• 제목/요약/키워드: rDNA sequencing

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Anti-inflammatory effects of Lactobacillus reuteri LM1071 via MAP kinase pathway in IL-1β-induced HT-29 cells

  • Kim, Tae-rahk;Choi, Kyoung-sook;Ji, Yosep;Holzapfel, Wilhelm H.;Jeon, Min-Gyu
    • Journal of Animal Science and Technology
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    • 제62권6호
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    • pp.864-874
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    • 2020
  • Lactic acid bacteria are well-known probiotics, conferring several health benefits. In this study, we isolated lactobacilli from human breast milk and identified Lactobacillus reuteri LM1071 (RR-LM1071) using 16S rDNA sequencing. We tested the hemolytic activity, biogenic amine production, and antibiotic susceptibility of this strain to assess its safety. RR-LM1071 was found to be negative for hemolytic activity and biogenic amine production, as well as was measured in susceptible level for antibiotics in the minimal inhibitory concentration (MIC) test. The adhesive properties of RR-LM1071 were higher than those of LGG in HT-29 cells, and showed a greater hydrophobicity than LGG in hexadecane solvent. Under inflammatory conditions, RR-LM1071 suppressed the mRNA expression of IL-6, TNF-α, and IL-4 produced in IL-1β-induced HT-29 cells. Our results suggest that RR-LM1071 is a safe and valuable probiotic that can be used for the treatment of inflammatory bowel disease.

Re-Identification on Korean Penicillium Sequences in GenBank Collected by Software GenMine

  • Chang Wan Seo;Sung Hyun Kim;Young Woon Lim;Myung Soo Park
    • Mycobiology
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    • 제50권4호
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    • pp.231-237
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    • 2022
  • Penicillium species have been actively studied in various fields, and many new and unrecorded species continue to be reported in Korea. Moreover, unidentified and misidentified Korean Penicillium species still exist in GenBank. Therefore, it is necessary to revise the Korean Penicillium inventory based on accurate identification. We collected Korean Penicillium nucleotide sequence records from GenBank using the newly developed software, GenMine, and re-identified Korean Penicillium based on the maximum likelihood trees. A total of 1681 Korean Penicillium GenBank nucleotide sequence records were collected from GenBank. In these records, 1208 strains with four major genes (Internal Transcribed Spacer rDNA region, b-tubulin, Calmodulin and RNA polymerase II) were selected for Penicillium reidentification. Among 1208 strains, 927 were identified, 82 were identified as other genera, the rest remained undetermined due to low phylogenetic resolution. Identified strains consisted of 206 Penicillium species, including 156 recorded species and 50 new species candidates. However, 37 species recorded in the national list of species in Korea were not found in GenBank. Further studies on the presence or absence of these species are required through literature investigation, additional sampling, and sequencing. Our study can be the basis for updating the Korean Penicillium inventory.

Characterization of Newly Recorded Talaromyces veerkampii Isolated from Field Soil in Korea based on Morphology and Multigene Sequence Analysis

  • Mahesh Adhikari;Hyun Seung Kim;Hyo Bin Park;Ki Young Kim;In Kyu Lee;Eun Jeong Byeon;Ji Min Woo;Hyang Burm Lee;Youn Su Lee
    • 한국균학회지
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    • 제50권4호
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    • pp.347-355
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    • 2022
  • A fungal isolate belonging to the phylum Ascomycota was isolated and identified as Talaromyces veerkampii in 2017 during a survey of fungal diversity in field soils in Korea. This fungal isolate was identified and described based on macro- and micromorphological and molecular characterization. The identification was also based on partial 18S-ITS1-5.8S-ITS2-28S rDNA and calmodulin (CaM)-encoding gene sequencing data. Talaromyces veerkampii has not been previously reported in Korea. Thus, we report here a newly discovered species from soil in Korea along with its morphological and molecular characteristics.

An Outbreak of Gray Mold Caused by Botrytis cinerea on Kenaf(Hibiscus cannabinus L.)

  • Kwon, Jin-Hyeuk;Kang, Dong-Wan;Lee, Seong-Tae;Shim, Chang-Ki;Kim, Min-Jeong;Kim, Jinwoo
    • 농업생명과학연구
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    • 제50권2호
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    • pp.25-32
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    • 2016
  • A severe outbreak of gray mold on kenaf (Hibiscus cannabinus L.) was observed on kenaf grown in the research field of Gyeongsangnam-do Agricultural Research and Extension Services, Jinju, Korea in 2014. Gray mold appeared on young plants as gray-brown velvety mold covering stems and leaves. Infections that girdled the stem caused wilting above the infected area and developed a canker. The casual fungus formed grayish brown colonies on potato dextrose agar. The conidia were one celled, mostly ellipsoid or ovoid in shape, colorless or pale brown in color, and 6-18 × 4-10 ㎛ in size. The conidiophores were 15-32 ㎛ in length. These measurements and taxonomic characteristics were most similar to those of Botrytis. DNA sequencing and phylogenetic analysis of the complete internal transcribed spacer rRNA gene region confirmed that the fungal isolates were indeed Borytis cinerea. Koch's postulates were supported by pathogenicity tests conducted on healthy plants. On the basis of mycological characteristics and pathogenicity test on host plants, the fungus was identified as Botrytis cinerea. To the best of our knowledge, this is the first report of a gray mold caused by B. cinerea on kenaf in Korea.

First Report of Pectobacterium aroidearum Causing Soft Rot on Zamioculcas zamiifolia

  • Kyoung-Taek Park;Soo-Min Hong;Leonid N. Ten;Chang-Gi Back;Seung-Yeol Lee;In-Kyu Kang;Hee-Young Jung
    • 식물병연구
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    • 제29권4호
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    • pp.445-451
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    • 2023
  • Zamioculcas zamiifolia is a popular indoor ornamental plant in Korea. In August 2021, a severe outbreak of soft rot disease affected Z. zamiifolia in Emseong, Chungcheongbuk-do, Korea. Infected plants displayed wilting, water-soaked lesions, stem collapse, and green-brown discoloration. The bacterial strain KNUB-05-21 was isolated from infected stems and identified as Pectobacterium aroidearum using 16S rRNA nucleotide sequencing and multilocus sequence analysis based on partial sequences of dnaX, leuS, and recA genes. Confirmation of its affiliation with P. aroidearum was also obtained through biochemical and morphological characterization. To confirm the pathogenicity of strain KNUB-05-21, its suspension was injected into Z. zamiifolia stems. Within a week, soft rot developed on the stems, exhibiting symptoms similar to those observed in field-infected plants. The reisolated strain was identical to those of P. aroidearum. Before this study, P. aroidearum was not reported as a causative pathogen of Z. zamiifolia soft rot in Korea.

First Report of Pectobacterium aroidearum Causing Soft Rot on Ficus carica in Korea

  • Kyoung-Taek Park;Leonid N. Ten;Soo-Min Hong;Song-Woon Nam;Chang-Gi Back;Seung-Yeol Lee;Hee-Young Jung
    • 식물병연구
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    • 제30권1호
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    • pp.88-94
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    • 2024
  • In July 2021, symptoms of soft rot were observed on the stems of Ficus carica in Yeongam, Jeollanamdo, Korea. To accurately diagnose the cause, infected stem was collected and bacterial strain was isolated. Among these, the pathogenic strain KNUB-08-21 was identified as Pectobacterium aroidearum through 16S rRNA gene sequencing and phylogenetic analysis based on the concatenated sequences of the dnaX, leuS, and recA genes. The affiliation of the isolate with this bacterial species was also confirmed by its biochemical characteristics obtained using API ID 32 GN system. Artificial inoculation confirmed the strain's pathogenicity in figs, causing significant damage to both stems and fruits. To our knowledge, this is the first report of P. aroidearum causing soft rot disease in F. carica in Korea.

Morphological and molecular characterization of root-lesion nematode Pratylenchus hippeastri from Korea

  • Sungchan Huh;Namsook Park;Heonil Kang;Changhwan Bae;Insoo Choi
    • 환경생물
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    • 제41권4호
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    • pp.657-665
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    • 2023
  • The root-lesion nematode Pratylenchus spp. is the most important plantparasitic nematode due to its worldwide distribution, wide host ranges, and migratory endoparasitic characteristics. One population of Pratylenchus collected from the giant pussy willow (Salix chaenomeloides Kimura) in the Andong area as part of a nematode survey in Korea was characterized morphologically and by molecular methods. The analysis of morphological measurements and morphometric characteristics, as well as DNA sequencing of the rRNA large subunit (LSU) D2/D3 expansion segments and the internal transcribed spacer (ITS) gene sequence, confirmed the identity of this population as P. hippeastri. This study is the first report of P. hippeastri associated with Salix chaenomeloides in Korea and worldwide. Further studies on distribution and pathogenicity in different P. hippeastri host crops, such as grapevines, strawberries, and apples, are necessary. The taxonomic keys to 16 Pratylenchus species in Korea are provided.

Development of an Enrichment Culture Growing at Low Temperature used for Ensiling Rice Straw

  • Yang, Hong Yan;Wang, Xiao Fen;Gao, Li Juan;Haruta, Shin;Ishii, Masaharu;Igarashi, Yasuo;Cui, Zong Jun
    • Journal of Microbiology and Biotechnology
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    • 제18권4호
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    • pp.711-717
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    • 2008
  • To speed up the conversion of rice straw into feeds in a low-temperature region, a start culture used for ensiling rice straw at low temperature was selected by continuous enrichment cultivation. During the selection, the microbial source for enrichment was rice straw and soil from two places in Northeast China. Lab-scale rice straw fermentation at $10^{\circ}C$ verified, compared with the commercial inoculant, that the selected start culture lowered the pH of the fermented rice straw more rapidly and produced more lactic acid. The results from denatured gradient gel eletrophoresis showed that the selected start culture could colonize into the rice straw fermentation system. To analyze the composition of the culture, a 16S rRNA gene clone library was constructed. Sequencing results showed that the culture mainly consisted of two bacterial species. One (A) belonged to Lactobacillus and another (B) belonged to Leuconostoc. To make clear the roles of composition microbes in the fermented system, quantitative PCR was used. For species A, the DNA mass increased continuously until sixteen days of the fermentation, which occupied 65%. For species B, the DNA mass amounted to 5.5% at six days of the fermentation, which was the maximum relative value during the fermentation. To the authors' best knowledge, this is the first report on ensiling rice straw with a selected starter at low temperature and investigation of the fermented characteristics.

Bacterial Diversity at Different Sites of the Digestive Tract of Weaned Piglets Fed Liquid Diets

  • Hong, Tran Thi Thu;Passoth, Volkmar;Lindberg, Jan Erik
    • Asian-Australasian Journal of Animal Sciences
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    • 제24권6호
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    • pp.834-843
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    • 2011
  • Bacterial diversity was studied using PCR-DGGE, cloning and sequencing. DNA was isolated from digesta samples from stomach, ileum and colon of 28 weaned piglets (Large White${\times}$Mong Cai) fed dry control feed, naturally fermented liquid feed (FE) and a liquid diet with inclusion of rice distiller's residue feed. General bacterial diversity was described using DGGE analysis of the V3 region of 16S rDNA. The microbial populations in the stomach and the ileum were considerably influenced by the diet, while only marginal effects were observed in the colon. There was a large variation of the microbial flora in the stomach between individuals fed non-fermented diets. In contrast, animals fed diet FE had a more uniform microbial flora in the stomach and the ileum compared to the other diets. In total 47 bands from the DGGE profiles were cloned. In stomach, most frequently lactic acid bacteria were found. Feeding diet FE resulted in the occurrence of Pediococcus species in stomach and ileum. In pigs fed the other diets, Lactobacillus gallinarum, Lactobacillus johnsonii and Lactobacillus fermentum were found in stomach and ileum. Most of the sequences of bands isolated from colon samples and several from ileum matched to unknown bacteria, which often grouped within Prevotellaceae, Enterobacteriaceae, Bacteroidaceae and Erysipelotrichaceae. This study demonstrates that fermented liquid feed affects bacterial diversity and the specific microflora in stomach and ileum, which provides a potential to modulate the gut microflora with dietary means to increase the abundance of beneficial bacteria and improve piglets' health.

Casein phosphopeptide를 생산하는 김치 유래 유산균의 분리 및 특성 연구 (Isolation and Characteristics of Lactic Acid Bacteria Producing Casein Phosphopeptides from Kimchi)

  • 이미경;권효정;변옥희;방보연;김유진;박정민;배동훈
    • 한국미생물·생명공학회지
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    • 제44권1호
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    • pp.68-73
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    • 2016
  • 본 연구에서는 전국 가정집에서 제조한 김치로부터 분리한 유산균들 중에서 protease 활성 및 CPP 생산 능력이 가장 높은 균주인 strain 0301을 동정하였고 해당 균주의 특성과 CPP 생산 최적 조건을 조사하였다. 0301은 $0.6-0.8{\mu}m$ 크기의 gram 양성 구균이며, 16s rDNA 염기서열 분석 결과 Enterococcus faecalis MG-379로 동정 명명하였다. E. faecalis MG-379 균주를 2% fructose를 첨가하고 pH 6.0인 배지에서 37oC에서 36시간 배양하였을 때 CPP 생산 능력이 최대였으며, ICP를 이용하여 측정한 결과 2227.5 mg/kg으로, 비교 균주인 Enterococcus faecalis KCCM 40450에 비해 약 2배 가량 높은 CPP 생산 능력을 갖는 것으로 나타내었다.