : The marker nucleotides, enough to distinguish Z. schinifolium, Z. piperitum, Z. bungeanum, and Z. simulans, were obtained at 30 SNP marker nucleotides from ITS2 sequences. These differences could be used to authenticate official Zanthoxylum Pericarpium from its adulterants as well as discriminating each four species.

Isolation and Identification of Stenotrophomonas maltophilia BW-13 Active Against Rhizoctonia solani Causing Crisphead Lettuce Bottom Rot (Rhizoctonia solani에 의한 결구상추 밑둥썩음병 방제균주 Stenotrophomonas maltophilia BW-13의 분리 및 동정)

  • Kim Han-Woo;Park Jong-Young;Kim Hyun-Ju;Lee Kwang-Youll;Lee Jin-Woo;Choi Woobong;Lee Seon-Woo;Moon Byung-Ju
    • Research in Plant Disease
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    • v.11 no.2
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    • pp.152-157
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    • 2005
  • In a course of searching for biofungicide to control crisphead lettuce bottom rot caused by Rhizoctonia solani, we have isolated an antagonistic bacterium from lettuce rhisophere soil. A total of 702 bacterial isolates were isolated and tested for in vitro growth inhibition of R. solani. Seven strains appeared to have strong antagonistic effect against R. solani in in vitro growth inhibition assay. In the pot experiments, a strain BW-13 showed the most potent disease control effect on the both lettuce seedlings and adults plants. Therefore, the BW-13 was selected as a biocotrol candidate against crisphead lettuce bottom rot. Based on its morphology, physiological characteristics, and 165 rRNA gene analysis, the BW-13 was finally identified as Stenotrophomonas maltophilia. This study indicated that S. maltophilia BW-13 could be used as a biocontrol agent to control crisphead lettuce bottom rot.

The Use of the Pathogen-specific Bacteriophage BCP8-2 to Develop a Rice Straw-derived Bacillus cereus-free Starter Culture (단일 박테리오파지를 이용한 볏짚 유래 Bacillus cereus free 스타터 컬쳐의 개발)

  • Bandara, Nadeeka;Chung, Seo-Jin;Jeong, Do-Youn;Kim, Kwang-Pyo
    • Korean Journal of Food Science and Technology
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    • v.46 no.1
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    • pp.115-120
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    • 2014
  • The purpose of this study was to develop a rice straw-derived Bacillus cereus (B. cereus)-free starter culture for traditional soybean fermented products using a B. cereus-specific bacteriophage, BCP8-2. To determine the optimal medium that supports the growth of rice straw-derived microorganisms and BCP8-2 activity, 5 different culture media were tested. The 5% ground bean (GB) medium was selected for further study. No B. cereus was detected in the BCP8-2-treated rice straw in GB medium, whereas B. cereus at a level of $10^7$ CFU/mL was recovered in the no-phage control. The total bacterial count reached approximately $10^9$ CFU/mL regardless of phage addition. When the 16S rRNA sequence-based microbial community was monitored using denaturing gradient gel electrophoresis (DGGE) and pyrosequencing, a similar microbial community was observed in the phage-treated and control samples. In conclusion, we demonstrate that phage can be used to prepare a rice straw-derived B. cereus-free starter culture with minimal effect on natural microflora.

Phylogenetic Diversity and Antibacterial Activity in Bacterium from Balloon Fish (Diodon holocanthus) of Jeju Island (제주 연안의 가시복(Diodon holoanthus)에서 분리된 세균의 다양성 및 항균활성 효과)

  • Moon, Chae-Yun;Ko, Jun-Cheol;Kim, Min-Seon;Heo, Moon-Soo
    • Microbiology and Biotechnology Letters
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    • v.48 no.1
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    • pp.57-63
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    • 2020
  • Over the past 20 years, global warming has transformed the marine ecosystem of the Jeju Island into a subtropical zone making it conducive to the production of tropical fishes. Recently, the balloon fish (Diodon holoanthus) has been found off the coast of the Jeju Island. In this study, we analyzed the diversity of its intestinal microorganisms as a representative for the surrounding environment. In addition, the isolates were evaluated for their antibacterial activity. A total of 161 strains of various species were identified and isolated using 16S ribosomal RNA gene sequence analysis. They were separated into three groups, of which Phylum Proteobacteria was found to be the most dominant with 91% sequence similarity. This includes the class γ-proteobacteria that is made up of twelve genera and twenty-four hundred species. The second group comprised strains of the genus Vibrio, made up of 35% Photobacteria, 32% Shewanella, and 6% Psychrobacter. It was also determined that 4% of the isolates were Acinetobacter, 3% were Enterovibrio, while Moraxella_g2 accounted for 1% of the total isolates. Class α-proteobactera includes five genera and five species; Brevundimonas, Allorhizobium, Pseudoceanicola and Erythrobcter, each accounting for 1% of the total isolates. The Firmicute strains belonged to six genera and ten species. 5% of the strains were Terribacillus, while Paenibacillus, Salinicoccus, Staphylococcus and Streptococcus accounted for 1% each of the total isolates. Actinobacteria accounted for the final phylum with strains belonging to three genera and ten species with Janibacter, Micrococcus and Isoptericola each accounting for 1% of the total isolates.

Paenibacillus donghaensis sp. nov., a Xylan-degrading and Nitrogen-fixing Bacterium Isolated from East Sea Sediment

  • Choi, Jeong-Hwa;Im, Wan-Taek;Yoo, Jae-Soo;Lee, Sang-Mahn;Moon, Deok-Soo;Kim, Hyeon-Ju;Rhee, Sung-Keun;Roh, Dong-Hyun
    • Journal of Microbiology and Biotechnology
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    • v.18 no.2
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    • pp.189-193
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    • 2008
  • A Gram-positive and endospore-forming strain, $JH8^T$, was isolated from deep-sea sediment and identified as a member of the genus Paenibacillus on the basis of 16S rRNA gene sequence and phenotypic analyses. According to a phylogenetic analysis, the most closely related species was Paenibacillus wynnii LMG $22176^T$ (96.9%). Strain $JH8^T$ was also facultatively anaerobic and grew optimally at $20-25^{\circ}C$. The major cellular fatty acid was anteiso-$C_{15:0}$, and the DNA G+C content was 53.1mol%. The DNA-DNA relatedness between the isolate and Paenibacillus wynnii LMG $22176^T$ was 7.6%, indicating that strain $JH8^T$ and P. wynnii belong to different species. Based on the phylogenetic, phenotypic, and chemotaxonomic characteristics, strain $JH8^T$ would appear to belong to a novel species, for which the name Paenibacillus donghaensis sp. novo is proposed (type strain=KCTC $13049^T=LMG\;237S0^T$).

Production and Characterization of Antifungal Chitinase of Bacillus licheniformis Isolated from Yellow Loess (황토로부터 분리한 Bacillus licheniformis의 항진균 chitinase 생산과 효소 특성)

  • Han, Gui Hwan;Bong, Ki Moon;Kim, Jong Min;Kim, Pyoung Il;Kim, Si Wouk
    • KSBB Journal
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    • v.29 no.3
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    • pp.131-138
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    • 2014
  • In this study, we isolated two novel chitinase producing bacterial strains from yellow loess samples collected from Jullanamdo province. The chitinase producing bacteria were isolated based on the zone size of clearance in the chitin agar plates. Both of them were gram positive, rod ($2{\sim}3{\times}0.3{\sim}0.4{\mu}m$), spore-forming, and motility positive. They were facultative anaerobic, catalase positive and hydrolyzed starch, gelatin, and casein. From the 16s rRNA gene sequence analysis, the isolates were labeled as Bacillus licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02. The isolates showed higher extracellular chitinase activities than B. licheniformis ATCC 14580 as a control. The optimum temperature and pH for chitinase production were $40^{\circ}C$ and pH 7.0, respectively. Response Surface Methodology (RSM) was used to optimize the culture medium for efficient production of the chitinase. Under this optimal condition, 1.5 times higher chitinase activity of B. licheniformis KYLS-CU02 was obtained. Extracellular chitinases of the two isolates were purified through ammonium sulfate precipitation and anion-exchange DEAE-cellulose column chromatography. The specific activities of purified chitinase from B. licheniformis KYLS-CU01 and B. licheniformis KYLS-CU02 were 7.65 and 5.21 U/mg protein, respectively. The molecular weights of the two purified chitinases were 59 kDa. Further, the purified chitinase of B. licheniformis KYLS-CU01 showed high antifungal activity against Fusarium sp.. In conclusion, these two bacterial isolates can be used as a biopesticide to control pathogenic fungi.

Biological Efficacy of Streptomyces sp. Strain BN1 against the Cereal Head Blight Pathogen Fusarium graminearum

  • Jung, Boknam;Park, Sook-Young;Lee, Yin-Won;Lee, Jungkwan
    • The Plant Pathology Journal
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    • v.29 no.1
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    • pp.52-58
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    • 2013
  • Fusarium head blight (FHB) caused by the filamentous fungus Fusarium graminearum is one of the most severe diseases threatening the production of small grains. Infected grains are often contaminated with mycotoxins such as zearalenone and trichothecences. During survey of contamination by FHB in rice grains, we found a bacterial isolate, designated as BN1, antagonistic to F. graminearum. The strain BN1 had branching vegetative hyphae and spores, and its aerial hyphae often had long, straight filaments bearing spores. The 16S rRNA gene of BN1 had 100% sequence identity with those found in several Streptomyces species. Phylogenetic analysis of ITS regions showed that BN1 grouped with S. sampsonii with 77% bootstrap value, suggesting that BN1 was not a known Streptomyces species. In addition, the efficacy of the BN1 strain against F. graminearum strains was tested both in vitro and in vivo. Wheat seedling length was significantly decreased by F. graminearum infection. However, this effect was mitigated when wheat seeds were treated with BN1 spore suspension prior to F. graminearum infection. BN1 also significantly decreased FHB severity when it was sprayed onto wheat heads, whereas BN1 was not effective when wheat heads were point inoculated. These results suggest that spraying of BN1 spores onto wheat heads during the wheat flowering season can be efficient for plant protection. Mechanistic studies on the antagonistic effect of BN1 against F. graminearum remain to be analyzed.