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KMID : 0903619960370010158
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Journal of the Korean Society for Horticultural Science
1996 Volume.37 No. 1 p.158 ~ p.165
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Detection of Cymbidium Mosaic Virus from Orchids by Reverse Transcription and Polymerase chain Reaction Amplification of the Viral coat Protein and 3 ´ - noncoding Region Sequence
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윤경은/Yoon, Kyung Eun
정소영/류기현/박원목/Chung, So Young/Ryu, Ki Hyun/Park, Won Mok
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Abstract
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The 3´-terminal part encoding coat protein gene and also 3´-noncoding region of cymbidium mosaic virus (CymMV), a species of the Potexvirus genus, was selected for the design of oligonucleotide primers based upon the nucleotide sequence of CymMV Korean strain (CymMV-K). A combined assay of reverse transcription and polymerase chain reaction (RT PCR) was performed with a set of 20-mer CymMV coat protein-specific primers (CYP1-2 for downstream and CYP2 for upstream primers) to amplify a 759 by DNA fragment from crude nucleic acid extracts of virus-infected orchid leaf tissue as well as purified CymMV RNA. The lowest concentration of template viral RNA required to detect of the virus was 10 fg of purified CymMV RNA. No PCR product was obtained when odontoglossum ringspot Tobamovirus (ORSV) RNA or extracts of healthy Cymbidium sp. were used as templates in RT PCR using the same primers. The RT PCR required 5 hrs for detection of CymMV in various orchid plants. The PCR product would be useful as a template of plant transformation for producing virus-resistant plant as well as molecular probe.
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