• Title/Summary/Keyword: onion yellow dwarf virus

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An Inexpensive System for Rapid and Accurate On-site Detection of Garlic-Infected Viruses by Agarose Gel Electrophoresis Followed by Array Assay

  • Kazuyoshi Furuta;Shusuke Kawakubo;Jun Sasaki;Chikara Masuta
    • The Plant Pathology Journal
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    • v.40 no.1
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    • pp.40-47
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    • 2024
  • Garlic can be infected by a variety of viruses, but mixed infections with leek yellow stripe virus, onion yellow dwarf virus, and allexiviruses are the most damaging, so an easy, inexpensive on-site method to simultaneously detect at least these three viruses with a certain degree of accuracy is needed to produce virus-free plants. The most common laboratory method for diagnosis is multiplex reverse transcription polymerase chain reaction (RT-PCR). However, allexiviruses are highly diverse even within the same species, making it difficult to design universal PCR primers for all garlic-growing regions in the world. To solve this problem, we developed an inexpensive on-site detection system for the three garlic viruses that uses a commercial mobile PCR device and a compact electrophoresis system with a blue light. In this system, virus-specific bands generated by electrophoresis can be identified by eye in real time because the PCR products are labeled with a fluorescent dye, FITC. Because the electrophoresis step might eventually be replaced with a lateral flow assay (LFA), we also demonstrated that a uniplex LFA can be used for virus detection; however, multiplexing and a significant cost reduction are needed before it can be used for on-site detection.

RT-PCR-Based Detection of Six Garlic Viruses and Their Phylogenetic Relationships

  • PARK KWANG-SOOK;BAE YOUNG-JOO;JUNG EUN-JEONG;KANG SOON-JA
    • Journal of Microbiology and Biotechnology
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    • v.15 no.5
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    • pp.1110-1114
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    • 2005
  • Six viruses of the genera Carlavirus (Garlic mosaic virus, GarMV, and Garlic latent virus, GarLV), Allexivirus (Garlic virus X, GarV-X, and Garlic mite-borne filamentous virus, GarMbFV) and Potyvirus (Leek yellow stripe virus, LYSV, and Onion yellow dwarf virus, OYDV) from Korean garlic plants with mosaic symptoms were simultaneously detected by multiplex RT-PCR and subsequently sequenced. An immunocapture RT-PCR for the detection of GarLV, LYSV, and OYDV was also performed. The coat protein phylogenetic analysis of the garlic viruses showed that the Korean isolates were most closely related to the isolates from China, Japan, Brazil, and Argentina. This study is the first report for the differentiation of six garlic viruses in Korea by simultaneous detection using multiplex RT-PCR.