• Fisheries and Aquatic Sciences
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• 제15권2호
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• pp.157-162
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• 2012

To analyze nucleotide sequence encoding internal transcribed spacer (ITS) regions specific to the Laminariaceae family, genomic DNA was isolated from six brown algae species distributed along the east coast of Korea. These included three species from the Laminariaceae family (Agarum clathratum Dumortier, Costaria costata [C. Agardh] Saunders, and Saccharina japonica Areschoug) and two species from the Alariaceae family (Undaria pinnatifida [Harvey] Suringer and Ecklonia cava Kjellman), both in the order Laminariales, and one species from the family Sargassaceae in the order Fucales (Sargassum serratifolium). Based on a sequence analysis of ITS-1 and ITS-2 for A. clathratum, C. costata, and E. cava, oligonucleotides were designed from the regions that showed sequence conservation in Laminariaceae. Following polymerase chain reaction using three sets of primers, amplification of ITS-1 and ITS-2 was detected in reactions using genomic DNA isolated from the species belonging to Laminariaceae, but not from the species belonging to the other families. The results indicate that this method can be used for the detection and identification of Laminariaceae species.

• Parasites, Hosts and Diseases
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• 제42권3호
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• pp.145-148
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• 2004

We compared the DNA sequence difference of isolates of Clonorchis sinensis from one Korean (Kimhae) and two Chinese areas (Guangxi and Shenyang), The sequences of nuclear rDNA (18S, internal transcribed spacer 1 and 2: ITS1 and ITS2) and mitochondrial DNA (cytochrome c oxidase subunit 1: cox1) were compared. A very few intraspecific nucleotide substitution of the 18S, ITS1, ITS2 and cox1 was found among three isolates of C. sinensis and a few nucleotide insertion and deletion of ITS1 were detected. The 18S, ITS1, ITS2 and cox1 sequences were highly conserved among three isolates. These findings indicated that the Korean and two Chinese isolates are similar at the DNA sequence level.

• 한국균학회지
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• 제44권1호
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• pp.1-7
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• 2016

우리나라 토양에서 효모 종 분포특성을 조사하기 위해 먼저 대전광역시를 포함하는 충청남도 전 지역의 밭 토양 61점을 2015년 6월부터 8월까지 수집하여 97균주 20종의 야생효모들을 분리하였고, polymerase chain reaction (PCR)을 통해 internal transcribed spacer (ITS) 부위와 26S rDNA의 D1/D2 부위의 염기서열 상동성 비교법을 이용하여 종을 동정하였다. 이들 가운데 Cryptococcus podzolicus가 11균주를 포함하는 Cryptococcus 속 균이 전체 분리 균주 중 16균주(16%)로 가장 많이 분리되었고, 다음으로 Debaryomyces hansenii와 Trichorsporon asahii도 각각 6주씩 분리되었다.

• 한국균학회지
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• 제44권4호
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• pp.252-258
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• 2016

본 연구는 제주도 사려니숲과 동백동산에서 편백나무(Chamaecyparis obtusa), 삼나무(Cryptomeria japonica), 비자나무(Torreya nucifera), 꽝꽝나무(Ilex crenata), 동백나무(Camellia japonica) 등 5종의 목본 식물의 잎에서 내생균을 분리하고, 형태 및 분자생물학적 특징 통해 동정하였다. 이를 위해 primer인 ITS1F와 ITS4를 이용하여 internal transcribed spacer (ITS) rDNA영역과 primer LR0R과 LR16을 이용하여 large subunit rDNA 영역을 그리고 primer Bt2a과 Bt2b를 이용하여 ${\beta}$-tubulin 영역의 염기서열을 동시에 분석하였다. 그 결과 Mycosphaerella aleuritidis, Neofusicoccum eucalyptorum, Neofusicoccum parvum, Phyllosticta citrichinensis, Phyllosticta cryptomeriae, Phomopsis cotoneastri, Sphaerulina rhododendricola, Guignardia mangiferae, Lophodermium jiangnanense, Lophodermium minus 등 10종의 미기록 내생균을 동정하였고, 이를 보고하고자 한다.

• 한국균학회지
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• 제44권4호
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• pp.365-370
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• 2016

본 연구에서는 잣나무, 눈측백의 침엽과 자란의 뿌리에서 식물에 공생하는 내생균을 분리하여 동정하였다. 이를 위해 specific primer인 ITS1F와 ITS4를 이용하여 internal transcribed spacer (ITS) rDNA영역과 primer LR0R과 LR 16을 이용하여 large subunit rDNA 영역을, primer Bt2a과 Bt2b를 이용하여 ${\beta}$-tubulin 영역을, 그리고 EF-1 primer를 이용하여 translation elongation factor 지역의 염기서열을 동시에 분석하였다. 연구 결과 Colletotrichum simmondsii, Fusarium sterilihyphosum, Diatrypella pulvinata, Ochroconis globalis, Sphaeria chrysosperma 등 국내 미기록 균주 5종을 분리하여 형태적, 분자생물학적 동정 결과를 서술하였다.

• 한국약용작물학회지
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• 제23권6호
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• pp.439-445
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• 2015

Background : Correct identification of Panax species is important to ensure food quality, safety, authenticity and health for consumers. This paper describes a high resolution melting (HRM) analysis based method using internal transcribed spacer (ITS) and 5.8S ribosomal DNA barcoding regions as target (Bar-HRM) to obtain barcoding information for the major Panax species and to identify the origin of ginseng plant. Methods and Results : A PCR-based approach, Bar-HRM was developed to discriminate among Panax species. In this study, the ITS1, ITS2, and 5.8S rDNA genes were targeted for testing, since these have been identified as suitable genes for use in the identification of Panax species. The HRM analysis generated cluster patterns that were specific and sensitive enough to detect small sequence differences among the tested Panax species. Conclusion : The results of this study show that the HRM curve analysis of the ITS regions and 5.8S rDNA sequences is a simple, quick, and reproducible method. It can simultaneously identify three Panax species and screen for variants. Thus, ITS1HRM and 5.8SHRM primer sets can be used to distinguish among Panax species.

• Journal of Ginseng Research
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• 제36권2호
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• pp.211-217
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• 2012

Endophytic fungi were isolated from various tissues (root, stem, petiole, leaf, and flower stalk) of 3- and 4-year-old ginseng plants (Panax ginseng Meyer) cultivated in Korea. The isolated endophytic fungi were identified based on the sequence analysis of the internal transcribed spacer (ITS), 1-5.8-ITS 2. A morphological characterization was also conducted using microscopic observations. According to the identification, 127 fungal isolates were assigned to 27 taxa. The genera of Phoma, Alternaria and Colletotrichum were the most frequent isolates, followed by Fusarium, Entrophospora and Xylaria. Although 19 of the 27 taxa were identified at the species level, the remainder were classified at the genus level (6 isolates), phylum level (Ascomycota, 1 isolate), and unknown fungal species (1 isolate). Endophytic fungi of 13 and 19 species were isolated from 3- and 4-year-old ginseng plants, respectively, and Phoma radicina and Fusarium solani were the most frequently isolated species colonizing the tissues of the 3- and 4-year-old ginseng plants, respectively. The colonization frequency (CF%) was dependant on the age and tissue examined: the CFs of the roots and stems in the 3-year-old ginseng were higher than the CF of tissues in the 4-year-old plants. In contrast, higher CFs were observed in the leaves and petioles of 4-year-old plants, and endophytic fungi in the flower stalks were only detected in the 4-year-old plants. In conclusion, we detected diverse endophytic fungi in ginseng plants, which were distributed differently depending on the age and tissue examined.

• Parasites, Hosts and Diseases
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• 제52권5호
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• pp.471-478
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• 2014

Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.

• Mycobiology
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• 제29권3호
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• pp.121-131
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• 2001

This study was carried out to identify the phylogenetic relationships among several caterpillar fungi by comparing the sequences of internal transcribed spacer regions(ITS1 and ITS2) and 5.8S ribosomal DNA(rDNA) repeat unit. The sequences of ITS1, ITS2, and the 5.8S rDNA from 10 strains of Cordyceps species, 12 strains of Paecilomyces, 3 strains of Beauveria, 2 strains of Metarhizium and 1 strains of Hirsutella were amplified, determined and compared with the previously known Cordyceps species. The sequences of 5.8S rDNA were more conserved in length and variation than those of ITS regions. Although the variable ITS sequences were often ambiguously aligned, the conserved sites could be found. In the phylogenetic tree, the species generally divided into three clusters, supported by their morphology and/or host ranges. The 5.8S rDNA and TTS1 sequences among 10 species of Cordyceps militaris were identical and only one base pair in ITS2 sequence was different. Cordyceps sinensis and Cordyceps ophioglossoides were also clearly different, although they belonged to the same cluster. The Geniank database search of species revealed sister taxa of an entomogenous fungus. Metarhizium was used as an putgroup in all taxa.

• 한국균학회지
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• 제43권1호
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• pp.58-63
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• 2015

진균에 의한 오염은 재배사에서 톱밥 배지를 이용한 표고 재배에 중대한 영향을 미치기 때문에 표고 재배사 실내 공기에 존재하는 진균에 대한 모니터링을 수행하던 중 여러 진균을 분리하였다. 분리된 진균 중에는 국내 미기록인 Aspergillus pulverulentus와 Cosmospora butyri 등 2종을 비롯하여 기록은 있으나 균학적 확증이 부족한 Lecanicillium psalliotae와 L. antillanum 등 2종이 존재하였다. 본 논문에서는 이들 균류에 대한 형태적 특성과 더불어 internal transcribed spacer (ITS) rDNA region 또는 ${\beta}$-tubulin 유전자 염기서열에 기반한 계통학적 분석 결과를 기술하였다.