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http://dx.doi.org/10.7744/kjoas.20220075

Development of an RT-PCR assay and its positive clone for plant quarantine inspection of American plum line pattern virus in Korea  

Da-Som Lee (Plant Quarantine Technology Center, Department of Plant Quarantine, Animal and Plant Quarantine Agency)
Junghwa Lee (Plant Quarantine Technology Center, Department of Plant Quarantine, Animal and Plant Quarantine Agency)
Seong-Jin Lee (Plant Quarantine Technology Center, Department of Plant Quarantine, Animal and Plant Quarantine Agency)
Seungmo Lim (Plant Quarantine Technology Center, Department of Plant Quarantine, Animal and Plant Quarantine Agency)
Jaeyong Chun (Plant Quarantine Technology Center, Department of Plant Quarantine, Animal and Plant Quarantine Agency)
Publication Information
Korean Journal of Agricultural Science / v.49, no.4, 2022 , pp. 821-831 More about this Journal
Abstract
American plum line pattern virus (APLPV), a member of the genus Ilarvirus in the family Bromoviridae, is one of the plant quarantine pathogens in Korea. In this study, 15 candidate primer sets were designed and examined to develop a reverse transcription polymerase chain reaction (RT-PCR) assay for plant quarantine inspection of APLPV. Using APLPV-infected and healthy samples, the primer sets were assessed for APLPV detection. To confirm the occurrence of nonspecific reactions, six ilarviruses (Apple mosaic virus, Asparagus virus 2, Blueberry shock virus, Prune dwarf virus, Prunus necrotic ringspot virus, and Tobacco streak virus) and 10 target plants (Prunus mume, P. yedoensis, P. persica, P. armeniaca, P. dulcis, P. tomentosa, P. avium, P. glandulosa, P. salicina, and P. cerasifera) were examined. Finally, two primer sets were selected. These primer sets could generate the expected amplicons even with at least 1 ng of the total RNA template in concentration-dependent amplifications. In addition, a positive clone was developed for use as a positive control in the abovementioned RT-PCR assay.
Keywords
APLPV (American plum line pattern virus); diagnosis; plant quarantine; plant virus; RT-PCR (reverse transcription polymerase chain reaction);
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