• Title/Summary/Keyword: spore analysis

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A new record of epiphytic red alga Madagascaria erythrocladioides (Erythropeltidales, Rhodophyta) in Korea

  • Wen, Xianying;Lee, Ji Woong;Shim, Eunyoung;Kim, Gwang Hoon
    • Korean Journal of Environmental Biology
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    • v.39 no.3
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    • pp.383-389
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    • 2021
  • The Erythropeltidales are a common group of small, mostly epiphytic, marine red algae. However, they are little known in Korea. Many of the described species of Erythropeltidales differ subtly in morphology, and often the morphological differences are due to the substrate or environmental changes. Integration of molecular data with standardized culture conditions has been recommended to account for these algae. A Madagascaria species was first collected from the western coast of Korea and was identified as Madagascaria erythrocladioides based on the morphological and molecular characteristics. Morphological characteristics conformed well with its original description, and the phylogenetic analysis based on rbcL sequence showed Korean M. erythrocladioides nests in the same clade with the original species described in Japan with a genetic distance of 0.0-0.1%. This species was isolated from a red alga, Pterocladiella capillacea, in laboratory culture. The thallus ontogeny and host preference were examined by a co-culture with 13 different species of algae. Results showed a relatively broad host preference in mono-spore attachment and epiphyte development of Madagascaria erythrocladioides. Mono-spores of M. erythrocladioides attached to most of the red algal hosts' surfaces but no crustose thalli developed on some of the algal hosts even after one month of co-culture.

Isolation and Characterization of a Weizmannia coagulans Bacteriophage Youna2 and Its Endolysin PlyYouna2

  • Bokyung Son;Youna Kim;Booyoung Yu;Minsuk Kong
    • Journal of Microbiology and Biotechnology
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    • v.33 no.8
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    • pp.1050-1056
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    • 2023
  • Weizmannia coagulans (formerly Bacillus coagulans) is Gram-positive, and spore-forming bacteria causing food spoilage, especially in acidic canned food products. To control W. coagulans, we isolated a bacteriophage Youna2 from a sewage sludge sample. Morphological analysis revealed that phage Youna2 belongs to the Siphoviridae family with a non-contractile and flexible tail. Youna2 has 52,903 bp double-stranded DNA containing 61 open reading frames. There are no lysogeny-related genes, suggesting that Youna2 is a virulent phage. plyYouna2, a putative endolysin gene was identified in the genome of Youna2 and predicted to be composed of a N-acetylmuramoyl-L-alanine amidase domain (PF01520) at the N-terminus and unknown function DUF5776 domain (PF19087) at the C-terminus. While phage Youna2 has a narrow host range, infecting only certain strains of W. coagulans, PlyYouna2 exhibited a broad antimicrobial spectrum beyond the Bacillus genus. Interestingly, PlyYouna2 can lyse Gram-negative bacteria such as Escherichia coli, Yersinia enterocolitica, Pseudomonas putida and Cronobacter sakazakii without other additives to destabilize bacterial outer membrane. To the best of our knowledge, Youna2 is the first W. coagulans-infecting phage and we speculate its endolysin PlyYouna2 can provide the basis for the development of a novel biocontrol agent against various foodborne pathogens.

Purification and Identification of Paenibacillus sp., Isolated from Diseased Larvae of Allomyrina dichotoma (Linnaeus, 1771) (Coleoptera: Scarabaeidae) in Insect Farms

  • Kang, Tae Hwa;Han, Sang Hoon;Weon, Hang Yeon;Lee, Young Bo;Kim, Namjung;Nam, Sung Hee;Park, Hae Chul
    • International Journal of Industrial Entomology and Biomaterials
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    • v.25 no.2
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    • pp.195-203
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    • 2012
  • In reared populations of Allomyrina dichotoma, commercial insects, the skin of last instar larvae was changed softer with opaque white, and infested grubs eventually died. To clarify the cause of the symptom, we collected the larvae of A. dichotoma from five farms and examined their intestinal bacterial florae using pyrosequencing technique. From those results, a member of Paenibacillus was found only in the larvae showing the symptom of disease. Through PCR analysis using a Paenibacillus specific primer set, we obtained the partial 16S rRNA gene sequence and confirmed the microbe as Paenibacillus sp. For clear identification, a whole guts was extracted from each larva showing the sign of the disease and incubated at $70^{\circ}C$ for 15 min to isolate spore forming bacteria. After then, each content of guts was cultured on $MYPGP_{NAL}$ agar medium($12.5{\mu}g/ml$ of nalidixic acid) at $30^{\circ}C$. The 16S rRNA gene sequence analysis for the isolated bacteria showed that they were closely related to P. rigui(97.9% similarity), to P. chinjuensis(96.1% similarity), and to P. soli(95.3% similarity). Additional tests including API test and cellular fatty acid composition analysis were performed, but the strain couldn't be identified at species level, suggesting it may represent novel species of the genus Paenibacillus.

A Yeast MRE3/REC114 Gene is Essential for Normal Cell Growth and Meiotic Recombination

  • Leem, Sun-Hee
    • Journal of Microbiology
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    • v.37 no.4
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    • pp.248-255
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    • 1999
  • We have analyzed the MRE3/REC114 gene of Saccharomyces cerevisiae, previously detected in isolation of mutants defective in meiotic recombination. We cloned the MRE3/REC114 gene by complementation of the meiotic recombination defect and it has been mapped to chormosome XIII. The DNA sequence analysis revealed that the MRE3 gene is identical to the REC114 gene. The upstream region of the MRE3/REC114 gene contains a T_4C site, a URS (upstream repression sequence) and a TR (T-rich) box-like sequence, which reside upstream of many meiotic genes. Coincidentally, northern blot analysis indicated that the three sizes of MRE3/REC114 transcripts, 3.4, 1.4 and 1.2 kb, are induced in meiosis. A less abundant transcript of 1.4 kb is detected in both mitotic and meiotic cells, suggesting that it is needed in mitosis as well as meiosis. To examine the role of the MRE3/REC114 gene, we constructed mre3 disruption mutants. Strains carrying an insertion or null deletion of the MRE3/REC114 gene showed slow growth in nutrient medium and the doubling time of these cells increased approximately by 2-fond compared to the wild-type strain. Moreover, the deletion mutant (${\delta}$mre3) displayed no meiotically induced recombination and no viable spores. The mre3/rec114 spore lethality can be suppressed by spo13, a mutation that causes cells to bypass reductional division. The double-stranded breaks (DSBs) which are involved in initiation of meiotic recombination were not detected in the analysis of meiotic chromosomal DNA from the mre3/rec114 disruptant. From these results we suggest that the MRE3/REC114 gene product is essential in normal growth and in early meiotic stages involved in meiotic recombination.

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Taxonomic Study of the Genus Pholiota (Strophariaceae, Basidiomycota) in Korea

  • Lee, Jun Won;Park, Myung Soo;Park, Ji-Hyun;Cho, Yoonhee;Kim, Changmu;Kim, Chang Sun;Jo, Jong Won;Lim, Young Woon
    • Mycobiology
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    • v.48 no.6
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    • pp.476-483
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    • 2020
  • The genus Pholiota (Strophariaceae, Basidiomycota) is made up of wood-rotting saprotrophic mushrooms characterized by a yellow or brown pileus with scales and/or slimy, and by a brownish smooth spore with a germ pore. However, these features are not enough to distinguish its species, or separate the genus Pholiota from other brown-spored wood-rotting genera such as Hypholoma and Stropharia. Although internal transcribed spacer (ITS) sequencebased identification has improved identification accuracy for species of Pholiota, most Pholiota species in Korea are reported based on morphological features. To evaluate the taxonomy of Pholiota species, we investigated 62 specimens collected from 1999 to 2019 in Korea using ITS sequence analysis and morphological observation. Twelve of the 16 recorded Pholiota species in Korea were identified. While eight species were clearly separated, the ITS analysis did not distinguish three in the Pholiota adiposa complex. Therefore, further investigation is required to distinguish these three species. ITS sequences deposited in GenBank confirm that P. highlandensis exists in Korea. The presence of the other four Pholiota species could not be confirmed through specimens or sequence information in GenBank. A taxonomic key and the ITS sequence data for Korean Pholiota species are included and can be good baselines for further research on Pholiota taxonomy and diversity.

Potential of Cross-infection of Colletotrichum Species Causing Anthracnose in Persimmon and Pepper

  • Kim, Hye-Ryoung;Lim, Tae-Heon;Kim, Joo-Hyung;Kim, Young-Ho;Kim, Heung-Tae
    • The Plant Pathology Journal
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    • v.25 no.1
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    • pp.13-20
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    • 2009
  • Ninety isolates of Colletotrichum species from new persimmon tree twigs and 50 isolates from pepper plant fruits were isolated via single-spore isolation. Of the 140 isolates, 26 were examined for mycelial growth, carbendazim sensitivity, and ITS sequence. Four of the isolates from the persimmon trees, which were cultivated exclusively in an orchard, showed fast mycelial growth and sensitivity to carbendazim, while five of the pepper isolates showed slower mycelial growth and were resistant to the fungicide. However, 17 isolates from persimmon trees cultivated with pepper plants in the same orchard showed slow mycelial growth like the pepper isolates and they were sensitive to carbendazim like the persimmon isolates. ITS sequence analysis of these 27 isolates led to the identification of the 22 persimmon isolates as C. gloeosporioides and the five pepper isolates as C. acutatum. PCR with species-specific primers confirmed that the 90 isolates from persimmon were C. gloeosporioides whereas the 50 isolates from pepper were C. acutatum. The 90 persimmon isolates of C. gloeosporioides and 50 pepper isolates of C. acutatum were compared by a wound inoculation test to determine their capacity for host cross-infection. All of the C. acutatum isolates from pepper caused typical symptoms of anthracnose on the fruits of pepper plants and twigs of persimmon; they differed from the C. gloeosporioides isolates from persimmon, more than 90% of which were able to infect only persimmon. Amplified fragment length polymorphism analysis revealed the existence of two groups (C. gloeosporioides and C. acutatum isolates group). At 80% genetic similarity, the C. gloeosporioides group was defined within four clusters, while the C. acutatum group was within three clusters. However, these clusterings were unrelated with the virulence of Colletotrichum species against pepper fruits.

The Holocene Environmental Change and Reconstruction of the Palaeogeography at Ilsan Area with the Special Reference to Pollen Analysis (花粉分析을 중심으로 본 一山지역의 홀로세 環境變化와 古地理復元)

  • Yoon, Soon-Ock
    • Journal of the Korean Geographical Society
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    • v.32 no.1
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    • pp.15-30
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    • 1997
  • This paper concerns the Holocene environmental change with vegetational history and sea-level fluctuation at Ilsan area by the analytical data of pollen, sedimentary facies and $C^14$-dating. The hypothetic palaeogeographic maps of the vegetation cover have been reconstructed with the reference to the periods of pollen zone. The environmental characteristics from the pollen zonation have been summerized as follows. 1)Pollenzone I(3.75~5.75m) showed the period of Alnus-and EMW-dominance. The study area was very humid under the influence of the transgression spreading widely from the rapid sea-level rise during the period(8,000~4,200y.BP). 2)Pollen zone II(5.75~6.35m) has been influenced by the fall of the sea-level and ground water surface. This zone(4,200~2,300y.BP) represented the period of spore~ and NAP-dominance with the increase of Pinus. 3) Pollen zone III(6.35~6.55m) has reflected the influence of the transgression and human interferences together. This zone(2,300~1,800y.BP) represented the period of NAP-dominance. The boundary between Subzone Ilb and Pollen zone III represents the same characteristics as what Weber says Grenzhorizont.

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Isolation and Characterization of Oligotrophic Bacteria Possessing Induced Systemic Disease Resistance against Plant Pathogens

  • Han, Song-Hee;Kang, Beom-Ryong;Lee, Jang-Hoon;Kim, Hyun-Jung;Park, Ju-Yeon;Kim, Jeong-Jun;Kim, Young-Cheol
    • The Plant Pathology Journal
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    • v.28 no.1
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    • pp.68-74
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    • 2012
  • Biocontrol microbes have mainly been screened among large collections of microorganisms $via.$ nutrient-rich $in$ $vitro$ assays to identify novel and effective isolates. However, thus far, isolates from only a few genera, mainly spore-forming bacilli, have been commercially developed. In order to isolate field-effective biocontrol microbes, we screened for more than 200 oligotrophic bacterial strains, isolated from rhizospheres of various soil samples in Korea, which induced systemic resistance against the soft-rot disease caused by $Pectobacterium$ $carotovorum$ SCC1; we subsequently conducted in $planta$ bioassay screening. Two oligotrophic bacterial strains were selected for induced systemic disease resistance against the $Tobacco$ $Mosaic$ $Virus$ and the gray mold disease caused by $Botrytis$ $cinerea$. The oligotrophic bacterial strains were identified as $Pseudomonas$ $manteilii$ B001 and $Bacillus$ $cereus$ C003 by biochemical analysis and the phylogenetic analysis of the 16S rRNA sequence. These bacterial strains did not exhibit any antifungal activities against plant pathogenic fungi but evidenced several other beneficial biocontrol traits, including phosphate solubilization and gelatin utilization. Collectively, our results indicate that the isolated oligotrophic bacterial strains possessing induced systemic disease resistance could provide useful tools as effective biopesticides and might be successfully used as cost-effective and preventive biocontrol agents in the field.

Culture-Independent Analysis of Microbial Succession During Composting of Swine Slurry and Mushroom Cultural Wastes

  • Cho, Kye-Man;Lee, Sun-Mi;Math, Renukaradhya K.;Islam, Shah Md. Asraful;Kambiranda, Devaiah M.;Kim, Jong-Min;Yun, Myoung-Geun;Cho, Ji-Joong;Kim, Jong-Ok;Lee, Young-Han;Kim, Hoon;Yun, Han-Dae
    • Journal of Microbiology and Biotechnology
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    • v.18 no.12
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    • pp.1874-1883
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    • 2008
  • Bacterial diversity and the composition of individual communities during the composting process of swine and mushroom cultural wastes in a field-scale composter (Hazaka system) were examined using a PCR-based approach. The composting process was divided into six stages based on recorded temperature changes. Phylogenetic analysis of eighty 16S rRNA sequences from uncultured composting bacterial groups revealed the presence of representatives from three divisions, including plant pathogenic bacteria, high-molecule-degrading bacteria and spore-forming bacteria. The plant pathogen A. tumefaciens gradually decreased in abundance during the composting process and eventually disappeared during the thermophilic and cooling stage. A bacterium homologous to Bacillus humi first appeared at the early thermophilic stage and was established at the intermediate thermophilic, post-thermophilic, and cooling stages. It was not possible to isolate the B. humi during any of the stages using general culture techniques.

Molecular Diversity of pagA Gene from Baciilus anthracis (탄저균 pagA 유전자의 분자적 다양성)

  • 김성주;조기승;최영길;채영규
    • Korean Journal of Microbiology
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    • v.37 no.1
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    • pp.49-55
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    • 2001
  • Bacillus anthracis is a gram-positive spore-forming bacterium that causes the disease anthrax. The anthrax toxin contains three components, including the protective antigen (PA), which binds to eucaryotic cell surface receptors and mediates the transport of toxins into the cell. In this study, the entire 2,294-nucleotide protective antigen gene (pag) was sequenced from 4 of B. anthracis strains to identify potential variation in the toxin and to further our understanding of B. anthracis evolution in Korea. Sequence alignment of the entire PA gene from 30 strains representative of the four B. anthracis diversity groups revealed mutations. The mutation of B. anthracis BAK are located adjacent to a highly antigenic region crossing the junction between PA domains 3 and 4 shown to be critical to LF binding. The different mutational combinations observed in this study give rise to 11 PA genotypes and 4PA phenotypes. Three-dimensional analysis of all the amino acid changes (Ala to Val) observed in BAK indicated that these changes are not only close sequentially but also very close in three-dimensional space to the antigenic region importan tfor LF binding. Phylogenetic (cladistic) analysis of the pag corresponded with previous strain grouping based on chromosomal variation, suggesting that plasmid evolution in B. anthracis has occurred with little or no horizontal transfer between the different strains.

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