• Title/Summary/Keyword: LSU rDNA

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Population analysis of the toxic dinoflagellate genus Alexandrium by novel molecular markers

  • Kim, Choong-jae;Kim, Sook-Yang;Kim, Kui-Young;Kang, Young-Sil;Kim, Hak-Gyoon;Kim, Chang-Hoon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.134-135
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    • 2003
  • The geographic expansion of the toxic dinoflagellates genus Alexandrium has been shown to be world wide ranging. The members of the genus Alexandrium ocnstituted of 20-30 species did not show substantial differences in their morphology, which is mostly referred in the 'tamarensis species complex', except some species. Though rDNA sequences variations are very few and pseudogene types are so diverse that it is difficult to use them as the specific markers. In this study, we outlined Korean and Japanese A, tamarense and A. catenella regional isolates by phylogenetic analysis inferred from no cutting alignments of LSU rDNA D1-D2 and SSU rDNA sequences to group these regional isolates. The results were compared to RFLP patterns of PCR products targeted chloroplast DNA. Lastly screening of highly repeated microsatellite DNA which is frequently used for population analysis in eukaryotes was conducted. A. catenella regional strains identified by the sequencing of rDNA D1-D2 domain were divided into at least 3 groups of type E, CMC and Chinese type, divergence root may not be deep comparing with that of A. tamarense whose pseudogenes are very variable. Results of RFLP pattern and the phylogeny of the unknown gene targeting chloroplast showed that Korean and Japanese A. catenella regional isolates were divided into 3 types: Korean, Japanese and the third CMC types. Population-specific PCR amplification with Japanese A. catenella type-specific PCR primers was useful method for population analysis of A. catenella. Various types of satellite sequences such as 5 nucleotides repeats were obtained from A. tamarense and A. catenella. The 5 nucleotides repeats were primed at the both 3'and 5' ends, and these repeats were prominent as longer repeated motifs. This repeated DNA was intercalated as internal sequences containing various types subrepeats. It is expected that these satellite DNA would be a useful molecular population marker through detail comparison among Alexandrium regional isolates to trace their transferring pathway and to prevent their human-associated their regional extents.

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Rapid and exact molecular identification of the PSP (paralytic shellfish poisoning) producing dinoflagellate genus Alexandrium

  • Kim, Choong-jae;Kim, Sook-Yang;Kim, Kui-Young;Kang, Young-Sil;Kim, Hak-Gyoon;Kim, Chang-Hoon
    • Proceedings of the Korean Aquaculture Society Conference
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    • 2003.10a
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    • pp.132-133
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    • 2003
  • The marine dinoflagellate genus Alexandrium comprise PSP producing A. acatenella, A. angustitabuzatum, A. catenella, A. fundyense, A. minutum, A. ostenfezdii, A. tamiyavanichii and A. tamarense. In monitoring toxic Alexandrium, rapid and exact species identification is one of the significant prerequisite work, however we have suffered confusion of species definition in Alexandrium. To surmount this problem, we chose DNA probing, which has long been used as an alternative for conventional identification methods, primarily relying on morphological approaches using microscope in microbial field. Oligonucleotide DNA probes targeting rRNA or rDNA have been commonly used in diverse studies to detect and enumerate cells concerned as a culture-indetendent powerful tool. Despite of the massive literature on the HAB species containing Alexandrium, application of DNA probing for species identification and detection has been limited to a few documents. DNA probes of toxic A. tamarense, A. catenella and A. tamiyavanichii, and non-toxic A. affine, A. fraterculus, A. insuetum and A. pseudogonyaulax were designed from LSU rDNA D1-D2, and applied to whole cell-FISH. Each DNA probes reacted only the targeted Alexandrium cells with very high species-specificity within Alexandrium. The probes could detect each targeted cells obtained from the natural sea water samples without cross-reactivity. Labeling intensity varied in the growth stage, this showed that the contents of probe-targeted cellular rRNA decreased with reduced growth rate. Double probe TAMID2S1 achieved approximately two times higher fluorescent intensity than that with single probe TAMID2. This double probe did not cross-react with any kinds of microorganisms in the natural sea waters. Therefore we can say that in whole-cell FISH procedure this double DNA probe successfully labeled targeted A. tamiyavanichii without cross-reaction with congeners and diverse natural bio-communities.

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First Report of Leptosphaerulina australis Isolated from Soil in Korea

  • Li, Weilan;Back, Chang-Gi;Lee, Seung-Yeol;Ten, Leonid N.;Jung, Hee-Young
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.369-374
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    • 2018
  • The fungal strain KNU16-004 was isolated from a field soil sample collected in Seoul. The isolate was identified as Leptosphaerulina australis based on morphological characterization and phylogenetic analysis using the internal transcribed spacer (ITS), large subunit (LSU) rDNA regions, and ${\beta}-tubulin$ (Tub2). This is the first report of Leptosphaerulina australis in Korea.

Notes on Five Endophytic Fungal Species Isolated from Needle Leaves of Conifers in Korea (침엽수의 잎에서 분리한 5종의 국내 미기록 내생균)

  • Lee, Bong-Hyung;Eo, Ju-Kyeong;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.44 no.1
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    • pp.51-56
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    • 2016
  • Diverse endophytic fungi were isolated from surface-sterilized leaves of three species of conifers inhabiting various sites in Korea: Abies nephrolepis, Pinus koraiensis and Taxus cuspidate. The isolates were identified based on morphological characteristics and sequences analysis of both internal transcribed spacer and large subunit regions of rDNA. In this paper, we report on five previously unreported species of endophytic fungi isolated from conifers: Biscogniauxia maritime, Nemania diffusa, Pezicula carpinea, Phomopsis juglandina and Sydowia polyspora.

New Species of Termitomyces (Lyophyllaceae, Basidiomycota) from Sabah (Northern Borneo), Malaysia

  • Seelan, Jaya Seelan Sathiya;Yee, Chong Shu;Fui, Foo She;Dawood, Mahadimenakbar;Tan, Yee Shin;Kim, Min-Ji;Park, Myung Soo;Lim, Young Woon
    • Mycobiology
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    • v.48 no.2
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    • pp.95-103
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    • 2020
  • The genus Termitomyces (Lyophyllaceae, Basidiomycota) is often associated with fungus-feeding termites (Macrotermitinae) due to their strong symbiotic relationships. The genus is widely found exclusively in certain regions of Africa and Asia. They are recognized as edible mushroom within Southeast Asia as well. But it is often misidentified based on morphology by the local communities especially in Malaysia for Chlorophyllum molybdites which is a highly poisonous mushroom. Thus, it is necessary to study the genus for Malaysia with the synergy of using both morphological and molecular identification. In this study, we aim to describe another new species as an addition to the genus Termitomyces found within Sabah, Malaysia. We generated two new sequences (nrLSU and mtSSU) for the new species and a total of 28 nrLSU and mtSSU sequences were retrieved from GenBank for the phylogenetic analysis using maximum likelihood and Bayesian inferences. We identified that the new collection from Sabah province is a new species and named as Termitomyces gilvus based on the termites found in the mound. A phylogeny tree made from the concatenated genes of LSU and mtSSU suggests that T. gilvus is closely related to T. bulborhizus from China. According to our results, the combination of molecular and morphology proved to be a robust approach to re-evaluate the taxonomic status of Termitomyces species in Malaysia. Additional surveys are needed to verify the species diversity and clarify their geographic distribution.

Molecular Characterization of Gastrothylax crumenifer (Platyhelminthes: Gastrothylacidae) from Goats in the Western Part of India by LSU of Nuclear Ribosomal DNA

  • Kumar, Ashwani;Chaudhary, Anshu;Verma, Chandni;Singh, Hridaya Shanker
    • Parasites, Hosts and Diseases
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    • v.52 no.6
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    • pp.701-705
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    • 2014
  • The rumen parasite, Gastrothylax crumenifer (Platyhelminthes: Gastrothylacidae), is a highly pathogenic trematode parasite of goat (Capra hircus). It sucks blood that causes acute disease like anemia, and severe economic losses occur due to morbidity and mortality of the ruminant infected by these worms. The study of these rumen paramphistomes, their infection, and public health importance remains unclear in India especially in the western part of state Uttar Pradesh (U.P.), Meerut, India, where the goat meat consumption is very high. This paper provides the molecular characterization of G. crumenifer recovered from the rumen of Capra hircus from Meerut, U.P., India by the partial sequence of 28S rDNA. Nucleotide sequence similarity searching on BLAST of 28S rDNA from parasites showed the highest identity with those of G. crumenifer from the same host Capra hircus. This is the first report of molecular identification of G. crumenifer from this part of India.

Unrecorded Endophytic Fungi Belonging to Genus Phyllosticta Isolated from Leaves of Woody Plants (목본식물 잎에서 분리된 Phyllosticta 속의 국내 미기록종 내생균)

  • Park, Hyeok;Lee, Jong-Chul;Gwon, Ju-Hui;Lee, Hyang Burm;Eom, Ahn-Heum
    • The Korean Journal of Mycology
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    • v.49 no.1
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    • pp.81-86
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    • 2021
  • We isolated endophytic fungi from Smilax china and Cryptomeria japonica. These fungal strains were identified based on their morphological characteristics and phylogenetic analyses of their internal transcribed spacer, large subunit rDNA, and translation elongation factor 1-α DNA. Among them, we discovered two fungi belonging to the genus Phyllosticta, which have not been previously recorded in Korea. We have described these two fungal strains, Phyllosticta ericarum 19E458 and Phyllosticta philoprina 19E012 in this study.

Morphology and molecular characterization of the epiphytic dinoflagellate Amphidinium massartii, isolated from the temperate waters off Jeju Island, Korea

  • Lee, Kyung Ha;Jeong, Hae Jin;Park, Kila;Kang, Nam Seon;Yoo, Yeong Du;Lee, Moo Joon;Lee, Jin-Woo;Lee, Soojin;Kim, Taekyung;Kim, Hyung Seop;Noh, Jae Hoon
    • ALGAE
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    • v.28 no.3
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    • pp.213-231
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    • 2013
  • Amphidinium massartii Biecheler is an epiphytic and toxic dinoflagellate. Prior to the present study, A. massartii has been reported in the waters off the Mediterranean, Australian, USA, and Canadian coasts. We isolated Amphidinium cells from the coastal waters of Jeju Island, Korea and their morphology and rDNA sequences indicated that they were A. massartii. Herein, we report for the first time the occurrence of A. massartii in the waters of the temperate region in the northwestern Pacific Ocean. The large subunit (LSU) rDNA sequences of the Korean strains were 0.7% different from those of an Australian strain of A. massartii CS-259, the closest species, but were 4.1-5.8% different from those of the other Australian strains and the USA strains of A. massartii and from those of Amphidinium sp. HG115 that was isolated from subtropical Okinawan waters. In phylogenetic trees based on LSU, internal transcribed spacer, small subunit rDNA, and cytochrome b sequences, the Korean strains belonged to the A. massartii clade, which was clearly divergent from the A. carterae clade. The morphology of the Korean A. massartii strains was similar to that of the originally described French strain and recently described Australian strain. However, we report for the first time here that scales were observed on the surface of the flagella. In conclusion, the Korean A. massartii strains have unique rDNA sequences, even though they have a very similar morphology to that of previously reported strains. This report extends the known range of this dinoflagellate to the temperate waters of the northwestern Pacific Ocean.

Confirmation of Coleosporium zanthoxyli on Zanthoxylum ailanthoides in Korea (머귀나무에서 녹병균 Coleosporium zanthoxyli의 확인)

  • Shin, Hyeon-Dong;Choi, Young-Joon;Lee, Chong-Kyu;Lee, Ho-Sang;Choi, Won-Il
    • The Korean Journal of Mycology
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    • v.47 no.1
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    • pp.83-88
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    • 2019
  • A previous record of Coleosporium phellodendri on Zanthoxylum ailanthoides in Korea by Korean researchers was regarded as C. zanthoxyli by foreign scientists without further studies. To clarify the identity of a Coleosporium species occurring on Z. ailanthoides in Korea, we examined the morphological characteristics of eight materials and analyzed the internal transcribed spacer (ITS) and 28S large subunit (LSU) rDNA regions of three representative specimens which were collected in Korea and deposited in the Mycological Herbarium at Korea University (Seoul, Korea). All specimens collected in Korea were morphologically and molecularly determined to be C. zanthoxyli. Therefore, this is the first confirmed report on the presence of C. zanthoxyli on Z. ailanthoides in Korea.

Eleven previously unrecorded fungal species isolated from hyphosphere soil supporting wild mushrooms in Jeju Island (제주도 야생버섯 hyphosphere 토양에서 분리된 국내 미기록 진균 11종 보고)

  • Hyeongjin Noh;Ye In Kim;Dong Hyeung Lee;Pyung Yeol Ko;Hye Sung Park;Kang-Hyo Lee;Seong Hwan Kim
    • Journal of Mushroom
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    • v.21 no.4
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    • pp.228-240
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    • 2023
  • To understand microorganism effects on wild mushroom fruiting bodies, we investigated the fungi in hyphosphere soil supporting wild mushroom species Cortinarius violaceus, Amanita hemibapha, Laccaria vinacelavellanea, and Amanita verna found in the Gotjawal area of Jeju Island. Fungal species identification based on morphological traits and molecular analysis of ITS, LSU rDNA, and β-tubulin gene sequences resulted in isolation and identification of eleven fungal species previously unrecorded in Korea. These newly-recorded species are: Arthrinium kogelbergensis, Kalmusia longisporum, Keithomyces carneum, Neopyrenochaeta cercidis, Penicillium ranomafanaense, Phomatodes nebulosa, Pyrenochaeta nobilis, Tolypocladium album, Talaromyces kendrickii, Talaromyces qii, and Umbelopsis gibberispora, and their morphological characteristics and phylogenetic positions are described.