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

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Identification and Phylogenetic Relationship of Dermatophytes Based on RFLP Analysis and Nucleotide Sequence of Internal Transcribed Spacer (ITS)1 in Nuclear Ribosome DNA (ITS-RFLP와 ITS1 염기서열 분석에 의한 피부사상균의 동정과 계통적 유연관계)

  • Choi, Yeon-Hwa;Lee, Yeong-Seon;Yoo, Jae-Il;Kim, Bong-Su
    • The Journal of the Korean Society for Microbiology
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    • v.35 no.1
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    • pp.49-60
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    • 2000
  • ITSI-5.8S-ITSII rDNA region was amplified from the reference strains and clinical isolates with ITS1 and ITS4 primers. These primers amplified DNA fragments of 550 bp in Microsporum audouinii and Trichophyton violaceum, 700 bp in Microsporum gypseum, Trichophyton mentagrophytes, Trichophyton rubrum, and Trichophyton tonsurans, and 750 bp in Microsporum ferreugineum and Microsporum canis. The restriction enzyme patterns of PCR products digested with 13 restriction enzyme including PstI were distint among the genera, whereas identical in the same species. Examination of the ITS (Internal Transcribed Spacers)1 nucleotide sequence revealed that there was the genetic difference in each genera and species. Phylogenetic relationship among each species showed that the Trichophyton mentagrophytes was more closely related Trichophyton tonsurans than Trichophyton rubrum, and Microsporum gypseum was less related than Microsporum spp..

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Biomass and Molecular Characteristics of Multi-tillering Miscanthus Mutants

  • Lee, Geung-Joo;Zhang, Lili;Choi, Young In;Chung, Sung Jin;Yoo, Yong Kweon;Kim, Dong Sub;Kim, Sang Hoon
    • Korean Journal of Plant Resources
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    • v.25 no.6
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    • pp.745-752
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    • 2012
  • Compared to wide ranges of genetic variation of natural populations, very limited Miscanthus cultivar has been released. This study was the first report on the development of Miscanthus cultivar by means of radiation breeding. Seeds of M. sinensis were initially exposed to gamma rays of 250 Gy for 24 h, generated from a $^{60}Co$ gamma-irradiator. The irradiated seeds were sown and then the highly tiller-producing mutants were selected for this study. Biomass-related parameters including tiller number, plant height, stem diameter, and leaf number were measured. Ploidy level and internal transcribed spacer (ITS) were investigated to characterize the mutants compared to wild type (WT) Miscanthus. Plant height and tiller number were negatively related, where multi-tillering mutants were relatively short after 4 month growth. However stem diameter and leaf number were greater in mutants. All the materials used in this study were diploid, implying that the mutants with greater tiller numbers and stem diameter were not likely related to polyploidization. Based on the sequence of ITS regions, the mutants demonstrated base changes from the gamma irradiation where G+C content (%) was decreased in the ITS1, but increased in ITS2 when compared to WT sequence. ITS2 region was more variable than in ITS1 in the mutants, which collectively allows identification of the mutants from WT. Those mutants having enhanced tillers and allelic variations might be used as breeding materials for enhanced biomass-producing Miscanthus cultivars.

Molecular Phylogenetic Classification of Dermatophytes Isolated from Dogs and Cats (개와 고양이 유래 피부사상균의 분자생물학적 계통 분석)

  • Kim, Doo;Jeoung, Seok-Young;Ahn, So-Jeo
    • Journal of Veterinary Clinics
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    • v.23 no.4
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    • pp.405-410
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    • 2006
  • Using internal transcribed spacer 1 (ITS1) region ribosomal DNA sequences from 9 strains of Microsporum canis and 5 strains of Microsporum gypseum isolated from dogs and a cat with dermatophytosis, we demonstrated the mutual phylogenetic relationship of these strains. Nucleotide sequence analysis of the ITS 1 gene fragments from the 9 strains of M canis had the 100% nucleotide sequence similarities and the 5 strains of M gypseum also had the 100% nucleotide sequence similarities. The phylogenetic analysis of the nucleotide sequences of the 9 strains of M canis formed a nested cluster with the reference strains of M canis originating from USA, Australia, Japan, and Europe. M canis were genetically distinct from the other reference strains of Microsporum spp, but M distortum, M equinum, and M. ferrugineum were genetically very close to M canis. M gypseum from a cluster in the phylogenetic tree with M canis as an outgroup. The molecular analysis of ITS 1 genes provided the useful information for the identification of these microsporum species and the understanding of their relationship.

Use of Restriction Fragment Length Polymorphism Analysis to Differentiate Fungal Strains in Sunchang Meju

  • Jung, Jong-Hyun;Seo, Dong-Ho;Bhoo, Sung-Hee;Ha, Suk-Jin;Kim, Jong-Sang;Kim, Jeong-Hwan;Kwon, Dae-Young;Cha, Jae-Ho;Park, Cheon-Seok
    • Food Science and Biotechnology
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    • v.17 no.4
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    • pp.888-891
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    • 2008
  • Twenty-three fungal strains were isolated from meju that had originated from the Sunchang province, the famous location for making fermented soybean foods in Korea. The restriction fragment length polymorphism (RFLP) of the internal transcribed spacer (ITS) region of the rDNA (ITS-RFLP) was applied to differentiate the isolated fungal strains. First, the ITS region by polymerase chain reaction (PCR) with specific primers was amplified and then cleaved the products with different restriction enzymes. Cleavage of the amplified fragments with the restriction enzymes AluI, HaeIII, HhaI, and TaqI revealed extensive polymorphisms. The ITS-RFLP results highly correlated with ITS sequence analysis. All of the 23 fungal strains were classified into 5 groups by ITS-RFLP analysis. Aspergillus oryzae was the major fungal strain isolated from Sunchang meju (12 out of 23), while Aspergillus fumigatus was the next most frequently isolated strain (7 out of 23). In contrast, it was found that Fusarium asiaticum, Aspergillus sydowii, and Arthrinium sp. were the minor fungal strains in meju.

Determination of false positives in PCR diagnostics based on the internal transcribed spacer (ITS) of Gyrodactylus salaris using RFLP (RFLP를 이용한 Gyrodactylus salaris의 internal transcribed spacer(ITS) PCR 위양성 판별)

  • Min Seong Kim;Hee Jung Choi;Ji-Min Jeong;Mun-Gyeong Kwon;Seong Don Hwang
    • Journal of fish pathology
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    • v.37 no.1
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    • pp.147-153
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    • 2024
  • The World Organization for Animal Health (WOAH) recommends two protocols (ITS and COI) for conventional PCR of G. salaris diagnosis. However, ITS PCR protocol may yield false-positive results, leading to unnecessary countermeasures. It's difficult to distinguish between G. salaris and false-positive by similar amplicon size of PCR, since the amplicon size of ITS PCR in G. salaris and false-positive was 1,300 and 1,187 bp, respectively. The nucleotide sequences of ITS false-positive in rainbow trout is 99.7% identical to previously reported host genome sequences of rainbow trout (Oncorhynchus mykiss) and 95.3 to 89.1% identical to those of other salmonid fish species. To reduce false-positive PCR band, PCR was performed by the different annealing temperature, but PCR bands were still detected. In RFLP analysis by HaeIII, the PCR product of G. salaris was digested into four bands of 512, 399, 234 and 154 bp, while the false-positive was digested into seven bands of 297, 263, 242, 144, 93, 80 and 68 bp. In the RFLP patterns digested by HindIII, G. salaris showed two bands of 659 and 640 bp, while false-positive had one fragment of 1,187 bp without any digestion. Therefore, the RFLP method of ITS PCR with HaeIII and HindIII can be used for differentiation between G. salaris and false-positive. These results might provide important information on the improvement of PCR diagnostic method of G. salaris.

Intrageneric Relationships of Trichoderma Based on Internal Transcribed Spacers and 5.8S rDNA Nucleotide Sequences

  • Kim, Gi-Young;Lee, Goang-Jae;Ha, Myung-Gyu;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.28 no.1
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    • pp.11-16
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    • 2000
  • The nucleotide sequences of the internal transcribed spacer (ITS) regions of the ribosomal DNA including the 5.8S ribosomal RNA gene (rDNA) have been determined for 11 species in order to analyze their intrageneric relationships. The total length of these sequences ranged from 530 nucleotides for Trichoderma reesei KCTC 1286 to 553 nucleotide for Trichoderma koningii IAM 12534. Generally speaking, the length of ITS1 region was about 30 nucleotides longer than that of the ITS2 region. Also, 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 were also found. Thus, a neighbor-joining tree was constructed using the full sequence data of the ITS regions and the 5.8S rDNA. The Trichoderma genus used to be grouped on the basis of the morphological features and especially the shape of phialides needs to be reexamined. The phylogenetic tree displayed the presence of monophylogeny in the species of Trichoderma. Therefore, it was difficult to distinguish the intrageneric relationships in the Trichoderma genus.

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Analysis of the ITS (Internal Transcribed Spacer) Region of Opuntia ficus-indica (백년초선인장의 ITS(internal transcribed spacer) 유전자 분석)

  • In Jun-Gyo;Lee Bum-Soo;Kim Eun-Jeong;Choi Kwan-Sam;Han Seung-Ho;Shin Cheol-Woo;Yang Deok-Chun
    • Korean Journal of Plant Resources
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    • v.19 no.1
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    • pp.161-168
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    • 2006
  • To investigate the origin of backyeoncho (Opuntia ficus-indica var. saboten), we isolated 685 bp clone using ITS primer pairs. The rDNA consists of the genes coding for the partial 54 bp 185, 162 bp 5.8S, and partial 56 bp 26S. The coding regions are interrupted by two internal transcribed spacers, 193 bp ITS1 and 220 bp ITS2. The ITS2 of backnyeoncho in length was shorter than that previously registered in Cucurbitoideae plants. The GC contents was 66.8% in ITS1, and 67.7% in ITS2. The rDNA of backnyeoncho matched to the previously reported genes and showed a high similarity with the 95% identity with Pereskiopsis porteri (L708037). In the phylogenetic analysis, the backnyeoncho rDNA was clustered with Pereskiopsis porteri (L708037).

Identification of Entomopathogenic Fungus, Beauveria spp. F-101 Isolated from Thecodiplosis japonensis Using Internal Transcribed Spacer Sequence

  • Shin, Sang-Chul;Roh, Jong-Yul;Kim, Chul-Su;Park, Il-Kwon;Jeon, Mun-Jang;Je, Yeon-Ho
    • International Journal of Industrial Entomology and Biomaterials
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    • v.8 no.1
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    • pp.77-80
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    • 2004
  • For the development of the alternative control system against the major forest pests, Beauveria spp. F-101, isolated from a dead larva of Thecodiplosis japonensis, was selected because this isolate showed high pathogenicities against T. japonensis and Acantholyda parki. Beauveria spp. F-101 had irregular clustered conidio-phores and conidia borne on a distinctive apical zigzag extension, and it showed typical characteristic of the genus, Beauveria in morphology. For molecular based-identification, the ribosomal ITS region of Beauveria spp. F-101 was amplified with ITS1 and ITS4 primers, and cloned into pGEM- T Easy vector. The amplified PCR product was 569 bp in size and completely sequenced. The similarities of the cloned ITS sequence were 99 % and 97% to those of B. bassiana and B. brongniartii, respectively. In comparison to other species among the genus Beauveria, the ITS region of Beauveria spp. F-101 showed a similarity of 95% to B. amorpha, 95% to B. tenella, 89% to B. vermiconia and 69% to B. alba, respectively. In addition, in comparison to different genus, it had 95% similarities to Cordyceps militaris and 91% to Paecilomyces tenuipes. Accordingly, the current result suggests that Beauveria spp. F-101 was a variant of B. bassiana and it seems to be a new isolate considering sequence variation in ITS region.

Characteristics of Plantlets Redifferentiated from F1 Hybrid between Panax ginseng and Panax quinquefolius (고려인삼과 미국삼의 종간잡종으로부터 재분화된 식물체의 특성)

  • Ahn, In-Ok;Lee, Sung-Sik;Lee, Jang-Ho;Lee, Bum-Soo;In, Jun-Gyo;Yang, Deok-Chun
    • Journal of Plant Biotechnology
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    • v.33 no.1
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    • pp.45-48
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    • 2006
  • The characteristics of plantlets redifferentiated from calli of F1 hybrid between Panax ginseng and Panax quinquefolius were investigated. Growth of plantlets redifferentiated from F1 hybrid was superior to the plants redifferentiated from Korean ginseng. Stem color of plantlets redifferentiated from F1 hybrid was more purple than that from Korean ginseng and leaf color of the former was also greener than that of the latter. Chunpoong, Yunpoong and Seonweon which are belonged to Korean ginseng showed same PCR band(A), while American ginseng showed different PCR band (B) in Internal Transcribed Spacer (ITS) region. F1 hybrid exhibited both A and B PCR band which belonged to Korean ginseng and American ginseng, respectively. F1 hybrid calli and plantlets redifferentiated from F1 hybrid calli showed same PCR band with that of F1 hybrid plant in ITS region. Therefore it was confirmed that piantlets redifferentiated from F1 hybrid exhibited genetic stability in ITS region.