• Title/Summary/Keyword: Ds-isolates

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RT-PCR Detection of dsRNA Mycoviruses Infecting Pleurotus ostreatus and Agaricus blazei Murrill

  • Kim, Yu-Jeong;Park, Sang-Ho;Yie, Se-Won;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.21 no.4
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    • pp.343-348
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    • 2005
  • The partial nucleotide sequences of the genomic dsRNA mycoviruses infecting Pleurotus ostreatus (isolates ASI2596, ASI2597, and Bupyungbokhoe) and Agaricus blazei Murrill were determined and compared with those of the other dsRNA mycoviruses. Partial nucleotide sequences of the purified dsRNA from ASI2596 and ASI2597 revealed RNA-dependent RNA polymerase sequences that are closely related to Oyster mushroom isometric virus 2, while nucleotide sequences and the deduced amino acid sequence from dsRNA mycovirus infecting Agaricus blazei did not show any significant homology to the other dsRNA mycoviruses. Specific primers were designed for RT-PCR detection of these dsRNA viruses and were found to specifically detect each dsRNA virus. Northern blot analysis confirmed the homogeneity of RT-PCR products to each purified dsRNA. Altogether, our results suggest that these virus-specific primer sets can be employed for the specific detection of each dsRNA mycovirus in infected mushrooms.

Identification and Characterization of Three Isolates of Cucumber mosaic virus Isolated from Weed Hosts (잡초에서 분리한 3종 Cucumber mosaic virus의 동정과 특성)

  • Lee, Hyeok-Geun;Kim, Sung-Ryul;Jeon, Yong-Woon;Kwon, Soon-Bae;Ryu, Ki-Hyun;Choi, Jang-Kyung
    • Research in Plant Disease
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    • v.14 no.1
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    • pp.15-20
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    • 2008
  • Three isolates of Cucumber mosaic virus (CMV) were isolated from weed hosts showing typical mosaic symptoms, and some properties of the viruses were investigated. CMV isolates, designated as Is-CMV, Jd-CMV and Pla-CMV from Isodon inflexus, Jeffersonia dubia and Phryma leptostachya var. asiatica, respectively, were identified and characterized by biological reaction in several host plants, serological property, dsRNA analysis, reverse transcription-polymerase chain reaction (RT-PCR), restriction fragment-length polymorphism (RFLP). All isolates systemically infected in Nicotiana benthamiana, Cucurbita pepo cv. Black beauty and Cucumis sativus, and did not reveal any differences in these host plants between the isolates. However, remarkable difference in the symptoms was found between the CMVs in Capsicum annuum. Is-CMV induced an asymptomatic symptoms, while Jd-CMV and Pla-CMV produced severe mosaic symptoms in C. annuum plants. In dsRNA analysis, all isolates revealed four major bands with estimated molecular size of 3.4, 3.2, 2.1 and 1.0 kbp. The cDNAs of coat protein gene of the isolates were amplified by RT-PCR using a genus-specific single pair primers that designed to amplify a DNA fragment of approximately ranging from 938 to 966 bp. By restriction mapping analysis using RFLP of the RT-PCR products as well as by serological properties of gel diffusion test, the CMV isolates belong to a typical members of CMV subgroup IA. This is the first report on the occurrence of CMV in the three weed hosts.

Antimicrobial activity by Paenibacillus elgii DS381 and its antimicrobial substances against microbial residents on human skin and pathogenic bacteria (인간 피부 상재균과 병원성 세균에 대한 Paenibacillus elgii DS381과 그 항균물질의 항균활성)

  • Lee, Da-Sol;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.54 no.3
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    • pp.244-253
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    • 2018
  • This study was carried out to evaluate effects of antimicrobial substances produced by isolated soil bacteria. Among two thousands of bacterial isolates Paenibacillus elgii DS381 exhibited high antimicrobial activities against several microbial residents on human skin and pathogenic bacteria. DS381 showed 15.3~26.0 mm inhibition zone diameter against all target bacteria and yeast in agar well diffusion test. Antimicrobial peptide produced by DS381 indicated low minimum inhibitory concentration (0.039-5.000 mg/ml). DS381 produced biosurfactant such as lipopeptide, and surface tension of culture supernatant of DS381 reduced from 60.0 to 40.3 mN/m. DS381 also showed $1.56{\pm}0.13U/ml$ of chitinase activity. These results suggest that Paenibacillus elgii DS381 may be utilized as an efficient biocontrol agent against some important human skin microbes and pathogenic bacteria.

Characterization and RT-PCR Detection of dsRNA Mycoviruses from the Oyster Mushroom, Pleurotus ostreatus

  • Seo, Jang-Kyun;Lim, Won-Seok;Jeong, Ji-Hye;Yoo, Young-Bok;Yie, Se-Won;Kim, Kook-Hyung
    • The Plant Pathology Journal
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    • v.20 no.3
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    • pp.200-205
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    • 2004
  • The partial nucleotide sequences of the genomic dsRNA mycovirus infecting Pleurotus ostreatus isolates ASI2223 and Suhan were determined and compared with those of mycoviruses belonging to partitiviruses and totiviruses. Partial nucleotide sequences of the purified dsRNA from ASI2223 and Suhan showed RNA-dependent RNA polymerase sequences that are closely related to those of partitiviruses, including Fusarium poae virus 1, Fusarium solani virus, Rhizoctoniasolani virus, Discula destructiva virus 2, and Oyster mushroom isometric virus 2. Specific primers were designed for RT-PCR detection of dsRNA viruses from the P. ostreatus isolate ASI2223 and Suhan. Two virus specific primer sets were found to specifically detect each virus among six sets of designed oligonucleotide primers. Collectively, these results suggest that dsRNA mycoviruses from P. ostreatus isolates ASI2223 and Suhan belong to the family Partitiviridae, although, they are not the same virus species. Our results also suggest that these virus-specific primer sets can be employed for the specific detection of each viral sequence in infected tissues.

Relationship between Virulence and Cultural Characteristics in Calonectria ilicicola (Calonectria ilicicola의 병원성과 배양적 특성간의 상호관계)

  • Kim, K.D.;Russin, J.S.;Snow, J.P.;Valverde, R.A.
    • The Korean Journal of Mycology
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    • v.26 no.3 s.86
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    • pp.387-395
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    • 1998
  • Mycelial growth, production of microsclerotia and perithecia, and presence of double-stranded RNA were examined in Calonectria ilicicola isolates from several hosts to detect morphological and/or genetic markers for comparison with levels of virulence. Variability in disease severity, production of microsclerotia and perithecia, and mycelial growth was observed across all isolates. None of 35 isolates of C. ilicicola examined contained detectable levels of double-stranded RNA. Disease severity on soybean cultivars correlated positively with production of both microsclerotia and perithecia. Mycelial growth correlated negatively with production of perithecia. Virulence on the cultivars and production of microsclerotia and perithecia were greater in isolates of C. ilicicola from soybean than those from peanut. These results suggest that the ability of isolates to produce microsclerotia and perithecia is a component of inoculum potential. Perithecia production may serve as a useful marker for characterizing virulence or host specialization in this homothallic fungus.

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Antimicrobial activities of Bacillus subtilis DS660 and Paenibacillus polymyxa DS842 (Bacillus subtilis DS660과 Paenibacillus polymyxa DS842의 항균활성)

  • Lee, Da-Sol;Song, Hong-Gyu
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.354-361
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    • 2018
  • This study was carried out to isolate soil bacteria with antimicrobial activity and evaluate antimicrobial substances produced by isolated bacteria. Among many isolates Bacillus subtilis DS660 and Paenibacillus polymyxa DS842 showed high antimicrobial activities against 6 species of microbial residents on human skin and 3 species of pathogenic bacteria. DS660 and DS842 showed 15.3~26.8 and 11.3~27.5 mm of inhibition zone diameter, respectively on nutrient agar medium against most target bacteria and fungi. DS660 and DS842 produced $57{\pm}8$ and $170{\pm}15{\mu}mol/ml$ of siderophore, respectively as an antimicrobial substance. Analysis of ethyl acetate extract of culture supernatants of DS660 and DS842 suggested production of glycolipid biosurfactant which reduced surface tension of culture supernatant of DS660 and DS842 from 60.0 to 40.3 and 30.3 mN/m, respectively. DS660 and DS842 also showed $169.2{\pm}9.9$ and $357.2{\pm}13.7nmol/min/mg$ protein of ${\beta}-1,3$-glucanase activity, respectively, and hydrolyzed cell wall components of 3 bacterial species. These results suggest that B. subtilis DS660 and P. polymyxa DS842 may be utilized as an environment-friendly biocontrol agent against some skin microbes and pathogenic bacteria.

Isolation of Cryphonechia parasitica from Cankers on Chestnut Trees in ]Korea

  • Ju, Young-Jik;Kim, Dae-Hyuk;Cha, Byeong-Jin
    • The Plant Pathology Journal
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    • v.18 no.1
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    • pp.23-29
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    • 2002
  • A total of 672 Cryphonectria parasitica was isolated from 2,536 blight lesions on chestnut twigs, which were collected from major chestnut plantations all over Korea. Isolation rates of each province ranged from 13.5% in Jeonbuk-ds to 37.4% in Gyeongnam-do, with an average rate of 25.6%. The isolates were classified into six groups according to color and shape of colony on PDA: smooth margin (S), irregular margin (I), yellow to brown (Y), white (W), and white with yellow center (C). Among these groups, IY was the most abundant with an isolation rate of 65%. On the other hand, SW, SC, IW, and SY were quite rare, with isolation rates ranging from 1.5% to 5.8%. When the 672 isolates were inoculated on the chestnut twigs,380 isolates (56.5%) caused lesions larger than the standard virulent isolate EPISS-2, while 158 isolates (23.4%) caused smaller lesions than the standard hypovirulent isolate UEP-1. However, 87.4% of the isolates belonged to the virulent group and only 12.6% belonged to the hypovirulent group based on Bavendamm test. In the provinces of Jeonnam-do, Jeonbuk-do, and Gyeongnamdo, which have high density of chestnut trees, the rates of hypovirulent-like isolates were over 20%.

Characteristics of ustilago maydis virus of SH14 killer strain isolated in Korea

  • Hwang, Seon-Hee;Jung, Cheong-Hwan;Yie, Se-Won
    • Journal of Microbiology
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    • v.33 no.2
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    • pp.154-159
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    • 1995
  • SH-14, a novel killer strain of Ustilago maydis was isolated in Korea. It has been reported in other papers that the toxin specificity and double-stranded RNA pattern of SH-14 strain were different from other laboratory strains. In this paper, we analyzed the biochemical characteristics of U. maydis SH-14 virus. Three distinctive peaks were isolated from CsCl density gradient, designated as top (T), intermediate (I) and bottom (B) components. We found that the densities of each components, 1.285, 1.408 g/cm$\^$3/, respectively, are very similar to those of other strains. As previously reported by the analysis of dsRNA in each component, the dsRNA segments are separately encapsidated. Capsid protein of SH-14 virus consists of two proteins about 70 Kd shown by SDS-PAGE analysis. Electron microscopic examination of the virus particles revealed that UmV particles are very similar in size and morphology to all isolates as well as all lab-strains. In order to test immunological cross reactivity of UmV, werstern bolt analysis was carriedout with antiserum against A8 virus. All capsid protein had positive reaction against A8 antibody which indicated that UmV are immunologically cross-reactive with all isolates from Korea. The results presented in this paper may show that UmV isolated from SH-14 strain has very similar biochemical characteristics to those of other UmV. However, the difference in the toxin specificity and the molecular weight of toxin protein from the SH-14 strain has us to conclude that U. maydis SH-14 strain is a new killer type.

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Characterization of Rice black-streaked dwarf virus in Maize (옥수수에 발생하는 벼검은줄오갈병의 유전자 비교)

  • Lee, Bong-Choon;Yoon, Young-Nam;Hong, Sung-Jun;Hong, Yeon-Kyu;Hwang, Jae-Bok;Song, Sek-Bo;Kang, Hwang-Won;Lee, Key-Woon
    • Research in Plant Disease
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    • v.14 no.3
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    • pp.223-225
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    • 2008
  • This study was carried out to identify the Rice black-streaked dwarf virus that infected maize plants collected from Gochang-gun in Jeollabukdo in 2005. The genomic dsRNA from infected plants was extracted and the genome pattern was analyzed by polyacrylamide gel electrophoresis. Results of the electrophoresis revealed the already known to-segment genome and the difference of mobility was confirmed in isolates by collected areas. The RBSDV was identified from the result of RT-PCR using the template of extracted dsRNA and specific primer. The results of S10 cloned to pGEM-T vector and the conducted in sequence analysis consisted of 1,801nt and 559aa. This was of the same size as the RBSDV S10 identified in rice, and the change was confirmed in 18 base and displayed homology of 99%.