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

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Monitoring of Fasciola Species Contamination in Water Dropwort by COX1 Mitochondrial and ITS-2 rDNA Sequencing Analysis

  • Choi, In-Wook;Kim, Hwang-Yong;Quan, Juan-Hua;Ryu, Jae-Gee;Sun, Rubing;Lee, Young-Ha
    • Parasites, Hosts and Diseases
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    • v.53 no.5
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    • pp.641-645
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    • 2015
  • Fascioliasis, a food-borne trematode zoonosis, is a disease primarily in cattle and sheep and occasionally in humans. Water dropwort (Oenanthe javanica), an aquatic perennial herb, is a common second intermediate host of Fasciola, and the fresh stems and leaves are widely used as a seasoning in the Korean diet. However, no information regarding Fasciola species contamination in water dropwort is available. Here, we collected 500 samples of water dropwort in 3 areas in Korea during February and March 2015, and the water dropwort contamination of Fasciola species was monitored by DNA sequencing analysis of the Fasciola hepatica and Fasciola gigantica specific mitochondrial cytochrome c oxidase subunit 1 (cox1) and nuclear ribosomal internal transcribed spacer 2 (ITS-2). Among the 500 samples assessed, the presence of F. hepatica cox1 and 1TS-2 markers were detected in 2 samples, and F. hepatica contamination was confirmed by sequencing analysis. The nucleotide sequences of cox1 PCR products from the 2 F. hepatica-contaminated samples were 96.5% identical to the F. hepatica cox1 sequences in GenBank, whereas F. gigantica cox1 sequences were 46.8% similar with the sequence detected from the cox1 positive samples. However, F. gigantica cox1 and ITS-2 markers were not detected by PCR in the 500 samples of water dropwort. Collectively, in this survey of the water dropwort contamination with Fasciola species, very low prevalence of F. hepatica contamination was detected in the samples.

Phylogenetic Relationships Among Pleurotus species Inferred from Sequence Data of PCR Amplified ITS II Region in Ribosomal DNA (rDNA의 ITS II 부위의 염기서열분석에 의한 느타리버섯 종간의 근연관계)

  • Bae, Shin-Churl;Seong, Ki-Young;Lee, Shin-Woo;Go, Seung-Joo;Eun, Moo-Young;Rhee, In-Koo
    • The Korean Journal of Mycology
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    • v.24 no.2 s.77
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    • pp.155-165
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    • 1996
  • This study was carried out to identify the phylogenetic relationship among several isolates of Pleurotus species by comparing ITS II region of ribosomal DNA(rDNA) repeat unit. Two primers from ribosomal DNA sequences were chosen to amplify the specific internal transcribed spacer (ITS) II region of Pleurotus spp. The exact ITS II region with an unique band from six species of Pleurotus genus could be amplified using the two primers taken from at the 3'-end of 5.8S rDNA and 5'-end of 28S rDNA. Six representative species of the Pleurotus genus were easily characterized according to the length differences of ITS II region. Furthermore, within P. ostreatus species, different sizes of ITS II region could be observed in the isolates of ASI 2025 and ASI 2095 although they were classified as P. ostreatus by the conventional observation. The nucleotide sequence analyses of PCR-amplified ITS II region indicated that the isolates ASI 2025 and ASI 2095 were different from other Pleurotus spp. When the nucleotide sequences of six Pleurotus species were compared, three typical ITS II regions were highly variable especially at both ends of this region. The phylogenetic tree obtained by the Neighbor program of Felsenstein PHYLIP package with all the nucleotide sequence of Pleurotus spp. indicated that P. ostreatus, P. florida, P. sajor-caju and P. eryngii were closely related to one phylogenetic branch and P. cystidious was related to other branch with P. cornucopiae. The isolates ASI 2025 and 2038, however, were not closely related to any other Pleurotus spp. and formed their own individual branches.

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Phylogenetic Study of Ganoderma spp. Based on the DNA Sequences in ITS II Region (ITS II 영역의 DNA 염기서열 분석에 의한 불로초(Ganoderma)속의 계통분류학적 고찰)

  • Park, Dong-Suk;Go, Seung-Joo;Ryu, Jin-Chang;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.27 no.1 s.88
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    • pp.39-43
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    • 1999
  • The internal transcribed spacer II regions (ITS II) of the ribosomal DNA gene repeat from Ganoderma spp. were amplified using polymerase chain reaction (PCR) and sequenced. Sequences from 9 species including Ganoderma lucidum, G. tsugae, G. pfeifferi, G. resinaceum, G. australe-applanatum, G. oregonense, G. neo-japonicum, G. applanatum and Inonotus xeranticus as an out-group were compared. The spacer regions of them were $247{\sim}257$ nucleotides in length and contained partial sequences of 5.8S and 25S gene. The reciprocal homologies of each ITS II sequence of the species were in the range of $70{\sim}100%$ except outgroup species, I. xeranticus. According to the analysis of ITS II sequences, Ganoderma spp. constructed 5 clusters. Ganoderma lucidum isolates were to be divided into two groups. One group was consisted of isolates from South Korea. The other group comprised isolates from UK. G. lucidum isolates belonging to the group I were closely related with G. tsugae. These results suggested that G. lucidum from Korea should be G. tsugae, otherwise G. tsugae was to be synonym of G. lucidum.

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Twig Blight on Chinese Magnolia Vain Caused by Botryosphaeria dothidea in Korea (Botryosphaeria dothidea에 의한 오미자 줄기마름병)

  • Park, Sangkyu;Kim, Seung-Han;Lee, Seung-Yeol;Back, Chang-Gi;Kang, In-Kyu;Jung, Hee-Young
    • Research in Plant Disease
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    • v.22 no.1
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    • pp.44-49
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    • 2016
  • The twig blight symptoms were observed in Chinese magnolia vine (Schisandra chinensis) at Mungyeong city, Gyeongbuk province, Korea in June 2015. The typical symptoms of infected plant were shriveled and wilted in leaves which led to blight resulted in death. Based on the morphological characteristics, the isolate was suspected as Botryosphaeria sp. Inoculation of isolated pathogen was performed to identify its pathogenicity according to Koch's postulates. Re-isolated fungi from the inoculated stem was showed same morphological characteristics with original pathogen. Phylogenetic analysis was performed using combined sequence of rDNA internal transcribed spacer region, EF1-${\alpha}$ and ${\beta}$-tubulin gene. The isolated pathogen was identified to the B. dothidea by phylogenetic analysis. This is the first report of twig blight on S. chinensis caused by B. dothidea in Korea.

Cellulose degrading basidiomycetes yeast isolated from the gut of grasshopper in Korea (한국의 메뚜기의 장에서 분리된 Cellulose를 분해하는 담자균 효모)

  • Kim, Ju-Young;Jang, Jun Hwee;Park, Ji-Hyun;Jung, Hee-Young;Park, Jong-Seok;Cho, Sung-Jin;Lee, Hoon Bok;Limtong, Savitree;Subramani, Gayathri;Sung, Gi-Ho;Kim, Myung Kyum
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.362-368
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    • 2018
  • Grasshoppers play vital role in the digestion of photosynthetically fixed carbons. With the aid of intestinal microflora, the grasshopper can degrade leaves constituents such as cellulose and hemicellulose. The purpose of this study was to examine cellulolytic yeast isolates from the gut of grasshoppers collected in Gyeonggi Province, South Korea. Among the yeast isolates, ON2, ON17 (two strains), and ON6 (one strain) showed positive cellulolytic activity in the CMC-plate assay. The sequence analyses of D1/D2 domains of the large subunit rDNA gene and the internal transcribed spacer (ITS) regions revealed that the strains ON2 and ON17 were most closely related to Papiliotrema aspenensis CBS $13867^T$ (100%, sequence similarity in D1/D2 domains; 99.4% sequence similarity in ITS) and strain ON6 related to Saitozyma flava (100% in D1/D2 domains; 99.0% in ITS). All these three yeast strains are capable of degrading cellulose; therefore, the members of endosymbiotic yeasts may produce their own enzymes for carbohydrate degradation and convert mobilized sugar monomers to volatile fatty acids. Thus, the endosymbiotic yeast strains ON2, ON17 (represents the genus Papilioterma) and ON6 (Saitozyma) belonging to the family Tremellomycetes, are unreported strains in Korea.

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 Relationships of Coprinus spp. on the Basis of Sequences in ITS II Region (먹물버섯류(Coprinus spp.)의 ITS II 영역 염기서열에 의한 유연관계 분석)

  • Park, Dong-Suk;Go, Seung-Joo;Kim, Yang-Sup;Seok, Soon-Ja;Song, Jae-Kyeong;Yeo, Yun-Soo;Ryu, Jin-Chang;Sung, Jae-Mo
    • The Korean Journal of Mycology
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    • v.27 no.1 s.88
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    • pp.27-31
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    • 1999
  • The internal transcribed spacer II regions (ITS II) of the ribosomal DNA gene repeat from Coprinus spp. were amplified using polymerase chain reaction (PCR) and sequenced. Sequences from 11 species including Coprinus comatus, C. atramentarius, C. micaceus, C. lagopus, C. cinereus, C. rhizophorus, C. flocculosus, C. radians, and C. echinosporus were compared. The spacer regions of them were $253{\sim}275$ nucleotide in length and partially contained 5.8S and 25S. The reciprocal homologies of each ITS II sequence among these strains were in the range of $50.6{\sim}100%$. According to the analysis of ITS II sequences, Coprinus spp. were classified into three clusters. Cluster I consisted of Coprinus lagopus, C. cinereus, C. echinosporus, C. rhizophorus, C. niveus, and C. atramentarius. Cluster II comprised C. micaceus, C. flocculosus, C. radians, and C. disseminatus. On the other hand C. comatus is in Cluster III even though this species is belonging to the section Coprinus in morphological aspect. These results suggest that Coprinus comatus, which was considered as a type species of the genus Coprinus in morphological classification, gives a doubt of monophyletic evolution and is assumed to be paraphyletic or polyphyletic.

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Insecticidal Activity of Ethyl Acetate Extracts from Culture Filtrates of Mangrove Fungal Endophytes

  • Abraham, Silva;Basukriadi, Adi;Pawiroharsono, Suyanto;Sjamsuridzal, Wellyzar
    • Mycobiology
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    • v.43 no.2
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    • pp.137-149
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    • 2015
  • In the search for novel potent fungi-derived bioactive compounds for bioinsecticide applications, crude ethyl acetate culture filtrate extracts from 110 mangrove fungal endophytes were screened for their toxicity. Toxicity tests of all extracts against brine shrimp (Artemia salina) larvae were performed. The extracts with the highest toxicity were further examined for insecticidal activity against Spodoptera litura larvae and acetylcholinesterase (AChE) inhibition activity. The results showed that the extracts of five isolates exhibited the highest toxicity to brine shrimp at 50% lethal concentration ($LC_{50}$) values of 7.45 to 10.24 ppm. These five fungal isolates that obtained from Rhizophora mucronata were identified based on sequence data analysis of the internal transcribed spacer region of rDNA as Aspergillus oryzae (strain BPPTCC 6036), Emericella nidulans (strains BPPTCC 6035 and BPPTCC 6038), A. tamarii (strain BPPTCC 6037), and A. versicolor (strain BPPTCC 6039). The mean percentage of S. litura larval mortality following topical application of the five extracts ranged from 16.7% to 43.3%. In the AChE inhibition assay, the inhibition rates of the five extracts ranged from 40.7% to 48.9%, while eserine (positive control) had an inhibition rate of 96.8%, at a concentration of 100 ppm. The extracts used were crude extracts, so their potential as sources of AChE inhibition compounds makes them likely candidates as neurotoxins. The high-performance liquid chromatography profiles of the five extracts differed, indicating variations in their chemical constituents. This study highlights the potential of culture filtrate ethyl acetate extracts of mangrove fungal endophytes as a source of new potential bioactive compounds for bioinsecticide applications.

Potential Reasons for Prevalence of Fusarium Wilt in Oriental Melon in Korea

  • Seo, Yunhee;Kim, Young Ho
    • The Plant Pathology Journal
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    • v.33 no.3
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    • pp.249-263
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    • 2017
  • This study aims to examine the potential reasons for the current prevalence of the fusarium wilt in the oriental melon. Twenty-seven Fusarium isolates obtained from oriental melon greenhouses in 2010-2011 were identified morphologically and by analysis of elongation factor-1 alpha gene (EF-$1{\alpha}$) and internal transcribed spacer (ITS) rDNA sequences as 6 Fusarium species (8 isolates of F. oxysporum, 8 F. commune, 5 F. proliferatum, 3 F. equiseti, 2 F. delphinoides, and 1 F. andiyazi), which were classified as same into 6 EF-$1{\alpha}$ sequence-based phylogenetic clades. Pathogenicity of the Fusarium isolates on the oriental melon was highest in F. proliferatum, next in F. oxysporum and F. andiyazi, and lowest in the other Fusarium species tested, suggesting F. proliferatum and F. oxysporum were major pathogens of the oriental melon, inducing stem rots and vascular wilts, respectively. Oriental melon and watermelon were more susceptible to F. oxysporum than shintosa and cucumber; and cucumber was most, oriental melon and watermelon, medially, and shintosa was least susceptible to F. proliferatum, whose virulence varied among and within their phylogenetic subclades. Severe root-knot galls were formed on all the crops infected with Meloidogyne incognita; however, little indication of vascular wilts or stem and/or root rots was shown by the nematode infection. These results suggest the current fungal disease in the oriental melon may be rarely due to virulence changes of the fusarium wilt pathogen and the direct cause of the severe root-knot nematode infection, but may be potentially from other Fusarium pathogen infection that produces seemingly wilting caused by severe stem rotting.

Pathogenicity and Occurrence of Pepper Seedling Anthracnose Caused by Colletotrichum acutatum (고추 유묘에 대한 Colletotrichum acutatum의 병원성과 탄저병 발생)

  • Han, Kyung-Sook;Park, Jong-Han;Han, You-Kyoung;Hwang, Jung-Hwan
    • Research in Plant Disease
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    • v.15 no.2
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    • pp.88-93
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    • 2009
  • In 2008, leaf rot and blight on pepper seeding ("Dokya-chungchung") occurred in a pepper farm at Hwaseong-si, Gyeonggi-do, Korea. The typical symptom is water-soaking and dark brown leaf blight at edges and tips of leaves. The fungal colonies isolated from infected tissues were pinkish at first and turned gradually to gray. Conidia were fusiform, non-septum, and $8.1-17.0{\times}2.0-3.8{\mu}m$ in size. Several specific PCR primers derived from the sequence of the internal transcribed spacer (ITS) region of the rDNA, such as CaINT, CgINT and CcINT were used for the identification of the fungal pathogen. The C. acutatum-specific primer CaINT was amplified single fragment of 496 bp that discriminated C. acutatum from the other species. The pathogenicity test was performed on seedlings and fruits of red pepper. On the basis of the morphological, molecular characteristics and pathogenicity test, we identified as Colletotrichum acutatum. This is the first report on leaf rot and blight on pepper seedling caused by C. acutatum in Korea.