• 제목/요약/키워드: spore analysis

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Two groups of S-layer proteins, SLP1s and SLP2s, in Bacillus thuringiensis co-exist in the S-layer and in parasporal inclusions

  • Zhou, Zhou;Peng, Donghai;Zheng, Jinshui;Guo, Gang;Tian, Longjun;Yu, Ziniu;Sun, Ming
    • BMB Reports
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    • 제44권5호
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    • pp.323-328
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    • 2011
  • We screened four B. thuringiensis strains whose parasporal inclusions contained the S-layer protein (SLP), and cloned two slp genes from each strain. Phylogenetic analysis indicated these SLPs could be divided into two groups, SLP1s and SLP2s. To confirm whether SLPs were present in the S-layer or as a parasporal inclusion, strains CTC and BMB1152 were chosen for further study. Western blots with isolated S-layer proteins from strains CTC and BMB1152 in the vegetative phase showed that SLP1s and SLP2s were constituents of the S-layer. Immunofluorescence utilizing spore-inclusion mixtures of strains CTC and BMB1152 in the sporulation phase showed that SLP1s and SLP2s were also constituents of parasporal inclusions. When heterogeneously expressed in the crystal negative strain BMB171, four SLPs from strains CTC and BMB1152 could also form parasporal inclusions. This temporal and spatial expression is not an occasional phenomenon but ubiquitous in B. thuringiensis strains.

입자추적기법을 이용한 바다숲 조성지의 확장에 관한 수치예측 (A Numerical Estimation on Extension of Marine Afforestation Area using Particle Tracking Method)

  • 조재권;김대권;이문옥;오태건
    • Journal of Advanced Marine Engineering and Technology
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    • 제34권6호
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    • pp.922-934
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    • 2010
  • 바다숲 조성용 시설물의 효율적인 배치를 위하여 수치실험을 통하여 바다숲 조성 예정지의 조석류에 의한 해수흐름을 분석하였으며, 분석된 해수흐름을 바탕으로 하여 해조포자의 거동을 파악하기 위하여 입자추적실험을 수행하였다. 수치실험에 사용된 수치모형은 EFDC(Environmental Fluids Dynamics Code)를 사용하였으며, 연직방향으로 총 5개 층으로 나누어 실험을 수행하였다. 실험결과, 바다숲 조성대상해역의 해수흐름패턴은 대상해역 바깥쪽의 주류에 크게 영향을 받고 있음을 알 수 있었고, 잔차류를 분석한 결과 크게 두 개의 순환 고리가 형성되어 있음을 확인하였다. 해수흐름의 패턴을 고려하여 3곳의 바다숲 시설물의 설치장소를 선정하고 저층에 입자를 투입하여 15일간 추적을 실시한 결과, 1번과 3번 정점은 해안선에 연하여 띠형태로 시설하며, 2번 정점은 연안에서 약 300 ~ 500m정도 떨어진 곳에 타원의 형태로 시설하는 것이 보다 효율적일 것으로 판단되었다.

Sphingopyxis granuli sp. nov., a $\beta$-Glucosidase-Producing Bacterium in the Family Sphingomonadaceae in $\alpha$-4 Subclass of the Proteobacteria

  • Kim Myung Kyum;Im Wan Taek;Ohta Hiroyuki;Lee Myung Jin;Lee Sung Taik
    • Journal of Microbiology
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    • 제43권2호
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    • pp.152-157
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    • 2005
  • Strain Kw07$^T$, a Gram-negative, non-spore-forming, rod-shaped bacterium, was isolated from granules in an Up-flow Anaerobic Sludge Blanket (UASB) bioreactor used in the treatment of brewery waste­water. 16S rRNA gene sequence analysis revealed that strain Kw07T belongs to the a-4 subclass of the Proteobacteria, and the highest degree of sequence similarity was determined to be to Sphingopyxis macrogoltabida IFO 15033T (97.8%). Chemotaxonomic data revealed that strain Kw07T possesses a quinone system with the predominant compound Q-I0, the predominant fatty acid C,s:, OJ7c, and sphingolipids, aU of which corroborated our assignment ofthe strain to the Sphingopyxis genus. The results of DNA-DNA hybridization and physiological and biochemical tests clearly demonstrated that strain Kw07T represents a distinct species. Based on these data, Kw07T (= KCTC 12209T = NBRC 100800T) should be classified as the type strain for a novel Sphingopyxis species, for which the name Sphingopyxis granuli sp. novo has been proposed.

Genome Sequence of Bacillus cereus FORC_021, a Food-Borne Pathogen Isolated from a Knife at a Sashimi Restaurant

  • Chung, Han Young;Lee, Kyu-Ho;Ryu, Sangryeol;Yoon, Hyunjin;Lee, Ju-Hoon;Kim, Hyeun Bum;Kim, Heebal;Jeong, Hee Gon;Choi, Sang Ho;Kim, Bong-Soo
    • Journal of Microbiology and Biotechnology
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    • 제26권12호
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    • pp.2030-2035
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    • 2016
  • Bacillus cereus causes food-borne illness through contaminated foods; therefore, its pathogenicity and genome sequences have been analyzed in several studies. We sequenced and analyzed B. cereus strain FORC_021 isolated from a sashimi restaurant. The genome sequence consists of 5,373,294 bp with 35.36% GC contents, 5,350 predicted CDSs, 42 rRNA genes, and 107 tRNA genes. Based on in silico DNA-DNA hybridization values, B. cereus ATCC $14579^T$ was closest to FORC_021 among the complete genome-sequenced strains. Three major enterotoxins were detected in FORC_021. Comparative genomic analysis of FORC_021 with ATCC $14579^T$ revealed that FORC_021 harbored an additional genomic region encoding virulence factors, such as putative ADP-ribosylating toxin, spore germination protein, internalin, and sortase. Furthermore, in vitro cytotoxicity testing showed that FORC_021 exhibited a high level of cytotoxicity toward INT-407 human epithelial cells. This genomic information of FORC_021 will help us to understand its pathogenesis and assist in managing food contamination.

국내 주요 수계 내 원생동물의 분석과 매질의 영향 (Cryptosporidium and Giardia (oo)cysts in Korean Rivers and the Effect of Sample Matrix)

  • 정현미;오덕화
    • 한국물환경학회지
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    • 제22권2호
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    • pp.271-276
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    • 2006
  • The outbreak cases of Cryptosporidium or Giardia from drinking water in abroad have drawn attentions on the public health. It is well known that Cryptosporidium is the most resistant organism against chlorine disinfection. To guesstimate the levels of Cryptosporidium and Giardia in Korean surface water, 1~2 samples from 22 drinking water sources in four Korean major rivers of Han, Keum, Nakdong, and Youngsan were monitored. In addition, two sites in Kyunganchon, a contaminated river were monitored for comparison. In source waters, detection rates of Cryptosporidium and Giardia were 15% (6/39) and 21% (5/24) with the range of 1~3 oocysts/10 L and 1~6 cysts/10 L, respectively. In Kyunganchon, they were 60% (6/10) and 70% (7/10) in the range of 1~9 oocysts/10 L and 10~72 cysts/10 L, respectively. When one of the source waters in Han river was monitored monthly, Cryptosporidium were found mostly in cold season. Matrix of the samples gave influence on the recoveries of the spiked protozoa. The recoveries of both Cryptosporidium and Giardia increased in the samples of Kyunganchon, known as contaminated area. However, protozoan recovery did not show significant relation with turbidity, the index of matrix contamination, which implies that there are additional unveiled features of matrix affecting the recoveries of the protozoa. The protozoan distribution in Kyunganchon showed significant relations with Cl. perfringens, anaerobic and spore forming indicator bacteria of fecal contamination by regression analysis, but not with turbidity, the general indicator of water quality.

Efficient Target-Site Assay of Chemicals for Melanin Biosynthesis Inhibition of Magnaporthe grisea

  • Kim, Jin-Cheol;Son, Mi-Jung;Kim, Heung-Tae;Park, Gyung-Ja;Hahn, Hoh-Gyu;Nam, Kee-Dal;Cho, Kwang-Yun
    • The Plant Pathology Journal
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    • 제16권3호
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    • pp.125-129
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    • 2000
  • A rapid and efficient assay to determine melanin biosynthesis inhibition of Magnaporthe grisea, a causal agent of the rice blast, by chemicals was developed. Wells in 24-well plates were loaded with spore suspension of the fungus and three known melanin biosynthesis inhibitors of KC10017, tricyclazole, and carpropamid. Subsequent color changes of mycelia and culture media in the wells were observed 7 days after incubation. The wells treated with KC10017 (an inhibitor of polyketide synthesis step and/or pentaketide cyclization step) became colorless, whereas tricyclazole (an inhibitor of 1, 3, 8-trihydroxynaphthalene reductase) or carpropamid (an inhibitor of scytalone dehydratase)-treated wells exhibited red color. They did not show any inhibitory effect on fungal growth. The inhibition of reaction steps prior to 1, 3, 6, 8-tetrahydroxynaphthalene formation was easily determined by colorless medium and mycelia. However, it was impossible to distinguish between inhibition of reduction steps and inhibition of dehydration steps by colors of the cultures. It was accomplished through HPLC analysis of the melanin biosynthesis-involving pentaketide metabolites accumulated by the inhibitors. Through screening of a number of synthetic chemicals using the in vitro assay, we could find a novel chemical group of melanin biosynthesis inhibitor.

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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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    • 제29권1호
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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.

Genotyping of Agaricus bisporus Strains by PCR Fingerprints

  • Min, KyongJin;Oh, YounLee;Kang, HeeWan
    • 한국균학회소식:학술대회논문집
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    • 한국균학회 2014년도 추계학술대회 및 정기총회
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    • pp.41-41
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    • 2014
  • Agaricus bisporus, commonly known as the button mushroom, is the most widely cultivated species of edible fungi. Low frequency of recombination ratio and homokaryotic or monokaryotic spore on meiotic basidia form obstacles for breeding programs. Since the first hybrid varieties for white button mushrooms were released in Europe, new varieties released afterwards were either identical of very similar to these first hybrids on morphologies. Therefore, different DNA markers have been used to define unique varieties of A. bisporus strains. Aim of this study is to assess the genetic diversity of different A. bisporus strains in Korea. Twelve UFP (Universal fungal primer, JK BioTech. Ltd), 12 simple sequence repeat (ISSR) and 30 SSR primers were used to assess genetic diversity of monokaryotic and dikaryotic Agaricus bisporus strains including other 19 Agaricus spp. Of them, four UFP, four SSR primers, $(GA)_8T$, $(AG)_8YC$, $(GA)_8C$ and $(CTC)_6$ and seven SSR markers produced PCR polymorphic bands between the Agaricus species or within A. bisporus strains. PCR polymorphic bands were inputted for UPGMA cluster analysis. Forty five strains of A. bisporus are genetically clustered into 6 groups, showing coefficient similarity from 0.75 to 0.9 among them. In addition, genetic variations of monokaryotic and dikaryotic Agaricus bisporus strains were partially detected by PCR technologies of this study. The varieties, Saea, saedo, Saejeong and Saeyeon that have recently been developed in Korea were involved in the same group with closely genetic relationship of coefficient similarity over 0.96, whereas, other strains were genetically related to A. bisporus strains that were introduced from USA, Eroupe and Chinese.

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Biocontrol Activity of Bacillus amyloliquefaciens CNU114001 against Fungal Plant Diseases

  • Ji, Seung Hyun;Paul, Narayan Chandra;Deng, Jian Xin;Kim, Young Sook;Yun, Bong-Sik;Yu, Seung Hun
    • Mycobiology
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    • 제41권4호
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    • pp.234-242
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    • 2013
  • A total of 62 bacterial isolates were obtained from Gomsohang mud flat, Mohang mud flat, and Jeju Island, Republic of Korea. Among them, the isolate CNU114001 showed significant antagonistic activity against pathogenic fungi by dual culture method. The isolate CNU114001 was identified as Bacillus amyloliquefaciens by morphological observation and molecular data analysis, including 16SrDNA and gyraseA (gyrA) gene sequences. Antifungal substances of the isolate were extracted and purified by silica gel column chromatography, thin layer chromatography, and high performance liquid chromatography. The heat and UV ray stable compound was identified as iturin, a lipopeptide (LP). The isolate CNU114001 showed broad spectrum activity against 12 phytopathogenic fungi by dual culture method. The semi purified compound significantly inhibits the mycelial growth of pathogenic fungi (Alternaria panax, Botrytis cinera, Colletotrichum orbiculare, Penicillium digitatum, Pyricularia grisea and Sclerotinia sclerotiorum) at 200 ppm concentration. Spore germ tube elongation of Botrytis cinerea was inhibited by culture filtrate of the isolate. Crude antifungal substance showed antagonistic activity against cucumber scleotiorum rot in laboratory, and showed antagonistic activity against tomato gray mold, cucumber, and pumpkin powdery mildew in greenhouse condition.

Heterologous Expression of Streptomyces albus Genes Linked to an Integrating Element and Activation of Antibiotic Production

  • Kwon, Hyung-Jin;Lee, Soon-Youl;Hong, Soon-Kwang;Park, Uhn-Mee;Suh, Joo-Won
    • Journal of Microbiology and Biotechnology
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    • 제9권4호
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    • pp.488-497
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    • 1999
  • Probing Streptomyces albus ATCC 21838 chromosomal DNA with a proline tRNA sequence resulted in an isolation of a putative integrating element in the 6.4-kb EcoRI fragment. It was found that Streptomyces lividans TK-24 transformed with a cloned DNA fragment on a multicopy plasmid, produced a higher level of spore pigment and mycelial red pigment on a regeneration agar. Furthermore, the transformant S. lividans TK-24 produced a markedly increased level of undecylprodigiosin in a broth culture. A nucleotide sequence analysis of the cloned region revealed several open reading frames homologous to the integrases of integrating plasmids or temperate bacteriophages, signal-transducing regulatory proteins with a conserved ATP-binding domain, oxidoreductases ($\beta$-ketoacyl reductase), and an AraC-like transcriptional regulator. To examine the effect on antibiotic production, each coding region was overexpressed separately from the other genes in the region in S. lividans TK-24 with; pJHS3044 for the expression of the signal-transducing regulatory protein homologue, pJHS3045 for the homologue of oxidoreductase, and pJHS3051 for the homologue of the AraC-like transcriptional regulator. Phenotypic studies of S. lividans TK-24 strains harboring plasmids for the overexpression of individual genes suggested the following effects of the genes on antibiotic production: The oxidoreductase homologue stimulated the production of actinorhodin and undecylprodigiosin, which was influenced by the culture conditions; the homologue of the AraC-like transcriptional regulator was the most effective factor in antibiotic production within all the culture conditions tested; the signal-transducing regulatory protein homologue repressed the effect due to the homologue of the AraC-like transcriptional regulator, however, the antibiotic production was derepressed upon entering the stationary phase.

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