• 제목/요약/키워드: internal transcribed spacer(ITS)

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대전 계족산과 충남 오서산 및 전북 백암산 주위 야생화들로부터 효모의 분리 및 동정 (Isolation and Identification of Yeasts from Wild Flowers in Gyejoksan, Oseosan and Beakamsan of Korea)

  • 민진홍;류진주;김하근;이종수
    • 한국균학회지
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    • 제41권1호
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    • pp.47-51
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    • 2013
  • 국내 자연환경의 효모 균총을 확인함으로써 유용한 효모자원을 확보하기 위한 연구의 일환으로 대전 계족산, 충남 오서산 그리고 전북 백암산 지역에 서식하는 야생화들을 채집하여 이들로부터 효모들을 분리, 동정하였다. 대전의 계족산에서 수집한 야생화로부터는 10종에 속하는 12균주의 효모들을 분리하였고, 충남의 오서산에서 수집한 야생화로부터는 10종의 효모들 17균주를 분리하였다. 또한 전북 백암산의 야생화로부터 모두 24종에 속하는 38균주의 효모들을 분리하였다. 계족산, 오서산, 그리고 백암산에서 모두 발견된 효모는 Cryptococcus 속, Pseudozygoma 속, Rhodotorula 속, Sporobolomyces 속에 속하는 9종의 효모들이었다. 특히 3개 지역에서 수집한 야생화들로부터 Cryptococcus 속에 속하는 Cryptococcus aureus만이 모두 분리되었는데, 이 결과는 각 지역마다 독특한 효모 다양성을 보이고 있음을 시사하는 것이다.

Screening of Selected Korean Sweetpotato (Ipomoea batatas) Varieties for Fusarium Storage Root Rot (Fusarium solani) Resistance

  • Lee, Seung-yong;Paul, Narayan Chandra;Park, Won;Yu, Gyeong-Dan;Park, Jin-Cheon;Chung, Mi-Nam;Nam, Sang-Sik;Han, Seon-Kyeong;Lee, Hyeong-Un;Goh, San;Lee, Im Been;Yang, Jung-Wook
    • 한국균학회지
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    • 제47권4호
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    • pp.407-416
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    • 2019
  • A common post-harvest disease of sweetpotato tuber is root rot caused by Fusarium solani in Korea as well as the other countries. Storage root rot disease was monitored earlier on sweetpotato (Ipomoea batatas) in storehouses of different locations in Korea. In the present study, an isolate SPL16124 was choosen and collected from Sweetpotato Research Lab., Bioenergy Crop Research Institute, NICS, Muan, Korea, and confirmed the identification as Fusarium solani by conidial and molecular phylogenetic analysis (internal transcribed spacer ITS and translation elongation factor EF 1-α gene sequences). The isolate was cultured on potato dextrose agar, and conidiation was induced. The fungus was screened for Fusarium root rot on tuber of 14 different varieties. Among the tested variety, Yenjami, Singeonmi, Daeyumi, and Sinjami showed resistant to root rot disease. Additionally, the pathogen was tested for pathogenicity on stalks of these varieties. No symptom was observed on the stalk, and it was confirmed that the disease is tissue specific.

Sensitivity of Colletotrichum spp. Isolated from Grapes in Korea to Carbendazim and the Mixture of Carbendazim Plus Diethofencarb

  • Hwang, Sook-Yung;Kim, Hye-Ryoung;Kim, Joo-Hyung;Park, Jong-Han;Lee, Sang-Bum;Cheong, Seung-Ryong;Kim, Heung-Tae
    • The Plant Pathology Journal
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    • 제26권1호
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    • pp.49-56
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    • 2010
  • Thirty-six isolates of Colletotrichum spp. were obtained from infected grapes in two different locations of Korea; 18 isolates from Cheonahn, where carbendazim (MBC) and the mixture of MBC and diethofencarb (NPC) had been applied to control grape ripe rot, and 18 isolates from Cheongju, where no fungicides had been used. Sequences analysis of the internal transcribed spacer (ITS) and the $\beta$-tubulin gene identified 34 of the 36 isolates as Colletotrichum gloeosporioides. The remaining two isolates from Cheongju were identified as C. acutatum. Of the 18 isolates from Cheonahn, 12 were resistant to both MBC and the mixture (MBC+NPC), and six were sensitive to them. All C. gloeosporioides isolates from Cheongju, but not the two C. acutatum isolates, were sensitive to these fungicides. Sequence analysis of the $\beta$-tubulin gene in all isolates revealed that C. gloeosporioides resistant to MBC and MBC+NPC had a tyrosine instead of phenylalanine at the amino acid position 200. The appearance of resistance to MBC and the mixture in C. gloeosporioides correlated with the history of fungicide application in Korea.

Application of LATE-PCR to Detect Candida and Aspergillus Fungal Pathogens by a DNA Hybridization Assay

  • Gopal, Dhayaalini Bala;Lim, Chua Ang;Khaithir, Tzar Mohd Nizam;Santhanam, Jacinta
    • 한국미생물·생명공학회지
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    • 제45권4호
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    • pp.358-364
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    • 2017
  • Asymmetric PCR preferentially amplifies one DNA strand for use in DNA hybridization studies. Linear-After-The-Exponential-PCR (LATE-PCR) is an advanced asymmetric PCR method which uses innovatively designed primers at different concentrations. This study aimed to optimise LATE-PCR parameters to produce single-stranded DNA of Candida spp. and Aspergillus spp. for detection via probe hybridisation. The internal transcribed spacer (ITS) region was used to design limiting primer and excess primer for LATE-PCR. Primer annealing and melting temperature, difference of melting temperature between limiting and excess primer and concentration of primers were optimized. In order to confirm the presence of single-stranded DNA, the LATE-PCR product was hybridised with digoxigenin labeled complementary oligonucleotide probe specific for each fungal genus and detected using anti-digoxigenin antibody by dot blotting. Important parameters that determine the production of single-stranded DNA in a LATE-PCR reaction are difference of melting temperature between the limiting and excess primer of at least $5^{\circ}C$ and primer concentration ratio of excess primer to limiting primer at 20:1. LATE-PCR products of Candida albicans, Candida parapsilosis, Candida tropicalis and Aspergillus terreus at up to 1:100 dilution and after 1 h hybridization time, successfully hybridised to respective oligonucleotide probes with no cross reactivity observed between each fungal genus probe and non-target products. For Aspergillus fumigatus, LATE-PCR products were detected at 1:10 dilution and after overnight hybridisation. These results indicate high detection sensitivity for single-stranded DNA produced by LATE-PCR. In conclusion, this advancement of PCR may be utilised to detect fungal pathogens which can aid the diagnosis of invasive fungal disease.

Five New Wood Decay Fungi (Polyporales and Hymenochaetales) in Korea

  • Kim, Nam Kyu;Park, Jae Young;Park, Myung Soo;Lee, Hyun;Cho, Hae Jin;Eimes, John A.;Kim, Changmu;Lim, Young Woon
    • Mycobiology
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    • 제44권3호
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    • pp.146-154
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    • 2016
  • The wood decay fungi are a diverse taxonomic group that plays a pivotal role in forest carbon cycling. Wood decay fungi use various enzymatic pathways to digest dead or living wood in order to obtain carbon and other nutrients and these enzymatic systems have been exploited for both industrial and medical applications. Over 600 wood decay fungi species have been described in Korea; however, the recent application of molecular markers has dramatically altered the taxonomy of many of these wood decay fungi at both the genus and species levels. By combining molecular methods, specifically sequences of the internal transcribed spacer region, with traditional morphological characters, this study identified five new species records for Korea in five genera: Aurantiporus, Favolus, Neofavolus, Loweomyces, and Hymenochaetopsis. Three of these genera (Aurantiporus, Favolus, and Loweomyces) were previously unknown in Korea. The relatively simple morphology of the wood decay fungi often leads to ambiguous taxonomic assignment. Therefore, molecular markers are a necessary component of any taxonomic or evolutionary study of wood decay fungi. Our study highlights the need for a more robust and multifaceted approach in investigating new wood decay fungi in Korea.

Parasitism of the protozoan Perkinsus atlanticus in Manila clams, Ruditapes philippinarum, in Gomso Bay (Korea) and Ariake Bay (Japan)

  • Park, Kyung-Il;Choi, Kwang-Sik;Ngo, Thao T.T.;Tsutsumi, Hiro;Hong, Jae-Sang
    • 한국양식학회:학술대회논문집
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    • 한국양식학회 2004년도 수산관련학회 공동학술대회 발표요지집
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    • pp.513-513
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    • 2004
  • Manila clam, Ruditapes philippinarum, is commercially and ecologically important marine bivalve in Korea and japan. However, clam landings in the two countries have dramatically declined since the 1980-1990's. In the present study, the protozoan parasite, Perkinsus sp., lectin (host's defense-related glycoprotein) and histopathological features were investigated in Manila clams collected from Gomso Bay in Korea and Ariake Bay in japan (one of the largest clam beds in each country) during summer and fall, 2002-2003. DNA sequences of non-transcribe spacer (NTS), internal transcribed space. (ITS) and 5.85 rRNA of Perkinsus sp. were identical to those of P. atlanticus that was reported in Europe and Korea. For diagnosis of Perkinsus, the fluid thioglycollate medium (FTM) and the 2 M NaOH lysis methods were used. Prevalence of the parasite varied from 92.5-98.7% in Gomso Bay and 35.5-37.9% in Ariake Bay. Infection intensity, in terms of the number of Perkinsuscells per gram tissue wet weight, in the clams of Gomso Bay in fall 2002 averaged 1,010,077-470,937 recording approximately100 times higher than that of Ariake Bay, and these were twice higher than those of summer samples in each location. Mean hemagglutination titer of the clams from Gomso Bay was approximately 60-folds higher than that of clams from Ariake Bay in 2002. In histological preparation of the clams from Gomso Bay in 2002, trophozoites of P. atlanticus were in groups and resulted in severe inflammatory response of host clam. Prevalence of the trematod, Cercaria tapes-like in the clams of Gomso Bay and Ariake Bay were 8.8 % and 10.5% respectively. In conclusion, the clams from Gomso Bay showed more severe pathologic symptoms and higher immune response than those of the clams from Ariake Bay.

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Prevalence and Molecular Characterization of Intestinal Trichomonads in Pet Dogs in East China

  • Li, Wen-Chao;Wang, Kai;Zhang, Wei;Wu, Jingjing;Gu, You-Fang;Zhang, Xi-Chen
    • Parasites, Hosts and Diseases
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    • 제54권6호
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    • pp.703-710
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    • 2016
  • The trichomonad species Tritrichomonas foetus and Pentatrichomonas hominis were recently detected in the feces of dogs with diarrhea. However, little information is available on the prevalence and pathogenicity of these parasites in the canine population. Therefore, the aim of this study was to determine the prevalence and molecular characterization of trichomonads infecting pet dogs in Anhui and Zhejiang provinces, east China. In total, 315 pet dogs, with or without diarrhea, from 7 pet hospitals were included in this epidemiological survey. Microscopy and PCR detected P. hominis in 19.7% (62/315) and 31.4% (99/315) of fecal samples, respectively. T. foetus infection was detected in 0% (0/315) of samples with microscopy and in 0.6% (2/315) with PCR. The prevalence of P. hominis was significantly higher in young dogs (${\leq}12months$) than in adult dogs (>12 months), and was significantly higher in diarrheic dogs (50.6%) than in non-diarrheic dogs (24.3%; P<0.05). Infection with T. foetus did not correlate with any risk factors evaluated in this study. A sequence analysis of the P. hominis PCR products showed minor allelic variations between our sequences and those of P. hominis strains from other hosts in different parts of the world. Type CC1 was the most common strain in dogs in east China. The internal transcribed spacer 1 (ITS1)-5.8S rRNA gene sequences from the 2 T. foetus isolates detected in this study displayed 100% identity and were homologous to the sequences of other strains isolated from domestic cats in other countries.

A molecular investigation of Saccharina sessilis from the Aleutian Islands reveals a species complex, necessitating the new combination Saccharina subsessilis

  • Starko, Samuel;Boo, Ga Hun;Martone, Patrick T.;Lindstrom, Sandra C.
    • ALGAE
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    • 제33권2호
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    • pp.157-166
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    • 2018
  • Cryptic species complexes are increasingly recognized in phycological research, obscuring taxonomy and raising questions about factors influencing speciation. A recent exploration of kelp genetic diversity on Haida Gwaii, British Columbia revealed the existence of a new species, Saccharina druehlii, which is cryptic with Saccharina sessilis. This suggests that molecular investigations further north may be required to elucidate the taxonomy and evolutionary history of this lineage. Although, for several decades, S. sessilis was considered a single highly variable species, its taxonomy has been far from straightforward. In particular, Hedophyllum subsessile (Areschoug) Setchell is now recognized as a synonym of S. sessilis in North America, but as a growth form of Saccharina bongardiana in Far East Russia. To resolve this taxonomic confusion, we sequenced mitochondrial (CO1-5P) and nuclear (internal transcribed spacer) markers of S. sessilis populations from the Aleutian Islands, Alaska, USA. Interestingly, none of our sequences matched S. sessilis sensu stricto. Instead, CO1-5P sequences from populations in the central and eastern Aleutians matched exactly S. druehlii with increasing sequence divergence occurring westward. Samples from Attu, the western-most island, composed a genetic group that clearly represents Kjellman's concept of Hafgygia bongardiana f. subsessilis and is distinct enough from S. druehlii and S. sessilis to potentially constitute a distinct species. Therefore, Saccharina subsessilis comb. nov. is proposed for this entity. Our results suggest the existence of a species complex at the crown node of S. sessilis and thus further investigation of Saccharina in Alaskan waters should be conducted to reconstruct the evolutionary history of this fascinating lineage.

Comparative Pathogenicity of Fusarium graminearum Isolates from Wheat Kernels in Korea

  • Shin, Sanghyun;Son, Jae-Han;Park, Jong-Chul;Kim, Kyeong-Hoon;Yoon, Young-mi;Cheong, Young-Keun;Kim, Kyong-Ho;Hyun, Jong-Nae;Park, Chul Soo;Dill-Macky, Ruth;Kang, Chon-Sik
    • The Plant Pathology Journal
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    • 제34권5호
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    • pp.347-355
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    • 2018
  • Fusarium head blight (FHB) caused by Fusarium species is a major disease of wheat and barley around the world. FHB causes yield reductions and contamination of grains with trichothecene mycotoxins including; nivalenol (NIV), deoxynivalenol (DON), 3-acetyldeoxynivalenol (3-ADON), and 15-acetyldeoxynivalenol (15-ADON). The objectives of this study were to identify strains of F. graminearum isolated in Korea from 2012-harvested wheat grain and to test the pathogenicity of these NIV- and DON-producing isolates. Three hundred and four samples of wheat grain, harvested in 2012 in Chungnam, Chungbuk, Gyeongnam, Jeonbuk, Jeonnam, and Gangwon provinces were collected. We recovered 44 isolates from the 304 samples, based on the PCR amplification of internal transcribed spacer (ITS) rRNA region and sequencing. Our findings indicate that F. asiaticum was the predominant (95% of all isolates) species in Korea. We recovered both F. asiaticum and F. graminearum from samples collected in Chungnam province. Of the 44 isolates recovered, 36 isolates had a NIV genotype while 8 isolates belonged to the DON genotype (3-ADON and 15-ADON). In order to characterize the pathogenicity of the strains collected, disease severity was assessed visually on various greenhouse-grown wheat cultivars inoculated using both NIV- and DON-producing isolates. Our results suggest that Korean F. graminearum isolates from wheat belong to F. asiaticum producing NIV, and both F. graminearum and F. asiaticum are not significantly different on virulence in wheat cultivars.

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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    • 제33권3호
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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.