title: Development of a Multiplex PCR Method to Detect Fungal Pathogens for Quarantine on Exported Cacti.
abstract: Major diseases in grafted cacti have been reported and Fusarium oxysporum, Bipolaris cactivora, Phytophthora spp. and Collectotrichum spp. are known as causal pathogens. These pathogens can lead to plant death after infection. Therefore, some European countries have quarantined imported cacti that are infected with specific fungal pathogens. Consequently, we developed PCR detection methods to identify four quarantined fungal pathogens and reduce export rejection rates of Korean grafted cacti. The pathogen specific primer sets F.oF-F.oR, B.CF-B.CR, P.nF-P.nR, and P.cF-P.CR were tested for F. oxysporum, B. cactivora, P. nicotinae, and P. cactorum, respectively. The F.oF-F.oR primer set was designed from the Fusarium ITS region; the B.CF-B.CR and P.nF-P.nR primers respectively from Bipolaris and Phytophthora ITS1; and the P.cF-P.CR primer set from the Ypt1protein gene region. The quarantine fungal pathogen primer pairs were amplified to the specific number of base pairs in each of the following fungal pathogens: 210-bp (F. oxysporum), 510-bp (B. cactivora), 313-bp (P. nicotinae), and 447-bp (P. cactorum). The detection limit for the mono- and multiplex PCR primer sets was 0.1 ng of template DNA under in vitro conditions. Therefore, each primer set successfully diagnosed contamination of quarantine pathogens in export grafted cacti. Consequently, our methodology is a viable tool to screen contamination of the fungal pathogen in exported grafted cacti.
authors.affiliation: Division of Applied Life Science, Gyeongsang National University, Jinju 660-701, Korea.
authors.forename: Hyun Ji
authors.initials: HJ
authors.lastname: Cho
authors.affiliation: Department of Plant Medicine and Research Institute of Life Science, Gyeongsang National University, Jinju 660-701, Korea.
authors.forename: Seong Won
authors.initials: SW
authors.lastname: Hong
authors.affiliation: Plant Quarantine Technology Center, Animal and Plant Quarantine Agency, Suwon 443-400, Korea.
authors.forename: Hyun-Ju
authors.initials: HJ
authors.lastname: Kim
authors.affiliation: Department of Plant Medicine and Research Institute of Life Science, Gyeongsang National University, Jinju 660-701, Korea.
authors.forename: Youn-Sig
authors.initials: YS
authors.lastname: Kwak
keywords: PCR detection; grafted cactus; quarantine pathogen