• Title/Summary/Keyword: ITS1 and ITS2

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Taxonomical Classification and Species-specific Detection of Genus Some Phellinus using Phylotype (Phylotype에 의한 수종의 Phellinus속의 분류체계 확립 및 종간구별을 위한 신속동정법 개발)

  • Kim, Cheng-Yun;Lee, Jae-Yun;Kim, Gi-Young;Lee, Ki-Won;Park, Jae-Min;Kim, Mun-Ok;Lee, Tae-Ho;Lee, Jae-Dong
    • The Korean Journal of Mycology
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    • v.31 no.3
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    • pp.121-128
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    • 2003
  • This study was carried out to identify the phylogenetic relationship of Phellinus species and to know its distribution by comparing the DNA sequences of internal transcribed spacer regions(ITS1 and IST2) and 5.8S ribosomal DNA (rDNA) repeat unit. The Phellinus species had their specific sequences in IST1 and 2 regions depending on suedes. The comparison of the ITS sequences of standard strains indicated that the sequences of ITS1 were more variable than those of ITS2. Nine strains of the commercial products of Phellinus species used in this study were identified as P. lintues, P. baumii, P. igniarius, and P. pini. Most of commercial species were P. pini and P. baumii, and P. gilvus was not found. Also, P. linteus was only found in form of mycelial culture rather than fruiting body. Moreover, the species-specific primers were designed based on ITS sequence data. Each species-specific primers were bound in P. lintues(ITSF-PL2R), P. baumii(PB1F-ITS4R), P. igniarius(IF1-IR3), P. pini(PF1-PR3), and P. gilvus(GF2-GR4), respectively. These primer sets would be useful fer the detection of specific-species among unidentified Phellinus species rapidly.

Genetic Diversity of Amylomyces rouxii from Ragi tapai in Java Island Based on Ribosomal Regions ITS1/ITS2 and D1/D2

  • Delva, Ega;Arisuryanti, Tuty;Ilmi, Miftahul
    • Mycobiology
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    • v.50 no.2
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    • pp.132-141
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    • 2022
  • Amylomyces rouxii is commonly found as amylolytic fungi in tapai fermentation. However, its diversity is rarely reported despite being often used for food production in Southeast Asia. This research aims to analyze the genetic diversity and the distribution pattern of A. rouxii from Ragi tapai in Java Island, Indonesia. We isolated the fungus from samples obtained from Ragi tapai producing centers in Bandung, Sumedang, Muntilan, Blora, Yogyakarta, and Bondowoso. The obtained isolates were molecularly identified based on the ribosomal regions ITS1/ITS2 and D1/D2, then analyzed for phylogenetic tree reconstruction, genetic distance, genetic variation, and haplotype networking. Six isolates showed specific morphological traits of A. rouxii. However, phylogenetic tree reconstruction on the ribosomal genes showed that the isolates were grouped into two different clades related to two species. Clade A included BDG, SMD, and MTL isolates related to A. rouxii, whereas clade B included YOG, BLR, and BDS isolates related to Mucor indicus. The genetic distances between clades for ITS1/ITS2 and D1/D2 were 0.6145 and 0.1556, respectively. In conclusion, we confirmed the genetic diversity of molds from Ragi tapai in Java Island and showed that the isolates are not only related to A. rouxii as reported before.

Phylogenetic Relationship of Ligularia Species Based on RAPD and ITS Sequences Analyses (RAPD 및 ITS 염기서열 분석을 이용한 곰취 속(Ligularia) 식물의 유연관계 분석)

  • Ahn, Soon-Young;Cho, Kwang-Soo;Yoo, Ki-Oug;Suh, Jong-Taek
    • Horticultural Science & Technology
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    • v.28 no.4
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    • pp.638-647
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    • 2010
  • The genetic relationships in 5 species of $Ligularia$ were investigated using RAPD (Randomly Amplified Polymorphic DNA) and ITS (Internal Transcribed Spacer) sequences analyses. In RAPD analysis, sixty three of 196 arbitrary primers showed polymorphism. The amplified fragments ranged from 0.2 to 1.6 kb in size. The dendrogram was constructed by the UPGMA clustering algorithm based on genetic similarity of RAPD markers. A total of 16 accessions were classified into 5 major groups corresponding each species at the similarity coefficient value of 0.77. In the ITS sequence analysis, the size of ITS 1 was varied from 248 to 256 bp, while ITS 2 was varied from 220 to 222 bp. The 5.8S coding region was 164 bp in lengths. Forty nine sites (10.2%) of the 478 nucleotides were variable, and the G+C content of ITS region ranged from 49.4 to 53.5%. In the ITS tree, five species of $Ligularia$ were monophyletic, and $L.$ $taquetii$ was the first branching within the clade. $Ligularia$ $intermedia$ formed a clade with $L.$ $fischeri$ var. $spiciformis$ (BS=79), and $L.$ $stenocephala$ and $L.$ $fischeri$ were also claded. Two data sets were congruent, except of the position of $L.$ $fischeri$ var. $spiciformis$.

Phylogenetic Analysis of Native Vigna sinensis in Korea Using DNA Sequence of Internal Transcribed spacer (ITS) Region (토종 갓끈동부의 ITS1, 5.8S 및 ITS2의 염기서열을 이용한 계통 분석)

  • Seo, Pil-Soo;Lee, Sook-Young;Shin, Yong Kook
    • Journal of Life Science
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    • v.27 no.3
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    • pp.351-354
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    • 2017
  • Cowpea (Vigna unguiculata (L.) Walp.) is recognized as a potential source of protein and other nutrients. The genus Vigna includes 100 wild species of plants. Especially, Vigna unguiculata includes annual cowpeas (ssp. unguiculata) and ten wild perennial subspecies. DNA sequence of internal transcribed spacer (ITS) region was determined for Vigna sinensis, one of native plant, which was found in recent but thought to have gone extinct in Korea. The seeds of Vigna sinensis used in this study were donated from Dong-Young Jo. The DNA sequence of ITS-5.8S-ITS2 for Vigna sinensis obtained from this study was deposited as Vigna sinensis AY195581 on GenBank of NCBI (National Center for Biotechnology Information). We investigated the sequence-based phylogenetic relationships of plants related and clarified its taxonomical position. DNA similarities among subspecies including Vigna unguiculata showed the range 98 to 100% in sequence-based phylogenetic analysis using total 507 base pairs of ITS1, 5.8S and ITS2. Vigna unguiculata and subspecies were grouped independently as one cluster from other Vigna species used in the phylogenetic analysis. In this study, based on the phylogenetic analysis using the ITS1-5.8S-ITS2 sequence of Vigna sinensis, it may be concluded to be classified to one of Vigna unguiculata substrains.

Phylogenetic Analysis of the Entomopathogenic Fungal Species and Taxonomical Positions of Their Commercial Products (동충하초의 계통분류 및 시판동충하초의 분류학적 위치)

  • 김순한;이영자;김인복;김미경;한정아;홍무기;이순호;이재동
    • Journal of Life Science
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    • v.13 no.4
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    • pp.400-411
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    • 2003
  • This study was carried out to identify the phylogenetic relationship and to know the distribution of the entomopathogenic fungi by comparing the DNA sequences of internal transcribed spacer regions (ITS1 and ITS2) and 5.8S ribosomal DNA (rDNA) repeat unit. The entomopathogenic fungi had their specific sequences in ITS1 and 2 regions depending on species. The comparison of the ITS sequences of standard strains indicated that the sequences ITS1 were more variable than those of ITS2. It seems that Paecilomyces tenuipes, Isaria japonicus and P. japonicus are the same species but called as different names because of very similar sequences, and unidentified Paecilomyces sp. KACC 40220 and KACC 40656 showed identical sequences to P. tenuipes. Thirty six strains of the commercial products of entomopathogenic fungi used in this study were divided into four groups by the phylogenetic analysis based on 5.85 rDNA and ITS regions. We found twenty-three strains were P. tenuipes / japonica, eleven strains were C. militaris, and other two strains were Beauveria bassiana and C. multiaxialis, respectively.

Phylogenetic and Chemical Analyses of Cirsium pendulum and Cirsium setidens Inhabiting Korea (국내에 자생하는 큰엉겅퀴와 고려엉겅퀴의 분자유전학적 및 화학적 분석)

  • Yoo, Sun-Kyun;Bae, Young-Min
    • Journal of Life Science
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    • v.22 no.8
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    • pp.1120-1125
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    • 2012
  • Cirsium pendulum plants were collected from Hongcheon, Pyeongchang, Wonju, Yangyang in Kangwondo, Gapyeong in Gyeongkido, and Choongju in Choongcheongbukdo. Cirsium setidens plants were collected from Taebaek in Kangwondo and Bonghwa in Kyeongsangbukdo. Genomic DNA was prepared from those plants and used for the amplification of 18S rDNA, ITS1, 5.8S rDNA, ITS2, and part of 28S rDNA. The PCR products were sequenced, and the sequence was deposited in the GenBank. The comparison of those sequences has revealed that the rDNA sequences are identical for all six C. pendulum plants, but that the ITS1 and ITS2 sequences contain variable nucleotides. The two C. setidens plants had different nucleotides in 18S rDNA, ITS1, and ITS2. The comparison of the DNA sequences of C. pendulum and C. setidens collected in this study with C. pendulum of Hokkaido in Japan and C. japonicum of Anhui in China indicated that the plants of those three species are clearly divided into three distinct groups. The silymarin content of the collected plants was analyzed and turned out to be quite high. Therefore, it has been found that both C. pendulum and C. setidens plants are producing large amounts of silymarin, which has been reported to have various medicinal effects.

Identification of Phellinus linteus by Morphological Characteristics and Molecular Analysis (형태적.분자생물학적 방법에 의한 Phellinus linteus의 동정에 관한 연구)

  • Kim, Sang-Hee;Kim, Soo-Ho;Sung, Jae-Mo;Harrington, Thomas C.
    • The Korean Journal of Mycology
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    • v.27 no.5 s.92
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    • pp.337-340
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    • 1999
  • The context and upper surface of Phellinus basidiocarp become blackened, rimose and woody. The basidiocarp is sessile, dimidiate and elongate. The basidiospores are pigmented and ovoid to globose. Hymenial setae are $17{\sim}35{\times}6{\sim}8{\mu}m$. Nineteen isolates of Phellinus species, including Phellinus linteus, were used for sequencing of the internal transcribed spacer (ITS) region of the nuclear rDNA. Based on these sequence data, specific primers were designed for identification of Phellinus linteus isolates in Korea. The specific primers were within the ITS1 and ITS2 regions and were nested within the universal primers flanking the spacer regions. A total of four primers (the universal primers ITS-1F and ITS-4, and the specific primers PL-F and PL-R) were used for detection of Phellinus linteus collected in Korea. The length of the four amplification products of Phellinus linteus DNA were 800 bp (ITS-1F/ITS-4), two bands of about 720 bp (ITS-1F/PL-R and PL-F/ITS-4), and 610 bp (PL-F/PL-R). Among 23 isolates of Phellinus species collected in Korea, Thirteen isolates were identified as Phellinus linteus based on the presence of the four bands. The other species produced only the single ITS-1F/ITS-4 product.

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A phylogenetic analysis of Korean Artemisia L. based on ITS sequences (ITS 염기서열에 의한 한국산 쑥속(Artemisia L.)의 계통분류학적 연구)

  • Lee, Jeong-Hoon;Park, Chung-Berm;Park, Chun-Geon;Moon, Sung-Gi
    • Korean Journal of Plant Resources
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    • v.23 no.4
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    • pp.293-302
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    • 2010
  • Taxa of Artemisia collected in Korea were constructed by molecular phylogenetic analysis based on the internal transcribed spacer(ITS) regions of nrDNA. The length of the ITS sequences aligned using the clustal X program was 636~643 bp, and the lengths of the ITS1 and ITS2 regions were 251~255 bp and 217~222 bp, respectively. The total number of variable sites was 95 for the entire sequence, and a parsimony- informative site represented an efficacious site in ITS1 rather than in ITS2. The maximum parsimony tree as calculated by the MEGA 4 program was clustered into five clades. The taxa(A. capillaris, A. japonica var. japonica, A. japonica var. hallaisanensis, A. japonica subsp. littoricora) degenerated ovary of clade 1 was supported as the subgenus Dracunculus by Ling's classification system. The results show that A. nakaii and A. fukudo were quite similar genetically(Boostrap 99%) and that the scientific name of Korean A. dubia should be reconsidered. A. sp. distributed in Ganghwa province was grouped with A. argyi(Boostrap 89%). These results suggest that the molecular techniques used in this study could be useful for the phylogenetic analysis of Korean Artemisia herbs having variations in their morphological characteristics.

Food nutritional characteristics of fruit of Cudrania tricuspidata in its various maturation stages (꾸지뽕나무 열매의 숙기별 식품학적 특성)

  • Jung, Gi-Tai;Ju, In-Ok;Choi, So-Ra;You, Dong-Hyun;Noh, Jae-Jong
    • Food Science and Preservation
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    • v.20 no.3
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    • pp.330-335
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    • 2013
  • This study was conducted to investigate the food value of Cudrania tricuspidata at its various maturation stages. The pH, total acid and reducing sugar contents of its fruit juice were determined to have been 4.2~5.1, 1.4~2.0% and 5.4~8.6%, respectively. The general chemical components of its fruit were observed as 76~80% moisture, 2.2~3.5% crude protein, 1.7~2.9% crude fat, 0.8~1.2% ash and 14.5~16.4% carbohydrate. Its free sugar, glucose and fructose contents were determined. The fructose contents of both its ripened and over-ripened fruits were higher than their glucose contents. Organic acids such as oxalic acid, citric acid, tartaric acid, malic acid and succinic acid were detected, and the concentration of the malic acid and the succinic acid were found to have been most abundant. The K content was higher than the amounts of other minerals, such as Ca, Fe, K, Mg, Na and P. Its vitamin C and the total amount of its dietary fiber were 127.5~149.2 mg% and 22.7~38.7%, respectively. Its insoluble dietary fiber content was higher than its soluble dietary fiber content. Its total polyphenol and flavonoid content were 18.9~19.6 mg% and 40.9~48.2 mg%, respectively.

A Stereoselective Synthesis of (Z,Z)-3,13-Octadecadien-1-yl Acetate, and Its (E,Z)-Isomer, the Sex Pheromone of the Cherry Tree Borer, Synanthedon hector Butler

  • Kang, Suk-Ku;Park, Sun-Ku
    • Bulletin of the Korean Chemical Society
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    • v.9 no.3
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    • pp.149-152
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    • 1988
  • A mixture of (Z,Z)-3,13-octadecadien-1-yl acetate(1) and its (E,Z)-isomer(2), the sex pheromone of the cherry tree borer, Synanthedon hector Butler was synthesized. (Z)-11-Octadecen-1-al(3) was prepared from 1,10-decandiol. The Wittig reaction the above aldehyde3 with carboethoxymethylenetriphenylphosphorane, or the Wadsworth-Emmons reaction of the above aldehyde3 with the anion of triethylphosphonoacetate gave ethyl (Z,Z)-2,13-octadecadienoate and its (E,Z)-isomer. Deconjugative protonation of ethyl (Z,Z)-2,13-octadecadienoate and its (E,Z)-isomer with potassium hexamethyldisilazide followed by aqueous ammonium chloride work-up afforded stereoselectiv디y ethyl (E,Z)-3,13-octadecadienoate and its (Z,Z)-isomer, respectively, of which stereoselectivity was adjusted to give the product in the required ratio. Exposure of the above deconjugated ester to excess lithium aluminium hydride resulted in formation of the penultimate (Z,Z)-3,13-octadecadien-1-ol and its (E,Z)-isomer. Acetylation of the desired alcohols afford the final products, (Z,Z)-3,13-octadecadien-1-yl acetate(1) and its (E,Z)-isomer(2).