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

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Ultrastructure and molecular phylogeny of Mesodinium annulatum sp. nov. (Mesodiniidae, Cyclotrichiida), a new member of the Mesodinium rubrum / Mesodinium major complex

  • Seung Won Nam;Miran Kim;Seok Won Jang;Myung Gil Park;Wonho Yih;Hyung Seop Kim;Woongghi Shin
    • ALGAE
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    • v.39 no.3
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    • pp.129-147
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    • 2024
  • The species complex Mesodinium rubrum / major, common red tide-forming ciliates, has been intensively studied with regards to its ecological roles in global marine ecosystems and the evolutionary aspects of its "stolen" organelles (kleptoplasty and karyoklepty). Nonetheless, the taxonomy of the species within the complex remains unclear. A new marine Mesodinium species isolated from Gomso Bay, Korea, was cultivated under mixotrophic conditions by providing Teleaulax amphioxeia, a red cryptomonad, as prey. Cells of the new isolate consisted of two portions separated by two types of polykinetids. The number of polykinetid associated with the equatorial ciliary belt was approximately 38, and each consisting of two rows of up to 18 alternating kinetosomes each. There was an equal number of cirral polykinetids, each consisting of 16 kinetosomes organized into four longitudinal rows having five, five, four, and two kinetosomes, respectively (in anti-clockwise direction). The two kinds of kinetids and their associated microtubules and fibers were structurally similar to those of M. rubrum from Denmark. However, the Korean Mesodinium species was characterized by its broad posterior portion, 20-22 tentacles, and a cytopharyngeal annulus. Molecular phylogeny based on internal transcribed spacer sequences placed the Korean isolate in clade B of the M. rubrum / major species complex, rather than in clade F representing the neotype of M. rubrum. Based on morphological, ultrastructural, and molecular data, we propose the Korean strain as a new marine Mesodinium species, M. annulatum.

Molecular Characterization of Enterocytozoon bieneusi in Domestic Rabbits (Oryctolagus cuniculus) in Northeastern China

  • Zhang, Xiao-Xuan;Jiang, Jing;Cai, Ya-Nan;Wang, Chun-Feng;Xu, Peng;Yang, Gui-Lian;Zha, Quan
    • Parasites, Hosts and Diseases
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    • v.54 no.1
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    • pp.81-85
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    • 2016
  • A study of 426 rabbits from 3 cities in Jilin province (Changchun City and Jilin City) and Liaoning province (Shenyang City) was conducted between May and June 2015. The overall prevalence of E. bieneusi in rabbits was 0.94% (4/426), with 0% (0/116), 1.72% (3/174), and 0.74% (1/136) in Jilin, Changchun, and Shenyang City, respectively. Only 3 farms (farm 1 and farm 3 in Changchun City, farm 8 in Shenyang City) were PCR-positive for E. bieneusi. Moreover, rabbits of more than 6 months (1.72%) had the highest E. bieneusi prevalence, followed by rabbits of 4-6 months (1.26%), 2-3 months (0.58%), and less than 1 month (0%). Analysis of ITS gene of E. bieneusi suggested that all 4 E. bieneusi isolates were genotype D, and were classified as group 1a. The present results first demonstrated the existence of zoonotic E. bieneusi in domestic rabbits in China. Effective control measures should be implemented to prevent E. bieneusi infection in domestic rabbits, other animals, and humans.

Genotype and Phenotype of Echinococcus granulosus Derived from Wild Sheep (Ovis orientalis) in Iran

  • Eslami, Ali;Meshgi, Behnam;Jalousian, Fatemeh;Rahmani, Shima;Salari, Mohammad Ali
    • Parasites, Hosts and Diseases
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    • v.54 no.1
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    • pp.55-60
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    • 2016
  • The aim of the present study is to determine the characteristics of genotype and phenotype of Echinococcus granulosus derived from wild sheep and to compare them with the strains of E. granulosus sensu stricto (sheep-dog) and E. granulosus camel strain (camel-dog) in Iran. In Khojir National Park, near Tehran, Iran, a fertile hydatid cyst was recently found in the liver of a dead wild sheep (Ovis orientalis). The number of protoscolices (n=6,000) proved enough for an experimental infection in a dog. The characteristics of large and small hooks of metacestode were statistically determined as the sensu stricto strain but not the camel strain (P=0.5). To determine E. granulosus genotype, 20 adult worms of this type were collected from the infected dog. The second internal transcribed spacer (ITS2) of the nuclear ribosomal DNA (rDNA) and cytochrome c oxidase 1 subunit (COX1) of the mitochondrial DNA were amplified from individual adult worm by PCR. Subsequently, the PCR product was sequenced by Sanger method. The lengths of ITS2 and COX1 sequences were 378 and 857 bp, respectively, for all the sequenced samples. The amplified DNA sequences from both ribosomal and mitochondrial genes were highly similar (99% and 98%, respectively) to that of the ovine strain in the GenBank database. The results of the present study indicate that the morpho-molecular features and characteristics of E. granulosus in the Iranian wild sheep are the same as those of the sheep-dog E. granulosus sensu stricto strain.

Occurrence of the Phytophthora Blight Caused by Phytophthora sansomeana in Atractylodes macrocephala Koidz. (Phytophthora sansomeana에 의한 큰꽃삽주 역병 발생 보고)

  • An, Tae Jin;Park, Myung Soo;Jeong, Jin Tae;Kim, Young Guk;Kim, Yong Il;Lee, Eun Song;Chang, Jae Ki
    • Korean Journal of Medicinal Crop Science
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    • v.27 no.6
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    • pp.404-411
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    • 2019
  • Background: In September 2017, wilting and rhizome rot symptoms were observed on Atractylodes macrocephala Koidz. in Jecheon-si and Eumseong-gun. This study was carried out to isolate hitherto unidentified pathogenic fungi from A. macrocephala and to test the pathogenicity of isolated fungi against Atractylodes spp. genus such as A. macrocephala, A. japonica, and their interspecific hybrids. Methods and Results: The diseased plants were washed with running tap water, and the boundary between the healthy area and the diseased area was cut while the pathogens were isolated by growing cultures from the diseased areas on Phytophthora semi-selective medium. The internal transcribed spacer (ITS) region of the isolates was used in this study for identification. Test plants were cultivated in the glasshouse at 20℃ - 30℃ for 4 months and then used for pathogenicity test. The pots with plants inoculated with mycelial plugs and zoospores were placed at 25℃ for 48 h in a dew chamber where relative humidity was above 95%, and then moved into the glasshouse at 20℃ - 30℃. The presence or absence of pathogenicity of the strains was determined by evaluating the symptom of plant wilting. The inoculation test was performed in three replicates with a non-treated control. Conclusions: On the basis of results of ITS sequence analysis, the strains isolated from the diseased plants was identified as Phytophthora sansomeana. Biological assay using test plants confirmed the pathogenicity of P. sansomeana against Atractylodes macrocephala. This is the first report of rhizome rot in A. macrocephala caused by P. sansomeana.

Morphological and cultural characteristics of a novel Phellinus linteus KACC93057P (신규 목질진흙버섯 KACC93057P의 배양적, 형태적 특성)

  • Min, Gyeong-Jin;Kwak, A-Min;Seok, Sun-Ja;Kang, Hee-Wan
    • Journal of Mushroom
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    • v.14 no.3
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    • pp.75-80
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    • 2016
  • Morphological and culture characteristics of the novel Phellinus linteus variant KACC93057P collected in Korea were characterized in this study. The surface of the was angular, sessile, tough-woody, concentrically zonate, and dark brown in color. Basidiocarppores were circular, with 5-7 pores per mm. The hyphal system was dimitic, and basidiospores were ellipsoid or oval, $4.5-6{\times}4-5$, exhibiting characteristics typical of P. linteus. The optimum temperature for mycelial growth was $25-30^{\circ}C$, and optimal pH for growth was 5-7. The mycelial growth rate of P. linteus KACC93057P was faster than that of other P. baumii isolates. On growth medium, KACC93057P formed aerial mycelia with density higher than that of other isolates. The internal transcribed spacer (ITS)-ribosomal DNA sequences were closely related to the sequences P. linteus complex.

Identification and Characterization of Gliocladium viride Isolated from Mushroom Fly Infested Oak Log Beds Used for Shiitake Cultivation

  • Kim, Jun-Young;Yun, Yeo-Hong;Hyun, Min-Woo;Kim, Myeong-Ho;Kim, Seong-Hwan
    • Mycobiology
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    • v.38 no.1
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    • pp.7-12
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    • 2010
  • A green mold species that has not previously been reported in Korea was isolated from oak log beds used for shiitake (Lentinula edodes) cultivation that were infested by mushroom flies. In this study, we identify the mold species as Gliocladium viride (an anamorph of Hypocrea lutea) and describe its mycological properties. The fungus was cottony on both potato dextrose agar (PDA) and Czapek yeast extract agar (CYA), but was colored white on PDA and became yellowish green and brown on CYA. Mycelial growth on PDA attained a diameter of 73 mm at $30^{\circ}C$ after 5 days. The fungus grew faster on malt extract agar (> 80 mm, 5 days at $25^{\circ}C$) compared to CYA and PDA (< 68 mm, 5 days at $25^{\circ}C$). Penicillate conidiophores of the fungus are hyaline, smooth walled, branching above typically in four stages, and $120\sim240\;{\mu}m$ in length. Club-shaped or slender phialides are formed on the metulae. Conidia of the fungus were ovate and elliptic, yellowish brown and green, and $2.5\sim3.0\;{\mu}m\times1.8\sim2.3\;{\mu}m$ in size. Typically, slimy conidia are formed in a mass and colored brown to dark green to almost black. The internal transcribed spacer rDNA and translation elongation factor 1 alpha gene sequences of the fungus isolated here show 99% identity with previously identified G. viride strains.

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.

Genotypic Identification of Cystoisospora in Immunocompromised Patients Using Tm-Variation Analysis

  • Basyoni, Maha M.A.;Elghobary, Hany Ahmed Fouad
    • Parasites, Hosts and Diseases
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    • v.55 no.6
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    • pp.601-606
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    • 2017
  • Cystoisospora is responsible for morbidity in immunocompromised patients. PCR is sensitive for diagnosing Cystoisospora; however, it needs reevaluation for differential molecular diagnosis of cystoisosporiasis. We aimed at evaluating melting curve analysis (MCA) after real-time PCR (qPCR) in diagnosis and genotyping of Cystoisospora as an alternative to conventional PCR. We included 293 diarrheic stool samples of patients attending the Department of Clinical Oncology and Nuclear Medicine of Cairo University Hospitals, Egypt. Samples were subjected to microscopy, nested PCR (nPCR), and qPCR targeting the internal transcribed spacer 2 region (ITS2) of the ribosomal RNA (r RNA) gene followed by melting temperatures ($T_ms$) analysis and comparing the results to PCR-RFLP banding patterns. Using microscopy and ITS2-nPCR, 3.1% and 5.8% of cases were Cystoisospora positive, respectively, while 10.9% were positive using qPCR. Genotyping of Cystoisospora by qPCR-MCA revealed 2 genotypes. These genotypes matched with 2 distinct melting peaks with specified $T_ms$ at $85.8^{\circ}C$ and $88.6^{\circ}C$, which indicated genetic variation among Cystoisospora isolates in Egypt. Genotype II proved to be more prevalent (65.6%). HIV-related Kaposi sarcoma and leukemic patients harbored both genotypes with a tendency to genotype II. Genotype I was more prevalent in lymphomas and mammary gland tumors while colorectal and hepatocellular tumors harbored genotype II suggesting that this genotype might be responsible for the development of cystoisosporiasis in immunocompromised patients. Direct reliable identification and differentiation of Cystoisospora species could be established using $qPCR-T_ms$ analysis which is useful for rapid detection and screening of Cystoisospora genotypes principally in high risk groups.

Seventeen Unrecorded Species from Gayasan National Park in Korea

  • Lee, Hyun;Park, Myung Soo;Park, Ji-Hyun;Cho, Hae Jin;Park, Ki Hyeong;Yoo, Shinnam;Lee, Jun Won;Kim, Nam Kyu;Lee, Jin Sung;Park, Jae Young;Kim, Changmu;Kim, Jae-Jin;Lim, Young Woon
    • Mycobiology
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    • v.48 no.3
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    • pp.184-194
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    • 2020
  • Macrofungi play important roles in forest ecology as wood decayers, symbionts, and pathogens of living trees. For the effective forest management, it is imperative to have a comprehensive overview of macrofungi diversity in specific areas. As a part of the National Institute of Biological Resources projects for discovering indigenous fungi in Korea, we collected macrofungi in Gayasan National Park from 2017 to 2018. These specimens were identified based on morphological characteristics and sequence analysis of internal transcribed spacer (ITS) or the nuclear large subunit rRNA (LSU) region. We discovered 17 macrofungi new to Korea: Butyrea japonica, Ceriporia nanlingensis, Coltricia weii, Coltriciella subglobosa, Crepidotus crocophyllus, Cylindrobasidium laeve, Fulvoderma scaurum, Laetiporus cremeiporus, Lentinellus castoreus, Leucogyrophana mollusca, Marasmius insolitus, Nidularia deformis, Phaeophlebiopsis peniophoroides, Phanerochaete angustocystidiata, Phlebiopsis pilatii, Postia coeruleivirens, and Tengioboletus fujianensis. We described their detailed morphological characteristics.

Phylogeny of Phellinus and Related Genera Inferred from Combined Data of ITS and Mitochondrial SSU rDNA Sequences

  • JEONG WON JIN;LIM YOUNG WOON;LEE JIN SUNG;JUNG HACK SUNG
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1028-1038
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    • 2005
  • To elucidate phylogenetic relationships of Phellinus and its related genera, nuclear internal transcribed spacer and mitochondrial small subunit ribosomal DNA sequences from 65 strains were determined and compared. The combined dataset of two sequences increased informative characters and led to the production of trees with higher levels of resolution. Phylogenetic analysis of the combined dataset revealed thirteen evolutionary lineages and several unresolved species that were together subdivided into two large clusters consisting of oligonucleate species and binucleate species. These results coincided with previous cytological, morphological, and molecular studies. It is newly recognized that the Phellinus linteus complex forms a sister clade to Inonotus, and that Fulvifomes is somehow related to Inocutis. The Phellinus linteus complex of dimitic perennial taxa made an independent clade from Inonotus and suggested that hyphal miticity and fruitbody permanence had enough phylogenetic significance to keep the complex within the traditional genus Phellinus. Taxa lacking setae were clustered into Fulvifomes, Phylloporia, Inocutis, and Fomitiporia, and the first three were closely related sister groups, but Fomitiporia was a genus distantly related to them. Several taxa with branched setae were shown among distantly related genera. Molecular evidence indicated that the ancestral nuclear type could be a binucleate feature, and that there might be parallel gains of branched setae and parallel losses of setae in the Hymenochaetales.