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Multiplex PCR Assay for Simultaneous Detection of Korean Quarantine Phytoplasmas
Kim, Young-Hwan,Win, Nang Kyu,Back, Chang-Gi,Yea, Mi-Chi,Yim, Kyu-Ock,Jung, Hee-Young The Korean Society of Plant Pathology 2011 Plant Pathology Journal Vol.27 No.4
Multiplex PCR assays were developed for the simultaneous detection of ten important Korean quarantine phytoplasmas. The species-specific primers were designed based on ribosomal protein, putative preprotein translocase Y, immunodominant protein, elongation factor TU, chaperonin protein and the 16S rRNA genes of 'Candidatus (Ca.) Phytoplasma' species. Three main primer sets were prepared from ten designed primer pairs to limit nonspecific amplification as much as possible. The multiplex PCR assay using the three primer sets successfully amplified the correct conserved genes for each 'Ca. Phytoplasma' species. In addition, ten important 'Ca. Phytoplasma' species could be easily determined by recognizing band patterns specific for each phytoplasma species from three primer sets. Moreover, a high sensitivity of multiplex PCR for each primer set was observed for samples containing a low DNA concentration (10 ng/${\mu}l$). This study provides the useful multiplex PCR assay as a convenient method to detect the presence of ten important quarantine phytoplasmas in Korea.
Eun Ju Hwang,Su Yeon Jeong,김민지,Jun Seong Jeong,Keon Hee Lee,Na Ra Jeong,Jeong Sun Park,Deuk-Soo Choi,Kyu-Ock Yim,Iksoo Kim 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.1
Korean greenhouse strawberries are mostly cultivated from October to May, which includes the cold winter season. During this time, the population size of the spotted-wing drosophila (SWD), Drosophila suzukii Matsumura (Diptera: Drosophilidae), is expected to decrease in the wild, and is also expected to decrease inside the greenhouses, as long as SWD are not already present inside. Field surveys of SWD have been extensively carried out for field-grown agricultural fruits, but no study has been conducted for greenhouse fruits, such as strawberries. In this study, SWD capture patterns were examined inside and outside of the greenhouse blocks, and in the nearby woodlands in a southwestern locality of Korea using selected traps and attractants for nearly 19 months—in addition to several greenhouse blocks—during the strawberry cultivating periods. The highest capture period was observed from October to mid-December in woodlands, whereas capture number subsequently and sharply decreased up to mid-April, resulting in mostly zero-captures or low captures (≤10). During this period, a zero-capture period was observed inside the greenhouse that lasted for nearly three months (late December to late February). An incubation of the fallen strawberries supported the results of trap capture from inside the greenhouses. Taken together, the occurrence of SWD in the strawberry greenhouses is likely to be highly dependent on that of the nearby woodlands. Thus, a sharp winter drop and the subsequent zero- or lowcapture periods in the woodland areas were likely responsible for the observed zero-capture periods inside the greenhouses.