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Rapaid Screening of Apple mosaic virus in Cultivated Apples by RT-PCR
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  • Rapaid Screening of Apple mosaic virus in Cultivated Apples by RT-PCR
저자명
Sun Hee Choi, Ki Hyun Ryu
간행물명
The Plant Pathology Journal KCI
권/호정보
2003년|19권 3호(통권85호)|pp.159-161 (3 pages)
발행정보
한국식물병리학회|한국
파일정보
정기간행물|ENG|
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영문초록

The coat protein (CP) gene of Apple mosaic virus (ApMV), a member of the genus Ilarvirus, was selected for the design of virus-specific primers for amplification and molecular detection of the virus in cultivated apple. A combined assay of reverse transcription and polymerase chain reaction (RT-PCR) was performed with a single pair of ApMV-specific primers and crude nucleic acid extracts from virus-infected apple for rapid detection of the virus. The PCR product was verified by restriction mapping analysis and by sequence determination. The lowest concentration of template viral RNA required for detection was 100 fg. This indicates that the RTPCR for detection of the virus is a 10(3) times more sensitive, reproducible and time-saving method than the enzyme-linked immunosorbent assay. The specificity of the primers was verified using other unrelated viral RNAs. No PCR product was observed when Cucumber mosaic virus (Cucumovirus) or a crude extract of healthy apple was used as a template in RT-PCR with the same primers. The PCR product (669 bp) of the CP gene of the virus was cloned into the plasmid vector and resultant recombinant (pAPCP1) was selected for molecule of apple transformation to breed virus-resistant transgenic apple plants as the next step. This method can be useful for early stage screening of in vitro plantlet and genetic resources of resistant cultivar of apple plants.

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