• Title/Summary/Keyword: rDNA internal transcribed spacer sequence

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Phylogenetic relationships of genera Polyporus on the basis of ITS region sequences (rDNA의 ITS 부위 염기서열 분석에 의한 겨울우산버섯(Polyporus)속 균주의 유전적인 유연관계 분석)

  • Lee, Chan-Jung;Jhune, Chang-Sung;Cheong, Jong-Chun;Kong, Won S.
    • Journal of Mushroom
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    • v.10 no.1
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    • pp.37-43
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    • 2012
  • This study was carried to identify a correct species and asses genetic diversity within the same species of Polyporus spp. preserved in Division of applied Microbiology. Contaminated isolates showed different growth rates, morphology and color of hyphae. We have reconstructed the phylogenetic tree of a select group of Polyporus spp. using nucleotide sequences of the internal transcribed spacer region(ITS) region. The phylogenetic tree was constructed by using the neighbor-joining method. PELF primers of 20-mer were used to assess genetic diversity of preserved isolates. Sequence analysis showed that three strains were different species and four strains were identified completely different nomenclature. According to the analysis of ITS sequences, the genus Polyporus clustered into five distinct group, most of which correlated with species-groups identified by RAPD method. Four isolates included strain PM02 showed high similarity with P. arcularius, four isolates included strain PM03 high similarity with P. alveolaris, three isolates included strain PM01 high similarity with P. tuberaster, and PM 06 and PM04 high similarity with P. brumalis and P. squamossus. Isolates were collected in the United States(PM10, PM11) was identified as P. alveolarius and P. arcularius. RAPD analysis of genetic polymorphisms of genus Polyporus showed a very different band patterns. As the result of RAPD and ITS region sequences analysis for preserved isolates, it seems likely that 6 isolates of Polyporus spp. may be need to reclassify or eliminate from preserved catalogue.

An assessment of the taxonomic reliability of DNA barcode sequences in publicly available databases

  • Jin, Soyeong;Kim, Kwang Young;Kim, Min-Seok;Park, Chungoo
    • ALGAE
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    • v.35 no.3
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    • pp.293-301
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    • 2020
  • The applications of DNA barcoding have a wide range of uses, such as in taxonomic studies to help elucidate cryptic species and phylogenetic relationships and analyzing environmental samples for biodiversity monitoring and conservation assessments of species. After obtaining the DNA barcode sequences, sequence similarity-based homology analysis is commonly used. This means that the obtained barcode sequences are compared to the DNA barcode reference databases. This bioinformatic analysis necessarily implies that the overall quantity and quality of the reference databases must be stringently monitored to not have an adverse impact on the accuracy of species identification. With the development of next-generation sequencing techniques, a noticeably large number of DNA barcode sequences have been produced and are stored in online databases, but their degree of validity, accuracy, and reliability have not been extensively investigated. In this study, we investigated the extent to which the amount and types of erroneous barcode sequences were deposited in publicly accessible databases. Over 4.1 million sequences were investigated in three largescale DNA barcode databases (NCBI GenBank, Barcode of Life Data System [BOLD], and Protist Ribosomal Reference database [PR2]) for four major DNA barcodes (cytochrome c oxidase subunit 1 [COI], internal transcribed spacer [ITS], ribulose bisphosphate carboxylase large chain [rbcL], and 18S ribosomal RNA [18S rRNA]); approximately 2% of erroneous barcode sequences were found and their taxonomic distributions were uneven. Consequently, our present findings provide compelling evidence of data quality problems along with insufficient and unreliable annotation of taxonomic data in DNA barcode databases. Therefore, we suggest that if ambiguous taxa are presented during barcoding analysis, further validation with other DNA barcode loci or morphological characters should be mandated.

ITS sequence variations in common ragweed and giant ragweed (돼지풀 및 단풍잎돼지풀의 ITS 염기서열 변이)

  • Kim, Young-Dong;Park, Chong-Wook;Sun, Byung-Yun;Kim, Ki-Joong;Lee, Eun-Ju;Kim, Sung-Hee
    • Korean Journal of Plant Taxonomy
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    • v.35 no.4
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    • pp.273-285
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    • 2005
  • Genetic variation of notorious invasive plants, common ragweed (Ambrosia artemisiifolia L.) and giant ragweed (Ambrosia trifida L.) were examined using the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA. A total of 18 different ITS types were detected from 156 individuals of common ragweed sampled mainly from the southern part of Korean peninsula whereas four types were identified from 46 individuals of giant ragweed. High sequence diversity observed from common ragweed in Korean populations was interpreted as multiple introduction. Genetic recombination was suggested as possible method for the production of some of the ITS types while point mutation was mainly responsible for the origin of the sequence diversity. This study provided some of basic genetic information needed for understanding of the evolutionary process in ragweed during invasion.

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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Re-Identification on Korean Penicillium Sequences in GenBank Collected by Software GenMine

  • Chang Wan Seo;Sung Hyun Kim;Young Woon Lim;Myung Soo Park
    • Mycobiology
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    • v.50 no.4
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    • pp.231-237
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    • 2022
  • Penicillium species have been actively studied in various fields, and many new and unrecorded species continue to be reported in Korea. Moreover, unidentified and misidentified Korean Penicillium species still exist in GenBank. Therefore, it is necessary to revise the Korean Penicillium inventory based on accurate identification. We collected Korean Penicillium nucleotide sequence records from GenBank using the newly developed software, GenMine, and re-identified Korean Penicillium based on the maximum likelihood trees. A total of 1681 Korean Penicillium GenBank nucleotide sequence records were collected from GenBank. In these records, 1208 strains with four major genes (Internal Transcribed Spacer rDNA region, b-tubulin, Calmodulin and RNA polymerase II) were selected for Penicillium reidentification. Among 1208 strains, 927 were identified, 82 were identified as other genera, the rest remained undetermined due to low phylogenetic resolution. Identified strains consisted of 206 Penicillium species, including 156 recorded species and 50 new species candidates. However, 37 species recorded in the national list of species in Korea were not found in GenBank. Further studies on the presence or absence of these species are required through literature investigation, additional sampling, and sequencing. Our study can be the basis for updating the Korean Penicillium inventory.

Morphological and molecular characterization of root-lesion nematode Pratylenchus hippeastri from Korea

  • Sungchan Huh;Namsook Park;Heonil Kang;Changhwan Bae;Insoo Choi
    • Korean Journal of Environmental Biology
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    • v.41 no.4
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    • pp.657-665
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    • 2023
  • The root-lesion nematode Pratylenchus spp. is the most important plantparasitic nematode due to its worldwide distribution, wide host ranges, and migratory endoparasitic characteristics. One population of Pratylenchus collected from the giant pussy willow (Salix chaenomeloides Kimura) in the Andong area as part of a nematode survey in Korea was characterized morphologically and by molecular methods. The analysis of morphological measurements and morphometric characteristics, as well as DNA sequencing of the rRNA large subunit (LSU) D2/D3 expansion segments and the internal transcribed spacer (ITS) gene sequence, confirmed the identity of this population as P. hippeastri. This study is the first report of P. hippeastri associated with Salix chaenomeloides in Korea and worldwide. Further studies on distribution and pathogenicity in different P. hippeastri host crops, such as grapevines, strawberries, and apples, are necessary. The taxonomic keys to 16 Pratylenchus species in Korea are provided.

Identification of Acremonium acutatum and Trichothecium roseum isolated from Grape with White Stain Symptom in Korea

  • Oh, Soh-Young;Nam, Ki-Woong;Yoon, Deok-Hoon
    • Mycobiology
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    • v.42 no.3
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    • pp.269-273
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    • 2014
  • During 2010 and 2012 grape harvest seasons in Gyeonggi-do, Korea, a white stain symptom was observed on the harvested grape fruits in 'Campbell-Early' and 'Kyoho' varieties. In samples collected from the infected vine, two different strains of pathogenic fungi have been found and identified as Acremonium acutatum and Trichothecium roseum based on fungal morphology and nucleotide sequence of internal transcribed spacer (ITS) and supported by the phylogenetic analysis of the rDNA-ITS region. The DNA homologies of the isolated strains were 99.8% and 99.6% identical with T. roseum (IFB-22133) and A. acutatum (CBS682.71), respectively. In the pathogenicity test, the spores of A. acutatum and T. roseum sprayed on the grapes caused white stain symptoms on the fruits in two weeks after the artificial inoculation, which is similar to observations in the field. To our knowledge, this is the first report of white stain symptoms caused by A. acutatum and T. roseum on the grapes in Korea.

Cultural Conditions for Mycelial Growth and Molecular Phylogenetic Relationship in Different Wild Strains of Schizophyllum commune

  • Alam, Nuhu;Cha, Youn-Jeong;Shim, Mi-Ja;Lee, Tae-Soo;Lee, U-Youn
    • Mycobiology
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    • v.38 no.1
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    • pp.17-25
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    • 2010
  • The common split-gilled mushroom, Schizophyllum commune is found throughout the world on woody plants. This study was initiated to evaluate conditions for favorable vegetative growth and to determine molecular phylogenetic relationship in twelve different strains of S. commune. A suitable temperature for mycelial growth was obtained at $30^{\circ}C$. This mushroom grew well in acidic conditions and pH 5 was the most favorable. Hamada, glucose peptone, Hennerberg, potato dextrose agar and yeast malt extract were favorable media for growing mycelia, while Lilly and glucose tryptone were unfavorable. Dextrin was the best and lactose was the less effective carbon source. The most suitable nitrogen sources were calcium nitrate, glycine, and potassium nitrate, whereas ammonium phosphate and histidine were the least effective for the mycelial growth of S. commune. The genetic diversity of each strain was investigated in order to identify them. The internal transcribed spacer (ITS) regions of rDNA were amplified using PCR. The size of the ITS1 and ITS2 regions of rDNA from the different strains varied from 129 to 143 bp and 241 to 243 bp, respectively. The sequence of ITS1 was more variable than that of ITS2, while the 5.8S sequences were identical. A phylogenetic tree of the ITS region sequences indicated that the selected strains were classified into three clusters. The reciprocal homologies of the ITS region sequences ranged from 99 to 100%. The strains were also analyzed by random amplification of polymorphic DNA (RAPD) with 20 arbitrary primers. Twelve primers efficiently amplified the genomic DNA. The number of amplified bands varied depending on the primers used or the strains tested. The average number of polymorphic bands observed per primer was 4.5. The size of polymorphic fragments was obtained in the range of 0.2 to 2.3 kb. These results indicate that the RAPD technique is well suited for detecting the genetic diversity in the S. commune strains tested.

Morphological and genetic characterization and the nationwide distribution of the phototrophic dinoflagellate Scrippsiella lachrymosa in the Korean waters

  • Lee, Sung Yeon;Jeong, Hae Jin;You, Ji Hyun;Kim, So Jin
    • ALGAE
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    • v.33 no.1
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    • pp.21-35
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    • 2018
  • The phototrophic dinoflagellate genus Scrippsiella is known to have a worldwide distribution. Here, we report for the first time, the occurrence of Scrippsiella lachrymosa in Korean waters. Unlike the other stains of S. lachrymosa whose cultures had been established from cysts in the sediments, the clonal culture of the Korean strain of S. lachrymosa was established from motile cells. When the sulcal plates of S. lachrymosa, which have not been fully described to date, were carefully examined using scanning electron microscopy, the Korean strain of S. lachrymosa clearly exhibited the anterior sulcal plate (s.a.), right sulcal plate (s.d.), left sulcal plate (s.s.), median sulcal plate (s.m.), and posterior sulcal plate (s.p.). When properly aligned, the large subunit (LSU) rDNA sequence of the Korean strain of S. lachrymosa was ca. 1% different from those of two Norwegian strains of S. lachrymosa, the only strains for which LSU sequences have been reported. The internal transcribed spacer (ITS) rDNA sequence of the Korean strain of S. lachrymosa was also ca. 1% different from those of the Scottish and Chinese strains and 3% different from those of the Canadian, German, Greek, and Portuguese strains. Thus, the Korean S. lachrymosa strain has unique LSU and ITS sequences. The abundances of S. lachrymosa in the waters of 28 stations, located in the East, West, and South Sea of Korea, were quantified in four seasons from January 2016 to October 2017, using quantitative real-time polymerase chain reaction method and newly designed specific primer-probe sets. Its abundances were >$0.1cells\;mL^{-1}$ at eight stations in January and March 2016 and March 2017, and its highest abundance in Korean waters was $26cells\;mL^{-1}$. Thus, S. lachrymosa has a nationwide distribution in Korean waters as motile cells.

Genetic Diversity and Pathogenicity of Cylindrocarpon destructans Isolates Obtained from Korean Panax ginseng

  • Song, Jeong Young;Seo, Mun Won;Kim, Sun Ick;Nam, Myeong Hyeon;Lim, Hyoun Sub;Kim, Hong Gi
    • Mycobiology
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    • v.42 no.2
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    • pp.174-180
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    • 2014
  • We analyzed the genetic diversity of Cylindrocarpon destructans isolates obtained from Korean ginseng (i.e., Panax ginseng) roots by performing virulence tests and nuclear ribosomal gene internal transcribed spacer (ITS) and mitochondrial small subunit (mt SSU) rDNA sequence analysis. The phylogenetic relationship analysis performed using ITS DNA sequences and isolates from other hosts helped confirm that all the Korean C. destructans isolates belonged to Nectria/Neonectria radicicola complex. The results of in vivo and ex vivo virulence tests showed that the C. destructans isolates could be divided into two groups according to their distinctive difference in virulence and the genetic diversity. The highly virulent Korean isolates in pathogenicity group II (PG II), together with foreign isolates from P. ginseng and P. quinquefolius, formed a single group. The weakly virulent isolates in pathogenicity group I, together with the foreign isolates from other host plants, formed another group and exhibited a greater genetic diversity than the isolates of PG II, as confirmed by the mt SSU rDNA sequence analysis. In addition, as the weakly virulent Korean isolates were genetically very similar to the foreign isolates from other hosts, they were likely to originate from hosts other than the ginseng plants.