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

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Suppression of Citrus Melanose on the Leaves Treated with Rhizobacterial Strains after Inoculation with Diaporthe citri (식물근권세균 처리에 의한 감귤 검은점무늬병에 대한 방제 효과)

  • Ko, Yun Jung;Kang, So Young;Jeun, Yong Chull
    • Research in Plant Disease
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    • v.18 no.4
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    • pp.331-337
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    • 2012
  • Citrus melanose is one of important disease in citrus cultivation, reducing quality of citrus fruits and resulting in economic loss. Like other diseases in citrus, melanose was mainly protected by chemical fungicide in the field. Recently, alternative method of disease control is highly required due to the side effect of the chemicals. In this study four rhizobacterial strains TRH423-3, MRL408-3, THJ609-3, and TRH415-2 are selected by dual-culture testing its antifungal activity against Diaporthe citri causing citrus melanose. To investigate the protection efficacy of the selected rhizobacterial strains to citrus melanose, the bacteria were pre-treated on citrus leaves following inoculation with melanose pathogen. Pre-treatment with all selected rhizobacterial strains showed disease suppression in which the levels of protection rates were different by the rhizobacterial strains. Additional treatment with the rhizobacterial strains after the pathogen inoculation enhanced protection rates in all cases. The strain MRL408-3 and TRH423-3 were identified as Burkholderia gladioli, TRH415-2 as Pseudomons fluorescens and THJ609-3 as Pseudomonas pudia as a result of analyzing the internal transcript spaces of the rhizobacterial strains rDNA. The selected rhizobacterial strains may be valuable as biological control agents in the environment-friendly citrus farm in which chemical application is limited.

Genetic Diversity of Culturable Endophytic Fungi Isolated from Halophytes Naturally Growing in Muan Salt Marsh (무안 염습지에 자생하는 염생식물(halophyte)의 뿌리로부터 분리된 내생진균(endophytic fungi)의 유전학적 다양성)

  • You, Young-Hyun;Yoon, Hyeok-Jun;Seo, Yeong-Gyo;Kim, Mi-Ae;Kang, Myung-Suk;Kim, Chang-Mu;Ha, Sang-Chul;Cho, Ga-Youn;Kim, Jong-Guk
    • Journal of Life Science
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    • v.22 no.7
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    • pp.970-980
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    • 2012
  • Native halophytes, such as Suaeda maritima, Limonium tetragonum, S. japonica, Zoysia sinica, and Phragmites australis were collected from the Muan salt marsh. Ninety endophytic fungi were isolated from the roots of the collected halophytes. Molecular insights inferred by internal transcribed spacer containing ITS1, 5.8s, and the ITS2 region showed that all the fungal strains belong to ten orders, i.e., Capnodiales (4.44%), Cystofilobasidiales (1.11%), Dothideales (3.33%), Eurotiales (53.33%), Glomerellales (3.33%), Hypocreales (8.89%), Mucorales (1.11%), Pleosporales (15.56%), Sordariales (1.11%), and Trichosphaeriales (1.11%). The rest (6.67%) of all fungal isolates were not identified. Ninety fungal strains were confirmed at the genus level, containing Acremonium, Alternaria, Aspergillus, Aureobasidium, Cephalosporium, Chaetomium, Cladosporium, Colletotrichum, Cryptococcus, Didymella, Dothideomycete, Emericellopsis, Epicoccum, Eupenicillium, Fusarium, Gibberella, Gongronella, Macrophoma, Microsphaeropsis, Nigrospora, Paecilomyces, Paraconiothyrium, Penicillium, Phaeomyces, Phoma, Pleosporales, Purpureocillium, and Talaromyces. Of all the endophytic fungi identified from the various halophytes, Aspergillus and Penicillium of Eurotiales had the highest abundance.

Identification of Korean Poaceae Weeds Based on DNA Sequences (DNA 염기서열에 기초한 벼과 잡초의 분자생물학적 동정)

  • Lee, Jeongran;Kim, Chang-Seok;Lee, In-Yong;Oh, Hyun-Ju;Kim, Jung Hyun;Kim, Sun Yu
    • Weed & Turfgrass Science
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    • v.4 no.1
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    • pp.26-34
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    • 2015
  • Korean Poaceae includes approximately 80 species of the agricultural weeds. Precise species identification is the first step for more effective weed management in the agricultural fields. However, the identification of species in Poaceae is not easy without the assistance of taxonomists or identification experts although they are relatively easy to distinguish from the plants of the other family by the unique characteristics of caryopsis. Thus, DNA barcode was suggested as an alternative powerful technique for species identification by using short sections of DNA from a specific region of the genome. Two standard barcode markers of vascular plants, chloroplast rbcL and matK, and a supplementary nuclear ribosomal Internal Transcribed Spacer (ITS) region were used for barcode of major Korean Poaceae weeds, 403 individuals of 84 taxa. All the barcode markers revealed a good level of sequencing success with the lowest 73.7% for matK and the highest 88.8% for rbcL. The barcode sequences were deposited to the National Center for Biotechnology Information (NCBI) database for public use. Combined matK and ITS showed very high resolving power with 92.9%. Besides the identification of weeds for weed managment, the generated DNA barcode data could be used for many other applications such as rapid biodiversity assessment and conservation prioritization.

First report of the photosynthetic dinoflagellate Heterocapsa minima in the Pacific Ocean: morphological and genetic characterizations and the nationwide distribution in Korea

  • Lee, Sung Yeon;Jeong, Hae Jin;Kwon, Ji Eun;You, Ji Hyun;Kim, So Jin;Ok, Jin Hee;Kang, Hee Chang;Park, Jae Yeon
    • ALGAE
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    • v.34 no.1
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    • pp.7-21
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    • 2019
  • The genus Heterocapsa is one of the major dinoflagellate groups, with some of its species having worldwide distributions. However, prior to the present study, the phototrophic species Heterocapsa minima has been reported only from the northeast Atlantic Ocean. Recently, H. minima was found in the Korean waters, and a clonal culture was established. This culture was used to examine the morphology of the Korean strain H. minima HMMJ1604 through light and scanning electron microscopy, as well as for its genetic characterization. Furthermore, to determine the nationwide distribution of H. minima in Korea, its abundance was quantified in the waters of 28 stations in all four seasons in 2016-2018 using the quantitative real-time polymerase chain reaction method. The overall morphology of H. minima HMMJ1604 was very similar to that of the Irish strain H. minima JK2. However, the Korean strain had five pores around the pore plate, whereas the Irish strain had six pores. When properly aligned, the sequences of the large subunit and internal transcribed spacer regions of the ribosomal DNA of the Korean strain were identical to those of the Irish strain. This species was detected in the waters of 26 out of 28 stations, but its abundance was greater than $1.0cells\;mL^{-1}$ at 8 stations. The highest abundance of H. minima was $44.4cells\;mL^{-1}$. Although this species was found in all seasons, its abundance was greater than $1.0cells\;mL^{-1}$ when the water temperature and salinity were $10.9-25.0^{\circ}C$ and 17.5-34.1, respectively. To the best knowledge, the present study reported for the first time that H. minima lives in the Pacific Ocean and is widely distributed in the Korean waters.

Antitumor and immunomodulatory effects of a hot water extract of Phellinus baumii using Sarcoma-180 in the mouse (Pheflinus baumii 자실체 추출물의 Sarcoma-180 에 대한 항암 및 면역효과)

  • Ha, Hyo-Cheol;Kim, Hyun-Pyo;Sim, Ji Young;Chang, Yun-Hee;Kim, Hyun Su
    • Journal of Mushroom
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    • v.2 no.4
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    • pp.169-174
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    • 2004
  • A hot water extract(HWE-P4) was separated from the fruit bodies of PMO-P4, and its antitumor and immunomodulatory activities against sarcoma-180 in ICR mice were investigated. The internal transcribed spacer(ITS) regions from PMO-P4 were amplified using polymerase chain reaction(PCR) and sequenced. The results revealed that PMO-P4 was belong to the Phellinus baumii. When oral administration at the dose of 160mg/kg/day in the mice until the end of the experiment with 2 week's pre-feeding of the HWE-P4, the survival rate of the mice was 152% for 50days after the inoculation of sarcoma-180 and the suppression rate of the tumor growth was 35.3%(p<0.05) for 28 days after inoculation of sarcoma-180. The HWE-P4 increased 71.4% of the CD4/CD8 ratio and 5-fold of the expression of CD25(IL-2 receptor chain) compared with the control. From these results, the antitumor activity of HWE-P4 is exerted through its immunomodulating activity on the host's immune system.

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Characteristics of Brown Rot Caused by Monilinia fructicola on Stone Fruit in Korea (핵과류 잿빛무늬병을 일으키는 Monilinia fructicola 병해 특성)

  • Oh, Hun-Tak;Choi, In-Young;Kim, Ju;Na, Young-Eun;Lee, Wang-Hyu;Lee, Kui-Jae;Shin, Hyeon-Dong
    • Research in Plant Disease
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    • v.23 no.4
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    • pp.322-333
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    • 2017
  • In June and July 2015 and 2017, typical signs and symptoms of brown rot were observed on the fruit of Japanese apricot, peach, apricot, Japanese plum, and sweet cherry with incidence levels of 2-5% in Jeonju and Imsil, Korea. Early symptoms were small, circular, light brown spots that eventually destroyed entire fruit. Small sporodochia later appeared on the surface. Conidia isolated from each host were one-celled, hyaline, lemon-shaped and borne in branched monilioid chains. The optimal temperature range for hyphal growth of all the isolates was $20-25^{\circ}C$. The growth of hyphae was faster on potato dextrose agar and oatmeal agar than others. Multiple alignments using the ITS sequences from different host showed that they matched each other (100%). The ITS sequences showed 100% identity to those of M. fructicola. Based on the morphological characteristics and phylogenetic analysis via internal transcribed spacer (ITS), all the isolate was identified as M. fructicola. Pathogenicity of representative isolates was proved by artificial inoculation, fulfilling Koch's postulates. This is the first confirmed report on brown rot caused by M. fructicola on stone fruit in Korea.

Investigation of Fungicides Inhibitory Effect of on Summer Patch Disease, Caused by Magnaporthiopsis poae, in Kentucky bluegrass (여름잎마름병(Summer patch) 병원균에 대한 살균제의 억제효과 조사)

  • Lee, Jung Han;Shim, Gyu Yul;Kim, Jeong Ho;Jeon, Chang Wook;Kwak, Youn-Sig
    • Weed & Turfgrass Science
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    • v.6 no.2
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    • pp.151-156
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    • 2017
  • Summer patch is the most serious disease at turfgrass field or golf course established with Kentucky bluegrass during high temperature season in Korea. Nevertheless, chemicals for the summer patch control are not yet registered in Korea. We isolated the pathogens from the turfgrass showing typical summer patch symptoms and identified as Magnaporthiopsis poae by using the internal transcribed spacer ITS1 and ITS4 sequences of rDNA. The inhibition rates of the pathogen were investigated for 10 fungicides. As results, the pathogen growth was suppressed when chemicals concentration increased and negatively correlated with incubation period with the chemicals. In triazole group, all chemicals (metconazole, myclobutanil, propiconazole and tebuconazole) treated showed the inhibition rates by 100%. Thiophanate-methyl showed the next highest inhibition effect against a summer patch pathogen. In strobilurin group, pyraclostrobin was the highest suppression effect compared with azoxystrobin and trifloxystrobin. Inhibition effect of fludioxonil and fluxapyroxad on pathogen was similar to the trifloxystrobin. Based on the results, triazole and carboxamide groups are strongly recommended due to the highest inhibition effect on the summer patch pathogen, Magnaporthiopsis poae.

Gibberellin A7 production by Aspergillus tubingensis YH103 and cultural characteristics of endophytic fungi isolated from Tetragonia tetragonoides in Dokdo islands (독도 번행초에서 분리된 내생균류의 배양적 특성과 Aspergillus tubingensis YH103의 gibberellin A7의 생산)

  • You, Young-Hyun;Park, Jong Myong;Lim, Sung Hwan;Kang, Sang-Mo;Park, Jong-Han;Lee, In-Jung;Kim, Jong-Guk
    • Korean Journal of Microbiology
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    • v.52 no.1
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    • pp.32-39
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    • 2016
  • Coastal plant species Tetragonia tetragonoides (Pall.) Kuntze native to the Dokdo islands was sampled and then 17 endophytic fungi were purely isolated based on morphological differences. The fungal isolates were characterized by their growth properties under NaCl concentration or pH gradient. Culture filtrates of the 17 fungal isolates were treated to Waito-c rice (WR) seedlings for verifying plant growth-promoting activity. As the results, YH103 strain showed the highest plant growth-promoting activity among them. Phylogenetic analysis of the isolates was done by the maximum likelihood method based on partial internal transcribed spacer region (ITS region: contaning ITS1, 5.8S, and ITS2), beta-tubulin (BenA), and calmodulin (CaM) gene sequences. Chromatographic analysis of the strain YH103 culture filtrate showed the existence of gibberellins ($GA_4$, $GA_7$, $GA_8$, and $GA_{19}$). Finally, the strain YH103 was identified as Aspergillus tubingensis by microscopic observation and molecular analysis and, to our knowledge, this is the first report of GAs producing A. tubingensis.

Identification and Characterization of Paraconiothyrium brasiliense from Garden Plant Pachysandra terminalis (가든식물 수호초(Pachysandra terminalis)로부터 Paraconiothyrium brasiliense의 분리 및 동정)

  • Choi, Min Ah;Park, Seung Jun;Ahn, Geum Ran;Kim, Seong Hwan
    • The Korean Journal of Mycology
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    • v.42 no.4
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    • pp.262-268
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    • 2014
  • A fungal isolate DUCC5000 from a garden plant Pachysandra terminalis was identified as Paraconiothyrium brasiliense based on the results of morphological and molecular studies. The fungus formed brown to black conidiomata of (0.2-0.7)-2(-3.5) mm singly or as a group on PDA. Conidia measured $2-5{\times}1.8-3{\mu}m$ in size, hyaline, ellipsoid to short-cylindrical, and rounded at both ends. The internal transcribed spacer (ITS) DNA of the isolate shared 100% nucleotide sequence homology with those of known P. brasiliense isolates. Phylogenetic tree inferred from the ITS sequence analysis showed that the DUCC5000 isolate formed a clade with known isolates of P. brasiliense. The fungal mycelia grew better on oatmeal agar than on MEA and PDA. On PDA media under various pH conditions, fungal mycelial growth was observed at pH 9. Colony morphology of the fungus tended to alter depending on the kinds of nutrient media and pH condition. On chromagenic media, the fungus demonstrated its ability to produce extracellular enzymes including amyalse, avicelase, ${\beta}$-glucosidase, protease, and xylanase. However, in pathogenicity testing, no disease symptoms were observed on the leaves of P. terminalis. This strain is the first report on P. terminalis in Korea.

Morphological and Molecular Identification of Spirometra Tapeworms (Cestoda: Diphyllobothriidae) from Carnivorous Mammals in the Serengeti and Selous Ecosystems of Tanzania

  • Ndosi, Barakaeli Abdieli;Park, Hansol;Lee, Dongmin;Choe, Seongjun;Kang, Yeseul;Nath, Tilak Chandra;Bia, Mohammed Mebarek;Eamudomkarn, Chatanun;Jeon, Hyeong-Kyu;Eom, Keeseon S.
    • Parasites, Hosts and Diseases
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    • v.58 no.6
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    • pp.653-660
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    • 2020
  • Spirometra tapeworms (Cestoda: Diphyllobothriidae) collected from carnivorous mammals in Tanzania were identified by the DNA sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 (cox1) and internal transcribed spacer 1 (ITS1), and by morphological characteristics. A total of 15 adult worms were collected from stool samples and carcasses of Panthera leo, Panthera pardus, and Crocuta crocuta in the Serengeti and Selous ecosystems of Tanzania. Three Spirometra species: S. theileri, S. ranarum and S. erinaceieuropaei were identified based on morphological features. Partial cox1 sequences (400 bp) of 10 specimens were revealed. Eight specimens showed 99.5% similarity with Spirometra theileri (MK955901), 1 specimen showed 99.5% similarity with the Korean S. erinaceieuropaei and 1 specimen had 99.5% similarity with Myanmar S. ranarum. Sequence homology estimates for the ITS1 region of S. theileri were 89.8% with S. erinaceieuropaei, 82.5% with S. decipiens, and 78.3% with S. ranarum; and 94.4% homology was observed between S. decipiens and S. ranarum. Phylogenetic analyses were performed with 4 species of Spirometra and 2 species of Dibothriocephalus (=Diphyllobothrium). By both ML and BI methods, cox1 and ITS1 gave well supported, congruent trees topology of S. erinaceieuropaei and S. theileri with S. decipiens and S. ranarum forming a clade. The Dibothriocephalus species were sisters of each other and collectively forming successive outgroups. Our findings confirmed that 3 Spirometra species (S. theileri, S. ranarum, and S. erinaceieuropaei) are distributed in the Serengeti and Selous ecosystems of Tanzania.