• Title/Summary/Keyword: ITS (internal transcribed spacer)

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Identification and Characterization of Paraconiothyrium brasiliense from Garden Plant Pachysandra terminalis (가든식물 수호초(Pachysandra terminalis)로부터 Paraconiothyrium brasiliense의 분리 및 동정)

  • Choi, Min Ah;Park, Seung Jun;Ahn, Geum Ran;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.42 no.4
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    • pp.262-268
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    • 2014
  • A fungal isolate DUCC5000 from a garden plant Pachysandra terminalis was identified as Paraconiothyrium brasiliense based on the results of morphological and molecular studies. The fungus formed brown to black conidiomata of (0.2-0.7)-2(-3.5) mm singly or as a group on PDA. Conidia measured $2-5{\times}1.8-3{\mu}m$ in size, hyaline, ellipsoid to short-cylindrical, and rounded at both ends. The internal transcribed spacer (ITS) DNA of the isolate shared 100% nucleotide sequence homology with those of known P. brasiliense isolates. Phylogenetic tree inferred from the ITS sequence analysis showed that the DUCC5000 isolate formed a clade with known isolates of P. brasiliense. The fungal mycelia grew better on oatmeal agar than on MEA and PDA. On PDA media under various pH conditions, fungal mycelial growth was observed at pH 9. Colony morphology of the fungus tended to alter depending on the kinds of nutrient media and pH condition. On chromagenic media, the fungus demonstrated its ability to produce extracellular enzymes including amyalse, avicelase, ${\beta}$-glucosidase, protease, and xylanase. However, in pathogenicity testing, no disease symptoms were observed on the leaves of P. terminalis. This strain is the first report on P. terminalis in Korea.

Morphological and Molecular Identification of Spirometra Tapeworms (Cestoda: Diphyllobothriidae) from Carnivorous Mammals in the Serengeti and Selous Ecosystems of Tanzania

  • Ndosi, Barakaeli Abdieli;Park, Hansol;Lee, Dongmin;Choe, Seongjun;Kang, Yeseul;Nath, Tilak Chandra;Bia, Mohammed Mebarek;Eamudomkarn, Chatanun;Jeon, Hyeong-Kyu;Eom, Keeseon S.
    • Parasites, Hosts and Diseases
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    • v.58 no.6
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    • pp.653-660
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    • 2020
  • Spirometra tapeworms (Cestoda: Diphyllobothriidae) collected from carnivorous mammals in Tanzania were identified by the DNA sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and internal transcribed spacer 1 (ITS1), and by morphological characteristics. A total of 15 adult worms were collected from stool samples and carcasses of Panthera leo, Panthera pardus, and Crocuta crocuta in the Serengeti and Selous ecosystems of Tanzania. Three Spirometra species: S. theileri, S. ranarum and S. erinaceieuropaei were identified based on morphological features. Partial cox1 sequences (400 bp) of 10 specimens were revealed. Eight specimens showed 99.5% similarity with Spirometra theileri (MK955901), 1 specimen showed 99.5% similarity with the Korean S. erinaceieuropaei and 1 specimen had 99.5% similarity with Myanmar S. ranarum. Sequence homology estimates for the ITS1 region of S. theileri were 89.8% with S. erinaceieuropaei, 82.5% with S. decipiens, and 78.3% with S. ranarum; and 94.4% homology was observed between S. decipiens and S. ranarum. Phylogenetic analyses were performed with 4 species of Spirometra and 2 species of Dibothriocephalus (=Diphyllobothrium). By both ML and BI methods, cox1 and ITS1 gave well supported, congruent trees topology of S. erinaceieuropaei and S. theileri with S. decipiens and S. ranarum forming a clade. The Dibothriocephalus species were sisters of each other and collectively forming successive outgroups. Our findings confirmed that 3 Spirometra species (S. theileri, S. ranarum, and S. erinaceieuropaei) are distributed in the Serengeti and Selous ecosystems of Tanzania.

Quantification of triterpenes in Centella asiatica cultivated in a smart farm, and their effect on keratinocyte activation (스마트팜 재배 병풀의 triterpenes 정량 및 각질형성세포 활성화 효과)

  • Jin Hong Park;Seong Min Jo;Da Hee Lee;Youngmin Park;Hwan Bong Chang;Tae Jin Kang;Kiman Lee
    • Food Science and Preservation
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    • v.30 no.3
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    • pp.483-491
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    • 2023
  • This study aimed to compare the bioactive compounds in Centella asiatica (C. asiatica) cultivated in a smart farm and a field and their effects on human keratinocyte cells. C. asiatica was collected in Jeju-do, Korea, and cultured in a smart farm and a field. The main bioactive compounds in the two differentially cultured C. asiatica were identified, and their activation in keratinocytes were assessed. Amplification and sequencing of the internal transcribed spacer (ITS) DNA in the nucleus and psbA-H DNA in the chloroplast were performed for species analysis. A comparison of DNA of plants reported in the NCBI GenBank was performed. The ITS DNA and psbA-H DNA sequences of C. asiatica cultivated in a smart farm and a field were consistent with No. MH768338.1 and No. JQ425422.1, respectively. Analysis of the triterpenes was performed using high performance liquid chromatography (HPLC) and as a result, C. asiatica cultured in a smart farm had more triterpenes than those cultured in a field. The effects of C. asiatica grown in a smart farm on cell proliferation and scratch recovery in HaCaT cells were greater than those grown in a field. These results suggest that C. asiatica cultivated in a smart farm can be effectively utilized as a health functional food.

Phylogeny of the Yeast Species Isolated from Wild Tiger Lily (Lilium lancifolium Thunb.) (야생 참나리(Lilium lancifolium Thunb.)로부터 분리한 효모의 분자계통학적 분석)

  • Kim, Jong-Shik;Kim, Dae-Shin
    • Korean Journal of Environmental Agriculture
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    • v.34 no.2
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    • pp.149-154
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    • 2015
  • BACKGROUND: Yeast isolates associated with the leaves, stems, and flowers of the tiger lily needed to be identified using isolation methods that have previously been used effectively in yeast biotechnology. A culture-based approach was necessary for the isolation of many yeast strains associated with tiger lily. METHODS AND RESULTS: In this study, the homogenized leaves, stems, and flowers of tiger lily were spreaded onto GPY medium containing chloramphenicol, streptomycin, Triton X-100, and L-sorbose. A total of 82 yeast strains from the leaves, 94 and 97 yeast strains from the stems and flowers were isolated, respectively. Yeast isolates were identified by phylogenetic analysis based on internal transcribed spacer region sequencing. The yeast species isolated from the leaves comprised of 31 isolates of the genus Pseudozyma, 28 of Aureobasidium pullulans, and 11 of the genus Cryptococcus. Those isolated from the stems comprised of 40 of A. pullulans and 11 of Cryptococcus, and 95 of A. pullulans While, 1 isolate each of the genera Rhodotorula and Metschnikowia were isolated from the flowers. CONCLUSION: We identified site-specific yeast communities associated with tiger lily. These yeast isolates may have high potential for application in the field of biotechnology.

Enhancement of the germination efficiency of Gastrodia elata seeds using a new Mycena species (신규 애주름버섯 NIFOS101을 이용한 천마 종자의 발아효율 증진)

  • Lee, Hyo-Ryeon;Han, Muho;Choi, Mi Na;Lee, Hyoshin;Lee, Seok-Woo;Park, Eung-Jun
    • Journal of Plant Biotechnology
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    • v.44 no.1
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    • pp.56-60
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    • 2017
  • This study investigated the in vitro symbiotic seed germination of the achlorophyllous orchid, Gastrodia elata, using a new Mycena species. A leaf-disc ($2{\times}2cm$) of Quercus accutissima was inoculated with either of the two fungal species, NIFOS101 (NCBI accession number KY449288) or KFRI1212 (HQ662845), following which it was placed on water agar medium, prior to sowing seeds of G. elata. After 20 days of symbiotic culture, NIFOS101 and KFRI1212 germinated 94% and 70% of G. elata seeds, respectively, although the initiation of protocorm development was induced much earlier by KFRI 1212 than by NIFOS101. Furthermore, the NIFOS101 mycelia grew much faster than KFRI 1212 at all temperatures tested. A phylogenetic analysis using the internal transcribed spacer (ITS) sequences showed that NIFOS101 belonged to a clade with M. purpureofusca, which completely differed from the clade with KFRI1212. This study not only identified a new fungal species, NIFOS101, which improved the rate of symbiotic seed germination up to 94% as compared to KFRI1212 (70%), but also revealed that G. elata required a broad taxonomic range of fungi for its symbiotic germination.

Insecticidal Effect of an Entomopathogenic Fungus, Beauveria bassiana ANU1 to Spodoptera exigua and Plutella xylostella by Different Temperature and Humidity Conditions (파밤나방과 배추좀나방에 대한 곤충병원성 곰팡이 Beauveria bassiana ANU1의 온도와 습도조건에 따른 살충효과)

  • Lee, Jung-Bok;Park, Youngjin
    • The Korean Journal of Pesticide Science
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    • v.19 no.2
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    • pp.125-133
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    • 2015
  • Entomopathogenic fungi have been studied to develop for biological control agents as an alternative to chemical control agents in insect pest management. Two Lepidopteran insects, Spodoptera exigua and Plutella xylostella, are serious insect pests infested various crops, but not effectively controlled by commercial chemical pesticides due to its high insecticide resistance. A fungal isolate was isolated from S. exigua larvae collected from green onion field in Andong, Korea. To identify the fungal isolate, 18srRNA sequence for internal transcribed spacer (ITS) and ${\beta}$-tubulin regions were sequenced. The ITS and ${\beta}$-tubulin sequence were highly matched to Beauveria bassiana and morphological characteristics also was fit to known B. bassiana. Finally, isolated fungus has identified as B. bassiana and named B. bassiana ANU1. The result of bioassay, median lethal concentrations were $2.7{\times}10^3$ and $0.9{\times}10^3conidia/ml$ and medial lethal times were 65.6 and 60.8 h to S. exigua and P. xylostella, respectively. B. bassiana ANU1 showed high pathogenicity to two insect pests from $20^{\circ}C$ to $30^{\circ}C$ at 50% relative humidity (RH) and more than 40% RH at $25^{\circ}C$ with $10^7conidia/ml$ of concentration.

Identification of Phellinus linteus by Comparison of Colony Shapes and Using PCR techniques (목질진흙버섯(Phellinus linteus)의 균총형태 비교 및 PCR 기법을 이용한 동정)

  • Kong, Won-Sik;Kim, Dong-Hyun;You, Chang-Hyun;Kim, Young-Ho;Kim, Kyung-Soo;Kim, Kwang-Ho
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.466-477
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    • 1998
  • Twenty-two Phellinus strains were characterized using colony morphologies and polymerase chain reaction (PCR) to divide into Phellinus linteus. There were some differences in mycelial growth and colony shapes among the strains when they were grown on various media such as PDA, MCM, MEA and YM. Phellinus linteus was slowly growing, formed golden-yellow colony, and produced blue pigment on PDA media. When the regions of internal transcribed spacer (ITS) were amplified from ribosomal RNA (rRNA) coding genes of P. igniarius and P. linteus strains by means of PCR, two types of band (700 bp and 800 bp) were appeared, respectively. For the amplified intergenic region I (IGRI), P. igniarius strains showed a different band among 500, 600, 700 and 800 bp according to the strains, whereas P. linteus strains did one specific band of 700 bp. By polymorphism analysis after digesting the amplified products with 6 different restriction enzymes, a band specific to P. linteus was generated when the products for ITS region were digested with HaeIII, suggesting that the enzyme digestion could provide effective method to distinguish between P. igniarius and P. linteus. And also, the analysis of genetic relationship showed that the genetic similarities were 89% and 95% in P. igniarius and P. linteus strains, respectively. Random amplification polymorphic DNA (RAPD) analysis using multiple primer sets and arbitrarily primed PCR (AP-PCR) with ITS3 primer could also result in a reproducible way to identify P. linteus strains.

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Occurrence of Sclerotium Rot of Cucumber Caused by Sclerotium rolfsii (Sclerotium rolfsii에 의한 오이 흰비단병 발생)

  • Kwon, Jin-Hyeuk;Lee, Sang-Dae;Choi, Okryun;Shen, Shun-Shan;Shim, Hong-Sik
    • Research in Plant Disease
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    • v.19 no.3
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    • pp.229-232
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    • 2013
  • Sclerotium rot of cucumber (Cucumis sativus L.) occurred at the experimental field of Gyeongsangnam-do Agricultural Research and Extension Services in July 2012. The typical symptoms included wilt, rot, and water-soaking on stems and fruits and severely infected plants eventually died. White mycelial mats spread over lesions, and then sclerotia were formed on fruit and near soil line. The sclerotia were globoid in shape, white to brown in color and 1-3 mm in size and the hyphal width was 4-8 ${\mu}m$. The optimum temperature for mycelial growth and sclerotia formation on PDA was $30^{\circ}C$. The typical clamp connections were observed in the hyphae of the fungus grown on PDA. For further identification, the complete internal transcribed spacer (ITS) rDNA region was amplified and sequenced. On the basis of mycological characteristics, ITS rDNA region comparison, and pathogenicity to host plants, this fungus was identified as Sclerotium rolfsii Saccardo. This is the first report of sclerotium rot on cucumber caused by S. rolfsii in Korea.

Mitochondrial Genome Sequence of Echinostoma revolutum from Red-Crowned Crane (Grus japonensis)

  • Ran, Rongkun;Zhao, Qi;Abuzeid, Asmaa M.I.;Huang, Yue;Liu, Yunqiu;Sun, Yongxiang;He, Long;Li, Xiu;Liu, Jumei;Li, Guoqing
    • Parasites, Hosts and Diseases
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    • v.58 no.1
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    • pp.73-79
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    • 2020
  • Echinostoma revolutum is a zoonotic food-borne intestinal trematode that can cause intestinal bleeding, enteritis, and diarrhea in human and birds. To identify a suspected E. revolutum trematode from a red-crowned crane (Grus japonensis) and to reveal the genetic characteristics of its mitochondrial (mt) genome, the internal transcribed spacer (ITS) and complete mt genome sequence of this trematode were amplified. The results identified the trematode as E. revolutum. Its entire mt genome sequence was 15,714 bp in length, including 12 protein-coding genes, 22 transfer RNA genes, 2 ribosomal RNA genes and one non-coding region (NCR), with 61.73% A+T base content and a significant AT preference. The length of the 22 tRNA genes ranged from 59 bp to 70 bp, and their secondary structure showed the typical cloverleaf and D-loop structure. The length of the large subunit of rRNA (rrnL) and the small subunit of rRNA (rrnS) gene was 1,011 bp and 742 bp, respectively. Phylogenetic trees showed that E. revolutum and E. miyagawai clustered together, belonging to Echinostomatidae with Hypoderaeum conoideum. This study may enrich the mitochondrial gene database of Echinostoma trematodes and provide valuable data for studying the molecular identification and phylogeny of some digenean trematodes.

Screening and Characteristics of Useful Fungi for Brewing from Commercial Nuruk in Chungcheong Provinces (충청지역 누룩에서 양조용 우수 곰팡이의 탐색 및 특성)

  • Baek, Seong-Yeol;Yun, Hye-Ju;Choi, Hye-Sun;Hong, Seung-Beom;Koo, Bon-Sung;Yeo, Soo-Hwan
    • Microbiology and Biotechnology Letters
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    • v.38 no.4
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    • pp.373-378
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    • 2010
  • Studies on standardization and quality upgrade of nuruk which is a basic component in brewing are required to increase the quality level of Korean traditional rice wines and to develop the technology for practical use of it. It is important to isolate best strains, to improve the properties and effectively preserve them for brewing industry. In this study, 16 commercial nuruk samples were obtained from the commercial markets located in Chungcheong areas in Korea. 174 fungal strains were isolated from the samples on DG18 medium using a dilution plating method and then screened for enzyme activity and acid production. The active strains were identified based on the morphological characteristics and ITS sequence analysis. Out of 174 strains, 12 strains showed high amylase activity. Especially, Rhizopus sp. CN084, CN174, Aspergillus sp. CN161 and Mycocladus sp. CN042 showed high saccharogenic power and dextrinogenic enzyme activity on cooked wheat bran medium. On the other hand, Aspergillus sp. CN010, CN161, Rhizopus sp. CN105, CN168 and Rhizomucor sp. CN088 produced high acid production on the same medium. Our results showed that the active strains may be used as microbial sources for nuruk starter with good quality in brewing.