• Title/Summary/Keyword: Cymbidium mosaic virus

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Studies on Virus Diseases of Orchids in Korea (우리나라 란 바이러스에 관한 연구)

  • Lee Hyun Sook;La Yong Joon
    • Korean journal of applied entomology
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    • v.15 no.3 s.28
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    • pp.137-145
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    • 1976
  • Orchids have been propagated vegetatively for a long time without adequate control measures against virus diseases in Korea. As a result, it is presumed that most of the orchid varieties in Korea may have been degenerated. Nevertheless there has been little work on the virus diseases of orchids in Korea. Therefore studies were initiated to isolate an4 characterize the orchid viruses occurring in Korea. The results obtained are summerized as follows. 1. Symptoms of virus diseases on orchid varieties can be grouped 1) mosaic, 2) necrotic streak with mosaic, 3) ring necrosis, 4) chlorotkc ring and 5) necrotic spot. 2. A total of 102 orchid plants representing 4 genera were investigated on the occurrence of Cymbidium mosaic virus and tobacco mosaic virus by serological agar-gel double diffusion test. The test revealed that approximately $45\%$ of the orchids were infected with Cymbidium mosaic virus. None of the plants were found to be infected with tobacco mosaic virus. 3. Local lesions appeared on the inoculated leaves of Chenopodium amaranticolor Cassia occidentalis and Datura stramonium 7-12 days after mechanical inoculation with Cymbidium mosaic virus. 4. Physical properties of the Cymbidium mosaic virus determined by inoculation on Chenopodium amaranticolor were as follows: Thermal inactivation Point; $75-80^{\circ}C$, dilution end Point; $10^{-5}-10^{-6}\%$ aging in vitro; 8 days. 5. Three different buffers at pH 7.0 and pH 9.0 were compared for the efficiency of agar-gel double diffusion test with Cymbidium mosaic virus. Phosphate, imidazol and tris buffer at pH 7.0 gave equally satisfactory results. 6. Electron microscopic examination of the Cymbidium mosaic virus revealed rod shaped particles measuring 460-580mu.

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Isolation of Cymbidium mild mosaic virus (Cymbidium mild mosaic virus의 분리동정)

  • Chang M. U.;Doi Y.;Yora K.
    • Korean journal of applied entomology
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    • v.17 no.3 s.36
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    • pp.131-138
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    • 1978
  • A virus named Cymbidium mild mosaic virus(Cy MMV), was mechanically transmitted to Chenopodium amaranticolor from the leaves of Cymbidium with mild mosaic symptoms. The virus was cultured in C. amaranticolor, in which it produced local chlorotic and ring spots, followed by systemic vein clearing with distortion. CyMMV infected 7 out of 35 species of plants. In C. amaranticolor juice infectivity was lost by heating at $90^{\circ}C$ for 10 miuntes, and by aging at$20^{\circ}C$ for 60 days, and by diluting at $10^{-6}$ when bioassayed on C. amaranticolor. CyMMV was not transmitted by Myzus persicae. The virus was purified after clarification of homogenized C. amaranticolor leaf tissues with chloroform, by differential centrifugation followed by sucrose density gradient centrifugation. Electron microscopic examination of purified preparation showed spherical particles of 28nm in diameter. The UV absorption spectrum of purified preparation was typical of u nucleoprotein (max. at 261nm. min. at 243nm), and showed 260/280=1.72 and max/min=1.26. The value of the sedimentation coefficient of the virus was S20.w=126. In gel-diffusion tests, CyMMV antiserum reacted with CarMV, but not with any of four other viruses (BBWV, CRSV, CMV, TBRV) having similar particles and properties in vitro. In ultra-thin sections of CyMMV infected tissues, a large number of virus particles were found in the cytoplasm of mesophyll cells and in xylem vessels.

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Viral Infection of Tissue Cultured Orchids and Evaluation of Damages

  • Chung, Bong-Nam;Yoon, Ju-Yeon;Kim, Mi-Sun
    • The Plant Pathology Journal
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    • v.26 no.2
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    • pp.194-197
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    • 2010
  • Most orchids are propagated by tissue culture. To survey the viral infection of tissue cultured Orchids, total RNA was extracted from in vitro Cymbridium and Phalaenopsis spp. collected from companies producing tissue-cultured orchids, and RT-PCR analysis was conducted with primer pairs specific to Cymbidium mosaic virus (CymMV) and Odontoglossum ring spot virus(ORSV), which are infecting wide range of orchid genera. The bulb size of Cymbidium infected with CymMV and ORSV was compared with healthy one at 10 months after planting in vitro orchids in the glasshouse. The CymMV or ORSV infection in 97 Cymbidium and 55 Phalaenopsis plants was 84.5 and 89.1 %, respectively. Mixed infection was found in 52.6 and 47.3% of Cymbidium and Phalaenopsis tested, whereas virus-free orchids were 15.5 and 10.9%, respectively. The CymMV and ORSV reduced the bulb size by 2.7-50% depending on the cultivars of Cymbidium. The both viruses caused yellowing, mottle and mosaic with or without necrosis in 4 Cymbidium cultivars.

Detection of Cymbidium Mosaic Virus and Odontoglosum Ringspot Virus by ELISA and RT-PCR from Cultivated Orchids in Korea (ELISA와 RT-PCR에 의한 국내재배난에서 심비디움 모자이크 바이러스와 오돈토글로섬 윤문 바이러스이 검정)

  • 박원목;심걸보;김수중;류기현
    • Korean Journal Plant Pathology
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    • v.14 no.2
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    • pp.130-135
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    • 1998
  • This study was carried out to detect cymbidium mosaic potexvirus (CymMV) and odontoglossum ringspot tobamovirus (ORSV) in cultivated orchid plants in Korea. The standard double antibody sandwich enzyme-linked immunosorbent assay (ELISA) and reverse transcription polymerase chain reaction (RT-PCR) were carried out for detection of the viruses in the collected orchid samples. ELISA was suitable for massive-scale diagnostic method for virus detection in orchids. RT-PCR was rapid, time-saving and reliable detective method, and detection limit data showed that RT-PCR was 103 times more sensitive than ELISA. Of the 321 individual orchids representing 5 orchids genera tested by the ELISA, CymMV and ORSV were detected in 15.6% and 22.4%, and mixed infection of the both viruses with 4.9%, respectively. Of the Cymbidium plants tested, cultivated plants showed 52.5% virus infection rate with either CymMV or ORSV and both viruses.

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Application of Rapid and Reliable Detection of Cymbidium Mosaic Virus by Reverse Transcription Recombinase Polymerase Amplification Combined with Lateral Flow Immunoassay

  • Do-Hyun, Kim;Rae-Dong, Jeong;Sena, Choi;Ho-Jong, Ju;Ju-Yeon, Yoon
    • The Plant Pathology Journal
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    • v.38 no.6
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    • pp.665-672
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    • 2022
  • Cymbidium mosaic virus (CymMV) is one of economically important viruses that cause significant losses of orchids in the world. In the present study, a reverse transcription recombinase polymerase amplification (RT-RPA) assay combined with a lateral flow immunostrip (LFI) assay was developed for the detection of CymMV in orchid plants. A pair of primers containing fluorescent probes at each terminus that amplifies highly specifically a part of the coat protein gene of CymMV was determined for RT-RPA assay. The RT-RPA assay involved incubation at an isothermal temperature (39℃) and could be performed rapidly within 30 min. In addition, no cross-reactivity was observed to occur with odontoglossum ringspot virus and cymbidium chlorotic mosaic virus. The RT-RPA with LFI assay (RT-RPA-LFI) for CymMV showed 100 times more sensitivity than conventional reverse transcription polymerase chain reaction (RT-PCR). Furthermore, the RT-PCR-LFI assay demonstrated the simplicity and the rapidity of CymMV detection since the assay did not require any equipment, by comparing results with those of conventional RT-PCR. On-site application of the RT-RPA-LFI assay was validated for the detection of CymMV in field-collected orchids, indicating a simple, rapid, sensitive, and reliable method for detecting CymMV in orchids.

Use of Triton X-100 and Sephacryl S-500 HR for the Purification of Cymbidium Mosaic Virus from Orchid Plants

  • Han, Jung-Heon;La, Yong-Joon;Lee, Cheol-Ho
    • The Plant Pathology Journal
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    • v.15 no.1
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    • pp.34-37
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    • 1999
  • Cymbidium mosaic virus (CyMV) was purified from CyMV infected orchid plant leaves by Sephacryl S-500 HR column chromatography. Partial purification was done by solubilization with Triton X-100 (alkylphenoxypolyethoxy ethanol) and precipitation with polyethylene glycol (PEG 6,000) followed by ultracentrifugation on 30% sucrose cushion. Based on the spectrophotometric analysis, 33 mg of CyMV could be obtained form 100 g of CyMV-infected orchid plant leaves. The purified CyMV represented one distinct homogeneous band by SDS-PAGE, and electron microscopy revealed that it was highly homogeneous and not fragmented. Bioassay demonstrated that the purified CyMV had a normal infectivity to Chenopodium amaranticolor and orchid plants. Based on these results, the purification method in this work could be served as an improved method for the purification of CyMV and similar viruses with good yield, high purity and native integrity.

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Survey of the Incidence of Viral Infections in Calanthe spp. and Characterization of a GW Isolate of Cymbidium mosaic virus in Korea

  • Park, Chung Youl;Baek, Da Some;Oh, Jonghee;Choi, Jong-Yoon;Bae, Dae Hyeon;Kim, Jeong-Seon;Jang, Gil-Hun;Lee, Su-Heon
    • Research in Plant Disease
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    • v.22 no.2
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    • pp.65-71
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    • 2016
  • Cymbidium mosaic virus (CymMV) is a major virus infecting orchid plants and causing economic loss. In this study, the incidence of viral infection in Calanthe spp. at the Korean Institute of Calanthe was investigated using reverse transcription polymerase chain reaction. The CymMV infection rate was 42%, and the two viruses Odontoglossum ringspot virus and Cucumber mosaic virus had frequencies of 8% and 2%, respectively. Additionally, we characterized an isolate of CymMV, CymMV-GW, using biological tests and examined the nucleotide sequence properties of its complete genome. CymMV-GW induced chlorotic ringspots and chlorotic spot symptoms in inoculated leaves of Chenopodium amaranticolor and Nicotiana benthamiana, respectively. In this study, we have for the first complete genome sequence of CymMV-GW in Korea. The CymMV-GW genome was 6,225 nucleotides in length, excluding the poly-(A) tail, and showed whole-genome nucleotide and amino acid sequence identities of 97.7% and 100%, respectively, with the NJ-1 isolate of CymMV. Here, we report the complete genome sequence of the CymMV-GW isolate and viral infection rates for Calanthe spp. in Korea.

Molecular Characterization and Survey of the Infection Rate of Orchid fleck virus in Commercial Orchids

  • Kim, Sung-Ryul;Yoon, Ju-Yoon;Choi, Gug-Sun;Chang, Moo-Ung;Choi, Jang-Kyung;Chung, Bong-Nam
    • The Plant Pathology Journal
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    • v.26 no.2
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    • pp.130-138
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    • 2010
  • Orchid fleck virus (OFV) is an unassigned plant virus in the family Rhabdoviridae. OFV was isolated from Cymbidium sp. showing oval necrotic lesions on their leaves in Korea, and designated as OFV-NHHS1. The complete nucleotide sequence of the RNA1 (6,413 nt) (GenBank accession no. AB516442) and RNA2 (6,001 nt) (GenBank accession no. AB516441) was determined in this study. RNA1 and RNA2 contained five and one ORF respectively. RNA1 encodes nucleocapsid (N) of 49 kDa, ORF2 of 26 kDa, ORF3 of 38 kDa, ORF4 of 20 kDa and glycoprotein (G) of 61 kDa proteins, whereas RNA2 encodes a single polymerase of 212 kDa. OFV-NHHS1 shared extremely high similarity of 98.6-100% and 98.9-99.6% in nucleotidle and amino acid sequences with a Japanese isolate, OFV-so, respectively. However, the N, G and L of OFV-NHHS1 revealed 6.9-19.3%, 7.3-12.0%, and 13.4-26.6% identities to those of 29 Rhabdoviruses, respectively. To survey the infection rate of OFV in commercial orchids in Korea, 51 Cymbidium sp., 10 Phalaenopsis sp., 22 Oncidium sp. and 21 Dendrobium sp. plants that showed typical viral symptoms were collected. RT-PCR with specific primers for detection of Cymbidium mosaic virus (CymMV), ORSV and OFV showed high infection rate by ORSV alone and double infection by ORSV and CymMV. One of the orchids tested was infected with OFV. This is the first report of the complete nucleotide sequences of OFV isolated in Korea.