title: Development of multiplex rt-PCR for simultaneous detection of garlic viruses and the incidence of garlic viral disease in garlic genetic resources.
abstract: Garlic generally becomes coinfected with several types of viruses belonging to the Potyvirus, Carlavirus, and Allexivirus genera. These viruses produce characteristically similar symptoms, they cannot be easily identified by electron microscopy (EM) or immunological detection methods, and they are currently widespread around the world, thereby affecting crop yields and crop quality adversely. For the early and reliable detection of garlic viruses, virus-specific sets of primers, including species-specific and genus-specific primers were designed. To effectively detect the twelve different types of garlic viruses, primer mixtures were tested and divided into two independent sets for multiplex polymerase chain reaction (PCR). The multiplex PCR assays were able to detect specific targets up to the similar dilution series with monoplex reverse transcription (RT)-PCR. Seventy-two field samples collected by the Gyeongbuk Agricultural Technology Administration were analyzed by multiplex RT-PCR. All seventy two samples were infected with at least one virus, and the coinfection rate was 78%. We conclude that the simultaneous detection system developed in this study can effectively detect and differentiate mixed viral infections in garlic.
authors.affiliation: School of Applied Biosciences, Kyungpook National University, Daegu 702-701, Republic of Korea ; Institute of Plant Medicine, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Moon
authors.initials: M
authors.lastname: Nam
authors.affiliation: Department of Functional Crop, National Institute of Crop Science, Miryang, Republic of Korea.
authors.forename: Yeong-Hoon
authors.initials: YH
authors.lastname: Lee
authors.affiliation: School of Applied Biosciences, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Chung Youl
authors.initials: CY
authors.lastname: Park
authors.affiliation: School of Applied Biosciences, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Min-A
authors.initials: MA
authors.lastname: Lee
authors.affiliation: School of Applied Biosciences, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Yang-Soo
authors.initials: YS
authors.lastname: Bae
authors.affiliation: Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Republic of Korea ; Biosystems and Bioengineering Program, University of Science and Technology, Daejeon 305-350, Republic of Korea.
authors.forename: Seungmo
authors.initials: S
authors.lastname: Lim
authors.affiliation: Institute for Bioresources Research, Gyeongsangbuk-do Agricultural Research and Extension Services, Andong 760-891, Republic of Korea.
authors.forename: Joong Hwan
authors.initials: JH
authors.lastname: Lee
authors.affiliation: Plant Systems Engineering Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Republic of Korea ; Biosystems and Bioengineering Program, University of Science and Technology, Daejeon 305-350, Republic of Korea.
authors.forename: Jae Sun
authors.initials: JS
authors.lastname: Moon
authors.affiliation: School of Applied Biosciences, Kyungpook National University, Daegu 702-701, Republic of Korea ; Institute of Plant Medicine, Kyungpook National University, Daegu 702-701, Republic of Korea.
authors.forename: Su-Heon
authors.initials: SH
authors.lastname: Lee
keywords: garlic; multiplex RT-PCR; plant virus; simultaneous detection; virus diagnosis