• Title/Summary/Keyword: rRNA genes

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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 Actinomycete Strain BK185 Possessing Antifungal Activity against Ginseng Root Rot Pathogens (인삼 뿌리썩음병균에 항균활성이 있는 방선균 BK185의 분리 및 특성)

  • Kim, Byung-Yong;Bae, Mun-Hyung;Ahn, Jae-Hyung;Weon, Hang-Yeon;Kim, Sung-Il;Kim, Wan-Kyu;Oh, Dong-Chan;Song, Jaekyeong
    • The Korean Journal of Pesticide Science
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    • v.18 no.4
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    • pp.396-403
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    • 2014
  • Ginseng (Panax ginseng C. A. Meyer) is an economically valuable pharmaceutical crop in Korea. In order to find promising biocontrol agents for soil-borne fungal pathogens which infect ginseng roots, we have isolated actinomycete, BK185 from soil. The isolate was investigated for the antifungal activity against to ginseng rot pathogens prior to testing genetic and chemical properties. The strain was identified as Streptomyces sp. using phylogenetic analysis based on 16S rRNA gene sequence. The most closely related species was S. sporoclivatus and S. geldanamycininus with high similarities (>99%). The isolate, BK185 showed positive reaction for PCR detection targeting biosynthetic gene clusters of PKS (Type-I polyketide synthase) and NRPS (Non-ribosomal polypeptide synthetase) genes. Major metabolite from the BK185 was analyzed by The LC/MS and identified to geldamycin, which was known to contained broad antibacterial, antifungal or anticancer activities. The results provide evidences that the strain, BK185 can be promising biocontrol agent for ginseng organic farming.

Characterization of Multifunctional Bacillus sp. GH1-13 (복합기능성 Bacillus sp. GH1-13 균주의 특징)

  • Kim, Sang Yoon;Sang, Mee Kyung;Weon, Hang-Yeon;Jeon, Young-Ah;Ryoo, Jae Hwan;Song, Jaekyeong
    • The Korean Journal of Pesticide Science
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    • v.20 no.3
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    • pp.189-196
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    • 2016
  • Several microorganisms in particular Bacillus subtilis group have been isolated from diverse places such as soils and the gastrointestinal tract of ruminants etc., and used as biocontrol agent against various plant pathogens and utilized as plant growth promoting agents. Among them, Bacillus is well known as one of the most useful bacteria for biocontrol and plant growth promotion. Bacterium GH1-13 was isolated from a reclaimed paddy field in Wando Island and identified as Bacillus velezensis using phylogenetic analysis on the basis of 16S rRNA and gyrB gene. It was confirmed that GH1-13 produced indole acetic acid (IAA) associated with promoted growth of rice root. GH1-13 showed characteristics of antagonization against the main pathogen of rice as well as diverse pathogenic fungi. GH1-13 had biosynthetic genes, bacillomycin, bacilycin, fengycin, iturin, and surfactin which are considered to be associated closely with inhibition of growth of pathogenic fungi and bacteria. This study showed that GH1-13 could be used as a multifunctional agent for biocontrol and growth promotion of crop.

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.

Antimicrobial and Biogenic Amine-Degrading Activity of Bacillus licheniformis SCK B11 Isolated from Traditionally Fermented Red Pepper Paste (전통고추장에서 유해균 억제 및 Biogenic Amines 분해 능력을 가지는 Bacillus licheniformis SCK B11의 분리)

  • Kim, Yong-Sang;Jeong, Jin-Oh;Cho, Sung-Ho;Jeong, Do-Yeon;Uhm, Tai-Boong
    • Korean Journal of Microbiology
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    • v.48 no.2
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    • pp.163-170
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    • 2012
  • In order to inhibit the growth of pathogens and degrade biogenic amines during the fermentation of soybean products, an isolate with antimicrobial activity against pathogens and biogenic amine-degrading property was obtained from 83 traditionally fermented soybean products. The morphological and biochemical tests and the phylogenetic relationship among 16S rRNA gene sequences indicated that the isolate named as the strain SCK B11 was most closely related to Bacillus licheniformis. The cell-free supernatant of two day cultures was active against several pathogens including Enterococcus faecalis, Listeria monocytosis, Micrococcus luteus, Pseudomonas aeruginosa, Bacillus cereus, and Staphylococcus aureus. PCR analysis was conducted to determine relatedness to antimicrobial lantibiotics and biosurfactants produced by Bacillus spp., but showed negative for the genes encoding surfactin, lichenysin, and lichenicidine. Electron microscopic observation indicated that the antimicrobial agent seemed to attack the membrane of the pathogens, leaving the ghost or shrunken cells. The strain was found to degrade histamine by 72% and tyramine by 66% in the cooked soybean containing 5.3% of biogenic amine over 10 days of fermentation time. The use of selected strain would be a potential control measure in manufacturing traditionally fermented soybean products that are difficult to control pathogens and biogenic amine levels.

Comparative Genomic Analysis of Staphylococcus aureus FORC_001 and S. aureus MRSA252 Reveals the Characteristics of Antibiotic Resistance and Virulence Factors for Human Infection

  • Lim, Sooyeon;Lee, Dong-Hoon;Kwak, Woori;Shin, Hakdong;Ku, Hye-Jin;Lee, Jong-eun;Lee, Gun Eui;Kim, Heebal;Choi, Sang-Ho;Ryu, Sangryeol;Lee, Ju-Hoon
    • Journal of Microbiology and Biotechnology
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    • v.25 no.1
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    • pp.98-108
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    • 2015
  • Staphylococcus aureus is an important foodborne pathogen that causes diverse diseases ranging from minor infections to life-threatening conditions in humans and animals. To further understand its pathogenesis, the genome of the strain S. aureus FORC_001 was isolated from a contaminated food. Its genome consists of 2,886,017 bp double-stranded DNA with a GC content of 32.8%. It is predicted to contain 2,728 open reading frames, 57 tRNAs, and 6 rRNA operons, including 1 additional 5S rRNA gene. Comparative phylogenetic tree analysis of 40 complete S. aureus genome sequences using average nucleotide identity (ANI) revealed that strain FORC_001 belonged to Group I. The closest phylogenetic match was S. aureus MRSA252, according to a whole-genome ANI (99.87%), suggesting that they might share a common ancestor. Comparative genome analysis of FORC_001 and MRSA252 revealed two non-homologous regions: Regions I and II. The presence of various antibiotic resistance genes, including the SCCmec cluster in Region I of MRSA252, suggests that this strain might have acquired the SCCmec cluster to adapt to specific environments containing methicillin. Region II of both genomes contains prophage regions but their DNA sequence identity is very low, suggesting that the prophages might differ. This is the first report of the complete genome sequence of S. aureus isolated from a real foodborne outbreak in South Korea. This report would be helpful to extend our understanding about the genome, general characteristics, and virulence factors of S. aureus for further studies of pathogenesis, rapid detection, and epidemiological investigation in foodborne outbreak.

Comparative Analysis of Soil Microbial Communities between Conventional and Organic Farming Systems in Pepper Cultivation (관행과 유기농 고추 재배지의 토양미생물 군집 비교)

  • Kim, Yiseul;Lee, Youngmi;Weon, Hang-Yeon;Sang, Mee Kyung;Song, Jaekyeong
    • Korean Journal of Organic Agriculture
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    • v.28 no.2
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    • pp.235-250
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    • 2020
  • Agricultural practices are known to have a crucial influence not only on soil physico-chemical properties but also on microbial communities. To investigate the effect of farming practices on soil microbial communities, a total of 10 soil samples were collected, including five conventional and five organic farming soils cultivated with peppers in plastic greenhouse. We conducted barcorded-pyrosequencing of V1-V3 regions of 16S rRNA genes to examine soil microbial communities of two different farming practices. Taxonomic classification of the microbial communities at the phylum level indicated that a total of 22 bacterial phyla were present across all samples. Among them, seven abundant phyla (>3%) including Proteobacteria, Actinobacteria, Firmicutes, Acidobacteria, Bacteroidetes, Chloroflexi, and Gemmatimonadetes were found, and Proteobacteria (33.0 ± 5.7%), Actinobacteria (19.9 ± 9.7%), and Firmicutes (13.6 ± 5.0%) comprised more than 66% of the relative abundance of the microbial communities. Organic farming soils showed higher relative abundances of Proteobacteria and Firmicutes, while Actinobacteria and Chloroflexi were more abundant in conventional farming soils. Notably, the genera Bacillus (higher in organic farming soils) and Streptomyces (higher in conventional farming soils) exhibited significant variation in relative abundance between organic and conventional farming soils. Finally, correlation analysis identified significant relationships (p<0.05) between soil chemical properties, in particular, pH and organic matter content and microbial communities. Taken together, this study demonstrated that the changes of soil physico-chemical properties by agricultural farming practices effected significantly (p<0.05) on soil microbial communities.

Bacterial Spot Disease of Green Pumpkin by Pseudomonas syringae pv. syringae (Pseudomonas syringae pv. syringae에 의한 애호박 세균점무늬병)

  • Park, Kyoung-Soo;Kim, Young-Tak;Kim, Hye-Seong;Lee, Ji-Hye;Lee, Hyok-In;Cha, Jae-Soon
    • Research in Plant Disease
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    • v.22 no.3
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    • pp.158-167
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    • 2016
  • A pathogen that causes a new disease on green pumpkin in the nursery and the field was characterized and identified. Symptoms of the disease on green pumpkin were water soaking lesions and spots with strong yellow halo on leaf, brown lesion on flower, and yellow spot on fruit. The bacterial isolates from the leaf spot were pathogenic on the 8 curcubitaceae crop plants, green pumpkin, figleaf gourd, wax gourd, young pumpkin, zucchini, cucumber, melon, and oriental melon, whereas they did not cause the disease on sweet pumpkin and watermelon. They were Gram-negative, rod shape with polar flagella, fluorescent on King's B agar and LOPAT group 1a by LOPAT test. Their Biolog substrate utilization patterns were similar to Pseudomonas syringae pv. syringae's in Biolog database. Phylogenetic trees with 16S rRNA gene sequences and multilocus sequence typing (MLST) with nucleotide sequences of 4 housekeeping genes, gapA, gltA, gyrB, rpoD and those of P. syringae complex strains in the Plant Associated and Environmental Microbes Database (PAMDB) showed that the green pumpkin isolates formed in the same clade with P. syringae pv. syringae strains. The clade in MLST tree was in the genomospecies 1 group. The phenotypic and genotypic characteristics suggested that the isolates from green pumpkin lesion were P. syringae pv. syringae.

Morphological and Molecular Characterization of Apharyngostrigea pipientis Faust, 1918 (Trematoda: Diplostomidea) from Ardeacinerea jouyi, Egretta intermedia and Nycticorax nycticorax in Korea

  • Kim, Hyeon-Cheol;Hong, Eui-Ju;Ryu, Si-Yun;Choi, Kyoung-Seong;Yu, Do-Hyeon;Cho, Jeong-Gon;Park, Jinho;Chae, Joon-Seok;Park, Bae-Keun
    • Journal of Veterinary Clinics
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    • v.37 no.5
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    • pp.242-248
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    • 2020
  • An intestinal strigeiid trematode, Apharyngostrigea pipientis, was detected from Ardea cinerea jouyi, Egretta intermedia and Nycticorax nycticorax between July, 2006 and June, 2020 in Daejeon metropolitan city and Kangwon-do, Republic of Korea. It was observed with a light and scanning electron microscope (SEM), and molecularly analyzed with 18S rRNA genes. The infection rate of the strigeiid trematode was 25.9% (7/27). The body is cylindrical in whole mount, greatly elongated in outline and shows a strong dorsal curvature. The overall measurements are: length 4.1-4.8 mm; fore-body 1.00-1.20 mm by 0.8-0.9 mm; hind-body 3.2-3.6 mm by 0.43-0.46 mm. The body is covered by a relatively thick cuticle, devoid of spines. The oral sucker leads into a short esophagus that is devoid of a muscular pharynx. The holdfast organ is enormously developed. The vitelline follicles distributed dorsal wall of fore-body and hind-body. This strigeiid trematode especially differs from other species of genus Apharyngostrigea in morphology. Although these A. pipientis are several morphological distinctions from Apharyngostrigea cornu, the partial sequence of 18S rRNA showed the sequence homology of A. pipientis (99.9%) and A. cornu (99.8%), and this result revealed their conspecific relationship between A. cornu and A. pipientis. This is the first report of A. pipientis in the Republic of Korea.

Isolation of bacteriophage-resistant Pseudomonas tolaasii strains and their pathogenic characters (박테리오파지 저항성을 갖는 Pseudomonas tolaasii 변이주 분리 및 이들의 병원특성)

  • Park, Soo-Jin;Han, Ji-Hye;Kim, Young-Kee
    • Journal of Applied Biological Chemistry
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    • v.59 no.4
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    • pp.351-356
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    • 2016
  • Bacterial blotch caused by Pseudomonas tolaasii is one of the major diseases of oyster mushroom, Pleurotus ostreatus. Application of bacteriophages is a very useful tool to decrease the density of pathogens and it has been successful to making disease-free cultivation area, known as phage therapy. Effect of phages on pathogen sterilization is very limited to the specific host strains. Minor variations of the host strains may cause changes in phage sensitivity. The phage-resistant strains of P. tolaasii were isolated and their pathogenic characters were investigated to improve the effectiveness of phage therapy. In the phylogenetic analysis, both phage-resistant strains and the corresponding host strains were identical based on the sequence comparison of 16S rRNA genes. The pathogenic characters, such as hemolytic activity and brown blotch formation, were measured on the phage-resistant strains and no correlation between phage-resistance and pathogenic characters was observed. Nevertheless, pathogenic characters were sometimes changed in the phage-resistant strains depending on the host strains. In order to make the phage therapy successful, the bacteriophages having a wide host range should be isolated.