• Title/Summary/Keyword: Nicotiana glutinosa

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Detection of Cucumber Mosaic Virus by RT-PCR Using a Simple and Rapid Crude Sap Extraction Method (간이 조즙액 추출법을 이용한 RT-PCR 방법에 의한 오이 모자이크 바이러스의 검정)

  • 이상용;홍진성;이진상;최장경
    • Korean Journal Plant Pathology
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    • v.12 no.4
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    • pp.432-436
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    • 1996
  • 역전사 중합효소 연쇄반응(RT-PCR)을 이용하여 담배(Nicotiana glutinosa)에 증식시킨 7종의 오이 모자이크 바이러스(CMV)를 검정하였다. RT-PCR을 위한 간단하고 신속한 바이러스 핵산의 조즙액 추출법이 개발되었으며, CMV 외피단백질 유전자 부위를 기초로 하여 제작한 20개의 염기로 구성된 primer를 사용하여 RT-PCR을 실시한 결과, 약 490 염기쌍의 DNA 단편들이 이병식물의 조즙액으로부터 증폭되었다. EcoRI 및 MspI을 이용한 RT-PCR 산물의 분석에 의하여, 공시한 7종의 바이러스는 모두 CMV subgroup I으로 동정되었다. Ouchterlony 한천젤 이중 확산법을 이용한 항혈청 검정에서도 7종의 바이러스 모두 CMV-Y의 항혈청과 단일의 침강선을 형성하였다.

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Characterization of an Isolate of Cucumber mosaic virus from Raphanus sativus L. (열무에서 분리한 오이모자이크바이러스 분리주의 특성)

  • Rhee, Sun-Ju;Hong, Jin-Sung;Choi, Jang-Kyung;Kim, Eun-Ji;Lee, Gung-Pyo
    • Research in Plant Disease
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    • v.17 no.2
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    • pp.211-215
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    • 2011
  • Cucumber mosaic virus (CMV)-like isolate was collected from Raphanus sativus (cv. Choon-hyang), which showed mosaic symptoms. The isolate was confirmed to a strain of CMV by host responses in Vigna unguiculata, Chenopodium amaranticolor and Gomphrena globosa, by viral genome composition with RT-PCR and PCR-RFLP, and by serological analysis. Symptom developed by the strain of CMV was severe in Nicotiana benthamiana, N. glutinosa, N. tabacum (cv. Samsun, cv. Xanthi), Cucumis melo (cv. Early hanover), Cucumis sativus (cv. White wonder), Capsicum annuum (cv. Chung-yang and cv. Geum-top), but mild symptom was developed in Raphanus sativus (cv. Choon-hyang), Brassica rapa ssp. pekinensis (cv. Bul-Am No. 3), and B. juncea (cv. Daenong Jukgot). Newly isolated strain of CMV could infect diverse crops including Solanaceae, Cucurbitaceae and Brassicaceae. We designated the new strain of CMV as Gn-CMV based on the novel infectivity of Brassicaceae. In double-stranded (ds) RNA analysis, Gn-CMV consisted of 3.3, 3.0, and 2.2 kb genomes likewise other strains of CMV. SDS-polyacrylamide gel electrophoresis (SDS-PAGE) showed 28 kDa of the CMV coat protein. By restriction enzyme mapping using Cac8I, ClaI and MspI of RT-PCR products indicated that Gn-CMV belongs to CMV subgroup I.

Inhibitory Effect of Chitosan and Phosphate Cross-linked Chitosan against Cucumber Mosaic Virus and Pepper Mild Mottle Virus

  • Gangireddygari, Venkata Subba Reddy;Chung, Bong Nam;Cho, In-Sook;Yoon, Ju-Yeon
    • The Plant Pathology Journal
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    • v.37 no.6
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    • pp.632-640
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    • 2021
  • Cucumber mosaic virus (CMV) and Pepper mild mottle virus (PMMoV) causes severe economic loss in crop productivity of both agriculture and horticulture crops in Korea. The previous surveys showed that naturally available biopolymer material - chitosan (CS), which is from shrimp cells, reduced CMV accumulation on pepper. To improve the antiviral activity of CS, it was synthesized to form phosphate cross-linked chitosan (PCS) and compared with the original CS. Initially, the activity of CS and PCS (0.01%, 0.05%, and 0.1% concentration) compound against PMMoV infection and replication was tested using a half-leaf assay on Nicotiana glutinosa leaves. The total number of local lesions represented on a leaf of N. glutinosa were counted and analyzed with phosphate buffer treated leaves as a negative control. The leaves treated with a 0.1% concentration of CS or PCS compounds exhibited an inhibition effect by 40-75% compared with the control leaves. The same treatment significantly reduced about 40% CMV accumulation measured by double antibody sandwich enzyme-linked immunosorbent assay and increased the relative expression levels of the NPR1, PR-1, cysteine protease inhibitor gene, LOX, PAL, SRC2, CRF3 and ERF4 genes analyzed by quantitative reverse transcriptase-polymerase chain reaction, in chili pepper plants.

Characterization of an Isometric virus Infecting Paprika (Capsicum annuum var. glosum) in Korea

  • Kim, Jae-Hyun;Kim, Jeong-Soo;Park, Gug-Seoun;Soo- Young Chae;Lee, Sin-Ho;Park, Jang-Kyung
    • Proceedings of the Korean Society of Plant Pathology Conference
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    • 2003.10a
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    • pp.140.1-140
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    • 2003
  • An Isometric virus was isolated from Paprika (Capsicum annuum var. glossum) showing necrosis spot and malformation on the fruit and the leaves, respectively, at yecheon in Korea. The virus could infect locally on Chenopodium amaranticolr, C. quinoa, Petunia x hybrida and Nicotiana glutinosa, but could not infect on Gomphrena globosa and Physalis floridana. The virus could infect systemically on red pepper and Lycopersicon esculentum. Datura stramonium, N. cleuarandii, N. rustim and N. tabacum cvs. were produced necrosis or necrotic ring spot lesions on the inoculated leaves and mosaic, vein necrosis or lethal death on the upper leaves. The virus was not related serologically to cucumber mosaic virus (CMV). In RT-PCR assay, it could not detected with specific primers of CMV and BBWV-II. The virions contain one molecule of genomic RNA, Which was approximately 3.8Kb and the coat protein (CP) of the purified virion migrated as a single band with molecular wight of about 29KDa in SDS-PAGE.

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Petunia Asteroid Mosaic Virus Isolated from Petunia hybrida Vilm. (폐츄니아에서 분리한 Petunia Asteroid Mosaic Virus)

  • 노궤미;최충원;최장경
    • Korean Journal Plant Pathology
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    • v.11 no.4
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    • pp.361-366
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    • 1995
  • A virus was isolated from petunia (Petunia hybrida Vilm.) plants showing chlorotic ring spots on the leaves and color breaking on the flowers, and was identified as petunia asteroid mosaic virus (PAMV). Identification of the PAMV was established by host range test, electron microscopy, serological reaction, and physical properties of the virus. In the host range test, Nicotiana glutinosa, N. rustica, N. clevelandii, P. hybrida, Gomphrena globosa, and Chenopodium amaranticolor were systemically infected with the virus. The virus produced local lesions on inoculated leaves of N. tabacum‘Samsun’, N. tabacum‘Xanthi nc’, Datura stramonium, Vigna unguiculata‘White eye’, C. quinoa, Capsicum annuum, Vicia faba, and Lycopersicon esculentum‘Rutgers’. However, Cucurbita sativus and C. moschata did not show any symptoms. PAMV particles were isometric with 30 nm in diameter. The crude sap from G. globosa infected with the virus reacted positively with antiserum to tomato bushy stunt virus (TBSV) in agar gel double diffusion test. Thermal inactivation point of the virus was 8$0^{\circ}C$ and the virus retained its infectivity at the dilution of 10-4. Longevity in vitro of the virus was estimated longer than 35 days.

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Isolation and Properties of Cucumber Mosaic Virus Inducing Mosaic Symptoms in Hippeastrum hybridum Hort (아마리리스에 모자익병을 일으키는 CMV에 관한 연구)

  • Kim J.S.;Kim H.B.;Lee S.H.
    • Korean journal of applied entomology
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    • v.20 no.2 s.47
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    • pp.76-82
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    • 1981
  • Cucumber Mosaic Virus (CMV) was isolated from naturally infected Hippenstrum hybridum. The virus caused mosaic symptoms on Nicotiana glutinosa and local lesions on Vignaunguiculata. The thermal inactivation point was 56C, dilution end point $10^{-3}$ and longevity in vitro was 2 days for CMV from Hippeastrum. Purified virus was obtained using citrate chloroform extraction procedure and polyethylene glycol precipitation followed by sucrose density gradient centrifugation. Purified virus had a typical absorption at 245nm. Electron micrographs of the purified virus from Hippeastrum showed spherical particles with 30nm in diameter. The purified virus reacted with CMV antiserum in agar gel double diffusion test.

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A Lectin with Mycelia Differentiation and Antiphytovirus Activities from the Edible Mushroom Agrocybe aegerita

  • Sun, Hui;Zhao, Chen Guang;Tong, Xin;Qi, Yi Peng
    • BMB Reports
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    • v.36 no.2
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    • pp.214-222
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    • 2003
  • A lectin named AAL has been purified from the fruiting bodies of the edible mushroom Agrocybe aegerita. AAL consisted of two identical subunits of 15.8 kDa, its pI was about 3.8 determined by isoelectric focusing, and no carbohydrate was discerned. Being treated by pyrogultamate aminopeptidase, the blocked N-terminus of AAL was sequenced as QGVNIYNI. AAL agglutinated human and animal erythrocytes regardless of blood type or animal species. Its hemagglutinating activity was unaffected by acid or alkali treatment and demetalization or addition of divalent metals $Mg^{2+}$, $Ca^{2+}$ and $Zn^{2+}$. AAL was toxic to mice: its LD50 was 15.85 mg per kilogram body weight by intraperitoneal injection. In this study, two novel activities of AAL were proved. It showed inhibition activity to infection of tobacco mosaic virus on Nicotiana glutinosa. The result of IEF suggested that AAL attached to TMV particles. Mycelia differentiation promotion was the other interesting activity. AAL promoted the differentiation of fruit body primordia from the mycelia of Agrocybe aegerita and Auricularia polytricha. AAL antiserum was prepared and immunologically cross-reactived with several proteins from five other kinds of mushrooms. These results suggested that AAL probably was a representative of a large protein family, which plays important physiological roles in mushroom.

Scopoletin Production Related to Induced Resistance of Tobacco Plants Against Tobacco mosaic virus

  • Kim, Young-Ho;Choi, Do-Il;Yeo, Woon-Hyung;Kim, Young-Sook;Chae, Soon-Yong;Park, Eun-Kyung;Kim, Sang-Seock
    • The Plant Pathology Journal
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    • v.16 no.5
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    • pp.264-268
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    • 2000
  • A fluorescent material was accumulated in inoculated leaves showing necrotic local lesions of tobacco plants with N gene, Nicotiana tabacum cvs. Xanthi-nc NN, Samsun NN, Burley 21 and KF 114, and N. glutinosa, and Datura stramonium at the early growth stages by the inoculation of Tobacco mosaic virus (TMV). It was identified as a coumarin phytoalexin, scopoletin. Although the material was most prominently produced in TMV-inoculated tobacco leaves with local necrotic lesions, its accumulation was also noted in uninoculated leaves of TMV-inoculated plants. Its accumulation was somewhat greater in high resistance-induced leaves than low resistance-induced and intact leaves. Scopoletin treatment induced the expression of a pathogenesis-related protein, PR-1, prominently at the concentration of 500 or 1000 ${\mu}$g/ml. This suggests that scopoletin is a phytoalexin abundantly accumulating in N gene-containing resistant plants in response to TMV infection, and may be related to hypersensitive responses (HR) and systemic acquired resistance (SAR) in the resistant tobacco plants.

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New Host Plants of Turnip Mosaic Potyvirus in Korea (순무 모자이크 바이러스(TuMV)의 새로운 기주식물 탐색)

  • 최준근;윤주연;이세원;최장경
    • Korean Journal Plant Pathology
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    • v.14 no.6
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    • pp.625-629
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    • 1998
  • Turnip mosaic potyviruses (TuMV) were isolated from Rorippa indica and Armoracia lapathifolia showing mosaic symptoms in field. Identification of the TuMVs were carried out by host reactions of indicator plants, electron micrograph, serological properties and reverse transcription-poly-merase chain reaction (RT-PCR). Both viruses systemically infected Chenopodium quinoa, Nicotiana clevelandii, Brassica rapa, B. campestris subsp. pekinensis, B. juncea and Raphanus sativus, and developed local infection on inoculated leaves of C. quinoa, C. amaranticola, C. album, N. tabacum cv. Xanthi nc and Gomphrena grobosa. However, the viruses did not infect on N. glutinosa, Cucumis sativus and Vigna unguiculata. The filamentous particles, about 720 nm in length, and inclusion bodies were observed from the infected leaf tissues by dipping on electron microscopy. Crude sap of leaf infected with the viruses was reacted positively with an antiserum of TuMV in agar gel double diffusion. For detection of the viruses, RT-PCR was carried out with TuMV--specfic oligonucleotide primer. The RT-PCR products, a 1,092 bp DNA fragment, were obtained from naturally infected leaves of R. indica and A. lapathifolia. In inoculation test to seven cruciferous weeds with TuMV, infection occurred in Arabis glabra, Barbarea orthoceras, Capsella bursa-pastoris, Draba nomorosa var. hebecarpa, Rorippa cantoniensis and Thlaspi arvense.

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Inhibitory Effect on Infection of Plant Viruses by Filtrate Powder from Culture Broth of Acinetobacter sp. and Its Mode of Action (Acinetobacter sp. 배양여과액 분말제제의 식물바이러스에 대한 감염억제 효과 및 작용)

  • Kim Mi-Soon;Jung Min-Young;Kim Yun-Sung;Jang Cheol;Hwang In-Cheon;Ryu Ki-Hyun;Choi Jang-Kyung
    • Research in Plant Disease
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    • v.12 no.2
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    • pp.91-98
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    • 2006
  • A filtrate powder, designated as KNF2022, produced from culture broth of Acinetobacter sp. KTB3 was tested for their inhibitory effects on Pepper mild mottle virus (PMMoV) infection to Nicotiana glutinosa or N. tabacum cv. Xanthi nc. When 1/100 dilution with distilled water was treated to the plants and PMMoV was inoculated, the inhibition was estimated to be 94.3 and 95.6%, respectively. The same concentrations of KNF2022 inhibited infections of Pepper mottle virus (PepMoV) and Cucumber mosaic virus (CMV) on Chenopodium amaranticolor by 97.1 and 92.5%, respectively. Duration of inhibitory activity of the filtrate powder from Acinetobacter sp. culture broth against PMMoV infection on N. glutinosa was maintained for 2 days at 80% inhibition level, however, the inhibitory effect was diminished from 4 days after treatment to 50% levels. To evaluate inhibitory effects on systemic host plants of the antiviral agent, symptom developments of PMMoV, PepMoV and CMV on KNF2022-treated pepper plants were investigated. Delayed symptom developments until 10 days after inoculation (DAI) were observed for all the three viruses when the viruses were inoculated individually, and these delayed symptom development effects were maintained until 30 DAI in case of PepMoV. Moreover, PepMoV was not detected by RT-PCR and ELISA until 30 DAI. These delayed symptom development effects were diminished in all combinations of three virus co-inoculations due to synergism of three viruses on symptom developments. Inhibitory effect of KNF2022 was verified under electron microscopic examinations using purified virus preparations. Particles of PMMoV and PepMoV were observed on specimens from 5 min after KNF2022 treatment, and the particle sizes were reached in the range of 200-250 nm and 400-600 nm, respectively. Furthermore, the viral particles were destructed and particle sizes were reached in the range of 100-150 nm and 300-500 nm, respectively, on 60 min after treatments. Reduction of local lesion numbers on N. tabacum cv. Xanthi nc and C. amaranticolor were accompanied with reduction of virus particle sizes. In the case of CMV destructed particle numbers were also increased according to incubation period after KNF2022 treatment and local lesions on C. amaranticolor were reduced.